Compositions and methods relating to pooled fetal support tissue

Abstract

Disclosed herein are compositions, methods comprising pooled fetal support tissue, the pooled fetal support tissue comprising a therapeutically effective amount of native HC-HA / PTX3 complex.

Description

[Technical field]

[0001] Cited literature This application claims the benefit of U.S. Patent Application No. 62 / 334,283, filed April 25, 2022, the entirety of which is incorporated herein by reference.

[0002] overview Provided herein are compositions and methods relating to pooled fetal support tissue. Fetal support tissue has shown great potential for numerous clinical applications, and thus there has been significant interest in developing fetal support tissue products. However, the clinical and commercial potential of fetal support tissue products depends on effective processing methods for the tissue products, as well as the uniformity and potency of the final product. Maintenance of bioactive components during processing of fetal support tissue is essential. For example, differences in processing methods can vary the structure and biochemical composition of fetal support tissue, potentially compromising the activity of essential signaling molecules that are crucial for the intended therapeutic activity. Disclosed herein are compositions and methods for isolating, processing, and producing fetal support tissue products and / or formulations that maintain the potency and stability of tissue bioactive factors.

[0003] Typically, conventional protocols do not utilize pooling of cells, tissues, or biological products from multiple human donors due to the risk of infectious diseases and the difficulty of establishing manufacturing and processing / production facilities that meet regulatory standards. Advantageously, the present disclosure describes pooling of donor fetal support tissue. First, pooling biological products from multiple donors reduces the inherent variability that is often present between different donors. Pooling donor tissues adjusts the potency of each individual donor, increasing uniformity and facilitating further qualification, sampling, and clinical trial advancement. Additionally, pooling donor samples enriches the components of the fetal support tissue. Disclosed herein are methods for generating pooled fetal support tissue from multiple donors. In some embodiments, lyophilizing the fetal support tissue significantly reduces the degradation of tissue bioactive components in the fetal support tissue, concentrating the bioactive tissue components, and increasing the stability and concentration of bioactive components contained in the fetal support tissue material, including, for example, HA complex (HC-HA / PTX3), cytokines, growth factors, and any other bioactive factors found in the fetal support tissue. Summary of the Invention

[0004] In one aspect, a composition is described herein that includes pooled fetal support tissue, the pooled fetal support tissue includes a therapeutically effective amount of native HC-HA / PTX3 complex. In some embodiments, the composition includes pooled fetal support tissue, the fetal support tissue includes a therapeutically effective amount of native HC-HA / PTX3 complex. In some embodiments, the pooled fetal support tissue increases the homogeneity of native HC-HA / PTX3 complex in the composition. In some embodiments, the pooled fetal support tissue includes fetal support tissue from multiple donors. In some embodiments, a composition is disclosed herein that includes fetal support tissue from at least 3 donors. In some embodiments, the pooled fetal support tissue includes fetal support tissue from at least 5 donors. In some embodiments, the pooled fetal support tissue includes fetal support tissue from at least 10 donors. In some embodiments, the compositions exhibit therapeutic efficacy as determined by an assay including one or more of an HA content assay, a BCA total protein assay, a PTX3 content assay, an ODI-TRAP assay, an M2 polarization assay, or a WST-1 assay.

[0005] In some embodiments, the therapeutic efficacy is determined by ODI-TRAP assay. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in the ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 200 μg / ml, 250 μg / ml, 300 μg / ml, 400 μg / ml, 500 μg / ml, 600 μg / ml, 700 μg / ml, 800 μg / ml, 900 μg / ml, 1,000 μg / ml.

[0006] In some embodiments, the positive control for ODI-TRAP is RANKL treatment. In some embodiments, the therapeutic efficacy is determined by M2 assay. In some embodiments, the therapeutic efficacy is determined by NO assay.

[0007] In some embodiments, therapeutic efficacy may be determined by various assays, including but not limited to, the WST-1 assay.

[0008] In some embodiments, the therapeutic efficacy is determined by a BCA protein assay. In some embodiments, the therapeutic efficacy is determined by a HA content assay. In some embodiments, the HA content of the composition is at least 75 μg / ml. In some embodiments, the HA content of the composition is at least 90 μg / ml. In some embodiments, the total protein content of the composition is at least 250 μg / ml. In some embodiments, the pooled fetal support tissue is lyophilized. In some embodiments, the pooled fetal support tissue is lyophilized before the fetal support tissue is pooled. In some embodiments, the pooled fetal support tissue is lyophilized after the fetal support tissue is pooled. In some embodiments, the pooled fetal support tissue composition is pre-frozen or cryopreserved. In some embodiments, the pooled fetal support tissue comprises a particle size of about 0.01 micrometers (μm) to about 240 μm in diameter. In some embodiments, the pooled fetal support tissue comprises an average particle size of about 0.5 μm. In some embodiments, the pooled fetal support tissue comprises a placenta, an umbilical cord, a placental amnion, an umbilical cord amnion, a Wharton's jelly, a chorion, or an amniotic chorion, or any combination thereof. In some embodiments, the pooled fetal support tissue comprises a placental amnion, an umbilical cord, or both. In some embodiments, the pooled fetal support tissue is pulverized before being pooled. In some embodiments, the pooled fetal support tissue is pulverized after being pooled. In some embodiments, the pooled fetal support tissue is pulverized. In some embodiments, the pooled composition is filtered or ultrafiltered. In some embodiments, the pooled fetal support tissue is decellularized. In some embodiments, the pooled fetal support tissue is devitalized. In some embodiments, the pooled fetal support tissue is a gel extract. In some embodiments, the pooled fetal support tissue is powdered. In some embodiments, the pooled fetal support tissue comprises 5% or more water by weight. In some embodiments, the pooled fetal support tissue comprises up to 15% water by weight.In some embodiments, the pooled fetal support tissue is obtained from frozen fetal support tissue or previously frozen fetal support tissue. In some embodiments, the pooled fetal support tissue is substantially free of veins or arteries. In some embodiments, substantially all cells of the pooled fetal support tissue are killed. In some embodiments, the pooled fetal support tissue is terminally sterilized, gamma irradiated, filtered, e-beam sterilized, or a combination thereof. In some embodiments, the composition is provided as a pharmaceutical composition and further comprises a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutically acceptable excipient is saline.

[0009] The present application further discloses a method of treating a degenerative disease in an individual comprising administering to the individual the pooled fetal support tissue described herein.

[0010] Described herein is a method of treating inflammation in an individual comprising administering to the individual the pooled fetal support tissue described herein. In yet another aspect, disclosed herein is a method of promoting wound healing in an individual comprising administering to the individual the pooled fetal support tissue described herein.

[0011] Further disclosed herein, in another aspect, is a method of treating osteoarthritis in an individual using a composition comprising pooled fetal support tissue, the method comprising administering the pooled fetal support tissue to an individual. In some embodiments, the osteoarthritis may be osteoarthritis of the knee. In some embodiments, the osteoarthritis may be osteoarthritis of the facet joint. In some embodiments, the osteoarthritis may be osteoarthritis of the ankle. In some embodiments, the osteoarthritis may be osteoarthritis of the hip. In some embodiments, the osteoarthritis may be osteoarthritis of the shoulder. In some embodiments, the osteoarthritis may be osteoarthritis of the elbow. In some embodiments, the osteoarthritis may be osteoarthritis of the wrist. In some embodiments, the osteoarthritis may be osteoarthritis of the small joints of the hand. In some embodiments, the osteoarthritis may be osteoarthritis of the small joints of the leg. In some embodiments, the osteoarthritis may be osteoarthritis of the cervical spine. Disclosed herein, in some embodiments, is a method of producing a composition of pooled fetal support tissue, comprising obtaining fetal support tissue from multiple donors and pooling the fetal support tissue from the multiple donors. In some embodiments, the fetal support tissue from the multiple donors is freeze-dried prior to pooling. In some embodiments, the fetal support tissue from the multiple donors is freeze-dried after pooling. In some embodiments, the method of producing a freeze-dried composition further comprises processing the fetal support tissue from the multiple donors by morselizing, pulverizing, micronizing, or a combination thereof. In some embodiments, the method further comprises devitalizing the fetal support tissue from the multiple donors. In some embodiments, the method further comprises decellularizing the fetal support tissue from the multiple donors. In some embodiments, the fetal support tissue from the multiple donors is not dehydrated. In some embodiments, the fetal support tissue from the multiple donors is not dehydrated to contain less than 20% water by weight.In some embodiments, the fetal support tissue from multiple donors is not dehydrated to contain less than 15% water by weight.

[0012] Disclosed herein, in certain embodiments, is a method for validating a product of pooled fetal support tissue, comprising performing a validation assay on the pooled fetal support tissue, the validation assay determining the amount and / or therapeutic efficacy of native HC-HA / PTX3 complex. In some embodiments, the native HC-HA / PTX3 is therapeutically active. In some embodiments, the validation assay comprises one or more of an HA content assay, a PTX3 content assay, an ODI-TRAP assay, a BCA protein assay, an M2 type polarization assay, an NO assay, or a WST-1 assay. In some embodiments, the validation assay comprises an ODI-TRAP assay. In some embodiments, the pooled fetal support tissue products exhibit statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 200 μg / ml, 250 μg / ml, 300 μg / ml, 400 μg / ml, 500 μg / ml, 600 μg / ml, 700 μg / ml, 800 μg / ml, 900 μg / ml, 1,000 μg / ml. In some embodiments, the positive control is RANKL treatment. In some embodiments, the therapeutic efficacy is determined by an M2 assay. In some embodiments, the therapeutic efficacy is determined by a NO assay. In some embodiments, the therapeutic potential is determined by a WST-1 assay. In some embodiments, the therapeutic efficacy is determined by a BCA protein assay. In some embodiments, the therapeutic potential is determined by an HA content assay. In some embodiments, the therapeutic potential is determined by a PTX3 content assay. In some embodiments, the HA content of the composition is at least 75 μg / ml. In some embodiments, the HA content of the composition is at least 90 μg / ml, hi some embodiments, the total protein content of the composition is at least 250 μg / ml. [Brief description of the drawings]

[0013] [Figure 1A]

[0023] Figure 1 shows a flow chart illustrating an example of a process flow or method for producing a fetal support tissue product. The process flow shows the processing steps from donor sample procurement, cleaning, and processing (Figure 1A) to primary packaging of processed powder or fine powder form by pooling multiple donors (Figure 1B), thereby reducing sample-to-sample variation, increasing consistency, increasing yield, increasing effective dose, and uniform potency as disclosed herein. [Figure 1B]

[0023] Figure 1 shows a flow chart illustrating an example of a process flow or method for producing a fetal support tissue product. The process flow shows the processing steps from donor sample procurement, cleaning, and processing (Figure 1A) to primary packaging of processed powder or fine powder form by pooling multiple donors (Figure 1B), thereby reducing sample-to-sample variation, increasing consistency, increasing yield, increasing effective dose, and uniform potency as disclosed herein. [Figure 2A] 2A-2C are flow charts illustrating an example of a process flow or method (drug substance processing) for producing a fetal support tissue product in liquid form by pooling multiple donors to increase yield and uniform potency. The process of sample receiving and washing (FIG. 2A), sample processing and pooling process flow (FIG. 2B), and drug product receiving, storage, and packaging (FIG. 2C) are shown. [Figure 2B] 2A-2C are flow charts illustrating an example of a process flow or method (drug substance processing) for producing a fetal support tissue product in liquid form by pooling multiple donors to increase yield and uniform potency. The process of sample receiving and washing (FIG. 2A), sample processing and pooling process flow (FIG. 2B), and drug product receiving, storage, and packaging (FIG. 2C) are shown. [Figure 2C]2A-2C are flow charts illustrating an example of a process flow or method (drug substance processing) for producing a fetal support tissue product in liquid form by pooling multiple donors to increase yield and uniform potency. The process of sample receiving and washing (FIG. 2A), sample processing and pooling process flow (FIG. 2B), and drug product receiving, storage, and packaging (FIG. 2C) are shown. [Diagram 3] An example of cell morphology analysis and ODI-TRAP assay of fetal support tissue products is shown. [Figure 4] An example of cell morphology analysis and M2 IL-12 assay of fetal support tissue products is shown. [Diagram 5] 1 shows an example of a cell morphology analysis of fetal support tissue products and an exemplary M2 NO assay. [Figure 6A] An example of a linear analysis of the dose-dependence of hyaluronan ("HA") concentration in three potency assays is shown: WST-1 assay (Figure 6A), TRAP assay (Figure 6B), and M2 IL-12 assay (Figure 6C). Abbreviations MS = MAU / saline, M / W = MAU / WFI, M / S dlyz = MAU / dialyzed into saline, MAU = minced amniotic membrane and umbilical cord, WFI = water for injection. [Figure 6B] An example of a linear analysis of the dose-dependence of hyaluronan ("HA") concentration in three potency assays is shown: WST-1 assay (Figure 6A), TRAP assay (Figure 6B), and M2 IL-12 assay (Figure 6C). Abbreviations MS = MAU / saline, M / W = MAU / WFI, M / S dlyz = MAU / dialyzed into saline, MAU = minced amniotic membrane and umbilical cord, WFI = water for injection. [Figure 6C] An example of a linear analysis of the dose-dependence of hyaluronan ("HA") concentration in three potency assays is shown: WST-1 assay (Figure 6A), TRAP assay (Figure 6B), and M2 IL-12 assay (Figure 6C). Abbreviations MS = MAU / saline, M / W = MAU / WFI, M / S dlyz = MAU / dialyzed into saline, MAU = minced amniotic membrane and umbilical cord, WFI = water for injection. [Figure 7]An example of hyaluronic acid (HA) quantification (μg / ml HA) of samples obtained from individual or single donor lots and pooled lots is shown. [Figure 8A] An example of a Western blot analysis of fetal support tissue products is shown. Sample lots from single donor and pooled lots are evaluated in a Western blot assay to quantitatively assess changes in polymers / macromolecules of heavy chain 1 (HC1; FIG. 8A) and pentraxin (PTX3; FIG. 8B) that make up the heavy chain 1-hyaluronic acid / pentraxin (HC-HA / PTX3) complex. [Figure 8B] An example of a Western blot analysis of fetal support tissue products is shown. Sample lots from single donor and pooled lots are evaluated in a Western blot assay to quantitatively assess changes in polymers / macromolecules of heavy chain 1 (HC1; FIG. 8A) and pentraxin (PTX3; FIG. 8B) that make up the heavy chain 1-hyaluronic acid / pentraxin (HC-HA / PTX3) complex. [Figure 9] An example of quantitative analysis of HA evaluated to assess changes in molecular weight in samples from agarose gel electrophoresis of single donor lots (SD-1–SD-4; unpooled fetal support tissue in lanes 3–7) and pooled donor lots (P1–P3; pooled fetal support tissue (pFST) in lanes 8–10) is shown. [Figure 10A] An example of an ODI-TRAP quantification assay performed on samples from single / individual donor lots (FIG. 10A) and pooled donor lots (FIG. 10B) is shown. [Figure 10B] An example of an ODI-TRAP quantification assay performed on samples from single / individual donor lots (FIG. 10A) and pooled donor lots (FIG. 10B) is shown. [Figure 11] An example of a bicinchoninic acid (BCA) quantification assay performed on samples from single / individual donor lots and pooled donor lots is shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] Specific Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs.

[0015] As used herein, in some embodiments, ranges and amounts are expressed as "about" a particular value or range. "About" includes the exact amount. Thus, "about 5 μg" also means "about 5 μg" and "5 μg." In general, the term "about" includes an amount that can be expected within experimental error.

[0016] As used herein, "fetal support tissue" refers to any isolated tissue derived from tissue used to support fetal development. Examples of fetal support tissue include, but are not limited to, (i) placental amnion (PAM) or substantially isolated PAM, (ii) umbilical cord amnion (UCAM) or substantially isolated UCAM, (iii) chorion or substantially isolated chorion, (iv) amniotic chorion or substantially isolated amniotic chorion, (v) placenta or substantially isolated placenta, (vi) umbilical cord or substantially isolated umbilical cord, or (vii) any combination thereof. In some embodiments, the fetal support tissue is selected from the group consisting of placental amnion (PAM), umbilical cord amnion (UCAM), chorion, amniotic chorion, placenta, umbilical cord, and any combination thereof. In some embodiments, the fetal support tissue comprises umbilical cord. Fetal support tissue includes any form of fetal support tissue, including fresh, cryopreserved, terminally sterilized, micronized, freeze-dried fetal support tissue, or a powder obtained from grinding fetal support tissue. In some embodiments, the fetal support tissue is a ground, pulverized, minced, graft, sheet, powder, gel, homogenate, extract, cryogenically ground, decellularized, sieved, freeze-dried, dehydrated or liquefied, evaporated, micronized, or terminally sterilized product.

[0017] Fetal support tissues, including amniotic membrane and umbilical cord, contain several innate biological factors that are useful for a number of purposes, including reducing inflammation and scarring to promote regenerative wound healing. Biological factors that may be found in fetal support tissues may include components such as extracellular matrix, growth factors, and cytokines. Among the extracellular matrices, hyaluronic acid (HA) and HA-containing complexes, namely native HC-HA / PTX3, are abundant and unique to amniotic membrane (AM) and umbilical cord (UC). The HC-HA / PTX3 complex (high molecular weight (HMW) hyaluronan (HA) covalently bound to the heavy chain (HC)1 of inter-α-trypsin inhibitor, further complexed with pentraxin 3 (PTX3)) is one important active component in AM and UC, and is responsible for the aforementioned effects of promoting regenerative wound healing. Thus, there is a need for the production of fetal support tissue products with high yields of HC-HA / PTX3, HA, and other proteins of interest. There is a need for the production of fetal support tissue products using processes that reduce or prevent the degradation of the HC-HA / PTX3 complex and other proteins of interest. It is important to prevent the degradation of HC-HA / PTX3, HA, and other proteins of interest, as this can render the fetal support tissue product unsuitable for use.

[0018] As used herein, "powder" means a substance in the form of dry particulates. In some embodiments, the particles are not uniform in size. In some embodiments, the particles are substantially uniform in size.

[0019] As used herein, "grinding" refers to any method that reduces fetal support tissue into small particles or powder. The term grinding includes pulverization, cryo-grinding, filing, milling, grinding, pounding, and crushing.

[0020] As used herein, "placenta" refers to the organ that connects the maternal uterine wall to the developing fetus, allowing for the intake of nutrients, the elimination of waste products, and the exchange of gases via the maternal blood supply. The placenta consists of three layers. The innermost placental layer that encases the fetus is called the amnion. The allantois is the middle layer of the placenta (derived from the embryonic hindgut) and blood vessels originating from the umbilicus traverse this membrane. The outermost layer of the placenta (the chorion) contacts the endometrium. The chorion and allantois fuse to form the chorioallantoic membrane.

[0021] As used herein, "chorion" refers to a membrane formed by two layers: the extraembryonic mesoderm and the trophoblast. The chorion consists of two layers, an outer layer formed by the trophoblast and an inner layer formed by the parietal mesoderm, to which the amnion abuts. The trophoblast is composed of an inner layer of three-dimensional or prismatic cells, the cytotrophoblast of Langhans, and an outer layer of abundantly nucleated protoplasm lacking cell borders, the syncytiotrophoblast. The avascular amnion is attached to the inner layer of the chorion.

[0022] As used herein, "amniotic chorion" refers to a product that includes the amniotic membrane and the chorion. In some embodiments, the amniotic membrane and the chorion are not separated (i.e., the amniotic membrane is naturally attached to the inner layer of the chorion). In some embodiments, the amniotic membrane is initially separated from the chorion and then combined with the chorion during processing.

[0023] As used herein, "umbilical cord" refers to the organ that connects the developing fetus to the placenta. The umbilical cord is made up of Wharton's jelly, a gelatinous substance made up mostly of mucopolysaccharides. The umbilical cord contains one vein that carries oxygenated, nutrient-rich blood to the fetus and two arteries that carry deoxygenated, nutrient-poor blood out.

[0024] As used herein, "placental amniotic membrane" (PAM) refers to the amniotic membrane derived from the placenta. In some embodiments, the PAM is substantially isolated.

[0025] As used herein, "umbilical cord amniotic membrane" (UCAM) refers to the amniotic membrane that extends to become part of the umbilical cord. The UCAM is a translucent membrane. The UCAM contains multiple layers: an epithelial layer (basement membrane), a stratum densa, a fibroblast layer, and a stratum spongiosum. The UCAM lacks blood vessels or a direct blood supply. In some embodiments, the UCAM contains Wharton's jelly. In some embodiments, the UCAM contains blood vessels and / or arteries. In some embodiments, the UCAM contains Wharton's jelly and blood vessels and / or arteries.

[0026] As used herein, "human tissue" refers to any tissue derived from the human body. In some embodiments, the human tissue is a fetal support tissue selected from the group consisting of placenta-amnion, umbilical cord, umbilical cord amnion, chorion, amnion-chorion, placenta, or any combination thereof.

[0027] As used herein, "minimal manipulation" means (1) with respect to structural tissue, processing that does not alter the original relevant properties of the tissue relevant to the utility of the tissue for reconstruction, repair, or replacement, and (2) with respect to cells or nonstructural tissue, processing that does not alter the relevant biological properties of the cells or tissue.

[0028] The term "fresh fetal support tissue" refers to fetal support tissue that is less than 72 hours postnatal and in substantially the same form as postnatal. In some embodiments, the fresh fetal support tissue comprises fetal support tissue cells. In some embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the biological activity in the cells of the cell support tissue is maintained.

[0029] "Substantially isolated" or "isolated," when used in the context of a fetal support tissue product, means that the fetal support tissue product is separated from most other non-fetal support tissue material (e.g., other tissues, red blood cells, veins, arteries) derived from the original source organism.

[0030] As used herein, the phrase "the biological and structural integrity of the isolated fetal support tissue product is substantially maintained" means that the biological activity and structural integrity of the isolated fetal support tissue is reduced by no more than about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 50%, or about 60% when compared to the biological activity and structural integrity of fresh fetal support tissue.

[0031] As used herein, "processing" refers to any activity performed on fetal support tissue or preparations containing native HC-HA / PTX3 other than recovery, donor screening, donor testing, storage, labeling, packaging, or delivery, such as microbial, viral testing, preparation, sterilization, processes for inactivation or removal of adventitious materials, storage for storage, and removal from storage. As used herein, "HC-HA / PTX3" refers to heavy chain 1-hyaluronic acid-pentraxin 3, "-" indicates a covalent bond, and " / " indicates a tight non-covalent bond. HC-HA / PTX3 is found in fetal support tissue birth tissues, i.e., AM and UC.

[0032] As used herein, the terms "purified" and "isolated" refer to a material that is substantially or essentially free from components that are normally associated with it in its natural state (e.g., native HC-HA / PTX3 complex). In some embodiments, "purified" or "isolated" refers to a material that is about 50% or more free from components that are normally associated with it in its natural state, e.g., about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% free from components that are normally associated with it in its natural state (e.g., native HC-HA / PTX3 complex).

[0033] As used herein, "biological activity" refers to the activity of the polypeptides and polysaccharides of fetal support tissue, including native HC-HA / PTX3 or HA (present as HC-HA / PTX3 in fetal support tissue), or any other components of fetal support tissue, and combinations thereof. In some embodiments, the biological activity of the polypeptides and polysaccharides found in the fetal tissue support product is anti-inflammatory, anti-scarring, or anti-adhesion. In some embodiments, biological activity refers to the in vivo activity of the native HC-HA / PTX3 complex in the fetal support tissue product, or the physiological response elicited when the fetal support tissue is administered in vivo. In some embodiments, the biological activity of the native HC-HA / PTX3 complex in the fetal support tissue is substantially maintained. In some embodiments, the activity of the polypeptides and polysaccharides found in the fetal support tissue product promotes regenerative wound healing. In some embodiments, the activity of the polypeptides and polysaccharides found in the fetal tissue support product prevents scarring. In some embodiments, the activity of the polypeptides and polysaccharides found in the fetal support tissue reduces inflammation. Thus, biological activity includes the therapeutic effects and pharmaceutical activities of fetal support tissue compositions, such as HA, natural HC-HA / PTX3 complexes, and combinations thereof, or any other compositions or components of fetal support tissue disclosed herein.

[0034] As used herein, "structural integrity" refers to the integrity of the stroma and basement membrane that make up the fetal support tissue.

[0035] As used herein, a "purified" native HC-HA / PTX3 (nHC-HA / PTX3) complex refers to a HC-HA / PTX3 complex purified from a biological source, such as a cell, tissue, or biological fluid. In some embodiments, a purified native HC-HA / PTX3 (nHC-HA / PTX3) complex refers to a HC-HA / PTX3 complex purified from human fetal support tissue. In some embodiments, the native HC-HA / PTX3 is purified from amniotic membrane. In some embodiments, the nHC-HA / PTX3 is purified from umbilical cord. In some embodiments, the purified native HC-HA / PTX3 complex is isolated and purified from fetal support tissue before the fetal support tissue is pooled. In some embodiments, the purified native HC-HA / PTX3 complex is isolated and purified from fetal support tissue after the fetal support tissue is pooled. In some embodiments, the nHC-HA / PTX3 is substantially isolated, and this substantial isolation occurs before or after the fetal support tissue is pooled. In some embodiments, the substantially isolated HC-HA complex is derived from placenta, umbilical cord, chorion, amniotic chorion, placental amniotic membrane (PAM), umbilical cord amniotic membrane (UCAM), or any combination thereof. In some embodiments, the substantially isolated HC-HA complex is derived from frozen placental amniotic membrane (PAM) or pre-frozen placental amniotic membrane (PAM), frozen umbilical cord amniotic membrane (UCAM) or pre-frozen umbilical cord amniotic membrane (UCAM), frozen placenta or pre-frozen placenta, frozen umbilical cord or pre-frozen umbilical cord, frozen chorion or pre-frozen chorion, frozen amniotic chorion or pre-frozen amniotic chorion, or any combination thereof. In all or any cases, isolation of the HC-HA complex may occur before or after the fetal support tissues described above are pooled.

[0036] As used herein, "hyaluronan," "hyaluronic acid," or "hyaluronate" (HA) are used interchangeably and refer to a fully non-sulfated linear glycosaminoglycan (GAG) containing repeating disaccharide units of D-glucuronic acid and N-acetylglucosamine (D-glucuronosyl-N-acetylglucosamine).

[0037] As used herein, the term "tissue with undesirable changes" refers to tissue that has differentiated due to, for example, a wasting disease (e.g., arthritis, multiple sclerosis, Parkinson's disease, muscular dystrophy, and Huntington's disease) or aging, scar tissue, or tissue that has been damaged by injury, such as a burn, wound, laceration, injury, ulcer, surgery, or ischemia.

[0038] As used herein, the term "high molecular weight" or "HMW" in high molecular weight hyaluronan (HMW HA) is intended to refer to HA having a weight average molecular weight of greater than about 500 kilodaltons (kDa), such as, for example, about 500 kDa to about 10,000 kDa, about 800 kDa to about 8,500 kDa, about 1100 kDa to about 5,000 kDa, or about 1400 kDa to about 3,500 kDa. In some embodiments, the HMW HA has a weight average molecular weight of 3000 kDa or more. In some embodiments, the HMW HA has a weight average molecular weight of 3000 kDa. In some embodiments, the HMW HA is Healon®, which has a weight average molecular weight of about 3000 kDa. In some embodiments, the HMW HA has a molecular weight of about 500 kDa to about 10,000 kDa. In some embodiments, the HMW HA has a molecular weight of about 800 kDa to about 8500 kDa, hi some embodiments, the HMW HA has a molecular weight of about 3,000 kDa.

[0039] As used herein, the term "low molecular weight" or "LMW" in low molecular weight hyaluronan (LMW HA) is intended to refer to HA having a weight average molecular weight of less than 500 kDa, e.g., less than about 400 kDa, less than about 300 kDa, less than about 200 kDa, less than about 100 kDa, less than about 50 kDa, less than about 40 kDa, less than about 30 kDa, less than about 20 kDa, about 200-300 kDa, about 1-300 kDa, about 15 to about 40 kDa, or about 8-10 kDa.

[0040] As used herein, pentraxin 3 or PTX3 protein or polypeptide refers to any PTX3 protein, including but not limited to natural PTX3 protein and PTX3 protein extracted from cells or tissues. PTX3 includes multimeric forms of PTX3 (e.g., homomultimers), including but not limited to dimers, trimers, tetramers, pentamers, hexamers, octamers, and other multimeric forms.

[0041] The term "effective amount" or "therapeutically effective amount" as used herein refers to a sufficient amount of an agent or compound administered to relieve to some extent one or more of the symptoms of the disease or disorder being treated. In some embodiments, the result is a reduction and / or amelioration of the signs, symptoms, or causes of the disease, or any other desired change in a biological system. For example, an "effective amount" in therapeutic applications is the amount of a compound disclosed herein that is required to clinically significantly reduce disease symptoms without causing undue adverse side effects. In some embodiments, the appropriate "effective amount" in each individual case is determined using techniques such as dose escalation studies. The term "therapeutically effective amount" includes, for example, an amount that is effective for prophylaxis. An "effective amount" of a compound disclosed herein is an amount that is effective to achieve a desired effect or therapeutic improvement without causing undue adverse side effects. In some embodiments, it is understood that an "effective amount" or "therapeutically effective amount" will vary from subject to subject, depending on changes in the metabolism of the composition, the age, weight, general condition of the subject, the disorder being treated, the severity of the disorder being treated, and the judgment of the prescribing physician. In some embodiments, an effective amount is an amount of a product or compound sufficient to promote angiogenesis or normal vascularization in a tissue.

[0042] As used herein, the terms "subject," "individual," and "patient" are used interchangeably. None of these terms are to be construed as requiring the supervision of a medical professional (e.g., doctor, nurse, physician assistant, nursing assistant, hospice worker). As used herein, a subject is any animal, including mammals (e.g., humans or non-human animals) and non-mammals. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0043] As used herein, the terms "treat", "treating" or "treatment" and other grammatical equivalents include alleviating, reducing or ameliorating one or more symptoms of a disease or condition, reducing, preventing or decreasing the appearance, severity or frequency of one or more additional symptoms of a disease or condition, improving or preventing the metabolic causes underlying one or more symptoms of a disease or condition, inhibiting a disease or condition, e.g., halting the progression of a disease or condition, relieving a disease or condition, regressing a disease or condition, alleviating a condition caused by a disease or condition, or prophylactically and / or therapeutically inhibiting a condition of a disease or condition. In a non-limiting example, a natural HC-HA / PTX3 complex or composition disclosed herein is administered for prophylactic benefit to an individual who is at risk of developing a particular disorder or has a predisposition to developing a particular disorder, or who reports one or more physiological symptoms of a disorder.

[0044] composition As used herein, the term "composition" refers to any combination of two or more substances, including any composite product, whether resulting from chemical bonding or physical mixing, or whether in gas, liquid, powder, or solid form. As used herein, a "composition" of fetal support tissue may refer to any form of fetal support tissue, including isolated fetal support tissue, for example, isolated HA, nHC-HA / PTX3, extracts, sheet forms, non-sheet forms, powders, gels, solutions, and bioactive components of fetal support tissue in any form. Fetal support tissue contains several inherent biological factors that are useful for a number of purposes, including wound healing and reducing inflammation and scarring. Characterization data have shown that the extracellular matrix (ECM) of AM and UC is rich in hyaluronic acid (HA) with an integral or critical component called native HC-HA / PTX3 (nHC-HA / PTX3) complex ("HC-HA / PTX3" indicates heavy chain 1-hyaluronic acid-pentraxin 3, "-" indicates covalent bond, " / " indicates tight non-covalent bond), which is unique to the extracellular matrix of birth tissues, i.e., AM and UC. Native HC-HA / PTX3 is isolated from fetal support tissue obtained from humans or animals. Disclosed herein is pooled fetal support tissue material containing native HC-HA / PTX3 (nHC-HA / PTX3) complex. The nHC-HA / PTX3 complex (high molecular weight (HMW) hyaluronan (HA) covalently bound to the heavy chain (HC) 1 of inter-α-trypsin inhibitor, further complexed with pentraxin 3 (PTX3)) is one important active component of umbilical cord and amniotic membrane, which is responsible for the therapeutic effect of promoting regenerative wound healing. Therefore, it is essential to generate fetal support tissue products (e.g., amniotic membrane and umbilical cord extracts for use in wound healing) with high yields of HA complex or nHC-HA / PTX3. Processes that can prevent the degradation of bioactive components such as HC-HA / PTX3 complex, extracellular matrix, cytokines, and growth factors while generating fetal support tissue products, compositions, extracts, or combinations thereof, are important for the generation of clinically relevant products.Due to the wide range of clinical applications, it is important to find a method for processing fetal support tissue extracts or compositions with significantly reduced donor-to-donor variability. Disclosed herein is a system, method, composition, or device for generating fetal support tissue with significantly reduced donor-to-donor variability. Disclosed herein is a pooled fetal support tissue from multiple donors. Pooling fetal support tissue from multiple donors reduces donor-to-donor variability. Disclosed herein is a pooled fetal support tissue processed before or after being freeze-dried. The freeze-drying process disclosed herein results in a fetal support tissue composition with reduced degradation. Freeze-drying as disclosed herein also increases the concentration of bioactive compositions, for example, increasing the yield of nHC-HA / PTX3, hyaluronan (HA), growth factors, and cytokines, other bioactive factors, while maintaining the stability and potency of the fetal support tissue extract, composition, or product.

[0045] Fetal support tissue is obtained from the donor after delivery. In some embodiments, the fetal support tissue may comprise at least 5 grams (g), 10 g, 15 g, 20 g (wet weight) of amniotic membrane per donor, or at least 0.3 grams (g), 0.4 g, 0.5 g, 0.6 g, 0.7 g, or 0.8 g dry weight (e.g., freeze-dried weight). In some embodiments, the fetal support tissue may comprise at least 20 grams (g), 25 g, 30 g, 35 g, 40 g, 45 g, 50 g (wet weight) of umbilical cord tissue per donor, or at least 0.7 grams (g), 0.8 g, 0.9 g, 1.0 g, 1.1 g, or 1.2 g dry weight (e.g., freeze-dried weight). The pooled fetal support tissue described herein may be processed to isolate or purify nHC-HA / PTX3 complex.

[0046] In some embodiments, the pooled fetal support tissue disclosed herein may have a therapeutically effective amount of nHC-HA / PTX3. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 is about 0.1 mg per 100 mg of pooled fetal support tissue to about 10 mg per 100 mg of pooled fetal support tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 is about 1 mg per 60 g of pooled fetal support tissue. For example, the therapeutically effective amount of nHC-HA / PTX3 is about 1 mg per 10 g of pooled amniotic membrane. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 is from about 0.1 mg per 100 mg pooled fetal support tissue to about 0.5 mg per 100 mg pooled fetal support tissue, from about 0.1 mg per 100 mg pooled fetal support tissue to about 1 mg per 100 mg pooled fetal support tissue, from about 0.1 mg per 100 mg pooled fetal support tissue to about 2 mg per 100 mg pooled fetal support tissue, from about 0.1 mg per 100 mg pooled fetal support tissue to about 3 mg per 100 mg pooled fetal support tissue, from about 0.1 mg per 100 mg pooled fetal support tissue to about 4 mg per 100 mg pooled fetal support tissue, from about 0.1 mg per 100 mg pooled fetal support tissue to about 4 mg per 100 mg pooled fetal support tissue, About 5 mg, about 0.1 mg per 100 mg pooled fetal support tissue to about 10 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 1 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 2 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 3 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 4 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 5 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 5 mg per 100 mg pooled fetal support tissue, about 0.5 mg per 100 mg pooled fetal support tissue to about 5 mg per 100 mg pooled fetal support tissue5mg to about 10mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 2mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 3mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 4mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 5mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 10mg per 100mg pooled fetal support tissue, about 2mg per 100mg pooled fetal support tissue to about 3mg per 100mg pooled fetal support tissue, about 2mg per 100mg pooled fetal support tissue to about 4mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 5mg per 100mg pooled fetal support tissue, about 1mg per 100mg pooled fetal support tissue to about 10mg per 100mg pooled fetal support tissue from about 2 mg per 100 mg pooled fetal support tissue to about 5 mg per 100 mg pooled fetal support tissue, from about 2 mg per 100 mg pooled fetal support tissue to about 10 mg per 100 mg pooled fetal support tissue, from about 3 mg per 100 mg pooled fetal support tissue to about 4 mg per 100 mg pooled fetal support tissue, from about 3 mg per 100 mg pooled fetal support tissue to about 5 mg per 100 mg pooled fetal support tissue, from about 3 mg per 100 mg pooled fetal support tissue to about 10 mg per 100 mg pooled fetal support tissue, from about 4 mg per 100 mg pooled fetal support tissue to about 5 mg per 100 mg pooled fetal support tissue, from about 4 mg per 100 mg pooled fetal support tissue to about 10 mg per 100 mg pooled fetal support tissue, or from about 5 mg per 100 mg pooled fetal support tissue to about 10 mg per 100 mg pooled fetal support tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 is about 0.1 mg per 100 mg of pooled fetal support tissue, about 0.5 mg, about 1 mg per 100 mg of pooled fetal support tissue, about 2 mg per 100 mg of pooled fetal support tissue, about 3 mg per 100 mg of pooled fetal support tissue, about 4 mg per 100 mg of pooled fetal support tissue, about 5 mg per 100 mg of pooled fetal support tissue, or about 10 mg per 100 mg of pooled fetal support tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 is at least about 0.1 mg per 100 mg of pooled fetal support tissue, about 0.5 mg per 100 mg of pooled fetal support tissue, about 1 mg per 100 mg of pooled fetal support tissue, about 2 mg per 100 mg of pooled fetal support tissue, about 3 mg per 100 mg of pooled fetal support tissue, about 4 mg per 100 mg of pooled fetal support tissue, or about 5 mg per 100 mg of pooled fetal support tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 is up to about 0.5 mg per 100 mg of pooled fetal support tissue, about 1 mg per 100 mg of pooled fetal support tissue, about 2 mg per 100 mg of pooled fetal support tissue, about 3 mg per 100 mg of pooled fetal support tissue, about 4 mg per 100 mg of pooled fetal support tissue, about 5 mg per 100 mg of pooled fetal support tissue, or about 10 mg per 100 mg of pooled fetal support tissue.

[0047] In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 comprises from about 0.1 g per 10 g wet tissue to about 2 g per 10 g wet tissue. In some embodiments, a therapeutically effective amount of nHC-HA / PTX3 comprises from about 0.1 g per 10 g wet tissue to about 0.2 g per 10 g wet tissue, from about 0.1 g per 10 g wet tissue to about 0.5 g per 10 g wet tissue, from about 0.1 g per 10 g wet tissue to about 1 g per 10 g wet tissue, from about 0.1 g per 10 g wet tissue to about 2 g per 10 g wet tissue, from about 0.2 g per 10 g wet tissue to about 0.5 g per 10 g wet tissue, from about 0.2 g per 10 g wet tissue to about 1 g per 10 g wet tissue, from about 0.2 g per 10 g wet tissue to about 2 g per 10 g wet tissue, from about 0.5 g per 10 g wet tissue to about 1 g per 10 g wet tissue, from about 0.5 g per 10 g wet tissue to about 2 g per 10 g wet tissue, or from about 1 g per 10 g wet tissue to about 2 g per 10 g wet tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 comprises about 0.1 g per 10 g wet tissue, about 0.2 g per 10 g wet tissue, about 0.5 g per 10 g wet tissue, about 1 g per 10 g wet tissue, or about 2 g per 10 g wet tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 comprises at least about 0.1 g per 10 g wet tissue, about 0.2 g per 10 g wet tissue, about 0.5 g per 10 g wet tissue, or about 1 g per 10 g wet tissue. In some embodiments, the therapeutically effective amount of nHC-HA / PTX3 comprises at most about 0.2 g per 10 g wet tissue, about 0.5 g per 10 g wet tissue, about 1 g per 10 g wet tissue, or about 2 g per 10 g wet tissue.

[0048] cryopreservation In some embodiments, the fetal support tissue is pooled and then frozen for cryopreservation. In some embodiments, the fetal support tissue is frozen for cryopreservation and pooled after freezing. In some embodiments, cryopreserving the fetal support tissue does not destroy the integrity of the extracellular matrix of the fetal support tissue. In some embodiments, the fetal support tissue is exposed to a liquid gas (e.g., liquid nitrogen or liquid hydrogen). In some embodiments, the fetal support tissue is exposed to liquid nitrogen. In some embodiments, the fetal support tissue is not contacted with a liquid gas. In some embodiments, the fetal support tissue is placed in a container and the container is contacted with a liquid gas. In some embodiments, the fetal support tissue is exposed to the liquid gas until the fetal support tissue is frozen.

[0049] As used herein, "morsel" refers to a particle of tissue obtained from a larger piece of tissue, ranging in size from about 0.1 mm to about 1.0 cm in length, width, or thickness. The "morsels" described herein maintain the properties of the tissue from which they are obtained and are identifiable as such upon inspection. As used herein, the terms "morselized," "morselizing," and "morselization" refer to the actions of the present application on "morsels." In some embodiments, the morselized fetal support tissue, generated before the fetal support tissue is pooled or after the fetal support tissue is pooled, is further processed into a solution, suspension, or emulsion by mixing the morselized fetal support tissue with a carrier. In some embodiments, the morselized fetal support tissue is formulated into a solution, suspension, paste, ointment, oil emulsion, cream, lotion, gel, patch, stick, film, paint, or combination thereof. In some embodiments, the morselized fetal support tissue is contacted with a patch or wound dressing. In some embodiments, the morselized fetal support tissue is formulated for parenteral injection, administered as a sterile solution, suspension, or emulsion, or formulated for inhalation.

[0050] Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of fetal support tissue, including components of fetal support tissue such as nHC-HA / PTX3, HA, cytokines, growth factors, or other bioactive components. Throughout this specification and claims, processing of fetal support tissue to maintain the efficacy of HC-HA / PTX3 during AM / UC is disclosed. In some embodiments, fetal support tissue, including "comminuted", also refers to "micronization". Hereinafter, the term "micronize" as used herein is used interchangeably and means to perform the following: mincing, grinding, mincing, blending, mixing, pulverizing, powdering, homogenizing, and any combination thereof, of fetal support tissue. In some embodiments, fetal support tissue, which may comprise a mixture of amniotic tissue and umbilical cord tissue in any ratio between 0.001:99.999% and 99.999:0.001% by weight, is micronized before the fetal support tissue harvested from fresh or frozen tissue is pooled and / or freeze-dried, or after the fetal support tissue is pooled and / or freeze-dried, through the use of any micronization tool known to one of skill in the art, such as a tissue grinder, sonicator, pan beater, freezer / mill, blender, mortar / pestle, rototate, kitchen chopper, grater, ruler, and scalpel, to yield pieces in a size range of about 0.1 mm to about 1.0 cm in length, width, or thickness. In some embodiments, the resulting pieces are micronized to obtain particles of consistent size. In some embodiments, the resulting strips are used wet, partially dehydrated, or substantially dehydrated by any means known to those of skill in the art, such as, for example, a centrifuge, an ultracentrifuge, or freeze-drying. In some embodiments, the resulting fetal support tissue is used immediately or stored for later use in any type of contained container known to those of skill in the art, such as, for example, pouches, jars, bottles, tubes, ampoules, and pre-filled syringes.In some embodiments, the micronized fetal support tissue is sterilized by any method known to those of skill in the art, such as, for example, gamma irradiation.

[0051] As used herein, "pulverized fetal support tissue" refers to fetal support tissue including tissue that has been broken down (or separated) into "micronized" and in some instances, pulverization is used interchangeably with "micronized". In some embodiments, the pulverized pooled fetal support tissue is a dry powder. In some embodiments, the pulverized pooled and / or lyophilized fetal support tissue is a dry powder that is pooled after being pulverized. In some embodiments, isolated fetal support tissue is used to generate the micronized fetal support tissue. In some embodiments, fetal support tissue is used to generate the micronized fetal support tissue before the fetal support tissue is pooled. In some embodiments, fetal support tissue is used to generate the micronized fetal support tissue after the fetal support tissue is pooled. In some embodiments, fetal support tissue is used to generate the micronized fetal support tissue before the fetal support tissue is pooled. In some embodiments, fetal support tissue is used to generate the micronized fetal support tissue before the fetal support tissue is pooled. In some embodiments, fetal support tissue is used to generate the micronized fetal support tissue before the fetal support tissue is lyophilized. In some embodiments, fetal support tissue is used to generate micronized fetal support tissue before / after the fetal support tissue is freeze-dried. In some embodiments, fetal support tissue is used to generate micronized fetal support tissue before the fetal support tissue is pooled, but before or after the fetal support tissue is freeze-dried. In some embodiments, fetal support tissue is used to generate micronized fetal support tissue after the fetal support tissue is pooled, but before or after the fetal support tissue is freeze-dried. In some embodiments, the processed micronized fetal support tissue before the fetal support tissue is pooled, or after the fetal support tissue is pooled and before or after the fetal support tissue is freeze-dried, is further processed into a solution, suspension, or emulsion by mixing the fetal support tissue powder with a carrier.In some embodiments, the micronized fetal support tissue (produced before or after pooling and / or before or after lyophilization and / or combinations thereof) is formulated into a solution, suspension, paste, ointment, oil emulsion, cream, lotion, gel, patch, stick, film, paint, or combinations thereof. In some embodiments, the micronized fetal support tissue is contacted with a patch or wound dressing. In some embodiments, the micronized fetal support tissue is formulated for parenteral injection, administered as a sterile solution, suspension, or emulsion, or formulated for inhalation.

[0052] Disclosed herein, in certain embodiments, are methods and processes for producing fetal support tissue while maintaining the biological activity of HC-HA / PTX3 from fetal support tissue. In certain embodiments, the process includes a pooling step, which reduces the variation in the amount or activity of HC-HA / PTX3. In some embodiments, the isolated fetal support tissue is micronized by any suitable method. In some embodiments, lyophilization of the fetal support tissue occurs before the pooled fetal support tissue is micronized. In some embodiments, lyophilization of the fetal support tissue may occur after the pooled fetal support tissue is micronized. In some embodiments, the isolated fetal support tissue is micronized by use of a grinder (e.g., Bessman Tissue Pulverizer, Biospec BioPulverizer, or Covaris CryoPrep). In some embodiments, the isolated fetal support tissue is micronized by use of a tissue grinder (e.g., Potter-Elvehjem grinder or Wheaton Overhead Stirrer). In some embodiments, the isolated fetal support tissue is pulverized by use of a sonicator. In some embodiments, the isolated fetal support tissue is pulverized by use of a pan beater. In some embodiments, the isolated fetal support tissue is pulverized by use of a freezer / mill (e.g., a SPEX® SamplePrep Freezer / Mill or a Retsch Ball Mill). In some embodiments, the isolated fetal support tissue is pulverized by use of a pestle and mortar. In some embodiments, the isolated fetal support tissue is pulverized by manual use of a pestle and mortar. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims.

[0053] Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims. In some embodiments, the fetal support tissue may be in sheet form or non-sheet form consisting of sheet products of various sizes. In some embodiments, the pooled fetal support tissue is micronized before the fetal support tissue is frozen. In some embodiments, the pooled fetal support tissue is micronized after the fetal support tissue is frozen. In some embodiments, the fetal support tissue is not pooled before being micronized. In some embodiments, the fetal support tissue is pooled after being micronized. In some embodiments, the fetal support tissue is micronized before being freeze-dried and before or after being pooled. In some embodiments, the fetal support tissue is micronized after being freeze-dried and before or after being pooled. In some embodiments, the pooled fetal support tissue is freeze-dried prior to pulverization, devitalization, decellularization, cryo-grinding, sterilization, cryopreservation, purification, or a combination thereof, of the dry or wet fetal support tissue extract. In some embodiments, the pooled fetal support tissue is freeze-dried prior to pulverization, sieving, centrifugation, filtration, devitalization, decellularization, cryo-grinding, sterilization, cryopreservation, purification, or a combination thereof, of the dry or wet fetal support tissue extract. In some embodiments, the non-pooled fetal support tissue is freeze-dried prior to sieving, micronization, devitalization, decellularization, cryo-grinding, sterilization, cryopreservation, purification, or a combination thereof, of the dry or wet fetal support tissue extract. In some embodiments, the non-pooled fetal support tissue is not freeze-dried prior to pulverization, devitalization, decellularization, cryo-grinding, sterilization, sieving, centrifugation, filtration, cryopreservation, purification, or a combination thereof, of the dry or wet fetal support tissue extract.In some embodiments, the pulverization, devitalization, decellularization, cryo-grinding, sterilization, cryopreservation, purification, or combinations thereof of dry or wet tissue occurs before the fetal support tissue is pooled, whether the fetal support tissue is fresh or frozen or not frozen, or previously frozen or frozen. In some embodiments, the pulverization, devitalization, decellularization, cryo-grinding, sterilization, cryopreservation, purification, or combinations thereof occurs on pooled fetal support tissue (i.e., after pooling) and is produced from fresh pooled tissue, pooled and frozen tissue, previously pooled and then frozen tissue, or pooled tissue just before the fetal support tissue is frozen (devitalized), or pooled tissue after the fetal support tissue is frozen (devitalized). In some embodiments, the pooled fetal support tissue may be used to produce an extract from which the native HC-HA / PTX3 complex (nHC-HA / PTX3) may be purified by any suitable method. In some embodiments, the pooling strategy described herein may be used to produce nHC-HA / PTX3 by purification. In some embodiments, the purified pooled fetal support tissue product may be an extract, composition, or any form of product disclosed herein, and the product or extract is purified by centrifugation. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of fetal support tissue, including components of fetal support tissue such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims.

[0054] Various forms of fetal support tissue In some embodiments, the fetal support tissue comprises micronized fetal support tissue. In some embodiments, the fetal support tissue may be a fetal support tissue graft, a fetal support tissue sheet, a fetal support tissue homogenate, or a fetal support tissue extract. In some embodiments, the fetal support tissue is lyophilized. In some embodiments, the fetal support tissue is subjected to centrifugation or ultracentrifugation. In some embodiments, the fetal support tissue is liquid. In some embodiments, the liquid fetal support tissue is subjected to filtration. In some embodiments, the fetal support tissue is a powder. In some embodiments, the powder fetal support tissue is sieved. In some embodiments, the fetal support tissue is devitalized or decellularized. In some embodiments, the fetal support tissue is terminally sterilized. In some embodiments, the fetal support tissue is a purified native HC-HA / PTX3 complex. In some embodiments, the fetal support tissue is an extract of fetal support tissue. In some embodiments, the fetal support tissue is placenta-amnion, umbilical cord, umbilical cord amnion, chorion, amniotic chorion, placenta, amniotic stroma, amniotic jelly, or any combination thereof.

[0055] In some embodiments, the fetal support tissue comprises an umbilical cord, an amnion, or an umbilical cord-amniotic membrane. In some embodiments, the umbilical cord comprises an umbilical cord, an amnion, and at least some Wharton's jelly. In some embodiments, the umbilical cord comprises an umbilical cord, an amnion, and ... is substantially free of Wharton's jelly. In some embodiments, the umbilical cord is free of an umbilical vein and an umbilical artery. In some embodiments, the umbilical cord comprises an umbilical vein and an umbilical artery.

[0056] In some embodiments, the fetal support tissue comprises an extract of fetal support tissue. In some embodiments, the fetal support tissue comprises a natural HC-HA / PTX3 complex (nHC-HA / PTX3). In some embodiments, the fetal support tissue consists essentially of nHC-HA / PTX3. In some embodiments, the fetal support tissue comprises other components, such as, for example, other extracellular matrices, growth factors, cytokines, and other bioactive components that can be isolated from the fetal support tissue. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as, for example, HA, HC-HA / PTX3, growth factors, cytokines, and other active biocomponents, such that the efficacy of HC-HA / PTX3 is observable to ensure sufficient maintenance as disclosed throughout this specification and claims. In certain embodiments, the HC-HA / PTX3 is biologically active.

[0057] Grinding / Cryogenic Grinding In some embodiments, the fetal support tissue is comminuted by any suitable method before or after pooling and before or after lyophilization. In some embodiments, pooling before or after lyophilization of the fetal support tissue occurs for fresh fetal support tissue, raw fetal support tissue, freshly frozen fetal support tissue, or previously frozen fetal support tissue. Exemplary procedures for pulverization or cryo-grinding of fetal support tissue are disclosed herein. In some embodiments, the fetal support tissue is pooled before being pulverized or cryo-grinded. In some embodiments, the fetal support tissue is pooled after being pulverized or cryo-grinded. In some embodiments, pulverizing the fetal support tissue comprises cryo-grinding of the fetal support tissue. In some embodiments, cryo-pulverizing the fetal support tissue comprises pulverizing, pulverizing, or otherwise comminuted the fetal support tissue while the fetal support tissue is in a frozen state (e.g., exposed to temperatures below 0° C., −20° C., −40° C., −50° C., −60° C., −70° C., −75° C., −80° C., −90° C., −100° C.) or cooled state. In some embodiments, cryo-pulverizing the fetal support tissue comprises pulverizing the fetal support tissue in a cryo-controlled environment. In some embodiments, cryo-pulverizing the fetal support tissue comprises pulverizing the fetal support tissue after the fetal support tissue has been immersed in or exposed (e.g., directly or indirectly) to liquid nitrogen. In some embodiments, cryo-pulverizing the fetal support tissue comprises pulverizing the fetal support tissue while the fetal support tissue has been immersed in or exposed (e.g., directly or indirectly) to liquid nitrogen. In some embodiments, cryo-grinding the fetal support tissue comprises placing the fetal support tissue in a grinding vessel and immersing the grinding vessel in liquid nitrogen prior to grinding. In some embodiments, the grinding vessel is immersed in liquid nitrogen for at least one minute of the grinding process. In some embodiments, cryo-grinding the fetal support tissue comprises exposing the frozen fetal support tissue to a hammer or rotating blades. In some embodiments, cryo-grinding the fetal support tissue comprises exposing the frozen fetal support tissue to an impactor.In some embodiments, the impactor is driven by an electromagnet. In some embodiments, the fetal support tissue is cryo-ground by use of a FreezerMill. In some embodiments, the fetal support tissue is cryo-ground by use of a mortar and pestle. In some embodiments, the fetal support tissue is cryo-ground by use of a blender. In some embodiments, the fetal support tissue is cryo-ground by use of a BioPulverizer. In some embodiments, cryo-grinding the fetal support tissue in liquid nitrogen avoids activation of proteases and / or hyaluronidases in the fetal support tissue, which may degrade proteins and hyaluronan in the fetal support tissue, as compared to grinding unfrozen fetal support tissue. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as, for example, HA, nHC-HA / PTX3, growth factors, cytokines, and other active biological components, such that the efficacy of HC-HA / PTX3 is observable to ensure sufficient retention as disclosed throughout this specification and claims.

[0058] In some embodiments, the fetal support tissue is reduced to a powder by cryogenic grinding. In some embodiments, the particles comprising the powder are of uniform size distribution. In some embodiments, the particles comprising the powder are not of uniform size distribution. In some embodiments, the fetal support tissue is reduced to a particle size of less than about 1000 μm, 500 μm, 400 μm, 300 μm, 200 μm, 100 μm, 50 μm, 40 μm, 30 μm, 20 μm, 10 μm, 5 μm, 1 μm, 0.5 μm, 0.4 μm, 0.3 μm, 0.2 μm, or 0.1 μm. In some embodiments, the fetal support tissue is reduced to a particle size of less than 500 μm. In some embodiments, the fetal support tissue is reduced to a particle size of less than about 0.5 μm. In some embodiments, the fetal support tissue is reduced to a particle size of less than 0.3 μm.

[0059] Freeze drying Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as nHC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims. In some embodiments, freeze-drying preserves the biological activity of HC-HA / PTX3. In some examples, the cryopreserved fetal support tissue or tissue product is placed in a vacuum chamber of a freeze-drying device until all or substantially all fluid (e.g., water) is removed. In some embodiments, the fetal support tissue is cryopreserved before or after pooling. In certain embodiments, the pooled fetal support tissue has a water content reduced to 20% or less by weight, 15% or less by weight, 10% or less by weight, or 5% or less by weight. In certain embodiments, the pooled fetal support tissue has a water content of 1% or more by weight, 2% or more by weight, 3% or more by weight, 5% or more by weight, or 10% or more by weight. In some embodiments, lyophilization is performed on fetal support tissue from individuals and / or small pools or donors (e.g., 2, 3, 4, or 5) prior to pooling. In some embodiments, the pooled fetal support tissue is freeze-dried after pooling. In some embodiments, the pooled fetal support tissue is fresh or freshly frozen or previously cryopreserved before or after freeze-drying. In some embodiments, the pooled fetal support tissue is freeze-dried followed by pulverization, sieving, cryopreservation, sterilization, purification, centrifugation, filtration, or any combination thereof to allow for different size yields. In some embodiments, the fetal support tissue is freeze-dried prior to pooling. In some embodiments, the fetal support tissue is not freeze-dried prior to pooling. In some embodiments, the fetal support tissue is not freeze-dried prior to pooling. In some embodiments, the fetal support tissue that has been micronized, sieved, or cryopreserved, sterilized, purified, centrifuged, filtered, or any combination thereof to allow for different size yields occurs prior to freeze-drying or after freeze-drying.In some embodiments, the fetal support tissue is freeze-dried and then pooled after pulverization, sieving or cryopreservation, sterilization, purification, centrifugation, filtration, or a combination thereof to allow for different size yields. In some embodiments, the pooled fetal support tissue is freeze-dried after being frozen. In some embodiments, the fetal support tissue is freeze-dried and then pooled after being frozen by any suitable method (e.g., exposure to liquid gas, placement in a freezer).

[0060] As disclosed herein, in some embodiments, the fetal support tissue is pooled fetal support tissue, where the fetal support tissue is pooled before or after being lyophilized, where the fetal support tissue is pooled before or after being micronized, sieved or cryogenically ground, cryopreserved, centrifuged, filtered, sterilized, decellularized, devitalized, frozen, thawed, cut, washed, or any other process disclosed herein, and / or combinations thereof.

[0061] In some embodiments, the pooled fetal support tissue is frozen by exposure to a temperature of less than about 0° C. In some embodiments, the pooled fetal support tissue is frozen by exposure to a temperature of less than about −20° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −40° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −50° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −60° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −70° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −75° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −80° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −90° C. In some embodiments, the fetal support tissue is frozen by exposure to a temperature of less than about −100° C. In some embodiments, the fetal support tissue is frozen by exposure to a liquid gas, such as liquid nitrogen gas. In some embodiments, the fetal support tissue is placed in a vacuum chamber of a freeze-drying apparatus until all or substantially all of the fluid (e.g., water) has been removed. In some embodiments, the cryopreserved fetal support tissue is freeze-dried.

[0062] Generation of fetal support tissue In some embodiments, the fetal support tissue is derived from umbilical cord (UC) tissue. In some embodiments, the fetal support tissue is derived from amniotic membrane (AM) tissue. In some embodiments, the fetal support tissue is derived from umbilical cord amniotic membrane tissue or a combination of any of the above. In some embodiments, the fetal support tissue comprises isolated fetal support tissue that does not include veins or arteries. In some embodiments, the fetal support tissue comprises isolated fetal support tissue that does not include veins or arteries, metabolically active cells, and the natural structural integrity of the fetal support tissue is substantially preserved after initial procurement. In some embodiments, the fetal support tissue comprises isolated fetal support tissue that includes veins or arteries. In some embodiments, the fetal support tissue comprises isolated fetal support tissue that includes veins or arteries, metabolically active cells, and the natural structural integrity of the fetal support tissue is substantially preserved after initial procurement. In some embodiments, the fetal support tissue comprises umbilical cord amniotic membrane and Wharton's jelly. In some embodiments, the biological activity of various bioactive components, such as, for example, HA, nHC-HA / PTX3 complex, or composition in the fetal support tissue is substantially maintained. In some embodiments, the biological activity of the bioactive component, HA, and / or nHC-HA / PTX3 complex in the fetal support tissue is substantially maintained when processed (washed, cut, centrifuged, etc.) and frozen 72 hours after delivery. The frozen tissue may be frozen for at least 1 day, 5 days, 10 days, 20 days, 30 days, or more. In some embodiments, the biological activity of the bioactive component, HA, or nHC-HA / PTX3 complex in the fetal support tissue is substantially maintained when processed (washed, cut, etc.) and frozen within 72 hours after delivery. In some embodiments, the biological activity of the bioactive component, HA, or nHC-HA / PTX3 complex in the fetal support tissue is substantially maintained for at least 15 days. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 20 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 25 days from initial procurement.In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 30 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 35 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 40 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 45 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 50 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 55 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 60 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 65 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 70 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 75 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 80 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 85 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 90 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 95 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 100 days from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least 180 days from initial procurement.In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least one year from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least two years from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least three years from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least four years from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least five years from initial procurement. In some embodiments, the biological and structural integrity of the fetal support tissue is substantially maintained for at least one day to at least five years from initial procurement. In some embodiments, the fetal support tissue is obtained from an animal, a mammal, e.g., a human or a non-human animal, e.g., a primate, a bovine, or a porcine.

[0063] In some embodiments, the fetal support tissue is kept below 0° C. until donor and specimen eligibility is determined. In some embodiments, the fetal support tissue is kept between 0° C. and −80° C. until donor and specimen eligibility is determined. In some embodiments, storing the fetal support tissue at −80° C. kills substantially all cells found in the fetal support tissue. In some embodiments, storing the fetal support tissue at −80° C. kills substantially all cells found in the fetal support tissue while maintaining or increasing the biological activity of the fetal support tissue (e.g., its anti-inflammatory, anti-scarring, anti-antigenic, and anti-adhesion properties) compared to fresh (i.e., not frozen) fetal support tissue. In some embodiments, storing the fetal support tissue at −80° C. results in loss of metabolic activity of substantially all cells found in the fetal support tissue. In some embodiments, loss of metabolic activity may result in cell death. In some embodiments, the cells may lose metabolic activity while maintaining the biological activity of the fetal support tissue or any biomolecules present in the fetal support tissue, including, but not limited to, HC-HA / PTX3. In some embodiments, the fetal support tissue is dried. In some embodiments, the fetal support tissue is not dehydrated. Disclosed herein are processes for producing fetal support tissue while retaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims.

[0064] Fetal Support Tissue Processing Disclosed herein, and throughout this disclosure, are fetal support tissue procurement and processing steps or protocols as shown in detail in Figures 1A-1B, and 2A, 2B, and 2C. Whenever fetal support tissue is disclosed herein, it should be taken, without limitation, that the fetal support tissue is pooled fetal support tissue, where the pooling may be pooled fetal support tissue that occurs prior to the processing steps of a given fetal support tissue as shown in Figures 1A-1B, and 2A-2B, and 2C, or where the pooling may be pooled fetal support tissue that occurs after the processing steps of a given fetal support tissue.

[0065] In some embodiments, processing is performed in accordance with Good Tissue Practices (GTPs) that prevent the introduction, transmission, contamination, or cross-contamination of diseases that may be transmitted from the donor.

[0066] In some embodiments, the fetal support tissue is tested for active viral infections, such as, for example, active HIV-1, HIV-2, HTLV-1, Hepatitis B and C, West Nile Virus, Cytomegalovirus, and Treponema pallidum, using FDA approved screening tests. In some embodiments, any indication that the tissue is contaminated with an active viral infection, such as HIV-1, HIV-2, HTLV-1, Hepatitis B and C, West Nile Virus, or Cytomegalovirus, will result in the immediate quarantine and subsequent destruction of the tissue specimen. In some embodiments, the donor's medical records are reviewed for risk factors and clinical evidence of Hepatitis B, Hepatitis C, or HIV infection.

[0067] In some embodiments, the fetal support tissue is frozen before being pooled. In some embodiments, the fetal support tissue is frozen after being pooled. In some embodiments, the pooled fetal support tissue is not frozen. In some embodiments, the not frozen fetal support tissue is not pooled. If the fetal support tissue is not frozen, it is first pooled and then processed as described below just before being frozen. In some embodiments, if the fetal support tissue is not frozen, it is processed just before being pooled or just before being frozen as described below.

[0068] In some embodiments, substantially all of the blood is removed from the fetal support tissue (e.g., from any arteries and veins found in the fetal support tissue, as well as from blood that has infiltrated the tissue) before pooling. In some embodiments, substantially all of the blood is removed before pooling and before the fetal support tissue is frozen. In some embodiments, substantially all of the blood is removed before pooling and after the fetal support tissue is frozen. In some embodiments, substantially all of the blood is not removed from the fetal support tissue (e.g., from any arteries and veins found in the fetal support tissue, or from blood that has infiltrated the tissue) before pooling. In some embodiments, pooling of the fetal support tissue occurs after substantially all of the blood has been removed from the fetal support tissue. In some embodiments, pooling of the fetal support tissue occurs before substantially all of the blood has been removed from the fetal support tissue. For example, blood is not removed before pooling and before the fetal support tissue is frozen. In some embodiments, substantially all of the blood is not removed before pooling or after the fetal support tissue is frozen. In some embodiments, blood is removed from the fetal support tissue before pooling. In some embodiments, blood is not removed from the fetal support tissue before pooling. In some embodiments, blood is removed from the fetal support tissue after pooling. In some embodiments, blood is not removed from the fetal support tissue after pooling. In some embodiments, blood is not removed from the fetal support tissue and may be pooled after the blood is removed. In some embodiments, blood is removed from the fetal support tissue before the fetal support tissue is pooled but before the fetal support tissue is frozen. In some embodiments, blood is not removed from the fetal support tissue after the fetal support tissue is pooled and before the fetal support tissue is frozen. In some embodiments, blood is substantially removed before the fetal support tissue is pooled and after the fetal support tissue is frozen. In some embodiments, blood is substantially removed after the fetal support tissue is pooled and after the fetal support tissue is frozen.

[0069] In some embodiments, the fetal support tissue is washed with a buffer solution, optionally under agitation, before being pooled, to remove excess blood and tissue. In some embodiments, after the fetal support tissue is pooled, the fetal support tissue is washed with a buffer solution, under agitation, to remove excess blood and tissue. In some embodiments, the fetal support tissue is first pooled before excess blood is removed by agitation with a buffer solution. In some embodiments, the fetal support tissue is not submerged with a buffer solution, under agitation, to remove excess blood and tissue, before being pooled. In some embodiments, after the fetal support tissue is pooled, the fetal support tissue is not submerged with a buffer solution, under agitation, to remove excess blood and tissue.

[0070] In some embodiments, the fetal support tissue is a fetal support tissue graft. In some embodiments, isolated fetal support tissue is used to generate the fetal support tissue graft before being pooled. In some embodiments, the fetal support tissue is cut into multiple pieces (e.g., using a scalpel). The size of the pieces depends on the desired use of the fetal support tissue derived fetal support tissue graft. In some embodiments, the cut fetal support tissue is optionally washed again with a buffer to further remove excess blood and tissue.

[0071] The umbilical cord (UC) contains two arteries (the umbilical artery) and one vein (the umbilical vein). In some embodiments, the vein and artery are removed from the UC. In some embodiments, the vein and artery are not removed from the UC. In certain cases, the vein and artery are enclosed (or suspended or embedded) within Wharton's jelly. In some embodiments, the vein and artery are removed at the same time that the Wharton's jelly is partially removed.

[0072] The desired thickness of the fetal support tissue determines how the fetal support tissue is processed. In some embodiments, the desired thickness of the fetal support tissue determines how much Wharton's jelly is removed. In some embodiments, the fetal support tissue is contacted with a buffer to facilitate separation of the Wharton's jelly and the umbilical cord amniotic membrane. In some embodiments, the Wharton's jelly is removed using abrasion, rotablator (i.e., a catheter attached to a drill with a diamond-coated burr tool), liposuction, liquid under high pressure, a brush (e.g., a brush mechanized to rotate under high speed), or a surgical dermatome. In some embodiments, the Wharton's jelly is not removed from the fetal support tissue. In some embodiments, the Wharton's jelly and the umbilical vein and umbilical artery are not removed. In some embodiments, the Wharton's jelly is not removed, and the umbilical vein and umbilical artery are removed from the fetal support tissue before or after pooling.

[0073] In some embodiments, the fetal support tissue comprises isolated umbilical cord amniotic membrane (UCAM). Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims. In some embodiments, the UCAM is isolated by partially removing Wharton's jelly and a portion of the umbilical blood vessels from the UC, leaving the UCAM. After the substantially pure UCAM is obtained, the UCAM is optionally washed with a buffer to remove excess blood and tissue. In some embodiments, the UCAM comprises Wharton's jelly. In some embodiments, the UCAM comprises Wharton's jelly and the umbilical vein and umbilical artery. In some embodiments, the UCAM comprises Wharton's jelly, but does not comprise the umbilical vein and umbilical artery.

[0074] In some embodiments, the fetal support tissue is any suitable shape (e.g., square, circular, triangular, rectangular). In some embodiments, the fetal support tissue is generated from a sheet of fetal support tissue. In some embodiments, the sheet is flat. In some embodiments, the sheet is tubular.

[0075] In some embodiments, the fetal support tissue is cut into multiple sections (e.g., using a scalpel). In some embodiments, the fetal support tissue is divided into sections of about 1.0 cm by about 0.25 cm, 0.5 cm, 0.75 cm, 1.0 cm, 2.0 cm, 3.0 cm, 4.0 cm, 5.0 cm, or 6 cm. In some embodiments, the fetal support tissue is divided into sections of about 2 cm by about 2 cm, 3 cm, 4 cm, 5 cm, or 6 cm. In some embodiments, the fetal support tissue is divided into sections of about 3 cm by about 3 cm, 4 cm, 5 cm, or 6 cm. In some embodiments, the fetal support tissue is divided into sections of about 4 cm by about 4 cm, 5 cm, or 6 cm. In some embodiments, the fetal support tissue is divided into sections of about 5 cm by about 5 cm or 6 cm. In some embodiments, the fetal support tissue is divided into sections of about 6 cm by about 6 cm. In some embodiments, the fetal support tissue is divided into sections of about 8 cm by about 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, or 8 cm. In some embodiments, the fetal support tissue is divided into sections of about 10 cm by about 10 cm. In some embodiments, the fetal support tissue is divided into sections of about 12 cm by about 10 cm. In some embodiments, the fetal support tissue is divided into sections of about 15 cm by about 10 cm. In some embodiments, the fetal support tissue is divided into sections of about 20 cm by about 10 cm. In some embodiments, the fetal support tissue is divided into sections of about 25 cm by about 10 cm. In some embodiments, the fetal support tissue is divided into sections of about 30 cm by about 10 cm.

[0076] In some embodiments, the fetal support tissue is contacted with the buffer under agitation to remove substantially all remaining red blood cells. In some embodiments, the fetal support tissue is contacted with the buffer for 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 40 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 18 hours, 24 hours, or more than 24 hours. In some embodiments, the UC product is contacted with the buffer for 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or more than 4 weeks. The fetal support tissue may be washed 1, 2, 3, 4, 5, or more times. The fetal support tissue may be translucent after observing that there is no visible trace of blood.

[0077] Multiple donor pooling In certain embodiments, a method for producing a pooled fetal support tissue product is provided herein. The process of pooling fetal support tissue leads to a reduction in sample-to-sample variation as seen in single donors due to the high variability in single donor products. Also, it is difficult to implement, develop, and validate potency assays on single donor products to ensure consistency. However, pooling donor samples has been shown to result in increased HA protein content, reduced sample-to-sample variation, and improved sample uniformity, suggesting that pooling donor fetal support tissue increases the consistency and uniformity of the fetal support tissue product or extract produced. In some embodiments, pooling fetal support tissue results in a reduction in sample-to-sample variation. For example, the resulting standard deviation or coefficient of variation rate after processing of single donor samples may be higher than that obtained from processing of pooled samples. In some embodiments, the variation between processing of single donor samples and processing of pooled donor tissue samples may be statistically significant. The variation obtained after single donor samples are processed may be one or more orders of magnitude higher than the variation after pooled tissue samples are processed. For example, assays performed using pooled donor samples may have at least 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, or more reduced variation than assays performed using single donor samples. As disclosed herein, pooling fetal tissue samples from multiple or multiple donors allows for efficient assessment and management of quality control processes while scaling up production processes and manufacturing without compromising product potency, consistency, stability, uniformity, or purity. In some embodiments, pooling of fetal support tissue at various stages of processing of fetal support tissue is disclosed herein, using parameters recommended for large-scale processing and commercialization. Pooling of fetal support tissue products occurs at each production stage to significantly eliminate challenges that may occur during single donor manufacturing and processing (some of which are described above and known in the art).

[0078] In some embodiments, the amount of HA obtained from pooled fetal support tissues may be, on average, higher than the average HA obtained from a collection of single donor samples. For example, the amount of HA in two or more pooled donor samples is at least 5 μg / mL, 10 μg / mL, 15 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 50 μg / ml, 60 μg / ml, 70 μg / ml, 80 μg / ml, 90 μg / ml, 100 μg / ml, 120 μg / ml, 130 μg / ml, 140 μg / ml, 150 μg / ml, 160 μg / ml, 170 μg / ml, 180 μg / ml, 190 μg / ml, 200 μg / ml, 210 μg / ml, 220 μg / ml, 230 μg / ml, 240 μg / ml, 250 μg / ml, 260 μg / ml, 270 μg / ml, 280 μg / ml, 290 μg / ml, 300μg / ml, 310μg / ml, 320μg / ml, 330μg / ml, 340μg / ml, 350μg / ml, 360μg / ml, 370μg / ml, 380μg / ml, 390μg / ml, 400μg / ml, 410μg / ml, 420μg / ml, 430μg / ml, 440μg / ml, 450μg / ml, 4 The range may be from 60 μg / ml, 470 μg / ml, 480 μg / ml, 490 μg / ml to about 500 μg / ml, 510 μg / ml, 520 μg / ml, 530 μg / ml, 540 μg / ml, 550 μg / ml, 560 μg / ml, 570 μg / ml, 580 μg / ml, or 590 μg / ml to about 600 μg / ml.In some embodiments, the amount of protein obtained from two or more pooled donor samples is at least 5 μg / mL, 10 μg / mL, 15 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 50 μg / ml, 100 μg / ml, 120 μg / ml, 130 μg / ml, 140 μg / ml, 150 μg / ml, 160 μg / ml, 170 μg / ml, 180 μg / ml, 190 μg / ml, 200 μg / ml, 210 μg / ml, 220 μg / ml, 230 μg / ml, 240 μg / ml, 250 μg / ml, 260 μg / ml, 270 μg / ml, 280 μg / ml, 290 μg / ml, 300 μg / ml μg / ml, 180μg / ml, 190μg / ml, 200μg / ml, 220μg / ml, 230μg / ml, 240μg / ml, 250μg / ml, 260μg / ml, 2 70μg / ml, 280μg / ml, 290μg / ml, 300μg / ml, 310μg / ml, 320μg / ml, 330μg / ml, 340μg / ml, 350μg / ml, 360μg / ml, 370μg / ml, 380μg / ml, 390μg / ml, 400μg / ml, 410μg / ml, 420μg / ml, 430μg / ml, 440μg / m l, 450μg / ml, 460μg / ml, 470μg / ml, 480μg / ml, 490μg / ml~approx. 500μg / ml, 510μg / ml, 520μg / ml, 530μ g / ml, 540μg / ml, 550μg / ml, 560μg / ml, 570μg / ml, 580μg / ml, 590μg / ml, 600μg / ml, 650μg / ml, 70 It may be 0 μg / ml, 750 μg / ml, 800 μg / ml, 850 μg / ml, 900 μg / ml, 950 μg / ml, 1000 μg / ml to about 2000 μg / ml.

[0079] In some embodiments, pooling fetal support tissues increases the amount of protein content produced from the fetal support tissue. In some embodiments, the amount of total protein obtained from pooled fetal support tissues may be higher on average than the average total protein of a single donor. In some embodiments, the amount of total protein obtained from two or more pooled donor samples may be higher than about 5 μg / mL, 10 μg / mL, 15 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, 50 μg / ml, 100 μg / ml, 120 μg / ml, 130 μg / ml, 140 μg / ml, 150 μg / ml, depending on the number of donor samples pooled. , 160μg / ml, 170μg / ml, 180μg / ml, 190μg / ml, 200μg / ml, 220μg / ml, 230μg / ml, 240μg / ml, 250μg / ml , 260μg / ml, 270μg / ml, 280μg / ml, 290μg / ml, 300μg / ml, 310μg / ml, 320μg / ml, 330μg / ml, 340μg / ml, 350μg / ml, 360μg / ml, 370μg / ml, 380μg / ml, 390μg / ml, 400μg / ml, 410μg / ml, 420μg / ml, 430μg / ml, 440μg / ml, 450μg / ml, 460μg / ml, 470μg / ml, 480μg / ml, 490μg / ml~about 500μg / ml, 510μg / ml, 520μg / ml, 530μg / ml, 540μg / ml, 550μg / ml, 560μg / ml, 570μg / ml, 580μg / ml, 590μg / ml, 600μg / ml, 650μg / ml, It may exceed 700 μg / ml, 750 μg / ml, 800 μg / ml, 850 μg / ml, 900 μg / ml, 950 μg / ml, 1000 μg / ml to about 2000 μg / ml.

[0080] In some cases, the method of producing a pooled fetal support tissue product includes any method of producing a fetal support tissue product described herein and a pooling step. In some cases, the pooling step includes pooling fetal support tissue derived from multiple donors. In some cases, the pooling step includes pooling fetal support tissue products derived from multiple donors to produce a pooled composition (e.g., a pooled drug substance). In some cases, the fetal support tissue product is produced by a method described herein, which may include processing including (a) micronizing or cryo-milling the fetal support tissue to produce a micronized or cryo-milled fetal support tissue, (b) extracting the micronized or cryo-milled fetal support tissue in an excipient to produce an extract, and (c) sterilizing the extract by filtration (liquid extract) or sieving (powder extract) to produce the fetal support tissue product. In some embodiments, the processing of the fetal support tissue (a) may occur before or after the fetal support tissue is pooled. In some embodiments, the processing (a) of the fetal support tissue may occur before the fetal support tissue is lyophilized. In some embodiments, the processing (a) of the fetal support tissue may occur after the fetal support tissue is lyophilized. In some embodiments, the lyophilization of the fetal support tissue may occur before or after it is micronized or cryo-milled, or before or after the donor fetal support tissue is pooled. In some cases, the donor lot is a placenta or any other fetal support tissue material coming from a single donor. In some cases, the pooling step includes pooling fetal support tissue products derived from multiple donor lots to generate a pooled composition (e.g., a pooled drug substance). In some cases, the pooling step includes pooling fetal support tissue (e.g., fetal support tissue products) from (e.g., derived from) at least 15 donors to generate a pooled composition. In some cases, the pooling step includes pooling fetal support tissue (e.g., fetal support tissue products) from at least 30 donors to generate a pooled composition.In some cases, the pooling step includes pooling fetal support tissue (e.g., fetal support tissue products) of at least 45 donors to generate a pooled composition. In some cases, the pooling step includes pooling fetal support tissue (e.g., fetal support tissue products) of at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or more than 100 donors to generate a pooled composition. In some cases, the pooling step includes pooling fetal support tissue (e.g., fetal support tissue products) of up to 5 donors. In some cases, the pooling step includes pooling fetal support tissue (e.g., fetal support tissue products) of up to 9, 8, 7, 6, 5, 4, 3, or 2 donors. In some cases, the pooling step includes pooling fetal support tissue products of two or more donors. In some cases, the pooling step includes pooling multiple batches of fetal support tissue products to generate a pooled composition. In some embodiments, the pooling step includes pooling 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 batches. In some embodiments, 3 or fewer batches are pooled to generate a pooled composition. In some embodiments, the batch is a pooled fetal support tissue composition from fetal support tissue products derived from multiple donors. In some cases, the batch may include fetal support tissue products derived from 15 donors. In some embodiments, the batch is pooled from at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or more than 100 donors. In some cases, batches are pooled from up to 9, 8, 7, 6, 5, 4, 3, or 2 donors. In some cases, the pooling includes pooling three batches, each batch containing fetal support tissue from 15 donors.In the steps of pooling fetal support tissue described herein, pooling may occur before or after the fetal support tissue is freeze-dried, with freeze-drying of the fetal support tissue occurring before or after pooling.

[0081] Pool process timing Disclosed herein are exemplary process illustrations (FIGS. 1A-1B and 2A-2C) showing the processing and pooling points of fetal support tissue material processed and manufactured under commercial production conditions. As shown in FIGS. 1A-1B and 2A-2C, potential donors are screened, raw material (fetal support tissue) is collected from the donors, transported to a processing facility, received and inspected, and donor eligibility is determined. In some embodiments, biological samples are frozen at -80°C before being pooled or processed by washing, cutting, and immersion in 0.9% saline. In some embodiments, biological samples are processed by washing, cutting, and immersion in 0.9% saline before freezing. Exemplary steps or procedures for processing and pooling of fetal support tissue, including parameters and in-process controls and critical quality attributes, are as shown in FIGS. 1A-1B and 2A-2C. After completing the assessment of the listed critical quality attributes, the biological sample (fetal support tissue material) becomes the regulatory starting material (RSM).

[0082] In some embodiments, the raw pharmaceutical starting material (RSM) or fetal support tissue material is pooled and processed to produce a product in sheet or non-sheet form. In some embodiments, the raw pharmaceutical starting material or fetal support tissue material is processed into a product in sheet form and non-sheet form before being pooled. In some embodiments, the pooled fetal support tissue is processed by micronization. In some embodiments, micronization is used to produce a gel product. In some embodiments, the pooled fetal support tissue that has been micronized into a dry powder, liquefied, and is further filtered before or after terminal sterilization. In some embodiments, the micronized fetal support tissue in liquid form may be centrifuged before or after terminal sterilization, for example, using gamma sterilization or a combination thereof. In some embodiments, filtration adjusts the sterilization of the fetal support tissue extract in liquid form, utilizing a filter pore size of 0.2 μm or less than about 0.2 μm. In some embodiments, the dry powder form obtained by micronization of the fetal support tissue is not liquefied, but is sieved and either used immediately or frozen to -80°C. In some embodiments, the fetal support tissue extract is micronized and sieved to control particle size. In some embodiments, the final sterilization of the dry powder extract occurs, for example, through a process of gamma sterilization. In some embodiments, the fetal support tissue may be cryo-milled to obtain a fine extract or liquefied. In some embodiments, the cryo-milled fetal support tissue, which is a powder extract, may be further sieved to obtain particles of different sizes. In some embodiments, the cryo-milled fetal support tissue, which is a fine powder, may be terminally sterilized, for example, by using gamma sterilization. In some embodiments, the fetal support tissue extract may be liquefied, and may be either filtered, centrifuged, or a combination thereof, sterilized (filter pore size of 0.2 μm or less than about 0.2 μm), or particle size controlled. In some embodiments, the fetal support tissue may be decellularized, devitalized, micronized, cryo-milled, or a combination thereof.In some embodiments, the micronized, cryo-grinded, decellularized, devitalized fetal support tissue extract may be further filtered, centrifuged (liquid form), or sieved (dry powder) in various (or combinations thereof) to obtain fetal support tissue products of various particle sizes. In some embodiments, the fetal support tissue may not be pooled before being processed (by micronization, cryo-grinding, decellularization, devitalization, centrifugation, sterilization, e.g., gamma sterilization, sieving, filtration, or combinations thereof) to obtain fetal support tissue products of various particle sizes.

[0083] In some embodiments, fetal support tissue may be freeze-dried, pulverized, cryo-grinded, decellularized, or devitalized, or a combination thereof, before being pooled. For example, fetal support tissue may be freeze-dried, pulverized, cryo-grinded, decellularized, or devitalized, or a combination thereof, before being pooled, and may be sieved (dry powder) or centrifuged, filtered (liquid form), terminally sterilized, such as gamma-irradiated, or a combination thereof, to obtain fetal support tissue products of various particle sizes. In some embodiments, fetal support tissue may be freeze-dried and pooled before being processed by pulverization, cryo-grinding, decellularization, devitalization, or a combination thereof. In some embodiments, the fetal support tissue may be freeze-dried and pooled prior to micronization, cryogenic grinding, decellularization, devitalization, or a combination thereof, may be subjected to sieving (dry powder) and terminal sterilization procedures to obtain a filter-sterilized fetal support tissue product or a fetal support tissue product of various particle sizes, or if in liquid form, may be centrifuged, filtered, or a combination thereof.

[0084] In some embodiments, the pooled fetal support tissue may be freeze-dried before being processed by pulverization, cryo-grinding, decellularization, devitalization, or a combination thereof, or may be freeze-dried before being sieved and / or sterilized from the dry powder fetal support tissue extract, or before being centrifuged, sterilized, filtered from the liquid form fetal support tissue extract / product, or a combination thereof, to provide a range of fetal support tissue products of different particle sizes. In some embodiments, the pooled fetal support tissue may not be freeze-dried before being processed by pulverization, cryo-grinding, decellularization, devitalization, or a combination thereof. In some embodiments, the fetal support tissue may not be freeze-dried before being sieved and / or sterilized from the dry powder fetal support tissue extract, or before being centrifuged, sterilized, filtered from the liquid form fetal support tissue extract / product, or a combination thereof, to provide a range of fetal support tissue products of different particle sizes.

[0085] In some embodiments, the particle size of the processed pooled fetal support tissue product may be less than 5 μm. In some embodiments, the particle size of the pooled processed fetal support tissue product may be less than about 5 μm. In some embodiments, the particle size of the pooled processed fetal support tissue may be about 5 to about 10 μm. In some embodiments, the particle size of the pooled processed fetal support tissue product may be less than 1 μm, less than about 1 μm, less than about 10 μm to about 15 μm. In some embodiments, the pooled fetal support tissue has a particle size in any range disclosed in the specification and claims of the present disclosure, as well as any combination thereof.

[0086] In some embodiments, the particle size of the fetal support tissue product before pooling may be less than 5 μm. In some embodiments, the particle size of the processed fetal support tissue product before pooling may be less than about 5 μm. In some embodiments, the particle size of the processed fetal support tissue before pooling may be about 5 to about 10 μm. In some embodiments, the particle size of the processed fetal support tissue product before pooling may be less than 1 μm, less than about 1 μm, less than about 10 μm to about 15 μm. In some embodiments, the fetal support tissue before pooling has a particle size in any range disclosed in the specification and claims of the present disclosure, and any combination thereof. In some embodiments, the fetal support tissue after the fetal support tissue is pooled has a particle size in any range disclosed in the specification and claims of the present disclosure, and any combination thereof. In some embodiments, the particle size of the pooled fetal support tissue product may be less than 5 μm. In some embodiments, the particle size of the pooled processed fetal support tissue product may be less than about 5 μm. In some embodiments, the particle size of the pooled processed fetal support tissue may be about 5 to about 10 μm. In some embodiments, the pooled, processed fetal support tissue product may have a particle size of less than 1 μm, less than about 1 μm, less than about 10 μm to about 15 μm, In some embodiments, the pooled fetal support tissue has a particle size in any of the ranges disclosed in the specification and claims of the present disclosure, and any combination thereof.

[0087] In some cases, the pooling step comprises a sieving step, hi some embodiments, the sieving step controls the maximum particle size of the pooled composition (e.g., bulk drug substance). In some embodiments, the sieve has an average pore size of about 0.1-0.2 μm or less, 0.2-0.3 μm or less, 0.3-0.4 μm or less, 0.4-0.5 μm or less, 0.5-0.6 μm or less, 0.6-0.7 μm or less, 0.7-0.8 μm or less, 0.8-0.9 μm or less, 0.9-1 μm or less, 1-2 μm or less, 2-3 μm or less, 3-4 μm or less, 4-5 μm or less, 5-10 μm or less, 10-20 μm or less, 20-30 μm or less, 30-40 μm or less, 40-50 μm or less, 50-100 μm or less, 100-150 μm or less, 150-200 μm or less, 200-250 μm or less, or 250-300 μm or less. In some cases, the pooling step produces compositions that have higher yields, improved stability, reduced variability, improved potency, uniformity, consistency, or a combination thereof, of HC-HA / PTX3, HA, and other proteins of interest, as compared to compositions produced by the same method but without undergoing the pooling step.

[0088] In some embodiments, particle size is adhered to when fetal support tissue extracts are administered by inhalation or intravenously as disclosed herein. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of fetal support tissue, including components of fetal support tissue such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims.

[0089] Preparation of Pooled Compositions In some aspects, as disclosed herein, pooled compositions may be prepared from pooling fetal support tissue from multiple donors, compared to non-pooled compositions (referred to as FLO). Prior to pooling, the micronized drug substance in each individual pool may be analyzed for hyaluronic acid content, total protein, and ODI-TRAP. Exemplary results from a procedure in which results are combined to produce a weighted average of each result prior to pooling are found in Figures 1A-1B, which show the pooling process from multiple donors. In some embodiments, stability testing may be performed with compositions produced by pooling donor fetal support tissue samples. As disclosed herein, results from this procedure met the acceptance criteria at T=0, 1, 3, and 6 months. Data ranges for each of the CQAs are shown in Tables 1 and 2.

[0090] Extraction and centrifugation In some embodiments, the extraction process produces either a dry powder or liquid form of the pooled fetal support tissue product, in some embodiments, the liquid form is centrifuged and purified to control particle size, e.g., to control the particle size of the fetal support tissue containing HC-HA / PTX3, HA, growth factors, cytokines, and other bioactive components. In some embodiments, the above extracts, liquids, or powders of fetal support tissues may be used to purify the nHC-HA / PTX3 complex by centrifugation (e.g., ultracentrifugation, gradient centrifugation), chromatography (e.g., ion exchange chromatography, affinity chromatography, size exclusion chromatography, and hydroxyapatite chromatography), gel filtration, or fractional lysis, ethanol precipitation, or other techniques available for protein purification (see, e.g., Scopes, Protein Purification Principles and Practice 2nd Edition, Springer-Verlag, New York, 1987; Higgins, SJ and Hames, BD (eds.), Protein Expression: A Practical Approach, Oxford Univ Press, 1999; and Deutscher, MP, Simon, MI, Abelson, JN (eds.), Guide to Protein Purification: Methods in Enzymology (Methods in Enzymology Series, Vol 182), Academic Press, 1997, all of which are incorporated herein by reference).

[0091] In some embodiments, nHC-HA / PTX3 is isolated from an extract of pooled fetal support tissue. In some embodiments, the extract is prepared from an amniotic membrane extract. In some embodiments, the extract is prepared from an umbilical cord extract. In some embodiments, the umbilical cord extract comprises umbilical cord stroma and / or Wharton's jelly, or a combination thereof. In some embodiments, the nHC-HA / PTX3 complex is included in an extract prepared by ultracentrifugation. In some embodiments, the nHC-HA / PTX3 complex is included in an extract prepared by ultracentrifugation using a gradient of CsCl / 4-6M guanidine HCl. In some embodiments, the extract is prepared by at least two rounds of ultracentrifugation. In some embodiments, the extract is prepared by more than two rounds of ultracentrifugation (i.e., second round of nHC-HA / PTX3). In some embodiments, the extract is prepared by at least four rounds of ultracentrifugation (i.e., fourth round of nHC-HA / PTX3). In some embodiments, the nHC-HA / PTX3 complex comprises small leucine-rich proteoglycans. In some embodiments, the nHC-HA / PTX3 complex comprises HC1, HA, PTX3, and / or a small leucine-rich proteoglycan.

[0092] In some embodiments, the extraction is performed on fetal support tissue. In some embodiments, the extraction comprises separating the protein of interest of the fetal support tissue from other components. In some embodiments, the extraction is performed on cryo-ground fetal support tissue. In some embodiments, the extraction comprises separating the protein of interest of the cryo-ground fetal support tissue from other components. In some embodiments, such extraction, with or without centrifugation, is aimed at maintaining or preserving the HA that is also present in the nHC-HA / PTX3.

[0093] In some embodiments, the extract is prepared by extraction in an excipient. In some embodiments, the excipient is saline, water, structured water, water for injection (WFI), or a combination thereof. In some embodiments, the excipient is WFI. In some embodiments, the use of WFI as an excipient produces higher recovery of HA and total protein compared to saline or structured water. In some embodiments, the majority of nHC-HA / PTX3 is extracted by extraction in saline or WFI.

[0094] In some embodiments, the extraction is performed for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12 hours-24 hours, about 24 hours-48 hours, or about 48 hours-72 hours. In some embodiments, the extraction is performed for about 1 hour. In some embodiments, the extraction is performed in WFI for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12 hours-24 hours, about 24 hours-48 hours, or about 48 hours-72 hours. In some embodiments, the extraction is performed in WFI for about 1 hour. In some embodiments, the extraction is carried out in saline for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12 hours to 24 hours, about 24 hours to 48 hours, or about 48 hours to 72 hours. In some embodiments, the extraction is carried out in saline for about 1 hour.

[0095] In some embodiments, the extraction is performed at a temperature of about 4° C. In some embodiments, the extraction is performed at a temperature of about 3° C., 4° C., 5° C., or 6° C. In some embodiments, the extraction is performed in WFI at a temperature of about 3° C., 4° C., 5° C., or 6° C. In some embodiments, the extraction is performed in WFI at a temperature of about 4° C. In some embodiments, the extraction is performed in WFI at a temperature of about 4° C. for about 1 hour. In some embodiments, the extraction is performed in WFI at a temperature of about 4° C. for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12 hours-24 hours, about 24 hours-48 hours, or about 48 hours-72 hours. In some embodiments, the extraction is performed in saline at a temperature of about 3° C., 4° C., 5° C., or 6° C. In some embodiments, the extraction is performed in saline at a temperature of about 4° C. In some embodiments, the extraction is carried out in saline for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12 hours to 24 hours, about 24 hours to 48 hours, or about 48 hours to 72 hours at a temperature of about 4° C. In some embodiments, the extraction is carried out in saline for about 1 hour at a temperature of about 4° C.

[0096] In some embodiments, the ratio of fetal support tissue to extraction vehicle is about 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 1:0.75, 1:0.5, or 1:0.25 (weight:volume). In some embodiments, the ratio of fetal support tissue to extraction vehicle is about 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 0.75:1, 0.5:1, or 0.25:1 (weight:volume). In some embodiments, the ratio of fetal support tissue to extraction vehicle is about 1:4. In some embodiments, the extraction is performed in WFI or saline. In some embodiments, the extraction is performed in WFI. In some embodiments, the extraction is performed for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12-24 hours, about 24-48 hours, or about 48-72 hours. In some embodiments, the extraction is performed for about 1 hour. In some embodiments, the extraction is performed at a temperature of about 3° C., 4° C., 5° C., or 6° C. In some embodiments, the extraction is performed at a temperature of about 4° C. in WFI with a fetal support tissue to excipient ratio of about 1:4 for 1 hour. In some embodiments, the extraction is performed in saline with a fetal support tissue to excipient ratio of about 1:4 for 1 hour at a temperature of about 4° C.

[0097] In some embodiments, the extraction is performed using a tube rotator. In some embodiments, the tube rotator rotates at a speed range of about 5-10 rpm, 10-20 rpm, 20-30 rpm, 30-40 rpm, or 40-50 rpm. In some embodiments, the tube rotator rotates at a speed of about 20 rpm. In some embodiments, the tube rotator rotates for about 0-1 hour, about 1-2 hours, about 2-3 hours, about 3-4 hours, about 4-5 hours, about 5-6 hours, about 6-12 hours, about 12 hours to 24 hours, about 24 hours to 48 hours, or about 48 hours to 72 hours. In some embodiments, the tube rotator rotates for about 1 hour. In some embodiments, the tube rotator rotates at a temperature of about 3° C., 4° C., 5° C., or 6° C. In some embodiments, the tube rotator rotates at a temperature of about 4° C. In some embodiments, the excipient used is saline or WFI. In some embodiments, the excipient used is WFI. In some embodiments, the ratio of fetal support tissue to the extraction excipient is about 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 1:0.75, 1:0.5, or 1:0.25 (weight:volume). In some embodiments, the ratio of fetal support tissue to the extraction excipient is about 1:4. In some embodiments, the ratio of fetal support tissue to the extraction excipient is about 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 0.75:1, 0.5:1, or 0.25:1 (weight:volume). In some embodiments, the extraction is performed in WFI at a temperature of about 4° C. using a tube rotator at a speed of about 20 rpm for about 1 hour. In some embodiments, the extraction is performed in saline at a temperature of about 4° C. using a tube rotator at a speed of about 20 rpm for about 1 hour.

[0098] In some embodiments, centrifugation is performed on the extract produced by the extraction process. In some embodiments, the extract is centrifuged for about 5-10 minutes, about 10-15 minutes, about 15-20 minutes, about 20-30 minutes, about 30 minutes to 1 hour, or about 1-2 hours. In some embodiments, the extract is centrifuged for about 30 minutes. In some embodiments, the extract is centrifuged at a speed of about 3,200 rcf to 10,000 rcf, about 10,000 to 14,000 rcf, about 14,000 to 32,000 rcf, or about 32,000 to about 48,000 rcf. In some embodiments, the extract is centrifuged at a speed of about 14,000 rcf or greater. In some embodiments, the extract is centrifuged at a speed of about 14,000 rcf or greater. In some embodiments, the extract is centrifuged for about 30 minutes at a speed of about 14,000 rcf or greater. In some embodiments, centrifugation has no or only a minimal effect on the content of the protein of interest in the extract.

[0099] In some embodiments, the extraction produces an extract that contains hyaluronan present in nHC-HA / PTX3. In some embodiments, extraction in saline or WFI results in less damage to the nHC-HA / PTX3 complex than extraction using a different excipient, such as structured water. In some embodiments, extraction in saline or WFI results in a greater yield of HA or nHC-HA / PTX3 complex than extraction using a different excipient, such as structured water. In some embodiments, the extract comprises greater than about 900 μg / g HA (wet weight) of the extract. In some embodiments, the extract comprises greater than about 1000 μg / g HA (wet weight) of the extract. In some embodiments, the extract comprises greater than about 1100 μg / g HA (wet weight) of the extract. In some embodiments, the extract comprises greater than about 1200 μg / g HA (wet weight) of the extract. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims.

[0100] Dilution Described herein, in certain embodiments, is a method of processing fetal support tissue, the method comprising a dilution step to produce different doses of fetal support tissue product. In some embodiments, the dilution step is performed on the fetal support tissue after it has undergone an extraction and centrifugation step. In some embodiments, the dilution step is performed on the fetal support tissue after it has undergone an extraction and centrifugation step and before it is sterile filtered. In some embodiments, diluting the extract (e.g., reducing the concentration of the protein of interest in the extract) is accomplished by mixing the extract with an excipient. In some embodiments, the excipient is saline, water, structured water, water for injection (WFI), or a combination thereof. In some embodiments, the excipient is WFI. In some embodiments, the extract is mixed with the excipient at a dilution factor of about 1-1.5, about 1.5-2, about 2-2.5, about 2.5-3, about 3-3.5, about 3.5-4, about 4-5, about 5-6, about 6-7, about 7-8, about 8-9, or about 9-10. In some embodiments, the dilution factor is greater than 5. In some embodiments, the dilution factor is greater than 10. In some embodiments, the dilution factor is about 2. In some embodiments, the excipient is WFI and the dilution factor is 2. In some embodiments, the excipient is saline and the dilution factor is 2. In some embodiments, the methods described herein further comprise mixing the extract and the excipient for about 10-30 minutes, about 30 minutes to 1 hour, or about 1 hour to 2 hours. In some embodiments, the extract and the excipient are mixed for about 30 minutes. In some embodiments, the extract and excipients are mixed at a speed of about 5-10 rpm, 10-20 rpm, 20-30 rpm, 30-40 rpm, or 40-50 rpm. In some embodiments, the extract and excipients are mixed at a speed of about 20 rpm. In some embodiments, the extract and excipients are mixed at a speed of about 20 rpm for about 1 hour. In some embodiments, the extract and excipients are mixed at a temperature of about 4° C. In some embodiments, the extract and excipients are mixed at a speed of about 20 rpm for about 1 hour at about 4° C. In some embodiments, the extract is diluted prior to the filtration step.In some embodiments, the extract is diluted after the filtration step.

[0101] In some embodiments, dilution increases the rate of filtration, or the recovery of one or more proteins of interest after filtration, or the potency of the extract (e.g., as measured by ODI-TRAP assay, M2 assay, NO assay, and / or WST-1 assay), or a combination thereof, compared to an undiluted extract containing the same fetal support tissue and excipients. In some embodiments, the diluted fetal support tissue comprises about 1 μg / ml to about 150 μg / ml of hyaluronan (HA). In some embodiments, the diluted fetal support tissue comprises about 1 μg / ml to about 90 μg / ml of hyaluronan (HA). In some embodiments, the diluted fetal support tissue comprises about 90 μg / ml to about 150 μg / ml of hyaluronan (HA).

[0102] Validation Assay: In some embodiments, the methods disclosed herein are used in an assay to validate a pooled fetal support tissue product. In some embodiments, the assay to validate a pooled fetal support tissue product, extract, composition, or combination thereof is an assay or method that includes performing a validation assay on the pooled fetal support tissue, the validation assay determining the amount of native HC-HA / PTX3 complex and / or its activity. In some embodiments, the assay disclosed herein verifies the concentration, potency, identity, range of retention of nHC-HA / PTX3, proteoglycans, cytokines, growth factors, and other biological components present in the fetal support tissue. In some embodiments, the method is for determining the degradation of nHC-HA / PTX3 complex or any other component present in the pooled fetal support tissue. In some embodiments, the method of observing the degradation of nHC-HA / PTX3 complex or any other factor present in the fetal support tissue may be performed, for example, when a degradation assay observes the degradation of nHC-HA / PTX3 or any component of the fetal support tissue. To observe stability or efficacy, such validation assays described herein may be used to observe the presence, potency, and / or activity of nHC-HA / PTX3 at any stage of processing, including, but not limited to, before pooling, after pooling, before lyophilization, after lyophilization, before micronization, after micronization, or after the finished product is stored, including ODI-TRAP (e.g., FIG. 3), NO production / release (e.g., FIG. 5), M2-IL-12 release (e.g., FIG. 4), or total HA content (e.g., FIG. 7).

[0103] Assay matrix: WST-1 assay: Potency of HC-HA / PTX3 complex and HC-HA / PTX3 extract against cancer In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 200 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 250 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 300 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 400 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 500 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 600 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 700 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 800 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 900 μg / ml. In some embodiments, the composition exhibits statistically significant inhibition of TRAP activity in an ODI-TRAP assay compared to a positive control when the HA content of the composition is at least 1,000 μg / ml.

[0104] Disclosed herein, in some embodiments, is a composition that exhibits significant inhibition of TRAP activity with an HA content of at least 50 μg / ml, 60 μg / ml, 70 μg / ml, 80 μg / ml, 90 μg / ml, 100 μg / ml, 150 μg / ml, 200 μg / ml, 250 μg / ml, 300 μg / ml, 350 μg / ml, 400 μg / ml, or at least 500 μg / ml, where inhibition is at least 70% inhibition of TRAP activity. Disclosed herein, in some embodiments, is a composition that exhibits significant inhibition of TRAP activity with at least 80% inhibition of TRAP activity. Disclosed herein, in some embodiments, is a composition that exhibits significant inhibition of TRAP activity with at least 85% inhibition of TRAP activity. In some embodiments, the HA content of the composition is at least 200 μg / ml. In some embodiments, the HA content of the composition is at least 250 μg / ml. In some embodiments, the HA content of the composition is at least 300 μg / ml. In some embodiments, the HA content of the composition is at least 350 μg / ml. In some embodiments, the HA content of the composition is about 300 μg / ml.

[0105] In some embodiments, the therapeutic efficacy is determined by WST-1 assay. For example, a pooled fetal support tissue extract, including a tissue extract containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as an umbilical cord extract, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at 1 μg / ml to 1,500 μg / ml (p<0.05). In some embodiments, a pooled fetal support tissue extract, including a tissue extract containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as an umbilical cord extract, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at least 1 μg / ml (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 10 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 50 μg / ml or less (p<0.05). In some embodiments, pooled fetal supportive tissue extracts, including HC-HA / PTX3 or tissue extracts containing HC-HA / PTX3 but not HA, such as umbilical cord extracts, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at at least 100 μg / ml or less (p<0.05).In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 200 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 250 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 300 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 350 μg / ml or less (p<0.05). In some embodiments, HC-HA / PTX3 but not HA or tissue extracts containing HC-HA / PTX3, such as pooled fetal support tissue extracts including umbilical cord extracts, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extracts are loaded at at least 400 μg / ml or less (p<0.05).In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 500 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 550 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 600 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 650 μg / ml or less (p<0.05). In some embodiments, pooled fetal supportive tissue extracts, including HC-HA / PTX3 or tissue extracts containing HC-HA / PTX3 but not HA, such as umbilical cord extracts, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at at least 700 μg / ml or less (p<0.05).In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 750 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 800 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 850 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or inhibit tumor metabolic acidity compared to a positive control when the extracts are loaded at least 900 μg / ml or less (p<0.05). In some embodiments, pooled fetal supportive tissue extracts, including HC-HA / PTX3 or tissue extracts containing HC-HA / PTX3 but not HA, such as umbilical cord extracts, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at at least 950 μg / ml or less (p<0.05).In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at least 1,000 μg / ml or less (p<0.05). In some embodiments, pooled fetal support tissue extracts, including tissue extracts containing HC-HA / PTX3 or HC-HA / PTX3, but not HA, such as umbilical cord extracts, can inhibit cancer or tumor metabolic acidity compared to a positive control when the extract is loaded at more than about 90 μg / ml, 100 μg / ml, 200 μg / ml, 300 μg / ml, 500 μg / ml, 500 μg / ml, or 1,500 μg / ml (p<0.05).

[0106] Identity and potency of fetal support tissue compositions by use of ODI-TRAP assay matrix. In one embodiment described herein, the composition of pooled fetal support tissue comprises a potency determined by ODI-TRAP. The cell-based ODI-TRAP assay is a quantitative bioassay that measures the specific activity of pooled fetal support tissue based on its activity in inhibiting TRAP activity. The inhibitory activity can be evaluated in a comparative assay, generally evaluating and detecting inhibition by ODI-TRAP (e.g., FIG. 6B), or can be configured to evaluate and detect TRAP inhibition in single donor sample lots (e.g., FLO, FIG. 10A) relative to inhibition detected in pooled sample lots (FIG. 10B). In certain embodiments, pooled fetal support tissue possesses sufficient potency if TRAP demonstrates a statistically significant difference compared to a positive control for fetal support tissue having an HA content of at least 5 μg / ml, 10 μg / ml, 15 μg / ml, 20 μg / ml, 30 μg / ml, 40 μg / ml, 50 μg / ml, 100 μg / ml, 150 μg / ml, 200 μg / ml, 250 μg / ml, 300 μg / ml, 350 μg / ml, 400 μg / ml, 450 μg / ml, 500 μg / ml, 600 μg / ml, 700 μg / ml, 800 μg / ml, 900 μg / ml, 1,000 μg / ml. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue with an HA content of at least 5 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue with an HA content of at least 10 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue with an HA content of at least 15 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue with an HA content of at least 20 μg / ml compared to the positive control.In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 30 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 40 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 50 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 100 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 150 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 200 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 250 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 300 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 350 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 400 μg / ml compared to the positive control.In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 500 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 600 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 700 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 800 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue will possess sufficient potency when a statistically significant difference is shown for fetal support tissue having an HA content of at least 900 μg / ml compared to the positive control. In certain embodiments, the pooled fetal support tissue possesses sufficient potency if a statistically significant difference is demonstrated compared to the positive control for fetal support tissue having an HA content of at least 1,000 μg / ml.

[0107] In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 60% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 65% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 70% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 75% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 80% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 85% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 90% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml of HA exerts at least 95% TRAP inhibitory activity. In some embodiments, pooled fetal support tissue demonstrates sufficient TRAP potency when at least about 100 μg / ml HA exerts at least 97% TRAP inhibitory effect.In some embodiments, pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml HA exerts at least 98% TRAP inhibitory activity, hi some embodiments, pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 100 μg / ml HA exerts at least 99% TRAP inhibitory activity.

[0108] In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 60% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 65% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 70% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 75% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 80% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 85% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 90% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts at least 95% TRAP inhibitory activity. In some embodiments, pooled fetal support tissue demonstrates sufficient TRAP potency when at least about 300 μg / ml HA exerts at least 97% TRAP inhibitory effect.In some embodiments, pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml HA exerts at least 98% TRAP inhibitory activity, hi some embodiments, pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml HA exerts at least 99% TRAP inhibitory activity.

[0109] In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 60% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 65% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 70% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 75% TRAP inhibitory effect. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 80% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 85% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 90% TRAP inhibitory activity. In some embodiments, the pooled fetal support tissue will demonstrate sufficient TRAP potency when at least about 300 μg / ml of HA exerts greater than 95% TRAP inhibitory activity.

[0110] HA Assay: Hyaluronic acid (HA) is the main extracellular matrix in both AM and UC and is a component of the HC-HA / PTX3 complex. The HA assay disclosed herein is validated and tested to assess the stability of fetal support tissue extracts such as HC-HA / PTX3. HA assay is important for the composition of fetal support tissue or drug release, and HA assay is a measure of identity and potency. In certain embodiments, the hyaluronic acid (HA) in the pooled fetal support tissue and / or product possesses sufficient potency when the resulting HA content is at least 90 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products possess sufficient HA potency if the HA test kit has a content of at least 50 μg / ml, 60 μg / ml, 70 μg / ml, 80 μg / ml, 90 μg / ml, 100 μg / ml, 110 μg / ml, 120 μg / ml, 130 μg / ml, 140 μg / ml, 150 μg / ml "no less than" (NLT). In some embodiments, the pooled fetal support tissue extracts and / or products have a HA test kit with a concentration of at least 50 μg / ml, 60 μg / ml, 70 μg / ml, 80 μg / ml, 90 μg / ml, 100 μg / ml, 110 μg / ml, 120 μg / ml, 130 μg / ml, 140 μg / ml, 150 μg / ml, 160 μg / ml, 170 μg / ml, 180 μg / ml, 190 μg / ml, Sufficient HA potency is considered to be possessed when the content is "no less than" (NLT) 200μg / ml, 250μg / ml, 300μg / ml, 350μg / ml, 400μg / ml, 450μg / ml, 500μg / ml, 550μg / ml, 600μg / ml, 650μg / ml, 700μg / ml, 750μg / ml, 800μg / ml, 850μg / ml, 900μg / ml, 950μg / ml, 1,000μg / ml, 2,000μg / ml, or 3,000μg / ml.In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 50 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 60 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 70 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 80 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 90 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 100 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 110 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 120 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 130 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 140 μg / ml.In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 150 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 160 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 170 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 180 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency if the HA test kit has a content of at least NLT 190 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 200 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 250 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 300 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 350 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 400 μg / ml.In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 450 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 500 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 550 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 600 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 650 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 700 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 750 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 800 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 850 μg / ml. In some embodiments, the pooled fetal support tissue extracts and / or products will have sufficient HA potency when the HA test kit has a content of at least NLT 900 μg / ml.In some embodiments, the pooled fetal support tissue extracts and / or products possess sufficient HA potency if the HA test kit has a content of at least NLT 1,000 μg / ml.

[0111] Bicinchoninic Acid (BCA) Protein Assay (BCA): The BCA assay is a quantitative and analytical assay for measuring the total protein content of fetal support tissue and / or products, including HC-HA / PTX3 complex, tissue matrix, growth factors / cytokines, and other bioactive factors found in tissue or drug substance. The BCA assay is one of the first product release tests to assess identity and potency. The BCA assay disclosed herein is validated and tested to assess the stability of fetal support tissue extracts, such as HC-HA / PTX3. The BCA assay is important for the release of fetal support tissue compositions or drugs, and the BCA assay is a measure of identity and potency. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is at least 20 μg / ml, 30 μg / ml, 40 μg / ml, 50 μg / ml, 60 μg / ml, 70 μg / ml, 80 μg / ml, 90 μg / ml, 100 μg / ml, 110 μg / ml, 120 μg / ml, 130 μg / ml, 140 μg / ml, 150 μg / ml, 160 μg / ml, 170 μg / ml, 180 μg / ml, 190 μg / ml, 200 μg / ml, 220 μg / ml, 230 μg / ml, 240 μg / ml, 250 μg / ml, 260 μg / ml, 270 μg / ml, 280 μg / ml, 290 μg / ml, 300 μg / ml, 310 μg / ml, 320 μg / ml, 330 μg / ml, 340 μg / ml, 350 μg / ml, 360 μg / ml, 370 μg / ml, 380 μg / ml, 390 μg / ml, 400 μg / ml, 410 μg / ml, 420 μg / ml, 430 μg / ml, 440 μg / ml, 450 μg / ml, 460 μg / ml, 470 μg / ml, 480 μg / ml, 490 μg / ml, 500 μg / ml, 510 μg / ml, 520 μg / ml, 530 μg / ml, 540 μg / ml, 550 μg / ml, 560 μg / ml, 570 μg / ml, ml, 290μg / ml, 300μg / ml, 310μg / ml, 320μg / ml, 330μg / ml, 340μg / ml, 350μg / ml, 360 μg / ml, 370μg / ml, 380μg / ml, 390μg / ml, 400μg / ml, 450μg / ml, 500μg / ml, 550μg / ml, 6 The potency is sufficient if the assay detects total protein contents of "no less than" (NLT) 00μg / ml, 650μg / ml, 700μg / ml, 750μg / ml, 800μg / ml, 850μg / ml, 900μg / ml, 950μg / ml, 970μg / ml, 1,000μg / ml, 2,000μg / ml, 3,000μg / ml, 4,000μg / ml, and 5,000μg / ml.In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 30 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 40 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 50 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 60 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 70 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient when the BCA assay detects a total protein content of at least NLT 80 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 90 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 100 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 110 μg / ml.In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 120 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 130 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 140 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 150 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 160 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 170 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 180 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 190 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 200 μg / ml.In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 210 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 220 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 230 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 240 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 250 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 260 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 270 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 280 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 290 μg / ml.In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 300 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 310 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 320 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 330 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficiently potent when the BCA assay detects a total protein content of at least NLT 340 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 350 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 360 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 370 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product is sufficient potency when the BCA assay detects a total protein content of at least NLT 380 μg / ml.In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 390 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 400 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 450 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 500 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 550 μg / ml. In certain embodiments, the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 550 μg / ml. The total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 600 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 650 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 700 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 750 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 800 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 850 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 900 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 950 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 970 μg / ml. In certain embodiments, the total protein content in the pooled fetal support tissue and / or product will be sufficiently potent if the BCA assay detects a total protein content of at least NLT 1,000 μg / ml.

[0112] M2 type polarization assay based on NO measurement: Lipopolysaccharide (LPS) and interferon gamma (IFNγ) stimulate mouse macrophage cell line RAW264.7 cells to polarize to M1 macrophages, which are pro-inflammatory type, and produce high nitric oxide (NO) by upregulating inducible NO synthase (iNOS), one of the pro-inflammatory mediators, while M2 macrophages, which are anti-inflammatory type, produce low NO. Therefore, inhibition of NO production indicates anti-inflammatory efficacy by downregulating M1 macrophages. In some embodiments herein, NO assay can demonstrate the anti-inflammatory effect of pooled fetal support tissue HC-HA / PTX3 or tissue extract containing HC-HA / PTX3 (e.g., FIG. 5). In some embodiments, pooled fetal support tissue possesses sufficient efficacy when it shows a statistically significant difference compared to the positive control for fetal support tissue.

[0113] Filtration of liquid extracts: sterilization and particle separation In some embodiments, the fetal support tissue is subjected to sterilization by any suitable method, such as by irradiation, e.g., gamma irradiation, exposure to chemical germicides, heating, filtration, exposure to ethylene oxide gas, membrane filtration with appropriate pore size, electron beam (E-beam) irradiation, or sterilization by any process that produces fetal support tissue free of viable microbial contamination while maintaining the biological activity of the biologically active components disclosed herein in some embodiments.

[0114] In some embodiments, filtration using different filters with different pore sizes may be used to adjust particle size or sterilize liquid tissue extracts. In some embodiments, the fetal support tissue is sterilized by filtration. In some embodiments, sterilizing the fetal support tissue by filtration includes passing the fetal support tissue through a filter. In some embodiments, the filter pore size is selected to prevent bacteria, yeast, mold, or viruses from passing through the filter. In some cases, such as when sterilization by filtration is used to sterilize liquid forms, the filtration process uses a filter with a pore size of 0.2 μm or less.

[0115] In some embodiments, the fetal support tissue extract is filtered to adjust the particle size. In some embodiments, the filter comprises pores having an average diameter of about 0.1-0.2 μm or less, 0.2-0.3 μm or less, 0.3-0.4 μm or less, 0.4-0.5 μm or less, 0.5-0.6 μm or less, 0.6-0.7 μm or less, 0.7-0.8 μm or less, 0.8-0.9 μm or less, 0.9-1 μm or less, 1-2 μm or less, 2-3 μm or less, 3-4 μm or less, 4-5 μm or less, 5-10 μm or less, 10-20 μm or less, 20-30 μm or less, 30-40 μm or less, 40-50 μm or less, or 50-100 μm or less. In some embodiments, the filter has an average pore diameter of about 0.05-0.2 μm. In some embodiments, the filter comprises pores having an average diameter of about 0.4 μm or less. In some embodiments, the filter comprises pores having an average diameter of about 0.3 μm or less. In some embodiments, the filter comprises pores having an average diameter of about 0.2 μm or less. In some embodiments, the filter comprises pores having an average diameter of about 0.2 μm or less. In some embodiments, the filter comprises pores having an average diameter of about 0.2 μm. In some embodiments, sterilization by filtration comprises passing the fetal support tissue through a first filter and a second filter. In some embodiments, the first filter comprises an average pore size larger than the average pore size of the second filter. In some embodiments, either the first filter or the second filter has an average pore size of about 0.1-0.2 μm or less, 0.2-0.3 μm or less, 0.3-0.4 μm or less, 0.4-0.5 μm or less, 0.5-0.6 μm or less, 0.6-0.7 μm or less, 0.7-0.8 μm or less, 0.8-0.9 μm or less, 0.9-1 μm or less, 1-2 μm or less, 2-3 μm or less, 3-4 μm or less, 4-5 μm or less, or 5-10 μm or less. In some embodiments, the first filter and the second filter each have an average pore size of about 0.05-0.2 μm. In some embodiments, the first filter has an average pore size of about 0.6 μm or less and the second filter has an average pore size of about 0.4 μm or less. In some embodiments, the first filter has an average pore size of about 0.5 μm or less and the second filter has an average pore size of about 0.3 μm or less.In some embodiments, the first filter has an average pore size of about 0.45 μm or less and the second filter has an average pore size of about 0.2 μm or less, In some embodiments, the first filter has an average pore size of about 0.45 μm and the second filter has an average pore size of about 0.2 μm.

[0116] In some embodiments, any of the filters described herein are housed in a sterilization device. In some embodiments, any of the filters described herein are membranes comprising polyethersulfone (PES), polyvinylidene fluoride (PVDF), polytetrafluoroethylene (PTFE), polypropylene, polyethylene, polyamide, cellulose, nitrocellulose, nylon, or combinations thereof. In some embodiments, any of the filters described herein are sterilized by gamma irradiation. In some embodiments, the filtration pressure during filtration is about 0-10 psi, about 10-20 psi, about 20-30 psi, about 30-40 psi, about 40-50 psi, about 50-60 psi, about 60-70 psi, about 70-80 psi, about 80-90 psi, or about 90-100 psi. In some embodiments, the effective filtration area of ​​any of the filters described herein is about 0-20 cm 2 , about 20~40cm 2 , about 40~60cm 2 , about 60~80cm 2 , about 80~100cm 2 , about 100~120cm 2 , about 120~140cm 2 , about 140~160cm 2 , about 160~180cm 2 , or about 180-200cm 2In some embodiments, the overall diameter of the filter is about 0-10 mm, about 20-20 mm, about 20-30 mm, about 30-40 mm, about 40-50 mm, about 50-60 mm, about 60-70 mm, about 70-80 mm, about 80-90 mm, about 90-100 mm, about 100-200 mm, about 200-300 mm, about 300-400 mm, about 400-500 mm, about 500-600 mm, about 600-700 mm, about 700-800 mm, about 800-900 mm, or about 900-1000 mm. In some embodiments, the overall diameter of the filter is about 67 mm. In some embodiments, the overall diameter of the filter is about 68 mm. In some embodiments, the overall height of the filter is about 0-10 mm, about 20-20 mm, about 20-30 mm, about 30-40 mm, about 40-50 mm, about 50-60 mm, about 60-70 mm, about 70-80 mm, about 80-90 mm, about 90-100 mm, about 100-200 mm, about 200-300 mm, about 300-400 mm, about 400-500 mm, about 500-600 mm, about 600-700 mm, about 700-800 mm, about 800-900 mm, or about 900-1000 mm. In some embodiments, the overall height of the filter is about 82 mm. In some embodiments, the overall height of the filter is about 83 mm. In some embodiments, the filter has successfully passed a forward flow test in production. In some embodiments, the diffusion flow limit of the filter is about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1.0, or greater than 1.0 mL / min. In some embodiments, the diffusion flow limit of the filter is about 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1.0, or greater than 1.0 mL / min, when fully wetted with water, at a test pressure of about 2400, 2450, 2500, 2550, 2600, 2650, 2700, 2750, 2800, 2850, or 2900, or greater than 2900 mbar. In some embodiments, the diffusion flow limit of the filter is about 0.58 mL / min, when fully wetted with water, at a test pressure of about 2760 mbar or greater.In some embodiments, the filter's diffusional flow limit, when fully wetted with water, is about 0.40-0.50 mL / min, 0.50-0.60 mL / min, or 0.60-0.70 mL / min at a test pressure of about 2700-2800 mbar. In some embodiments, the forward flow test limit is verified for bacterial removal by correlation of the diffusional flow limit with a microbial challenge test. In some embodiments, to verify bacterial retention of the filter, fetal support tissue is tested for retention of acceptable challenge microorganisms using procedures based on applicable U.S. Food and Drug Administration guidelines.

[0117] In some embodiments, the fetal support tissue powder products disclosed herein are subjected to terminal sterilization by any suitable (e.g., medically acceptable) method. In some embodiments, the fetal support tissue powder products disclosed herein are exposed to gamma irradiation for a period of time sufficient to sterilize the fetal support tissue powder products disclosed herein.

[0118] In some embodiments, the fetal support tissue powder products disclosed herein are exposed to about 10 to about 75 kilograys (kGy) of gamma radiation for a period of time sufficient to sterilize the fetal support tissue powder product. In some embodiments, the fetal support tissue powder products disclosed herein are exposed to about 10 to about 30 kGy of gamma radiation for a period of time sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue powder products disclosed herein are exposed to about 15 to about 30 kGy of gamma radiation for a period of time sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue powder products disclosed herein are exposed to about 25 kGy of gamma radiation for a period of time sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue powder products disclosed herein are exposed to about 17.5 kGy of gamma radiation for a period of time sufficient to sterilize the fetal support tissue powder product.

[0119] In some embodiments, the fetal support tissue powder product disclosed herein is subjected to electron beam (E-beam) sterilization. In some embodiments, the fetal support tissue disclosed herein is exposed to about 10 to about 75 kilograys of E-beam radiation for a period sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue disclosed herein is exposed to about 10 to about 30 kGy of E-beam radiation for a period sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue disclosed herein is exposed to about 15 to about 30 kGy of E-beam radiation for a period sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue disclosed herein is exposed to about 25 kGy of E-beam radiation for a period sufficient to sterilize the fetal support tissue. In some embodiments, the fetal support tissue disclosed herein is exposed to about 17.5 kGy of E-beam radiation for a period sufficient to sterilize the fetal support tissue.

[0120] In some embodiments, the fetal support tissue powder product disclosed herein is exposed to an electron beam for a period of time sufficient to sterilize the fetal support tissue powder product. In some embodiments, the fetal support tissue powder product disclosed herein is exposed to X-ray radiation for a period of time sufficient to sterilize the fetal support tissue powder product. In some embodiments, the fetal support tissue powder product disclosed herein is exposed to UV radiation for a period of time sufficient to sterilize the fetal support tissue powder product.

[0121] In certain embodiments, a method for preparing fetal support tissue is provided herein, which improves the percentage of HA recovered. In some embodiments, at least or about 50%, 55%, 60%, 65%, 70%, 75% 80%, 85%, 90%, 95%, 99%, or more than 99% of HA is recovered. In some embodiments, at least or about 50%, 55%, 60%, 65%, 70%, 75% 80%, 85%, 90%, 95%, 99%, or more than 99% of HA is recovered. In some embodiments, at least or about 50%, 55%, 60%, 65%, 70%, 75% 80%, 85%, 90%, 95%, 99%, or more than 99% of HA is recovered. In some embodiments, at least or about 50% of HA is recovered. In some embodiments, at least or about 55% of HA is recovered. In some embodiments, at least or about 60% of the HA is recovered. In some embodiments, at least or about 65% of the HA is recovered. In some embodiments, at least or about 70% of the HA is recovered. In some embodiments, at least or about 75% of the HA is recovered. In some embodiments, at least or about 80% of the HA is recovered. In some embodiments, at least or about 85% of the HA is recovered. In some embodiments, at least or about 85% of the HA is recovered. In some embodiments, at least or about 95% of the HA is recovered. In some embodiments, at least or about 99% or more of the HA is recovered. In some embodiments, the HA is HMW HA. In some embodiments, the method leads to the maintenance of nHC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 75%, 80%, 85%, 90%, 95%, 99%, or more than 99% of the HC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 75% of the HC-HA / PTX3.In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 80% of HC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 85% of HC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 90% of HC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 95% of HC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 99% of HC-HA / PTX3. In some embodiments, the potency of nHC-HA / PTX3 is maintained by recovering at least or about 99% or more of HC-HA / PTX3.

[0122] In certain embodiments, the methods described herein lead to the removal of particulates or degradants. In some embodiments, the methods described herein lead to at least or about 75% 80%, 85%, 90%, 95%, 99%, or greater than 99% of the particulates or degradants being removed. In some embodiments, the particulates or degradants include chloride.

[0123] rehydration In some embodiments, the fetal support tissue is partially or completely rehydrated so that the powder extract becomes a liquid extract. In some embodiments, the rehydration reconstitutes the dry powder fetal support tissue into a liquid form by using water or a suitable buffer. In some embodiments, the rehydrated fetal support tissue or tissue extract is further filtered for sterilization. The sterilization of the rehydrated fetal support tissue extract is completed by filtering the extract using a filter pore size of about 0.2 μm or less.

[0124] In some embodiments, the rehydrated tissue extract is further filtered to adjust particle size, e.g., the extract is further centrifuged differently to produce different particle sizes for administration or formulation of the fetal support tissue extract / composition. In some embodiments, the fetal support tissue is rehydrated by contacting the fetal support tissue with a buffer or water. In some embodiments, the fetal support tissue is contacted with an isotonic buffer. In some embodiments, the fetal support tissue is contacted with saline. In some embodiments, the fetal support tissue is contacted with PBS. In some embodiments, the fetal support tissue is contacted with Ringer's solution. In some embodiments, the Ringer's solution is lactated Ringer's solution. In some embodiments, the fetal support tissue is contacted with Hartmann's solution. In some embodiments, the fetal support tissue is contacted with TRIS buffered saline. In some embodiments, the fetal support tissue is contacted with HEPES buffered saline, 50% DMEM+50% glycerol, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% glycerol, and / or 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% propylene glycol.

[0125] In some embodiments, the fetal support tissue is contacted with the buffer for 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 40 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 18 hours, 24 hours, or more than 24 hours. In some embodiments, the UC product is contacted with the buffer for 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or more than 4 weeks. Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as HC-HA / PTX3, HA, growth factors, cytokines, or other active ingredients disclosed throughout this specification and claims. In some embodiments, the fetal support tissue is preserved for later use. In some embodiments, storing the fetal support tissue does not destroy the integrity of the extracellular matrix of the fetal support tissue. In some embodiments, the fetal support tissue is freeze-dried. In some embodiments, the fetal support tissue is preserved in any suitable preservation medium.

[0126] In some embodiments, the fetal support tissue is optionally contacted with a substrate (i.e., a supportive backing). In some embodiments, the fetal support tissue is not contacted with a substrate. In some embodiments, the fetal support tissue is oriented such that the fetal support tissue is in contact with a substrate. In some embodiments, the fetal support tissue is oriented such that the stroma is in contact with a substrate. In some embodiments, the fetal support tissue is oriented such that the epithelial side is in contact with a substrate.

[0127] In some embodiments, the fetal support tissue is attached to a substrate. In some embodiments, the substrate is trocellulose paper (NC). In some embodiments, the substrate is nylon membrane (NM). In some embodiments, the substrate is polyethersulfone membrane (PES).

[0128] Natural HC-HA / PTX3 (nHC-HA / PTX3) composition In some embodiments, the isolated nHC-HA / PTX3 complex is isolated from amniotic tissue. In some embodiments, the isolated nHC-HA / PTX3 complex is isolated from amniotic membrane or umbilical cord. In some embodiments, the isolated nHC-HA / PTX3 complex is isolated from fresh placental amniotic membrane (PAM), frozen placental amniotic membrane (PAM), or previously frozen placental amniotic membrane (PAM), fresh umbilical cord amniotic membrane (UCAM), frozen umbilical cord amniotic membrane (UCAM), or previously frozen umbilical cord amniotic membrane (UCAM), fresh placenta, frozen placenta, or previously frozen placenta, fresh umbilical cord, frozen umbilical cord, or previously frozen umbilical cord, fresh chorion, frozen chorion, or previously frozen chorion, fresh amniotic chorion, frozen amniotic chorion, or previously frozen amniotic chorion, or any combination thereof. Such tissues may be obtained from any mammal, such as, but not limited to, human, non-human primate, bovine, or porcine, or any combination thereof.

[0129] In some embodiments, nHC-HA / PTX3 is purified by any suitable method. In some embodiments, the nHC-HA / PTX3 complex is purified by centrifugation (e.g., ultracentrifugation, gradient centrifugation), chromatography (e.g., ion exchange chromatography, affinity chromatography, size exclusion chromatography, and hydroxyapatite chromatography), gel filtration, or fractional lysis, ethanol precipitation, or other techniques available for protein purification (see, e.g., Scopes, Protein Purification Principles and Practice 2nd Edition, Springer-Verlag, New York, 1987; Higgins, SJ and Hames, BD (eds.), Protein Expression: A Practical Approach, Oxford Univ Press, 1999; and Deutscher, MP, Simon, MI, Abelson, JN (eds.), Guide to Protein Purification: Methods in Enzymology (Methods in Enzymology Series, Vol 182), Academic Press, 1997, all of which are incorporated herein by reference).

[0130] In some embodiments, the nHC-HA / PTX3 is isolated from an extract. In some embodiments, the extract is prepared from an amniotic membrane extract. In some embodiments, the extract is prepared from an umbilical cord extract. In some embodiments, the umbilical cord extract comprises umbilical cord stroma and / or Wharton's jelly. In some embodiments, the nHC-HA / PTX3 complex is contained in an extract prepared by ultracentrifugation. In some embodiments, the nHC-HA / PTX3 complex is contained in an extract prepared by ultracentrifugation using a gradient of CsCl / 4-6M guanidine HC1.

[0131] In some embodiments, the extract is prepared by at least two rounds of ultracentrifugation. In some embodiments, the extract is prepared by more than two rounds of ultracentrifugation (i.e., a third round of nHC-HA / PTX3). In some embodiments, the extract is prepared by at least four rounds of ultracentrifugation (i.e., a fourth round of nHC-HA / PTX3). In some embodiments, the nHC-HA / PTX3 complex comprises small leucine-rich proteoglycans. In some embodiments, the nHC-HA / PTX3 complex comprises HC1, HA, PTX3, and / or small leucine-rich proteoglycans (SLRPs).

[0132] In some embodiments, the fetal support tissue comprises nHC-HA / PTX3 and a pharmaceutical excipient. In some embodiments, the fetal support tissue consists essentially of nHC-HA / PTX3 complex. In some embodiments, the fetal support tissue comprises a pharma- ceutically acceptable diluent, excipient, vehicle, or carrier. In some embodiments, the appropriate formulation depends on the route of administration selected.

[0133] In some embodiments, the nHC-HA / PTX3 complex is purified by immunoaffinity chromatography, affinity chromatography, or a combination thereof. In some embodiments, an anti-HC1 antibody, an anti-HC2 antibody, or both, are generated and attached to a stationary support. In some embodiments, the HC-HA complex binds to an antibody (e.g., by the action of (a) an anti-HC1 antibody and an HC1, (b) an anti-HC2 antibody and an HC2, (c) an anti-PTX antibody and a PTX3, (d) an anti-SLRP antibody and an SLRP, or (e) any combination thereof). In some embodiments, an HABP is generated and attached to a stationary support.

[0134] In some embodiments, the nHC-HA / PTX3 complex is purified from the insoluble fraction using one or more antibodies as described herein. In some embodiments, the nHC-HA / PTX3 complex is purified from the insoluble fraction using an anti-SLRP antibody as described herein.

[0135] In some embodiments, the nHC-HA / PTX3 complex is purified from the soluble fraction as described herein. In some embodiments, the nHC-HA / PTX3 complex is purified from the soluble fraction as described herein using an anti-PTX3 antibody.

[0136] In some embodiments, the fetal support tissue may include components such as extracellular matrix, growth factors, cytokines, and nHC-HA / PTX3 complexes. In some embodiments, one extracellular matrix is ​​a small leucine-rich proteoglycan (SLRP). In some embodiments, the nHC-HA / PTX3 complex includes type I, type II, or type II SLRPs. In some embodiments, the small leucine-rich proteoglycan is selected from among type I SLRPs, such as decorin and biglycan. In some embodiments, the small leucine-rich proteoglycan is selected from among type II SLRPs, such as fibromodulin, lumican, PRELP (proline arginine-rich terminal leucine-rich protein), keratocan, and osteoadherin. In some embodiments, the small leucine-rich proteoglycan is selected from among type III SLRPs, such as epipican and osteoglycin. In some embodiments, the small leucine-rich proteoglycan is selected from among bikunin, decorin, biglycan, and osteoadherin. In some embodiments, the small leucine rich protein comprises a glycosaminoglycan. In some embodiments, the small leucine rich proteoglycan comprises keratan sulfate.

[0137] In some embodiments, the pharmaceutical composition further comprises at least one pharma- ceutical acceptable carrier. In some embodiments, the pharmaceutical composition further comprises an adjuvant, excipient, preservative, agent for delaying absorption, filler, binder, adsorbent, buffer, and / or solubilizer. Exemplary pharmaceutical compositions formulated to contain the nHC-HA / PTX3 complex presented herein include, but are not limited to, gels, solutions, suspensions, emulsions, syrups, granules, powders, homogenates, ointments, tablets, capsules, pills, pastes, creams, lotions, patches, sticks, films, paints, aerosols, or combinations thereof. In some embodiments, the fetal support tissue containing nHC-HA / PTX3 is an implant or sheet.

[0138] Exemplary Biological Factors in Fetal Support Tissue Disclosed herein is a fetal support tissue that contains bioactive factors that can be maintained and utilized in various applications disclosed herein. Disclosed herein are fetal support tissues, such as amniotic membrane and umbilical cord, that contain several innate biofactors with numerous significant clinical efficacies disclosed herein. For example, nHC-HA / PTX3 complex (high molecular weight (HMW) hyaluronan (HA) covalently bound to inter-α-trypsin inhibitor heavy chain (HC) 1 and further complexed with pentraxin 3 (PTX3)) is an exemplary important active component of umbilical cord and amniotic membrane, which is responsible for the wound healing effect together with other extracellular matrix, growth factors, and cytokines, and also for some other therapeutic effects (components) of fetal support tissue. Therefore, it is essential to produce fetal support tissue products (e.g., amniotic membrane and umbilical cord extracts used in wound healing) with high yields of HC-HA / PTX3, HA, and other proteins of interest. The production of fetal support tissue products using processes that concentrate the HC-HA / PTX3 complex and other proteins of interest while reducing or preventing the degradation of these biomolecules is an important factor in processing fetal support tissue compositions. It is important to prevent the degradation of HA and other proteins of interest, as this can render the fetal support tissue product unsuitable for use.

[0139] Disclosed herein, in certain embodiments, is a method of preparing fetal support tissue, the method comprising (a) lyophilizing pooled or unpooled fetal support tissue to produce micronized, cryogenically ground (liquid or dry), devitalized, decellularized (frozen), or a combination thereof, or (b) extracting the processed fetal support tissue in an excipient to produce an extract, and (c) sterilizing the fetal support tissue by filtering the extract using a membrane having a pore size of about 0.2 μm or less. In some embodiments, the fetal support tissue is a placental amniotic membrane (PAM) or substantially isolated PAM, an umbilical cord amniotic membrane (UCAM) or substantially isolated UCAM, a chorion or substantially isolated chorion, an amniotic chorion or substantially isolated amniotic chorion, a placenta or substantially isolated placenta, an umbilical cord or substantially isolated umbilical cord, or any combination thereof.

[0140] In some embodiments, the diluted fetal support tissue contains about 1 μg / mL to about 1 mg / mL of HA, a portion of which is present as HC-HA / PTX3.

[0141] Described herein is a composition comprising a therapeutically effective amount of a pooled composition comprising fetal support tissue in certain embodiments. In some embodiments, the fetal support tissue comprises micronized fetal support tissue. In some embodiments, the pooled composition comprises fetal support tissue pooled from multiple donors. In some embodiments, the composition comprises fetal support tissue pooled from at least 15 donors. In some embodiments, the composition comprises fetal support tissue pooled from at least 30 donors. In some embodiments, the composition comprises fetal support tissue pooled from at least 45 donors. In some embodiments, the composition comprises fetal support tissue pooled from at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or more than 100 donors. In some embodiments, the composition is not pooled when it comprises tissue from up to 5 donors. In some embodiments, a composition is not pooled if it contains tissue from a maximum of 9, 8, 7, 6, 5, 4, 3, or 2 donors.

[0142] Pooling fetal support tissues reduces sample variability and improves sample consistency. Described herein, in certain embodiments, is a composition comprising pooled fetal support tissue or fetal support tissue extract or composition, comprising reduced inter-donor (or inter-subject) variability compared to non-pooled (individual donor) fetal support tissue extract compositions. In some embodiments, the pooled composition comprises reduced variability compared to a composition comprising fetal support tissue from up to 9, 8, 7, 6, 5, 4, 3, or 2 donors. In some embodiments, the pooled composition comprises reduced variability compared to a composition comprising fetal support tissue from up to 5 donors. In some embodiments, the pooled composition comprises reduced variability compared to a composition comprising fetal support tissue from one donor. Disclosed herein is a pooled fetal support tissue extract (in any form disclosed herein), which may comprise reduced variability between pooled samples compared to the variability between samples of non-pooled (individual) fetal support tissue extracts. For example, when the average coefficient of variation percentage (CV%) is calculated for pooled fetal support tissue products or extracts, the CV% of the pooled may be at least 2-fold, 3-fold, 4-fold, or more lower than the CV% of the non-pooled fetal support tissue products. Disclosed herein is any form of pooled fetal support tissue extract disclosed herein, including pooled product, which shows increased uniformity and improved sample consistency relative to the uniformity or sample consistency of unpooled fetal support tissue extract.For example, pooled fetal support tissue extract (any form, e.g., composition, kit, etc.) may show improved consistency, and pooled extract shows reduced variation in native HC-HA / PTX3 complex activity compared to unpooled fetal support tissue extract (product, composition, etc.) as determined by ODI-TRAP assay, BCA assay, or HA quantification assay.For example, improved consistency of pooled extract may be shown by at least 10% reduction in coefficient of variation.In some embodiments, the reduced coefficient of variation for pooled fetal support tissue extract compared to unpooled fetal support tissue extract may be 5% or less.In some embodiments, the reduced coefficient of variation for pooled fetal support tissue extracts compared to non-pooled fetal support tissue extracts may be 4% or less. In some embodiments, the reduced coefficient of variation for pooled fetal support tissue extracts compared to non-pooled fetal support tissue extracts may be about 3% or less. In some embodiments, the reduced coefficient of variation for pooled fetal support tissue extracts compared to non-pooled fetal support tissue extracts may be about 2% or less.

[0143] Regardless of the present disclosure of whether pooled fetal support donor samples or single donor samples, and pooled fetal support tissue extracts (compositions, kits, products, etc.) and non-pooled fetal support tissue extracts (compositions, kits, products, etc.) of any origin retain HA molecular weight (MW), pooling does not alter either HA MW or HC-HA binding. Pooled fetal support tissues and extracts or products obtained therefrom as disclosed herein provide higher yields of products with greater uniformity, which reduces variation between pooled samples. As disclosed above and throughout this disclosure, pooling of samples improves product consistency despite increased protein yield (e.g., HA protein content) due to reduced variation compared to that obtained from non-pooled single donor samples and / or extracts, as measured by a reduced coefficient of variation. As disclosed herein, the present disclosure produced by the pooling process improves the yield, uniformity, and consistency of pooled extracts while fully retaining the potency of fetal support tissue extracts (products / compositions, etc.), e.g., fully retaining the potency of native HC-HA / PTX3 complex. Thus, the teachings of this disclosure discuss pools of fetal support tissue and / or fetal support tissue products.

[0144] In some embodiments, the fetal support tissue is placental amnion, umbilical cord, umbilical amnion, chorion, amniotic chorion, placenta, amniotic stroma, amniotic jelly, or any combination thereof. In some embodiments, the fetal support tissue comprises the extracellular matrix (ECM) of amnion (AM) and umbilical cord (UC). The ECM of fetal support tissue, e.g., AM / UC, contains several innate biological factors useful for a number of purposes, including wound healing and reducing inflammation and scarring. Characterization data shows that fetal support tissue, e.g., AM and UC extracellular matrix (ECM), is rich in hyaluronic acid (HA), which has an essential or important matrix component called HC-HA / PTX3 complex (heavy chain 1-hyaluronic acid-pentraxin 3). In some embodiments, the fetal support tissue comprises AM and UC, and the HC-HA / PTX3 is isolated, and the HC-HA / PTX3 is natural (nHC-HA / PTX3). In some embodiments, the fetal support tissue comprises micronized fetal support tissue.

[0145] In some embodiments, the variation is determined by the average particle size in the pooled composition. In some embodiments, the pooled composition comprises about 10,000 or less particles with a diameter of more than about 10 μm. In some embodiments, the pooled composition comprises about 10,000, 9,000, 8,000, 7,000, 6,000, 5,000, 4,000, 3,000, 2,000, 1,000, 500, 250, or 100 or less particles with a diameter of more than about 10 μm or less than 10 μm, less than 5 μm, less than 1 μm, or about 1 μm, about 1 μm to about 5 μm, or about 5 μm to about 10 μm, or combinations thereof. In some embodiments, the pooled composition comprises no more than about 10,000, 9,000, 8,000, 7,000, 6,000, 5,000, 4,000, 3,000, 2,000, 1,000, 500, 250, or 100 particles with diameters greater than about 10 μm or less than 10 μm, less than 5 μm, less than 1 μm, or about 1 μm, between about 1 μm and about 5 μm, or between about 5 μm and about 10 μm, or combinations thereof. In some embodiments, the pooled composition comprises no more than about 1,000 particles with diameters greater than about 25 μm. In some embodiments, the pooled composition comprises no more than about 1000, 900, 800, 700, 600, 500, 400, 300, 200, 100, or 100 particles that comprise a diameter greater than about 10, 15, 20, 25, 30, 35, 40, 45, or 50 μm. In some embodiments, the pooled composition comprises no more than about 10000, 9000, 8000, 7000, 6000, 5000, 4000, 3000, 2000, 1000, 500, 250, or 100 particles that comprise a diameter greater than about 10, 15, 20, 25, 30, 35, 40, 45, or 50 μm.

[0146] Described herein, in certain embodiments, is a composition comprising a therapeutically effective amount of a pooled composition comprising fetal support tissue, the fetal support tissue comprising a particle size of a particular diameter. Further described herein, in certain embodiments, is a composition comprising a therapeutically effective amount of a pooled composition comprising fetal support tissue, the fetal support tissue comprising a particle size of about 20 μm to about 240 μm in diameter. In some embodiments, the fetal support tissue comprises micronized fetal support tissue.

[0147] In some embodiments, the fetal support tissue comprises an average particle size of about 10 μm to about 300 μm, about 10 μm to about 250 μm, about 10 μm to about 200 μm, about 10 μm to about 150 μm, about 10 μm to about 100 μm, or about 10 μm to about 50 μm. In some embodiments, the fetal support tissue comprises an average particle size of about 20 μm to about 200 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 20 μm to about 40 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 20 μm to about 300 μm, about 20 μm to about 250 μm, about 20 μm to about 200 μm, about 20 μm to about 150 μm, about 20 μm to about 100 μm, about 20 μm to about 50 μm, about 50 μm to about 300 μm, about 50 μm to about 250 μm, about 50 μm to about 200 μm, about 50 μm to about 150 μm, or about 50 μm to about 100 μm. In some embodiments, the fetal support tissue comprises an average particle size of about 60 μm to about 120 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 140 μm to about 240 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of at least or about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, or 300 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 100 μm in diameter.

[0148] In some embodiments, the fetal support tissue comprises an average particle size of about 0.010 μm to about 0.3 μm, about 0.01 μm to about 0.25 μm, about 0.01 μm to about 0.2 μm, about 0.01 μm to about 0.15 μm, about 0.01 μm to about 0.1 μm, about 0.01 μm to about 0.5 μm, 0.10 μm to about 0.3 μm, about 0.1 μm to about 0.25 μm, about 0.1 μm to about 0.2 μm, about 0.1 μm to about 0.15 μm, about 0.1 μm to about 0.25 μm, or about 0.1 μm to about 0.5 μm. In some embodiments, the fetal support tissue comprises an average particle size of about 0.20 μm to about 2.0 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 0.20 μm to about 4.0 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 0.20 μm to about 3.0 μm, about 0.20 μm to about 2.5 μm, about 0.20 μm to about 2.0 μm, about 0.20 μm to about 1.5 μm, about 0.20 μm to about 1.0 μm, about 0.20 μm to about 5.0 μm, about 0.50 μm to about 3.0 μm, about 0.50 μm to about 2.5 μm, about 0.50 μm to about 2.0 μm, about 0.50 μm to about 1.5 μm, or about 0.50 μm to about 1.0 μm. In some embodiments, the fetal support tissue comprises an average particle size of about 0.60 μm to about 1.2 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of at least about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.15, 0.20, 0.25, 0.3, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 1.0, 1.5, 2.0, 2.5, 3.0, or greater than 3.0 μm in diameter. In some embodiments, the fetal support tissue comprises an average particle size of about 0.50 μm in diameter. Disclosed herein is a process for producing fetal support tissue while retaining the biological activity of the fetal support tissue, including components of the fetal support tissue such as HA (present as nHC-HA / PTX3 in AM / UC) whose potency is maintained, for example, as disclosed throughout the specification and claims.

[0149] Dosage form Provided below are formulations of the compositions described herein, including pooled compositions that include fetal support tissue.

[0150] In some embodiments, the pooled compositions described herein are administered as an aqueous suspension. In some embodiments, the aqueous suspension comprises water, Ringer's solution, and / or sodium chloride solution. In some embodiments, the Ringer's solution is lactated Ringer's solution. In some embodiments, the aqueous suspension comprises a sweetener or flavoring agent, a colorant or dye, and optionally an emulsifying or suspending agent together with a diluent, water, ethanol, propylene glycol, glycerin, or a combination thereof. In some embodiments, the aqueous suspension comprises a suspending agent. In some embodiments, the aqueous suspension comprises sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and / or gum arabic. In some embodiments, the aqueous suspension comprises a dispersing or wetting agent. In some embodiments, the aqueous suspension comprises a natural phospholipid, such as lecithin, or a condensation product of an alkylene oxide with a fatty acid, such as polyoxyethylene stearate, or a condensation product of ethylene oxide with a long chain aliphatic alcohol, such as heptadecaethylene-oxycetanol, or a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol, such as polyoxyethylene sorbitol monocreate, or a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol anhydride, such as polyethylene sorbitan monocreate. In some embodiments, the aqueous suspension comprises a preservative. In some embodiments, the aqueous suspension comprises ethyl or propyl n-paraoxybenzoate. In some embodiments, the aqueous suspension comprises a sweetener. In some embodiments, the aqueous suspension comprises sucrose, saccharin, or aspartame.

[0151] In some embodiments, the pooled compositions described herein are administered as oil suspensions. In some embodiments, oil suspensions are formulated by suspending the active ingredient in vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or mineral oil (e.g., liquid paraffin). In some embodiments, oil suspensions contain a thickening agent (e.g., beeswax, hard paraffin, or acetyl alcohol). In some embodiments, oil suspensions contain a sweetener (e.g., those described above). In some embodiments, oil suspensions contain an antioxidant (e.g., butylated hydroxyanisole or alpha-tocopherol).

[0152] In some embodiments, the pooled compositions disclosed herein are formulated for administration by injection, such as parenteral injection (e.g., by infusion or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled compositions are administered by intra-articular injection. In some embodiments, the pooled compositions are administered as a sterile solution, suspension, or emulsion. In some embodiments, the pooled compositions are formulated for topical administration, local administration, or inhalation. In some embodiments, the bioactive components in the pooled compositions disclosed herein, such as HA, nHC-HA / PTX3, and other bioactive components disclosed, diffuse from the pooled composition into the surrounding tissue. In some embodiments, the pooled fetal support tissue product formulated for topical administration further comprises a penetration enhancer, a gelling agent, a patch, an adhesive, an emollient, or a combination thereof.

[0153] In some embodiments, the formulations to be injected are presented in unit dosage form, eg, in ampoules or in multi-dose containers, with a preservative.

[0154] In some embodiments, the pooled compositions described herein are formulated for topical administration. Topical formulations include, but are not limited to, ointments, creams, lotions, solutions, pastes, gels, films, sticks, liposomes, microparticles, microspheres, lipid complexes, nanoparticles. In some embodiments, the topical formulations are administered by use of a patch, bandage, or wound dressing. In some embodiments, the pooled fetal tissue products, compositions, extracts, methods, or devices are formulated for use in treating or preventing tissue damage, such as, for example, ulcers, wounds, perforations, burns, surgery, injuries, or fistulas. In some embodiments, the methods or compositions, etc. disclosed herein prevent or treat tissue necrosis or ischemic diseases. In some embodiments, the ischemic diseases include cardiac ischemia, ischemic colitis, mesenteric ischemia, cerebral ischemia, acute limb ischemia, cyanosis, and gangrene, and the treatment method may include contacting the ischemic tissue with the pooled fetal support tissue products disclosed herein. Further provided herein, in some embodiments, is a method of treating a neuropathic disease in an individual in need thereof, comprising contacting ischemic tissue with a pooled fetal support tissue product.

[0155] In some embodiments, the pooled composition described herein is formulated as a composition in the form of a solid, a cross-linked gel, or a liposome. In some embodiments, the fetal tissue indicating product comprising the nHC-HA / PTX3 complex is formulated as an insoluble cross-linked hydrogel. In some embodiments, the pooled fetal support tissue composition is formulated as a gel or topical preparation.

[0156] In some embodiments, the topical formulation comprises a gelling (or thickening) agent. Suitable gelling agents include, but are not limited to, cellulose and cellulose derivatives, cellulose ethers (e.g., carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose, methylcellulose), guar gum, xanthan gum, locust bean gum, alginates (e.g., alginic acid), silicates, starch, tragacanth gum, carboxyvinyl polymers, carrageenan, paraffin, petrolatum, acacia (gum arabic), agar, magnesium aluminum silicate, sodium alginate, and the like. thorium, sodium stearate, bladderwrack, bentonite, carbomer, carrageenan, carbopol, xanthan, cellulose, microcrystalline cellulose (MCC), ceratonia, chondrus, dextrose, furcellaran, gelatin, ghatti gum, guar gum, hectorite, lactose, sucrose, maltodextrin, mannitol, sorbitol, honey, corn starch, wheat starch, rice starch, potato starch, gelatin, sterculia gum gum), polyethylene glycol (e.g., PEG 200-4500), tragacanth gum, ethyl cellulose, ethyl hydroxyethyl cellulose, ethyl methyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl cellulose, poly(hydroxyethyl methacrylate), oxypolygelatin, pectin, polygeline, povidone, propylene carbonate, methyl vinyl ether / maleic anhydride copolymer (PVM / MA), poly(methoxyethyl methacrylate), poly(methoxyethoxyethyl methacrylate), hydroxypropyl cellulose, hydroxypropyl methyl cellulose (HPMC), sodium carboxymethyl cellulose (CMC), silicon dioxide, polyvinylpyrrolidone (PVP: povidone), or combinations thereof.

[0157] In some embodiments, the topical formulations disclosed herein include an emollient, including, but not limited to, castor oil esters, cocoa butter esters, safflower oil esters, cottonseed oil esters, corn oil esters, olive oil esters, cod liver oil esters, almond oil esters, avocado oil esters, palm oil esters, sesame oil esters, squalene esters, kukui oil, and the like. oil esters, soybean oil esters, acetylated monoglycerides, ethoxylated glyceryl monostearate, hexyl laurate, isohexyl laurate, isohexyl palmitate, isopropyl palmitate, methyl palmitate, decyloleate, isodecyl oleate, hexadecyl stearate, decyl stearate, isopropyl isostearate, methyl isostearate, diisopropyl adipate, diisohexyl adipate, dihexyldecyl adipate, diisopropyl sebacate, lauryl lactate, myristyl lactate, and cetyl lactate, oleyl myristate, oleyl stearate, and oleyl oleate, pelargonic acid, lauric acid, myristic acid, palmitic acid, Acids such as stearic acid, isostearic acid, hydroxystearic acid, oleic acid, linoleic acid, ricinoleic acid, arachic acid, behenic acid, erucic acid, lauryl alcohol, myristyl alcohol, cetyl alcohol, hexadecyl alcohol, stearyl alcohol, isostearyl alcohol, hydroxystearyl alcohol, oleyl alcohol, ricinoleyl alcohol, behenyl alcohol, erucyl alcohol, 2-octyldodecanyl alcohol, lanolin and lanolin derivatives, beeswax, spermaceti, myristyl myristate, stearyl stearate, carnauba wax, candelilla wax, lecithin, and cholesterol.

[0158] In some embodiments, the pooled compositions disclosed herein are formulated with one or more natural polymers. In some embodiments, the pooled compositions disclosed herein are formulated with natural polymers that are fibronectin, collagen, laminin, keratin, fibrin, fibrinogen, hyaluronic acid, heparan sulfate, chondroitin sulfate. In some embodiments, the pooled compositions disclosed herein are formulated with polymer gels formulated from natural polymers. In some embodiments, the pooled compositions disclosed herein are formulated with polymer gels formulated from natural polymers, such as, but not limited to, fibronectin, collagen, laminin, keratin, fibrin, fibrinogen, hyaluronic acid, heparan sulfate, chondroitin sulfate, and combinations thereof.

[0159] In some embodiments, the pooled fetal support tissue comprises a pharma- ceutically acceptable excipient, carrier, or combination thereof. In some embodiments, the pooled fetal support tissue is formulated as a non-solid dosage form. In some embodiments, the pooled fetal support tissue is formulated as a solid dosage form.

[0160] In some embodiments, the pooled compositions described herein are formulated for administration to the eye or tissues associated therewith. Formulations suitable for administration to the eye include, but are not limited to, solutions, suspensions (e.g., aqueous suspensions), ointments, gels, creams, liposomes, niosomes, pharmacosomes, nanoparticles, or combinations thereof. In some embodiments, the pooled compositions described herein for topical administration to the eye are administered by spraying, irrigation, or combinations thereof. In some embodiments, the pooled compositions described herein are administered to the eye by injectable depot formulations.

[0161] As used herein, a "depot formulation" is a controlled release formulation that is implanted (e.g., subcutaneously, intramuscularly, intravitreally, or subconjunctivally) in the eye or tissues associated therewith (e.g., the sclera). In some embodiments, the depot formulation is formulated by forming a microencapsulation matrix (also known as a microencapsulation matrix) of the pooled compositions described herein in a biodegradable polymer. In some embodiments, the depot formulation is formulated by entrapping the pooled compositions described herein in a liposome or microemulsion.

[0162] The formulations for administration to the eye have an ophthalmically acceptable tonicity. In certain cases, tears have an isotonicity value equivalent to a 0.9% sodium chloride solution. In some embodiments, isotonicity values ​​from about 0.6% to about 1.8% sodium chloride equivalent are suitable for topical administration to the eye. In some embodiments, the formulations for administration to the eye disclosed herein have an osmolality of about 200 to about 600 mOsm / L. In some embodiments, the formulations for administration to the eye disclosed herein are hypotonic and therefore require the addition of any suitable substance to reach a suitable tonicity range. Ophthalmologically acceptable substances that adjust tonicity include, but are not limited to, sodium chloride, potassium chloride, sodium thiosulfate, sodium sulfite, and ammonium sulfate.

[0163] Formulations for administration to the eye have an ophthalmologically acceptable clarity. Examples of ophthalmologically acceptable clarifying agents include, but are not limited to, polysorbate 20, polysorbate 80, or combinations thereof.

[0164] In some embodiments, the formulation for administration to the eye includes an ophthalmologically acceptable viscosity enhancer. In some embodiments, the viscosity enhancer increases the time that the formulations disclosed herein remain on the eye. In some embodiments, increasing the time that the formulations disclosed herein remain on the eye results in increased drug absorption and action. Non-limiting examples of mucoadhesive polymers include carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0165] In some embodiments, the composition is formulated for injection into the eye, for example, by injection into deeper tissues or connective spaces in tissues and surrounding areas near the eye, or by injection into a tumor or near a tumor. In some embodiments, the composition is administered by intravitreal injection into the eye. In some embodiments, the composition is administered by intraocular injection, subretinal injection, intravitreal injection, periocular administration, subconjunctival injection, retrobulbar injection, intracameral injection (including injection into the anterior chamber or vitreous chamber), or subtenon injection. In some embodiments, the formulation for administration to the eye is administered or delivered to the posterior part of the eye (e.g., the retina, choroid, vitreous, and optic nerve). In some embodiments, the topical formulation for administration to the eye disclosed herein for delivery to the posterior part of the eye comprises a solubilizing agent, for example, glucan sulfate and / or cyclodextrin. In some embodiments, glucan sulfates used include, but are not limited to, dextran sulfate, cyclodextrin sulfate, and β-1,3 glucan sulfate, both natural and derivatives, or any compound that can be temporarily bound to or retained in tissues containing fibroblast growth factor (FGF), improves drug stability and / or solubility, and / or improves the penetration and ocular absorption of topical formulations for administration to the eye as disclosed herein. In some embodiments, cyclodextrin derivatives used as solubilizers include, but are not limited to, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxyethyl β-cyclodextrin, hydroxypropyl γ-cyclodextrin, hydroxypropyl β-cyclodextrin, sulfated α-cyclodextrin, sulfated β-cyclodextrin, sulfobutyl ether β-cyclodextrin.

[0166] Dosage The amount of the composition administered depends, to some extent, on the individual being treated. In the case where the composition is administered to a human subject, the daily dosage is usually determined by the prescribing physician, since the dosage generally varies according to the individual's age, sex, diet, weight, overall health and response, the severity of the individual's symptoms, the specific disease or illness being treated, the severity of the disease or illness being treated, the time of administration, the route of administration, the nature of the composition, the rate of excretion, the drug combination, and the discretion of the prescribing physician.

[0167] In some embodiments, the fetal support tissue is in a lyophilized, water-soluble form. Both the powder and liquid forms of the fetal support tissue contain the nHC-HA / PTX3 complex. For the preparation of the powder form, fresh or frozen fetal support tissue that has been screened for donor eligibility is washed, cut, soaked, and made into drug substance starting material (RSM). The RSM is then subjected to a particulate and micronization process under sterile ambient temperature conditions to be pulverized into fine particles (Figure 1A). In some embodiments, the micronization may be performed with or without freeze-drying into a liquid or powder form. For the powder form, the micronized fetal support tissue is sieved to classify particle size, and then subjected to in-process and quality control assessments, made into bulk drug substance, and may be packaged and subjected to various terminal sterilization processes to become a finished drug product (Figure 1B). In some embodiments, the powdered tissue containing nHC-HA / PTX3 may be reconstituted in a suitable excipient or the powder may be frozen and stored at -80°C. In some embodiments, the liquid form of the fetal support tissue is prepared by rehydration after the micronization process. The liquid form of the fetal support tissue may also be prepared after cryo-grinding. For example, to cryo-grind the fetal support tissue into liquid form, the tissue is aseptically processed under conditions of 4°C (Figures 2A-2C). The fetal support tissue is subjected to cryo-grinding, extraction, centrifugation, and filtration. In some embodiments, the particle size is adjusted by centrifugation and filtration of the cryo-grinded tissue extract based on different centrifugation and filtration parameters. In some embodiments, filtration is performed to sterilize the extract (filter pore size equal to about 0.2 μm to less than 0.2 μm). After cryo-grinding of the tissue, processed as shown in Figures 2A-2C, the tissue composition is pooled and variously diluted, and after the end of the in-process control, the extract containing nHC-HA / PTX3 is packaged as a finished drug product. In some embodiments, the dosage of the HC-HA / PTX complex is about 0.001 to about 1000 mg / kg body weight / day. In some embodiments, the amount of the HC-HA / PTX complex disclosed herein is in the range of about 0.5 to about 50 mg / kg / day.In some embodiments, the amount of HC-HA / PTX complex disclosed herein is about 0.0001 to about 7 g / day. In some embodiments, the amount of HC-HA / PTX complex disclosed herein is about 0.01 to about 7 g / day. In some embodiments, the amount of HC-HA / PTX complex disclosed herein is about 0.02 to about 5 g / day. In some embodiments, the amount of HC-HA / PTX complex disclosed herein is about 0.05 to about 2.5 g / day. In some embodiments, the amount of HC-HA / PTX complex disclosed herein is about 0.1 to about 1 g / day.

[0168] In some embodiments, a composition comprising an nHC-HA / PTX3 complex disclosed herein is administered before, during, or after the onset of undesirable changes in tissue. In some embodiments, an nHC-HA / PTX3 composition disclosed herein is administered with a combination therapy before, during, or after the onset of a disease or disorder. In some embodiments, the timing of administration of an nHC-HA / PTX3 composition disclosed herein is variable. Thus, in some examples, an nHC-HA / PTX3 composition described herein is used as a prophylactic and is administered continuously to a subject prone to the development of undesirable changes in tissue to prevent undesirable changes in tissue from occurring. In some embodiments, an nHC-HA / PTX3 composition disclosed herein is administered to a subject during the onset of undesirable changes or as soon as possible after the onset of undesirable changes. In some embodiments, administration of an nHC-HA / PTX3 composition described herein is initiated within the first 48 hours of onset of undesirable changes, preferably within the first 48 hours of onset of symptoms, more preferably within the first 6 hours of onset of symptoms, and most preferably within 3 hours of onset of symptoms. In some embodiments, the initial administration is via any practical route, such as, for example, intravenous infusion, bolus infusion, infusion over 5 minutes to about 5 hours, pill, capsule, transdermal patch, buccal delivery, or combinations thereof. The nHC-HA / PTX3 compositions disclosed herein are preferably administered as soon as practicable after the onset of undesirable changes is detected or suspected, for the period required for treatment, such as, for example, from about 1 month to about 3 months. In some embodiments, the treatment period varies for each subject, and the period is determined using known criteria. In some embodiments, the compositions described herein are administered for at least 2 weeks, preferably from about 1 month to about 5 years, more preferably from about 1 month to about 3 years.

[0169] In some embodiments, the nHC-HA / PTX3 compositions described herein are administered once daily in a single dose. In some embodiments, the compositions described herein are administered more than once per day in multiple doses. In some embodiments, the compositions described herein are administered twice daily. In some embodiments, the compositions described herein are administered three times per day. In some embodiments, the compositions described herein are administered four times per day. In some embodiments, the compositions comprising nHC-HA / PTX3 complexes described herein are administered more than four times per day.

[0170] If the individual's disease does not improve, at the physician's discretion, the nHC-HA / PTX3 compositions described herein are administered chronically, i.e., for an extended period of time, including the individual's lifespan, to ameliorate or otherwise control or limit the symptoms of the individual's disease or disorder.

[0171] In some embodiments, if the individual's condition improves, at the discretion of the physician, the nHC-HA / PTX3 complex disclosed herein may be administered continuously or the dose of the administered drug may be temporarily reduced or temporarily stopped for a period of time (i.e., a "drug holiday"). In some embodiments, the length of the drug holiday may vary from 2 days to 1 year, including, by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. In some embodiments, the dose reduction during the drug holiday is from 10% to 100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

[0172] Once improvement of the individual's disease has occurred, a maintenance dose is administered as needed. In some embodiments, the dosage or frequency of administration, or both, is then reduced, depending on the symptoms, until improvement of the disease, disorder, or condition is maintained. In some embodiments, the individual requires intermittent treatment on a long-term basis upon the recurrence of any symptoms.

[0173] In some embodiments, the pharmaceutical compositions described herein are in unit dosage form suitable for single administration of precise dosage amounts. In unit dosage form, the formulation is divided into unit doses containing appropriate amounts of the nHC-HA / PTX3 complex disclosed herein. In some embodiments, the unit dose is in a packaged form containing discrete amounts of the formulation. Non-limiting examples are packaged tablets or capsules, and powders in vials or ampoules. In some embodiments, the aqueous suspension composition is packaged in a non-reclosable single-dose container. In some embodiments, reclosable multi-dose containers are used, in which case a preservative is typically included in the composition. In some embodiments, the formulation for parenteral injection is provided in unit dosage form, including but not limited to ampoules or multi-dose containers, together with a preservative.

[0174] Suitable daily dosages of the nHC-HA / PTX3 complexes disclosed herein are, for example, about 0.01-2.5 mg / kg of body weight. Indicated daily dosages for large mammals, including but not limited to humans, are in the range of about 0.5 mg to about 100 mg, conveniently administered in multiple doses up to four times a day or in sustained release form. Suitable unit dosage forms for oral administration contain about 1 mg to 50 mg of active ingredient. Due to the large variables associated with individual treatment regimens, the foregoing ranges are merely suggestive, and some deviation from these recommendations is not uncommon. In some embodiments, dosages vary depending on many variables, including but not limited to the activity of the nHC-HA / PTX3 complexes used, the disease or condition being treated, the method of administration, the requirements of the individual subject, the severity of the disease or condition being treated, and the physician's judgment.

[0175] In some embodiments, the compositions described herein are packaged as an article of manufacture comprising packaging material, a pharmaceutical composition effective for the prevention and / or treatment of a disease or disorder, and a label indicating that the pharmaceutical composition is used to reprogram fibroblasts in tissues that have undergone undesirable changes due to a disease or disorder. In some embodiments, the pharmaceutical composition is packaged in a unit dosage form and contains an amount of the pharmaceutical composition for a single or multiple administrations. In some embodiments, the packaged composition comprises a lyophilized powder of the pharmaceutical composition that is reconstituted (e.g., with water or saline) prior to administration.

[0176] Medical Devices and Biomaterial Compositions In some embodiments, the compositions described herein are directly assembled to the surface of an implantable medical device or prepared as a coating on an implantable medical device, hi some embodiments, the compositions described herein are directly assembled to the surface of an implantable medical device or portion thereof.

[0177] Exemplary implantable medical devices include, but are not limited to, artificial joints, orthopedic devices, bone implants, contact lenses, sutures, surgical staples, surgical clips, catheters, angioplasty balloons, sensors, surgical instruments, electrodes, needles, syringes, wound drains, shunts, urethral inserts, metal or plastic implants, heart valves, artificial organs, lap bands, annuloplasty rings, guide wires, K-wires or Denham pins, stents, stent grafts, vascular grafts, pacemakers, pellets, wafers, medical tubing, infusion sleeves, implantable defibrillators, neurostimulators, glucose sensors, cerebrospinal fluid, implantable drug pumps, spinal cages, artificial discs, eye implants, cochlear implants, breast implants, replacement devices for the nucleus pulposus, ear tubes, intraocular lenses, drug delivery systems, microparticles, nanoparticles, and microcapsules.

[0178] In some embodiments, the compositions, products, or extracts comprising various forms of pooled fetal support tissue described herein, or the compositions, products, or extracts, reduce pain, reduce body inflammation, and reduce scarring. In certain embodiments, the compositions, products, or extracts comprising various forms of pooled fetal support tissue described herein, or the compositions, products, or extracts, promote regenerative wound healing. In some embodiments, regenerative healing includes, for example, nerve regeneration, or articular cartilage regeneration, and various body regenerative healing. In some embodiments, nerve regeneration reduces sensitization / irritation. In some embodiments, the compositions comprising, for example, cryopreserved amniotic membrane (AM) and umbilical cord (UC) microparticles, or the compositions, products, compositions, or extracts may be disclosed herein, and the pooled fetal support products, compositions, or extracts may be used in various clinical applications, including, for example, knee osteoarthritis, facet osteoarthritis, plantar fasciitis, chronic wound healing, complex wound healing, and plantar injury healing.

[0179] In some embodiments, complex injuries may involve chronic internal ulcers, ischemic wounds, or may involve exposed bone, muscle, tendons, joint capsules, bone loss (e.g., due to necrosis of soft tissue, bone, etc.), which may be difficult to heal and more susceptible to skin, muscle, and tendon infections. In some embodiments, patients with complex injuries may be prone to bone infections, including osteomyelitis. In some embodiments, the pooled fetal supportive tissue extract of the present disclosure may be a useful treatment for complex injuries to effect healing through vascular remodeling, re-epithelialization, and re-keratinization from the peripheral edge of the injury to the granulation tissue. In some embodiments, complex injuries may correct congenital anomalies, such as, for example, the use of the present disclosure in performing spina bifida repair in utero. In yet other embodiments, the present disclosure may be useful in treating complex injuries resulting from diabetes, diabetic neuropathy, e.g., complex gangrenous injuries that may require amputation, where use of the present disclosure following sharp debridement, bone biopsy, and / or open decortication, etc., can preclude the amputation process (e.g., amputation of gangrenous legs, toes, limbs, etc.).

[0180] In some embodiments, the method of using a composition or product comprising, for example, cryopreserved amniotic membrane (AM) and umbilical cord (UC) from pooled fetal support tissue, such as clarix flo and microparticles from cryopreserved AM composition, or the composition or product can provide faster recovery of mechanical hypersensitivity and reduced thermal hypersensitivity. In certain embodiments, the combination of micronized and freeze-dried AM and UC can be a fetal support tissue product, composition, or extract, including sheet and non-sheet forms of any of the fetal support tissue products disclosed herein, and such compositions or methods of using such compositions can be used as drugs, treatments, or any of the methods of treatment disclosed herein. In some embodiments, such drugs, treatments, or methods of using such can be administered for the relief of physical pain, injury, inflammation, scarring, infection, or disease using any of the methods of administration disclosed herein. In some embodiments, the compositions described herein are directly incorporated into scaffolds, microparticles, microcapsules, or microcarriers used for the delivery of biomaterials, such as stem cells or insulin-producing cells. In some embodiments, the compositions described herein are attached to or directly incorporated into microcapsules.

[0181] How to use Disclosed herein is a method of using the compositions described herein by the methods described herein in certain embodiments. In some embodiments, the composition comprises a pooled composition comprising fetal support tissue in a therapeutically effective amount. In some embodiments, the composition comprises a pooled composition comprising micronized fetal support tissue in a therapeutically effective amount. In some embodiments, the composition comprises a pooled composition comprising pulverized fetal support tissue in a therapeutically effective amount. In some embodiments, the composition comprises a pooled composition comprising micronized fetal support tissue in a therapeutically effective amount. In some embodiments, the fetal support tissue is a placental amniotic membrane (PAM) or substantially isolated PAM, an umbilical cord amniotic membrane (UCAM) or substantially isolated UCAM, a chorion or substantially isolated chorion, an amniotic chorion or substantially isolated amniotic chorion, a placenta or substantially isolated placenta, an umbilical cord or substantially isolated umbilical cord, or any combination thereof. In some embodiments, the fetal support tissue produced by the methods disclosed herein comprises a fetal support tissue and a pharma- tically acceptable carrier. In some embodiments, the fetal support tissue products disclosed herein are formulated for topical administration or administration by injection. In some embodiments, the fetal support tissue products disclosed herein are formulated as a solution, suspension, or emulsion.

[0182] In some embodiments, the pooled compositions disclosed herein are used to inhibit at least one of scarring, inflammation, adhesion, and angiogenesis, but promote vasculogenesis. In some embodiments, the pooled fetal support tissue products, such as products comprising isolated nHC-HA / PTX3 compositions, are used in inhibiting the regeneration of cancer cells in a tumor, comprising contacting the area surrounding the tumor after surgery with an isolated nHC-HA / PTX3 complex to inhibit the regeneration of cancer cells in the area surrounding the tumor. In some cases, the method of killing cancer cells in a tumor comprises contacting the tumor or the area surrounding the tumor before, during, or after surgery with an isolated nHC-HA / PTX3 complex to kill the cancer cells. In some embodiments, a composition, product, or extract comprising isolated nHC-HA / PTX3 may be utilized as a method for inhibiting, reducing, or treating or reducing epithelial cell proliferation, cell migration, or epithelial-mesenchymal transition (EMT) in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of the pooled composition or product and a pharma- ceutically acceptable diluent, excipient, vehicle, or carrier to prevent or reduce epithelial cell proliferation, cell migration, or EMT. In some embodiments, any form of pooled composition, extract, or product disclosed herein may be administered as a treatment, medicament, or method for inhibiting regeneration of cancer cells in a tumor in an individual in need thereof, comprising contacting the area surrounding the tumor after surgery with isolated nHC-HA / PTX3 complex to inhibit regeneration of cancer cells in the area surrounding the tumor.

[0183] A method for killing cancer cells in a tumor in an individual in need thereof includes contacting the tumor or the area surrounding the tumor with an isolated nHC-HA / PTX3 complex before, during, or after surgery, thereby killing the cancer cells.

[0184] In some embodiments, fetal tissue products, compositions, such as nHC-HA / PTX3, may be used as bone implants comprising an outer surface coated with substantially isolated nHC-HA / PTX3 complex.

[0185] In some embodiments, the pooled compositions disclosed herein may be used to promote wound healing. In some embodiments, the pooled fetal support tissue products, such as the nHC-HA / PTX3 compositions, may be used to promote vascularization of tissues containing endothelial cells and pericytes, comprising contacting the tissue with a fetal support tissue product to reprogram the pericytes to a first progenitor cell phenotype, and contacting the tissue with a fetal support tissue product to reprogram the endothelial cells to a second progenitor cell phenotype. In some embodiments, such healing-promoting vascularization is involved in the treatment of ischemic tissues containing endothelial cells and pericytes, comprising contacting the tissue with a fetal support tissue product to reprogram the pericytes to a first progenitor cell phenotype, and contacting the tissue with a fetal support tissue product to reprogram the endothelial cells to a second progenitor cell phenotype. In some embodiments, the use is a pooled use. In some embodiments, the pooled fetal compositions disclosed herein may be used to promote neural regeneration. In some embodiments, methods comprising the use of the pooled compositions, extracts, or products disclosed herein promote, repair, reconstruct, replace, augment, or supplement damaged or reduced soft tissue in an individual in need. In some embodiments, contacting damaged or reduced soft tissue with the aforementioned products, extracts, or compositions reduces or prevents epidural fibrosis, adhesions, or scarring following spinal surgery in an individual in need. In some embodiments, the fetal support tissue products may be used in treating a disease or disorder, the treatment comprising contacting fibroblasts in a tissue affected by the disease in a subject in need with a composition comprising soluble nHC-HA / PTX3 for a period of time sufficient to reprogram the fibroblasts into cells having a different phenotype, thereby treating the disease or disorder.In some embodiments, the methods and / or compositions comprising pooled fetal support tissue products disclosed herein inhibit osteoclast differentiation, bone resorption, bone formation, bone remodeling, treat alveolar bone deterioration, treat Paget's disease. In some embodiments, the pooled tissue extracts, products formulations comprising nHC-HA / PTX3 disclosed herein, methods or compositions, devices formulated from pooled fetal support tissue, are formulated and administered for the treatment of bone tumors, or for the treatment of diseases, disorders, or conditions defined by deficient or defective bone formation, or for the treatment of osteoporosis, or to administer the treatment as an implant or patch, where the patch or implant is placed on the osteolytic bone or osteolytic joint of an individual in need. In some embodiments, the compositions are formulated into orthopedic prostheses. In some embodiments, the pooled fetal support tissue products, extracts, compositions, or methods may be administered to promote bone healing or to inhibit diseases or disorders characterized by bone injury, bone defects, bone deficiencies. In some embodiments, the pooled compositions disclosed herein are minimally manipulated. In some embodiments, the pooled compositions disclosed herein do not contain water, crystalloid fluids, or other formulated products except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled compositions disclosed herein do not have a systemic effect and do not rely on the metabolic activity of living cells for their primary function.

[0186] In some embodiments, the pooled composition or product comprises pooled fetal support tissue, such as umbilical cord amniotic membrane (UCAM), obtained from fresh, frozen, or previously frozen umbilical cords, and a storage medium, where substantially all of the cells of the UCAM are dead, and the UCAM is not dehydrated. In some embodiments, the pooled composition or product from the pooled fetal support tissue comprises a method of repairing, reconstructing, replacing, augmenting, or supplementing damaged or reduced soft tissue of an individual in need, comprising contacting the damaged or reduced soft tissue of the individual with an umbilical cord product from which water has not been removed.

[0187] In some embodiments, the pooled fetal support tissue product or composition may be utilized as a method for reducing or preventing epidural fibrosis, adhesions, or scarring following spinal surgery in an individual in need thereof, such method comprising contacting the umbilical cord product with the spinal surgery site.

[0188] In some embodiments, the pooled fetal support tissue product may be used as a composition or product or method for treating ocular wounds or repairing damaged ocular tissue in an individual in need. In some embodiments, the pooled composition, product, or extract disclosed herein may be used as a medicament, treatment, or method for treating or preventing proliferative vitreoretinopathy (PVR) in an individual with PVR or for preventing PVR in an individual who has experienced retinal detachment, which includes administering to an individual who has PVR or has experienced retinal detachment, and the treatment may include administering substantially isolated native heavy chain hyaluronic acid / pentraxin 3 complex (HC-HA / PTX3), reconstituted HC-HA / PTX3, or a combination thereof using the methods disclosed herein.

[0189] In some embodiments, the pooled compositions, products, extracts disclosed herein may be used as a drug, treatment, or method for treating or preventing proliferative vitreoretinopathy (PVR) in individuals with PVR or for preventing PVR in individuals who have experienced retinal detachment, and the pooled products are administered to individuals who have PVR or have experienced retinal detachment as described herein to provide a substantially isolated natural HC-HA / PTX3 product and an additional therapeutic agent for treating or preventing PVR. In this disclosure, providing in a therapeutically effective amount should be taken to mean any dosage or treatment that provides relief or cure according to any popular procedure or dosage that is considered effective for such treatment, prevention, or relief of the affliction, pain, or disease for which treatment or prevention is provided.

[0190] In some embodiments, the pooled compositions disclosed herein are used as dressings (e.g., wound dressings). In some embodiments, the use is pooled use. In some embodiments, the pooled compositions are minimally manipulated. In some embodiments, the pooled compositions do not contain water, crystalloids, or other excipients except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled compositions have no systemic effect and are not dependent on the metabolic activity of living cells for their primary function.

[0191] In some embodiments, the pooled compositions disclosed herein are used to promote wound repair. In some embodiments, the use is pooled use. In some embodiments, the pooled composition is minimally manipulated. In some embodiments, the pooled composition does not contain water, crystalloids, or other excipients except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled composition does not have a systemic effect and does not rely on the metabolic activity of living cells for its primary function.

[0192] In some embodiments, the pooled compositions disclosed herein are used as adhesion barriers. In some embodiments, the pooled compositions are minimally manipulated. In some embodiments, the pooled compositions do not contain water, crystalloids, or other excipients except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled compositions have no systemic effects and are not dependent on the metabolic activity of living cells for their primary function.

[0193] Repairing and supplementing damaged tissue In some embodiments, the pooled compositions disclosed herein are used as wound dressings or to promote wound repair. In some embodiments, the pooled compositions are minimally manipulated. In some embodiments, the pooled compositions do not contain water, crystalloids, or other excipients except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled compositions do not have systemic effects and do not rely on the metabolic activity of living cells for their primary function.

[0194] In some embodiments, the tissue is damaged, damaged, or lost due to injury (e.g., burns, incisions, necrotic areas caused by infection, trauma, or toxins, lacerations). In some embodiments, the tissue is damaged, damaged, or lost due to burns. In some embodiments, the tissue is damaged, damaged, or lost due to wounds (e.g., incisions, lacerations, abrasions). In some embodiments, the tissue is damaged, damaged, or lost due to necrosis. In some embodiments, the tissue is damaged, damaged, or lost due to ulceration. In some embodiments, the pooled compositions are administered by parenteral infusion (e.g., by infusion or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled compositions are administered epidurally, intrathecally, by inhalation, intravenously, or a combination thereof.

[0195] In some embodiments, the pooled compositions disclosed herein comprise HA, nHC-HA / PTX3 complexes, and various bioactive factors found, for example, in the stroma of AM / UC that promote tissue repair, including anti-angiogenic and anti-inflammatory proteins. In some embodiments, the HA, nHC-HA / PTX3 complexes in the pooled compositions disclosed herein diffuse from the pooled composition into the surrounding tissue.

[0196] Burns In some embodiments, the pooled composition disclosed herein is applied to a burn. In some embodiments, the pooled composition disclosed herein is applied to a first degree burn. In some embodiments, the pooled composition disclosed herein is applied to a second degree burn. In some embodiments, the pooled composition disclosed herein is applied to a third degree burn. In some embodiments, the pooled composition is applied to a substrate before being placed on the burn.

[0197] wound In some embodiments, the pooled compositions disclosed herein are applied to a skin wound (e.g., an incision, a laceration, an abrasion, an ulcer, a puncture wound, a penetration). In some embodiments, the pooled composition is applied to a substrate before being placed on the wound. In some embodiments, the pooled composition is administered by parenteral infusion (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravesical, subconjunctival, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0198] In some embodiments, the pooled compositions disclosed herein are applied to an incision scar of an organ (e.g., skin, brain, stomach, kidneys, liver, intestines, lungs, bladder, trachea, esophagus, vagina, ureters, and blood vessel walls). In some embodiments, the pooled compositions disclosed herein are applied to an incision scar. In some embodiments, the pooled compositions disclosed herein are applied to a site of a colectomy. In some embodiments, the pooled compositions disclosed herein are applied to a site of a gastrectomy. In some embodiments, the pooled compositions disclosed herein are applied to a site of breast surgery (e.g., breast reduction surgery, breast augmentation surgery, and mastectomy). In some embodiments, the pooled compositions are applied to a substrate prior to being placed over a wound.

[0199] In some embodiments, the pooled compositions disclosed herein are used as dressings to cover skin incision scars (e.g., epidermis, dermis, and / or subcutaneous tissue incision scars). In some embodiments, the pooled compositions disclosed herein are used to repair or supplement skin after hemorrhoid surgery. In some embodiments, the pooled compositions are applied to a substrate before being placed on a wound.

[0200] Necrosis In some embodiments, the pooled compositions disclosed herein are used as a protective graft to cover an area of ​​necrotic tissue (e.g., from infection). In some embodiments, the pooled compositions disclosed herein are used as a protective graft to cover an area of ​​necrotic skin. In some embodiments, the pooled compositions disclosed herein are placed on an area of ​​necrotic tissue. In some embodiments, the pooled compositions are applied to a substrate before being placed on the necrotic tissue. In some embodiments, the pooled compositions are administered by parenteral injection (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled compositions are administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0201] ulcer In some embodiments, the pooled compositions disclosed herein are used as a protective dressing to cover an ulcer. In some embodiments, the pooled compositions are applied to a substrate before being placed on the ulcer. In some embodiments, the pooled compositions are administered by parenteral infusion (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled compositions are administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0202] In some embodiments, the ulcer is a foot ulcer (e.g., a diabetic foot ulcer or an arterial insufficiency ulcer). In some embodiments, treating the foot ulcer comprises (a) preparing the wound (e.g., by debriding the wound), and (b) placing a pooled composition disclosed herein on the wound. In some embodiments, treating the foot ulcer comprises (a) preparing the wound (e.g., by debriding the wound), and (b) placing a pooled composition disclosed herein on the wound, and (c) covering the pooled composition with a protective barrier (e.g., a Silvercell dressing, metipel, gauze, or bandage). In some embodiments, the pooled composition is applied to a substrate before being placed on the ulcer. In some embodiments, the ulcer is a venous stasis (VS) ulcer.

[0203] In some embodiments, treating a VS ulcer comprises (a) preparing the wound (e.g., by debriding the wound) and (b) placing a pooled composition disclosed herein over the wound. In some embodiments, treating a VS ulcer comprises (a) preparing the wound (e.g., by debriding the wound), (b) placing a pooled composition disclosed herein over the wound, and (c) covering the pooled composition with a protective barrier (e.g., a wound veil, antimicrobial dressing, gauze, or bandage). In some embodiments, the pooled composition is applied to a substrate prior to being placed over the wound.

[0204] In some embodiments, the ulcer is a corneal ulcer (i.e., ulcerative keratitis). In some embodiments, treating a corneal ulcer includes (a) preparing a wound (e.g., by debriding the wound) and (b) placing a pooled composition disclosed herein on the wound. In some embodiments, treating a corneal ulcer includes (a) preparing a wound (e.g., by debriding the wound) and (b) placing a pooled composition disclosed herein on the wound and (c) covering the pooled composition or the pooled composition with a protective barrier (e.g., a contact lens or a bandage). In some embodiments, the pooled composition is applied to a substrate before being placed on the wound.

[0205] Soft Tissue Use Disclosed herein, in certain embodiments, is the use of the pooled composition disclosed herein to repair, reconstruct, replace, or replenish damaged, impaired, or lost recipient soft tissue (e.g., tendons). In certain embodiments, the pooled composition, product, or extract described herein, in sheet or non-sheet form, may be used to treat or as a method for reprogramming cells having a first phenotype, comprising contacting cells having a first phenotype with a composition comprising soluble NHC-HA / PTX3 for a sufficient period of time to reprogram the cells having a first phenotype into a second cell having a second phenotype. In some embodiments, the pooled composition may be used to treat or as a method for treating a disease or disorder in a subject in need thereof, comprising contacting fibroblasts in a tissue affected by the disease in a subject in need thereof with a composition comprising soluble nHC-HA / PTX3 for a sufficient period of time to reprogram the fibroblasts into a cell having a different phenotype, thereby treating the disease or disorder. In certain embodiments, any form of pooled composition, extract, or product described herein may be used as a treatment, medicament, or method for regenerating tissue comprising contacting cells having a first differentiated phenotype with a composition comprising soluble nHC-HA / PTX3 for a period of time sufficient to reprogram the cells having a first differentiated phenotype into progenitor cells having a progenitor cell phenotype, and differentiating the progenitor cells into a second differentiated phenotype.

[0206] In some embodiments, the use is a pooled use. In some embodiments, the pooled composition is minimally manipulated. In some embodiments, the pooled composition does not contain water, crystalloids, or other formulated products except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled composition has no systemic effects and does not depend for its primary function on the metabolic activity of living cells.

[0207] In some embodiments, the pooled compositions disclosed herein comprise HA, nHC-HA / PTX3 complexes, and various bioactive factors found, for example, in the stroma of AM / UC that promote tissue repair, including anti-angiogenic and anti-inflammatory proteins. In some embodiments, the HA, nHC-HA / PTX3 complexes in the pooled compositions disclosed herein diffuse from the pooled compositions into the surrounding tissue.

[0208] In some embodiments, the pooled compositions described and disclosed herein are used as a dressing to cover a soft tissue incision scar (e.g., the eyelid forms a tissue plane between different layers of soft tissue). In some embodiments, the pooled compositions are applied to a substrate and then used as a dressing to cover a soft tissue incision scar (e.g., the eyelid forms a tissue plane between different layers of soft tissue). In some embodiments, the pooled compositions are administered by parenteral injection (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled compositions are administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0209] In some embodiments, the pooled fetal support tissue compositions / extracts, methods, kits, devices, etc. disclosed herein are useful treatments or therapies for bladder disorders. In some embodiments, the bladder disorder may be a bladder disease, bladder syndrome, bladder disease, and / or bladder infection. For example, the bladder disorder may be, for example, urinary incontinence. For example, the bladder disorder may be, for example, bladder cancer. In some embodiments, the bladder disease may be associated with cancer treatment or tumor treatment, for example treatment for cancer of the lower bowel or lower urinary tract. In some embodiments, the present disclosure may be administered to the soft tissue of the bladder of patients with interstitial cystitis, overactive bladder, or other neurourological dysfunction of the lower urinary tract.

[0210] In some embodiments, the pooled compositions disclosed herein are used as structural (plastic) supports for soft tissue.

[0211] In some embodiments, the pooled compositions disclosed herein prevent adhesions in bone or joint repair.

[0212] In some embodiments, the pooled compositions disclosed herein are used in tendon or joint repair, such as rotator cuff repair, Achilles tendinopathy, and hand tendon repair. In some embodiments, the pooled compositions disclosed herein are used to strengthen tendons or joints. In some embodiments, the pooled compositions disclosed herein are used to prevent healing tendons from adhering to surrounding tissues, tendons, or joints. In some embodiments, the pooled compositions disclosed herein are used to prevent scar tissue formation in tendons.

[0213] In some embodiments, the pooled compositions disclosed herein are applied to a matrix and the matrix / pooled composition is used to reinforce small tendons and ligaments of the foot and ankle, including the posterior tibial tendon, peroneal tendon, flexor and extensor tendons, and lateral ankle ligament. In some embodiments, the pooled compositions disclosed herein are applied to a matrix and the matrix / pooled composition is used to reinforce primary repair of the quadriceps and patellar tendons around the knee. In some embodiments, the pooled compositions disclosed herein are applied to a matrix and the matrix / pooled composition is used as a periosteal patch for bone grafts in joint replacement. In some embodiments, the pooled compositions disclosed herein are applied to a matrix and the matrix / pooled composition is used to reinforce defective hip and knee capsule tissue following total joint revision surgery.

[0214] In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used in repairing a torn rotator cuff. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a patch to cover a rotator cuff muscle or tendon (e.g., supraspinatus tendon). In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used to reconstruct a rotator cuff muscle or tendon (e.g., supraspinatus tendon). In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used to augment a rotator cuff muscle or tendon (e.g., supraspinatus tendon). In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used to strengthen a rotator cuff muscle or tendon (e.g., supraspinatus tendon). In some embodiments, a pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used to prevent soft tissue adhesions to the rotator cuff muscles or tendons (e.g., supraspinatus tendon).

[0215] In some embodiments, the pooled compositions disclosed herein are used in the repair of gums. In some embodiments, the pooled compositions disclosed herein are used in the repair of gum recession. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and used as a patch to cover the gums. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and used as a patch to cover exposed root surfaces. In some embodiments, the pooled compositions disclosed herein are used to reconstruct gums. In some embodiments, the pooled compositions disclosed herein are used to augment gums. In some embodiments, the pooled compositions disclosed herein are used to strengthen gums. In some embodiments, the pooled compositions disclosed herein are used to prevent soft tissue adhesion to gums.

[0216] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a protective graft to cover a scar or tear in the fascia. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the fascia. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement or supplement for the fascia. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair a hernia (e.g., to repair the fascia). In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair an inguinal hernia. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair a femoral hernia. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair an umbilical hernia. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair incisional hernias. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair diaphragmatic hernias. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair Cooper's hernia, epigastric hernia, hiatal hernia, little hernia, lumbar hernia, meidl's hernia, obturator hernia, pantaloon hernia, paraesophageal hernia, paraumbilical hernia, perineal hernia, preperitoneal hernia, richter's hernia, sliding hernia, sciatic hernia, spigelian hernia, sports hernia, velpeau hernia, or amiand hernia.

[0217] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to repair a herniated disc. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a protective implant to cover a scar or tear in the disc. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a protective implant to cover a scar or tear in the annulus. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the disc. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the annulus. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement or replacement for the disc. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the intervertebral disc. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement or supplement for the annulus fibrosus.

[0218] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to cover the incision scar of the brain or one (or all) of the meninges (i.e., dura, pia, and / or arachnoid). In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for one (or all) of the meninges (i.e., dura, pia, and / or arachnoid). In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement for one (or all) of the meninges (i.e., dura, pia, and / or arachnoid).

[0219] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to cover the incision in the lung or pleura. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the pleura. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement for the pleura.

[0220] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to cover the incision scar of the tympanic membrane. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the tympanic membrane. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement for the tympanic membrane.

[0221] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a protective implant to cover an incision or tear in the heart or pericardium. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the pericardium. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement for the pericardium.

[0222] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a protective implant to cover an incision or tear in the peritoneum. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for the peritoneum. In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used as a replacement for the peritoneum.

[0223] Ocular Use Disclosed herein in certain embodiments is the use of the pooled compositions disclosed herein to repair, reconstruct, replace, or replenish damaged, impaired, or lost recipient ocular tissue. In some embodiments, the pooled compositions are administered by parenteral injection (e.g., by injection or infusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled compositions are administered by epidural, intrathecal, inhalation, intravenous, or combinations thereof.

[0224] In some embodiments, the use is a pooled use. In some embodiments, the pooled composition is minimally manipulated. In some embodiments, the pooled composition does not contain water, crystalloids, or other formulated products except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled compositions disclosed herein have no systemic effect and are not dependent on the metabolic activity of living cells for their primary function.

[0225] In some embodiments, the pooled compositions disclosed herein comprise HA, nHC-HA / PTX3 complexes, and various bioactive factors found, for example, in the stroma of AM / UC that promote tissue repair, including anti-angiogenic and anti-inflammatory proteins. In some embodiments, the HA, nHC-HA / PTX3 complexes in the pooled compositions disclosed herein diffuse from the pooled compositions into the surrounding tissue.

[0226] Treatment of glaucoma As used herein, "glaucoma" refers to a disorder characterized by the loss of retinal ganglion cells in the optic nerve.In certain cases, glaucoma is partially or completely caused by the increase in intraocular pressure in the anterior chamber (AC).Intraocular pressure fluctuates according to the production of aqueous humor by the ciliary process of the eye and the outflow of aqueous humor through the trabecular meshwork.

[0227] Glaucoma drainage devices (GDDs) are medical devices implanted in the eye to relieve intraocular pressure by providing an alternative route for drainage of aqueous humor. If left uncovered, GDD tubes can corrode, making the eye more susceptible to intraocular infections. Therefore, it is necessary to cover GDD tubes. Currently, patches used to cover GDD tubes are made of pericardium, sclera, and cornea. These patches are approximately 400-550 microns in thickness. Such thin patches can result in 25% dissolution over a two-year period, leaving the shunt tubes exposed again.

[0228] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used to coat the GDD tubing. In some embodiments, the substrate / pooled composition is 300-600 microns thick. In some embodiments, the substrate / pooled composition does not dissolve by more than 25% over a two year period. In some embodiments, the pooled composition is administered by parenteral injection (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0229] Treatment of eye ulcers In some embodiments, the pooled nHC-HA / PTX3 (complex) composition disclosed herein is applied to a substrate and the substrate / pooled composition is used to cover ocular epithelial defects and / or ulcers. In some embodiments, the pooled composition is administered in patients with defects or ulcers caused by dry eye, graft-versus-host disease, Graves' disease, traumatic corneal injury, corneal surgery, or neuroparalytic keratopathy. The present disclosure may be administered in combination with contact lenses, such as, but not limited to, protective contact lenses (BCLs) or scleral lenses. The utility of combination therapy has been demonstrated, for example, by administering Clarix / Neox Flo products in combination with BCLs to treat subjects with mild to moderate dry eye disease and when subjects are refractory to standard treatments. In such cases, the Flo-BCL combination may be administered for 2-3 days. In some embodiments, the present disclosure may be reconstituted in saline and applied within the concave surface of a BCL, which is further administered or applied to the corneal surface of a subject with mild to moderate dry eye disease (e.g., graded as DEWS (international dry eye workshop) severity 1-2) who may exhibit superficial epithelial keratopathy and / or remain symptomatic despite standard treatment. In some embodiments, the present disclosure may be administered in combination with a BCL, which may be a brand of BCL certified by an internationally recognized authority such as the United States Federal Drug Administration (FDA). For example, the BCL may be any trademark, such as, for example, ACUVUE OASYS®, AIR OPTIX® NIGHT and DAYR® AQUA, Bausch and Lomb PureVision®, Bausch and Lomb PureVision® 2, among other brands.In certain embodiments, the present disclosure reconstituted in saline and applied to the concave surface of the BCL may remain on the subject's corneal surface for an extended period of time of 3 days, 4 days, 5 days or more. In certain embodiments, the length of time may be 5 days or more. In some embodiments, the treatment is placed on the subject's cornea for at least 1 day, 2 days, 3 days, 4 days, or 5 days. In some embodiments, the fetal support tissue product may include a minced product, which may be a product or composition that includes pooled or unpooled fetal support tissue. For example, the minced product may include pooled fetal support tissue, including, for example, amniotic membrane and umbilical cord tissue, which is a minced, centrifuged, sterile filtered fetal support tissue product / extract or composition (hereinafter referred to as the minced product or TTBT01 drug product). In some embodiments, the TTBT01 drug product may include fetal support tissue from multiple donors and / or donor lots. In some embodiments, the TTBT01 drug product may include fetal support tissue from multiple donors and / or donor lots. In some embodiments, the TTB01 drug product or shredded product may be packaged in a single-use unit dose vial. In some embodiments, the TTBT01 drug product or shredded product may be applied to the concave spherical ocular surface of the PROSE or BostonSight SCLERAL lens in patients with severe dry eye, typically DEWS severity 3-4. The PROSE and BostonSight SCLERAL lenses are Class II FDA 510(k) cleared medical devices. Such products, combining TTBT01 with a selected scleral lens, may be administered to a subject as specified in the user guide. For example, TTBT01 may be loaded into the scleral lens on a daily basis and applied to the eye for at least 4 hours. In some embodiments, the TTBT01 drug product may be loaded into the scleral lens every other day, every 2 days, or more days, and applied intraocularly for a length of time as specified by a trained ophthalmologist or trained medical practitioner (physician).Administration of the TTBT01 drug product to the scleral lens and application to the eye may be done periodically during the treatment period, for example, daily for 5 to 7 days, for best results. In other cases, administration may be for less than 5 days or for 8 days or more. Generally, the drug product in the scleral lens may be reconstituted or administered daily, or as determined by the response of the physician and the subject being treated. In some embodiments, the pooled composition is administered by parenteral infusion (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof. In some embodiments, the substrate is a protective contact lens or a scleral lens.

[0230] In some embodiments, the base of the ulcer is debrided with a surgical sponge and poorly attached epithelium adjacent to the edge of the ulcer (e.g., in the portion of the eye where the epithelium becomes fully attached) is removed. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is transferred to the recipient's eye. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is then secured to the eye by embedded suture knots (e.g., interrupted 10-0 nylon sutures or continuous 10-0 nylon sutures). In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is secured to the eye by the use of fibrin glue. In some embodiments, a protective layer is applied over the pooled composition / substrate or the entire eye (e.g., a contact lens). In some embodiments, the substrate / pooled composition further comprises an antibiotic (e.g., neomycin, polymyxin b sulfate, and dexamethasone).

[0231] In some embodiments, the pooled compositions disclosed herein are applied to a substrate and the substrate / pooled composition is used in the reconstruction of conjunctival, scleral, eyelid, and orbital rim surfaces. In some embodiments, the conjunctival surface damage results from lysis of blepharophimosis, surgical removal of tumors, lesions, and / or scar tissue, excimer laser photorefractive keratectomy and therapeutic keratectomy, or a combination thereof.

[0232] Coronary Artery Use Disclosed herein, in certain embodiments, is the use of the pooled nHC-HA / PTX3 (complex) compositions disclosed herein to repair, reconstruct, replace, or replenish damaged, compromised, or lost coronary artery tissue. In some embodiments, the pooled compositions are administered by parenteral infusion (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous), hi some embodiments, the pooled compositions are administered epidurally, intrathecally, by inhalation, intravenously, or a combination thereof.

[0233] In some embodiments, the use is a pooled use. In some embodiments, the pooled composition is minimally manipulated. In some embodiments, the AM does not contain water, crystalloids, or other formulations except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled composition has no systemic effect and does not depend on the metabolic activity of living cells for its primary function.

[0234] In some embodiments, the pooled compositions disclosed herein comprise HA, nHC-HA / PTX3 complexes, and various bioactive factors found, for example, in the stroma of AM / UC that promote tissue repair, including anti-angiogenic and anti-inflammatory proteins. In some embodiments, the HA, nHC-HA / PTX3 complexes in the pooled compositions disclosed herein diffuse from the pooled compositions into the surrounding tissue.

[0235] Coronary artery bypass grafting Disclosed herein is the use of the pooled NHC-HA / PTX3 (complex) composition described herein in coronary artery bypass surgery. In some embodiments, the pooled composition disclosed herein is applied to a matrix, and the matrix / pooled composition is implanted into a coronary artery to bypass a portion of the artery characterized by atherosclerosis. In some embodiments, the pooled composition is administered by parenteral injection (e.g., by infusion or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0236] Heart valves In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a substrate and the substrate / pooled composition is applied over a heart valve. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for a heart valve. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a replacement for a heart valve. In some embodiments, the pooled composition is administered by parenteral injection (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0237] Veins and Arteries In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a vein or artery. In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a matrix and the matrix / pooled composition is applied to a vein or artery. In some embodiments, the pooled composition disclosed herein is applied to a matrix and the matrix / pooled composition is used as a structural (plastic) support for a vein or artery. In some embodiments, the pooled composition is administered by parenteral injection (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0238] Neurological Use In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a nerve or nerve tissue. In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a dressing to cover a nerve (e.g., peripheral nerve). In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a dressing to cover a nerve graft, nerve transfer, or repair nerve. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a dressing to cover a nerve (e.g., peripheral nerve) incision scar. In some embodiments, the pooled composition disclosed herein is applied to a substrate and the substrate / pooled composition is used as a structural (plastic) support for a nerve (e.g., peripheral nerve). In some embodiments, the pooled composition disclosed herein prevents adhesions in nerve repair.

[0239] Disclosed herein in certain embodiments is the use of pooled nHC-HA / PTX3 (complex) compositions to repair, reconstruct, replace, or replenish damaged, impaired, or lost recipient nerve tissue. Various diseases are associated with neuralgia and nerve damage. Exemplary diseases associated with neuralgia include, but are not limited to, pain in the mammalian musculoskeletal or locomotor systems, which may include skeletal, muscular, and connective tissues, including tendons, cartilage, and ligaments. Musculoskeletal pain may result from injury, which may be a temporary or lifelong condition, or injury, which may result in ongoing functional limitations, impairment, and pain from bone or joint deformities, which may include, for example, pulled muscles, repetitive overuse, sprains, dislocations, fractures, or scoliosis, which is a curvature of the spine, or combinations thereof. In some embodiments, musculoskeletal diseases may affect joints, including, for example, osteoarthritis, rheumatoid arthritis, psoriatic arthritis, gout, ankylosing spondylitis, and other diseases. In some embodiments, the musculoskeletal disease may affect bone, including, for example, osteoporosis, osteopenia and associated diseases such as fragility fractures, traumatic fractures, and amputations. In some embodiments, the musculoskeletal disease may affect muscle, including, for example, sarcopenia, or pain, including, for example, back pain and neck pain. In some embodiments, the musculoskeletal pain or disease may affect multiple body regions or systems, including, for example, localized body or system pain, widespread pain disorders, inflammatory diseases including systemic lupus erythematosus, connective tissue diseases including bursitis, and vasculitis, among other diseases that may result from musculoskeletal pain, infection, or injury. In some embodiments, the pooled composition is administered to treat diseases including neuropathic pain, such as musculoskeletal pain.

[0240] In some embodiments, the pooled nHC-HA / PTX3 (complex) composition is administered by parenteral infusion (e.g., by injection or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered epidurally, intrathecally, by inhalation, intravenously, or a combination thereof.

[0241] In some embodiments, the use is a pooled use. In some embodiments, the pooled composition is minimally manipulated. In some embodiments, the pooled composition does not contain water, crystalloids, or other formulated products except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled composition has no systemic effects and does not depend for its primary function on the metabolic activity of living cells.

[0242] Disclosed herein is a process for producing fetal support tissue while maintaining the biological activity of the fetal support tissue, including components of the fetal support tissue, such as, for example, HA, HC-HA / PTX3, growth factors, cytokines, and other active biological components, such that the efficacy of HC-HA / PTX3 is observable to ensure sufficient retention as disclosed throughout this specification and claims. In some embodiments, nHC-HC / PTX3 and other biologically active components found in the pooled compositions disclosed herein diffuse out of the pooled compositions into the surrounding tissues.

[0243] In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a substrate, and the substrate / pooled composition is used as a non-contractile encasement for injured nerves. In some embodiments, the pooled composition described herein prevents or minimizes scar formation, encapsulation, chronic compression, nerve tethering, and nerve encasement. In some embodiments, the pooled composition described herein prevents or minimizes neuroma formation. In some embodiments, the pooled composition described herein prevents or minimizes the migration of endogenous growth factors (e.g., nerve growth factor) present during nerve repair.

[0244] Spinal Use Disclosed herein, in certain embodiments, is the use of the pooled compositions described herein during spinal surgery.

[0245] In some embodiments, the pooled compositions described herein are used during laminectomy. In some embodiments, the use is pooled use. In some embodiments, the pooled composition is minimally manipulated. In some embodiments, the pooled composition does not contain water, crystalloids, or other excipients except for disinfectants, antiseptics, or preservatives. In some embodiments, the pooled composition has no systemic effects and does not rely on the metabolic activity of living cells for its primary function.

[0246] In some embodiments, the pooled composition disclosed herein comprises HA, nHC-HA / PTX3 complexes, and various bioactive factors found, for example, in the interstitium of AM / UC, that promote tissue repair, including anti-angiogenic and anti-inflammatory proteins. In some embodiments, the aforementioned HA, nHC-HA / PTX3 complexes in the pooled composition disclosed herein diffuse from the pooled composition into the surrounding tissue. In some embodiments, the pooled composition is administered by parenteral infusion (e.g., by infusion or infusion, including intra-arterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or a combination thereof.

[0247] In some embodiments, the pooled compositions described herein are used to reduce or prevent epidural fibrosis and / or scar adhesions after spinal surgery (e.g., laminectomy). In some embodiments, the pooled compositions described herein are implanted between the dura mater and the overlying tissue after spinal surgery (e.g., laminectomy). In some embodiments, implanting the pooled compositions described herein between the dura mater and the overlying tissue after spinal surgery (e.g., laminectomy) reduces or prevents fibroblast migration into the dura mater and collagen deposition in the dura mater.

[0248] In some embodiments, the pooled compositions described herein are used to reduce or prevent the progression of proliferative scarring after spinal surgery (e.g., laminectomy). In some embodiments, the pooled compositions described herein are used to reduce or prevent the progression of epidural / peridural / perineural scarring after surgery (e.g., laminectomy). In some embodiments, the pooled compositions described herein are used to reduce or prevent the progression of proliferative scarring after spinal surgery (e.g., laminectomy). In some embodiments, the pooled compositions disclosed herein are used to reduce or prevent the progression of epidural / peridural / perineural scarring after laminectomy.

[0249] In some embodiments, the pooled compositions described herein are used to reduce or prevent the progression of epidural compression or dural tethering after spinal surgery (e.g., laminectomy). In some embodiments, the pooled compositions described herein are used to reduce or prevent the progression of nerve root tethering after spinal surgery (e.g., laminectomy). In some embodiments, the pooled compositions described herein are used to reduce or prevent the progression of arachnoiditis after spinal surgery (e.g., laminectomy).

[0250] In some embodiments, the pooled composition disclosed herein further comprises morselized bone tissue. In some embodiments, the pooled composition disclosed herein comprising morselized bone tissue is used during spinal fusion surgery. In some embodiments, the pooled composition disclosed herein comprising morselized bone tissue is embedded between adjacent vertebrae. In some embodiments, embedding the pooled composition disclosed herein comprising morselized bone tissue between two adjacent vertebrae promotes vertebral fusion.

[0251] In some embodiments, the pooled compositions disclosed herein are used as a protective implant to cover the dura incision scar. In some embodiments, the pooled compositions disclosed herein are applied to a substrate, and the substrate / pooled composition is used as a structural (plastic) support for the dura. In some embodiments, the pooled compositions disclosed herein are applied to a substrate, and the substrate / pooled composition is used as a replacement for the dura.

[0252] Other Uses of Pooled Compositions In some embodiments, the pooled NHC-HA / PTX3 (complex) composition disclosed herein is applied to a patch or wound dressing. In some embodiments, the pooled composition is administered by parenteral injection (e.g., by injection or infusion, including intraarterial, intracardiac, intradermal, intraduodenal, intramedullary, intramuscular, intraosseous, intraperitoneal, intrathecal, intravascular, intravenous, intravitreal, epidural, and / or subcutaneous). In some embodiments, the pooled composition is administered by epidural, intrathecal, inhalation, intravenous, or combination thereof.

[0253] In some embodiments, the pooled compositions disclosed herein are used as dermal fillers. In some embodiments, the pooled compositions disclosed herein are injected into the facial skin sub-structure. In some embodiments, the pooled compositions disclosed herein are injected under facial wrinkles and age lines (e.g., nasolabial folds, cheek folds, "crow's feet", and forehead wrinkles). In some embodiments, the pooled compositions disclosed herein are used for lip augmentation. In some embodiments, the pooled compositions disclosed herein are injected into the lips. In some embodiments, the pooled compositions disclosed herein are applied after or in conjunction with a skin rejuvenation therapy, such as laser cauterization.

[0254] In some embodiments, the pooled compositions disclosed herein are used to treat arthritis (e.g., osteoarthritis, rheumatoid arthritis, septic arthritis, ankylosing spondylitis, spondylosis). In some embodiments, the pooled compositions disclosed herein are injected into an arthritic joint (e.g., the knee).

[0255] In some embodiments, the pooled nHC-HA / PTX3 (complex) composition disclosed herein is used to inhibit bone resorption in an individual in need thereof. In some embodiments, the individual suffers from arthritis, osteoporosis, alveolar bone deterioration, Paget's disease, or bone tumor. In some embodiments, the pooled compositi...

Claims

1. A composition comprising pooled umbilical cord and amniotic (MAU) tissue, wherein the pooled MAU tissue comprises MAU tissue from a plurality of donors, the pooled MAU tissue is freeze-dried, the freeze-dried pooled MAU tissue comprises a natural HC-HA / PTX3 (nHC-HA / PTX3) complex, the nHC-HA / PTX3 complex possesses therapeutic efficacy, and the freeze-dried pooled MAU tissue is richer in total protein than unpooled MAU tissue.

2. The composition according to claim 1, comprising umbilical amniotic membrane, Howardon's jelly, or any combination thereof.

3. The composition according to claim 1, wherein the composition is ultimately sterilized, gamma irradiated, filtered, membrane filtered, electron beam sterilized, or any combination thereof.

4. The composition according to claim 1, wherein the pooled MAU tissue is cut, pulverized, pulverized, finely ground, crushed, or a combination thereof.

5. The composition according to claim 1, wherein the pooled MAU tissue is substantially free of metabolic activity.

6. The composition according to claim 1, wherein the MAU structure comprises an average particle size of about 0.01 micrometers (μm) to about 240 μm in diameter.

7. The composition according to claim 1, wherein the MAU structure comprises an average particle size of about 0.5 μm.

8. The composition according to claim 1, wherein the composition is formulated as a gel, powder, or liquid.

9. The composition according to claim 1, wherein, as determined by an ODI-TRAP assay, the freeze-dried pooled MAU tissue shows reduced variability of the natural HC-HA / PTX3 complex compared to the unpooled MAU tissue, and the pooled MAU tissue and the unpooled MAU tissue are processed in the same manner and obtained from the same type of tissue source.

10. The composition according to claim 9, wherein the reduction in the variation of the natural HC-HA / PTX3 complex includes a reduction in variation determined by the coefficient of variation.

11. The composition according to claim 10, wherein the coefficient of variation is 3.0% or less.

12. The composition according to claim 1, wherein the total protein is determined by a bicinchoninic acid (BCA) assay.

13. The composition according to claim 1, wherein the composition inhibits TRAP activity by at least about 25% compared to RANKL 50 μm.

14. The composition according to claim 1, wherein the MAU tissue is stored at a low temperature or is pre-frozen.

15. A method for producing the composition described in Claim 1, the method comprising obtaining MAU tissue from a plurality of donors and producing pooled MAU tissue by pooling the MAU tissue from the plurality of donors, wherein the pooled MAU tissue is freeze-dried.

16. The method according to claim 15, wherein the MAU tissue is not dehydrated.

17. The method according to claim 15, wherein the MAU tissue is freeze-dried before or after the MAU tissue is pooled.

18. The method according to claim 15, further comprising processing the MAU structure by flaking, pulverizing, micronizing, or a combination thereof.

19. A method for treating a disease of an individual, wherein the disease includes an inflammatory disorder, bladder disease, bone disorder, wound, eye disorder, scar disease, pain disorder, nerve disorder, spina bifida, or any combination thereof of the individual requiring treatment, and the method comprises treating the disease by administering the composition according to claim 1 to the individual.

20. The method according to claim 19, wherein the inflammatory disorder includes dry eye.

21. The method according to claim 19, wherein the pain disorder includes shoulder pain.

22. The method according to claim 19, wherein the bone disorder includes osteoarthritis.

23. The method according to claim 19, wherein the bone disorder includes osteoporosis.

24. The method according to claim 19, wherein the eye disorder includes dry eye, ocular surface inflammation, or ulcerative keratitis.

25. The method according to claim 19, wherein the wound includes a compound wound.

26. A method for determining the efficacy of the freeze-dried pooled MAU tissue composition according to Claim 1, wherein the method is: Perform an efficacy cell-based assay on the composition described in claim 1. The efficacy cell-based assay includes, (i) Osteoclast differentiation inhibition assay using tartrate-resistant acid phosphatase (ODI-TRAP), (ii) Nitric oxide (NO assay), or (iii) Macrophage polarization (M2) assay A method comprising the lyophilized pooled MAU tissue composition, wherein the composition exhibits statistically significant inhibition of biological activity in the potency cell-based assay of (i), (ii), or (iii) compared to the assay of a positive control.

Images