System for producing fat-containing skin graft treatment composition, fat-containing skin graft treatment composition, and method for producing same

Abstract

A system for preparing a wound treatment product is provided, including an applicator (110) configured to apply a fat material to skin graft material (150). A wound treatment kit is provided, including skin graft material and an applicator configured to apply a fat material to the skin graft material. A wound treatment composition is provided, including a skin graft material and a fat material, such as for promoting cellular regeneration growth into a wound. A method is provided for preparing a wound treatment, including providing a fat material, providing a skin graft material, and applying the fat material to the skin graft material. A method is provided for treating a wound, including providing a skin graft material, applying the fat material to the skin graft material, and applying the combined fat and skin graft material to the wound.

Description

[Technical Field]

[0001] The present disclosure relates generally to wound care, wound care products for stabilizing, protecting, and / or healing damaged tissue, and methods and systems for manufacturing wound care products. [Background technology]

[0002] Healthy skin serves several distinct functions, including protecting underlying tissues from abrasion, microorganisms, water loss, and damage caused by ultraviolet light. A healthy, normal nervous system in the skin also provides tactile sensations (touch, pressure, and vibration), thermal sensations (hot and cold), and pain sensations. Body thermoregulation depends on the skin's ability to control blood flow to the skin through sweating to increase or decrease heat loss. Healthy skin contains three distinct tissue layers: a thin outer layer of cells called the epidermis, a thicker middle layer of connective tissue called the dermis, and an inner subcutaneous layer. The thin outer layer of the epidermis is composed of flattened, keratinized, dead keratinocytes, which form a barrier against water loss and microbial invasion. Dead keratinocytes derive from living keratinocytes in the basal layer above the dermis and are responsible for skin re-epithelialization. The epidermis does not contain nerves or blood vessels and obtains water and nutrients by diffusion from the dermis. The dermis, located below the epidermis, is composed primarily of collagen fibers and some elastic fibers, which together are produced by fibroblasts and, together with water and large proteoglycan molecules, constitute the extracellular matrix (ECM). This epidermal layer provides mechanical strength and a substrate for the diffusion of water and nutrients. It contains blood vessels, nerves, sweat glands, hair follicles, and cells involved in immune function, growth, and repair. The subcutaneous layer is composed of adipocytes, which form a thick layer of adipose tissue.

[0003] A wound may be considered to be a disruption in the structural and functional integrity of the skin. Thus, a "wound" may include, for example, a laceration, abrasion, incision, perforation, abrasion, burn, or other such injury that causes the skin to cut, tear, and / or break.

[0004] After hemostasis, which often follows wound healing, wounds go through three main stages as they heal: inflammation, proliferation, and remodeling. Chronic wounds can be thought of as wounds that have failed to undergo the normal healing process in an orderly and timely manner. Chronic wounds often remain in the inflammatory stage.

[0005]

[0003] Often, in cases of severe wounds, such as extensive or deep wounds, or large or severe burns, or in cases of chronic wounds, skin substitutes are often used to aid the wound healing process and more quickly restore at least some of the above-mentioned functions of healthy skin. Skin substitutes can be broadly thought of as a group of elements or materials that allow for temporary or permanent closure of a wound. Skin substitutes can generally be classified as biological skin substitutes, synthetic skin substitutes, or hybrid skin substitutes that include biological and synthetic skin substitutes.

[0006] While biological skin substitutes often have a more intact extracellular matrix structure, synthetic skin substitutes can be synthesized on demand and tailored to specific purposes. Both biological and synthetic skin substitutes have their advantages and disadvantages. Biological skin substitutes allow for a more natural construction of new dermis and, due to the presence of a basement membrane, allow for superior re-epithelialization properties. Synthetic skin substitutes can be chemically synthesized, offering the advantage of greater control over scaffold composition. Synthetic skin substitutes include, for example, synthetic collagen, protein-based matrices, or synthetic biolayers containing collagen- or protein-based components combined with silicone components. Hybrid skin substitutes can be partially synthetic or generated by living cells, and partially chemically synthesized.

[0007] Biological skin substitutes may include, but are not limited to, skin grafts such as autologous skin grafts, syngeneic skin grafts, allogeneic skin grafts, xenogeneic skin grafts such as porcine skin grafts, cadaveric skin allografts, and amniotic membrane tissue grafts.

[0008] Whether a biological, synthetic, or hybrid skin substitute is used, the goal of using a skin substitute is to provide effective, timely, and scar-free wound healing while restoring the skin to a similar degree of function as it was before the wound occurred.

[0009] Failure to maintain tissue integrity in traumatic wounds often results in tissue desiccation and deterioration. Furthermore, many current treatments fail to provide an adequate barrier against subsequent infection and / or fail to contain the wound from dirt and harmful pathogens. Furthermore, current treatments are inadequately adapted for long-term in-field care. In short, there is a need for improved approaches to stabilizing and / or protecting wounds, particularly in the field, to protect the patient and the wound for further care. Summary of the Invention

[0010] In view of the above, the present inventors have determined that there is a need for a wound treatment method that is rugged and durable, lightweight, compact, easy to carry and handle, has low dependency on external power sources or specialized equipment, is modular, and is interoperable with current approaches to treatment. There is also a need for a wound treatment method that is sustainable, scalable, and safe for human use.

[0011] A system for preparing a wound treatment is provided, the system including an applicator configured to apply a fat material to a skin graft material, the applicator including one or more of a syringe, a plunger, a roller, a screw, a container, a preparation chamber, a loading chamber, an extrusion press, a vacuum source, a mesh screen, an input channel, an output channel, a blade, a frame, a holding element, and a handle.

[0012] A wound treatment kit is provided that includes a skin graft material and an applicator configured to apply a fat material to the skin graft material. The kit may further include one or more of a syringe, a plunger, a roller, a screw, a container, a preparation chamber, a loading chamber, an extrusion press, a vacuum source, a mesh screen, an input channel, an output channel, a blade, a frame, a holding element, a handle, and a sterile container.

[0013] Wound treatment compositions are provided that include a skin graft material and an adipose material, such as for promoting regenerative cell ingrowth in a wound. The adipose material can include material obtained from an area of ​​the body that has adipose tissue. The skin graft material can include biological and / or synthetic skin substitute materials.

[0014] A method for preparing a wound treatment is provided that includes providing a fat material, providing a skin graft material, and applying the fat material to the skin graft material. Applying the fat material to the skin graft material can include infiltrating, infusing, impregnating, joining, and / or combining the fat material with the skin graft material.

[0015] A method for treating a wound is provided that includes the steps of providing a skin graft material, applying a fat material to the skin graft material, and applying the combined fat and skin graft material to the wound.

[0016] Additionally, embodiments of the present disclosure further apply to wound treatment dressings. Accordingly, wound treatments, dressings, kits, and methods for stabilizing, protecting, and / or healing wounds are disclosed.

[0017] This Summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an index of the scope of the claimed subject matter.

[0018] Additional features and advantages of the present disclosure will be set forth in the description that follows, and in part will be obvious from the description, or may be learned by the practice of the present disclosure. The features and advantages of the present disclosure may be realized and obtained by means of the instruments and combinations particularly pointed out in the appended claims. These and other features of the present disclosure will become more fully apparent from the following description and appended claims, or may be learned by the practice of the disclosure as set forth hereinafter. [Brief explanation of the drawings]

[0019] These and other features, aspects, and advantages of the present disclosure will become better understood with regard to the following description, appended claims, and accompanying drawings.

[0020] [Figure 1] 1 illustrates a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 2]1 illustrates an extrusion die used in a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 3] 1 is a diagram of a system for preparing a wound treatment in the form of a skin graft material with a fat material according to an embodiment of the present disclosure. FIG. [Figure 4] FIG. 1 illustrates a brewing chamber according to an embodiment of the present disclosure. [Figure 5] 1A-1C illustrate a method for preparing a wound treatment in the form of a skin graft material using adipose material according to an embodiment of the present disclosure. [Figure 6A] 1 illustrates a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 6B] 1 illustrates a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 7] 1A-1C illustrate a method for preparing a wound treatment in the form of a skin graft material impregnated with a fat material according to an embodiment of the present disclosure. [Figure 8] 1 illustrates a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 9A] 1 illustrates an embodiment of a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 9B] FIG. 1 illustrates another embodiment of a system for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. [Figure 9C] FIG. 1 illustrates another embodiment of a system for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. [Figure 9D] FIG. 1 illustrates another embodiment of a system for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. [Figure 9E]FIG. 1 illustrates another embodiment of a system for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. [Figure 9F] FIG. 1 illustrates another embodiment of a system for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. [Figure 10] 1 illustrates a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 11] 1A-1C illustrate a method for preparing a wound treatment in the form of a skin graft material using adipose material according to an embodiment of the present disclosure. [Figure 12] 1 illustrates a system for preparing a wound treatment in the form of a skin graft material with adipose material according to an embodiment of the present disclosure. [Figure 13] 1A-1D illustrate a system and corresponding method for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. [Figure 14A] 10 is a hematoxylin and eosin stained image of unannealed skin graft material. [Figure 14B] 10 is a hematoxylin and eosin stained image of unannealed skin graft material. [Figure 15] 1A-1C illustrate a method for preparing a wound treatment according to an embodiment of the present disclosure, the method including processing the skin graft material before applying the fat material. [Figure 16A] 10 is a hematoxylin and eosin stained image of an annealed skin graft material according to an embodiment of the present disclosure. [Figure 16B] 10 is a hematoxylin and eosin stained image of an annealed skin graft material according to an embodiment of the present disclosure. [Figure 17] 1A-1C illustrate a method for preparing a wound treatment according to an embodiment of the present disclosure, the method including processing the skin graft material before applying the fat material. [Figure 18]FIG. 1 illustrates another embodiment of a system for preparing a wound treatment in the form of a skin graft material with fat material according to an embodiment of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0021] Various embodiments of the present disclosure may be better understood from the following description read in conjunction with the accompanying drawings, in which like reference numerals refer to like elements and in which:

[0022] While the present disclosure is susceptible to various modifications and alternative constructions, specific illustrative embodiments are shown in the drawings and are described below. It should be understood, however, that there is no intention to limit the disclosure to the particular embodiments disclosed, but rather, the intention of this disclosure is to cover all modifications, alternative constructions, combinations, and equivalents falling within the spirit and scope of the present disclosure.

[0023] Unless a term is expressly defined in this application to have a stated meaning, it will be understood that no intention is intended, explicitly or implicitly, to limit the meaning of such term beyond its plain or ordinary meaning.

[0024] Any element in a claim that does not expressly recite a "means for" performing a particular function or a "step for" performing a particular function should not be construed as a "means" or "step" clause as defined in 35 U.S.C. 112.

[0025] As used herein, the term "treatment" is intended to be understood according to its common dictionary definition. That is, the term "treatment" broadly includes medical care and / or medicines given to a patient for illness or injury. "Treatment" can include the use of chemical, physical, or biological agents to preserve something or to impart certain properties to something. Thus, "treatment" can be medical care provided (i.e., in the form of a method or series of prescribed actions), or it can refer to medicines used to preserve something or to impart certain properties to something.

[0026] The term "wound" as used herein is intended to encompass tissue damage generally. Accordingly, the term "wound" includes, for example, lacerations, abrasions, incisions, perforations, abrasions, or other such injuries that cause cutting, tearing, and / or breaking of the skin. Wounds can be described by either the size, shape, or magnitude of the wound. For example, a paper cut represents a small, straight incision of relatively little significance, while a concussive blast resulting in a large laceration across one or more body parts represents a wound of greater significance. However, each of the above examples falls within the scope of the term "wound" as used herein.

[0027] The term "wound" further includes damage to underlying tissues, such as caused by traumatic injury. Thus, the term "wound" is intended to include combinations of multiple different wounds. For example, a traumatic cut caused by an explosive blast may be generally referred to as a wound, even though it is a complex combination of numerous different lacerations, abrasions, avulsions, and perforations. Furthermore, any damage to underlying tissues resulting from the aforementioned explosive blast may also be encompassed within the understanding of a reference to a wound. The term "wound" is also intended to encompass tissue damage caused by burns (e.g., thermal and / or chemical burns). Furthermore, the term "wound" is also intended to encompass damage resulting from, for example, diabetic foot ulcers, venous leg ulcers, surgery, pressure ulcers, and other causes.

[0028] Additionally, wounds that can be treated with the wound treatments and methods disclosed herein include injuries that can be located anywhere, including internal, interface, external, interstitial, extracorporeal, and / or intracorporeal. Examples of wounds suitable for covering with the scaffold material include incisions, long and deep gashes, open wounds, tissue ruptures, pressure sores, dermatitis, lesions, chronic wounds, battlefield wounds, necrotic wounds, acute, chronic, traumatic lacerations, abrasions, contusions, necrotizing fasciitis, toxic epidermal necrolysis, pressure wounds, venous insufficiency ulcers, arterial ulcers, diabetic or neuropathic ulcers, bed sores, mixed ulcers, burn wounds, mucormycosis, vasculitic wounds, pyoderma, gangrene, and the like, and / or combinations thereof. Treatment of wounds in human and animal subjects is contemplated.

[0029] As used herein, "traumatic wound" refers to any wound resulting from physical injury that injures both the skin and underlying tissue. A gunshot wound is a non-limiting example of a traumatic wound because it perforates (i.e., brakes) the skin and ruptures or otherwise injures the underlying tissue. As another non-limiting example, a percussive or explosive blast typically results in traumatic wound(s). Many, but not all, wounds sustained during wartime can be described as traumatic wounds due to the nature of warfare and war-related injuries. "Traumatic wounds" can include bleeding wounds, wounds exposing bone or tendons, severe burns, deep tissue wounds (e.g., asymmetric deep tissue wounds), and / or wounds with a large surface area.

[0030] As mentioned above, many different types of skin substitutes or skin graft materials can be used to aid the wound healing process and restore at least some of the functions of healthy skin more quickly. Skin substitutes can be broadly thought of as a group of elements or materials that allow for temporary or permanent closure of a wound. Skin substitutes can generally be classified as biological skin substitutes, synthetic skin substitutes, or hybrid skin substitutes that include biological and synthetic skin substitutes.

[0031] For purposes of this application, the terms "skin substitute," "skin graft material," "scaffold material," "graft product," or similar terms shall mean any material that is manufactured by Kerecis, Inc. TM 0mega3 Wound, Kerecis from Atlantic cod (Gadus morhua) TMThe skin substitute may include acellular fish skin, or any other skin graft material known for use in wound treatment, or similar to those described above. As used herein, the term "extracellular matrix" or "ECM" refers to the non-cellular tissue material present in fish skin that provides structural support to skin cells as well as performing a variety of other important functions. The ECM described herein does not necessarily include matrix materials composed entirely or reconstituted from extracted, purified, or isolated ECM components (e.g., collagen). However, in some embodiments, the ECM used as a skin substitute may include matrix materials composed entirely or reconstituted from extracted, purified, or isolated ECM components (e.g., collagen).

[0032] The extracellular matrix (ECM) of vertebrates is a complex structure that surrounds and supports cells. The ECM is composed of a complex mixture of structural proteins, the most abundant of which is collagen, as well as other specialized proteins and proteoglycans. The scaffold material described here is a largely intact acellular scaffold composed of natural biological ECM components derived from fish skin. The scaffold may also contain lipids naturally occurring in fish skin. The original three-dimensional structure, composition, and function of the dermal ECM remain substantially unchanged, providing a scaffold to support cell migration, adhesion, proliferation, and differentiation, thus facilitating tissue repair and / or replacement.

[0033] As used herein, the terms "acellular," "decellularized," "decellularized fish skin," and the like refer to biological skin material from which a substantial amount of cellular and nucleic acid content has been removed, leaving behind a complex three-dimensional stromal structure of ECM. In embodiments, "decellularized fish skin" may further comprise fish skin that contains omega-3 polyunsaturated fatty acids (PUFAs) in addition to a complex three-dimensional stromal structure of ECM that lacks a substantial amount of cellular and nucleic acid content.

[0034] Other scaffold or skin graft materials may include those derived from mammalian skin / membranes, reconstituted from collagen materials, or artificially prepared. Generally, scaffold materials are approximately 0.1-4.0 mm thick (i.e., in cross section), such as 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, or 3.5 mm thick. The thickness may depend on numerous factors, such as the starting material used, processing, lyophilization, and / or rehydration. Of course, when the product includes more than one layer of scaffold material, the thickness will be proportionally greater.

[0035] In embodiments, the scaffold material may be or may include decellularized fish skin. Decellularized fish skin is configured to provide a scaffold material for supporting cell migration, adhesion, proliferation, and differentiation to promote tissue repair and / or replacement, as described in U.S. Patent No. 8,613,957, granted December 24, 2013. The U.S. patent application was filed October 6, 2010, the contents of which are incorporated herein by reference in their entirety. The decellularized fish skin product described in U.S. Patent No. 8,613,957 serves as a scaffold material that provides an intact scaffold for supporting endothelial and / or epithelial cell ingrowth. The decellularized fish skin scaffold material is biocompatible and therefore capable of being integrated by the host. 0mega3 Wound is a commercially available skin substitute made from the minimally processed skin of wild Atlantic cod native to Iceland. Fish skin is structurally similar to human skin, with three basic layers including the epidermis, dermis, and hypodermis, and contains proteins, lipids, fatty acids, and other bioactive compounds homologous to human skin.

[0036] Decellularized fish skin can be granulated, ground, or otherwise processed into various sizes and shapes, as described in U.S. Patent Application No. 17 / 704,539, filed March 25, 2022, the contents of which are incorporated herein by reference in their entirety. The size of the individual ground particles can vary depending on the type and / or method of grinding. For example, decellularized fish skin particles can be produced by a jet milling process designed to output particles smaller than a specified size. In some embodiments, the decellularized fish skin is cut, chopped, or ground into particles, which may be done in a measured manner to produce uniform particles, or may be done roughly to generate particles of various different sizes.

[0037] The scaffold material according to the present invention can be obtained from intact fish skin. Any species of fish, including bony fish and cartilaginous fish, can be used as the source of fish skin. For example, the source can be round fish such as cod, haddock, and catfish; flatfish such as halibut, plaice, and sole; salmonids such as salmon and trout; mackerel such as tuna; or small fish such as herring, anchovies, mackerel, and sardines. In certain embodiments, the fish skin is obtained from oily cold-water fish and / or fish known to contain large amounts of omega-3 oil. Examples of fish rich in omega-3 oil include salmon, pilchard, tuna, herring, cod, sardine, mackerel, codfish, smelt, whitefish, hoki, and some varieties of trout.

[0038] Fish skin is removed from fish before processing. If the fish skin is from a scaled fish species, the fish skin must be descaled so that a substantial portion of the scales are removed or at least the hydroxyapatite is removed from the scales. The phrases "substantially descaled" or "substantially scale-free" mean that at least 95%, preferably at least 99%, and more preferably 100% of the scales on the fish skin are removed. "Substantially scale-free" fish skin can also refer to fish skin from scale-free fish species. To wash away the scales, the scales are removed simply by mechanical pressure (e.g., by a knife, shaking with abrasives, water pressure, special scale removal devices that use the same mechanical force as a knife, or other pressure devices such as polishing with ceramic or plastic) before any processing, or by mechanical pressure after some chemical treatment (e.g., decellularization). If the fish skin is first treated chemically and / or enzymatically (e.g., with TRITON® X-100), the mechanical pressure generally needs to be lessened, as the skin is more susceptible to tearing after decellularization. Scales can be removed in multiple steps, e.g., by removing some before decellularization, followed by further removal during and / or after decellularization. Alternatively, scales can be removed by chemical treatment alone.

[0039] After the scales are removed, the fish skin is optionally frozen prior to decellularization. The fish skin can be rapidly frozen to preserve the collagen structure of the scaffold by incubating the skin in liquid nitrogen or using other specialized freezing equipment that can freeze the skin to temperatures below -70°C. The freezing process can lyse or partially lyse the cells that make up the intact fish skin, helping to facilitate the decellularization of the fish skin. If the fish skin is frozen, it can be thawed later for further processing.

[0040] The fish skin, whether frozen or not, can be washed with a buffer solution before further processing. For example, the fish skin can be washed one to three times with a buffer solution optionally containing one or more antioxidants (e.g., ascorbic acid (e.g., 50 mM ascorbic acid), vitamins A, C, E, and beta-carotene), antibiotics (e.g., streptomycin and penicillin), proteases (e.g., dispase II), and protease inhibitors (e.g., antipain, aprotinin, benzamidine, bestatin, DFP, EDTA, EGTA, leupeptin, pepstatin, phosphoramidon, and PMSF) to promote disinfection and stabilization of the fish skin. The buffer solution can have a pH of at least 5.5, such as 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, or higher. In certain embodiments, the pH is 7.0 to 9.0, e.g., 7.5 to 8.5. The buffer can also be used as a medium in which the fish skin can be stored for days to weeks or longer, hi certain embodiments, the fish skin is stored in the buffer at a temperature of about 4°C.

[0041] After freezing and / or washing and / or storage in a buffer, the fish skin is treated with one or more decellularization solutions to remove cellular material, including antigenic material, from the fish skin with minimal or no damage to the mechanical and structural integrity and biological activity of the native extracellular matrix.

[0042] A "decellularizing agent" is an agent effective in removing a substantial amount of cellular and nucleic acid content from the ECM. An ECM is "decellularized" or "substantially free" of cellular and nucleic acid content (i.e., a "substantial amount" has been removed) when at least 50% of the viable and non-viable nucleic acids and other cellular material has been removed from the ECM. In certain embodiments, approximately 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% of the viable and non-viable nucleic acids and cellular material are removed. Decellularization can be confirmed, for example, by testing treated fish skin for DNA content. Removal of nucleic acids from the ECM can be determined, for example, by histological examination of the ECM and / or by biochemical assays such as the PICOGREEN® assay, diphenylamine assay, or by PCR.

[0043] Decellularization disrupts cell membranes and releases cellular contents. Decellularization may involve one or more physical treatments, one or more chemical treatments, one or more enzymatic treatments, or any combination thereof. Examples of physical treatments include sonication, mechanical agitation, mechanical massage, mechanical pressure, and freeze / thaw. Examples of chemical decellularization agents include ionic salts (e.g., sodium azide), salts, acids, detergents (e.g., non-ionic and ionic detergents), oxidizing agents (e.g., hydrogen peroxide and peracids), hypotonic solutions, hypertonic solutions, chelating agents (e.g., EDTA and EGTA), organic solvents (e.g., tri(n-butyl)phosphate), ascorbic acid, methionine, cysteine, maleic acid, and polymers that bind DNA (e.g., poly-L-lysine, polyethylimine (PEI), and polyamidoamine (PAMAM)). Non-ionic detergents include 4-(1,1,3,3-tetramethylbutyl)phenyl-polyethylene glycol, t-octylphenoxypolyethoxyethanol, polyethylene glycol tert-octylphenyl ether (TRITON® X-100) (Dow Chemical Company). Ionic detergents include sodium dodecyl sulfate (SDS), sodium deoxycholate, TRITON® X-200, and zwitterionic detergents (e.g., CHAPS). Other suitable decellularization detergents include polyoxyethylene (20) sorbitan monooleate and polyoxyethylene (80) sorbitan monooleate (Tween 20 and 80), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, octylglucoside, and sodium dodecyl sulfate. Examples of enzymatic decellularization agents include proteases, endonucleases, and exonucleases. Proteases include serine proteases (e.g., trypsin), threonine proteases, cysteine ​​proteases, aspartic acid proteases, metalloproteases (e.g., thermolysin), and glutamic acid proteases. Decellularization is generally carried out at a pH of at least 5.5, such as 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, or higher.In certain embodiments, the pH is between 7.0 and 9.0, for example, between 7.5 and 8.5.

[0044] One example of a decellularization step is to incubate fish skin in a solution containing 1 M NaCl, 2% deoxycholic acid, 0.02% sodium azide, and 500 ppm streptomycin. In another example, fish skin is incubated in a first decellularization solution containing a protease (e.g., 2.5 U / mL Dispase II) and other ingredients (e.g., 0.02% sodium azide). The first decellularization solution is poured off, and the fish skin is then treated with a second decellularization solution, such as a solution containing a detergent (e.g., 0.5% TRITON® X-100) and other ingredients (e.g., 0.02% sodium azide). In another example, fish skin is first treated with a decellularization solution containing a detergent (e.g., 0.5% TRITON® X-100) and other ingredients (e.g., 0.02% EDTA, sodium azide, and / or deoxycholic acid), and then cultured in a second decellularization solution containing a detergent such as SDS.

[0045] The decellularization step(s) can be repeated as necessary by pouring off any remaining decellularization solution, optionally washing the fish skin with a buffer (e.g., Hank's Balanced Salt Solution), and then treating the fish skin again with another decellularization step. Once a sufficient amount of cellular material has been removed, the decellularization solution can be removed (e.g., by aspirating or by gently pouring off the solution).

[0046] After decellularization, the fish skin can be optionally washed with water, buffer, and / or salt solution. Examples of suitable wash solutions include Dulbecco's phosphate-buffered saline (DPBS), Hank's balanced salt solution (HBSS), Medium 199 (M199, SAFC Biosciences, Inc.), and / or L-glutamine. The wash step(s) are generally performed at a pH of at least 5.5, such as 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, or higher. In certain embodiments, the pH is 7.0-9.0, e.g., 7.5-8.5.

[0047] To improve the appearance of the final product, the fish skin can optionally be bleached. Bleaching can be performed before, after, and / or simultaneously with decellularization. For example, one or more bleaching agents can be incorporated into one or more decellularization solutions and / or one or more buffer solutions. Examples of bleaching agents include sodium sulfite, hydrogen peroxide, ammonium persulfate, potassium persulfate, and sodium persulfate. In certain embodiments, if a strong bleaching agent such as persulfate(s) is used, bleaching and decellularization can be combined into a single step involving incubating the fish skin in a mixture of one or more bleaching agents, a thickener, and a peroxide source. For example, a dry bleaching mixture can be prepared (see, e.g., "Bleaching Mixture" in Example 5) followed by the addition of water, hydrogen peroxide, or a combination thereof to form a bleaching solution that may also be sufficient for decellularization. The bleaching agent (e.g., sodium sulfite, hydrogen peroxide, ammonium persulfate, potassium persulfate, sodium persulfate) should be approximately 40-60% w / w of the above dry mixture. To promote bleaching as well as decellularization, a combination of EDTA and persulfate can be added to the above mixture.

[0048] In certain embodiments, the concentration of EDTA in the dry mixture is about 0.25-5% w / w. Hydrogen peroxide can be about 15-25% of the mixture. The peroxide source can be sodium percarbonate and potassium percarbonate. Sodium phosphate perhydrate and sodium carbonate, or magnesium metasilicate and silicon silicate, can also be used as peroxide sources. The dry mixture can also contain, for example, 1-10% w / w of silica and hydrated silica, and optionally one or more stearates (e.g., ammonium stearate, sodium stearate, and / or magnesium stearate). Additionally, to increase the viscosity of the bleaching / decellularization solution and protect protein fibers from damage, the dry mixture can optionally contain a thickener, such as hydroxypropylmethylcellulose, hydroxyethylcellulose, algin (i.e., alginates), organic gums (e.g., cellulose, xanthan gum), sodium metasilicate, and combinations thereof. Bleaching and / or bleaching and decellularization is generally carried out at a pH of at least 5.5, such as 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0 or higher. In certain embodiments, the pH is 7.0-9.0, e.g., 7.5-8.5. After bleaching and / or bleaching and decellularization, the fish skin is optionally washed with a solution containing L-glutamine at the above pH conditions.

[0049] In certain embodiments, the fish skin is treated with digestion enzymes. Similar to bleaching, digestion can be performed before, after, and / or simultaneously with decellularization. Suitable enzymes include proteases, such as serine proteases, threonine proteases, cysteine ​​proteases, aspartic acid proteases, metalloproteases, and glutamic acid proteases. In certain embodiments, the digestion enzyme is a serine protease, such as trypsin. The digestion enzyme can be an enzyme that functions in an alkaline environment to limit cross-linking within the ECM and soften the fish skin. Digestion is generally performed at a pH of at least 5.5, such as 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, or higher. In certain embodiments, the pH is 7.0-9.0, e.g., 7.5-8.5.

[0050] The decellularized fish skin can be optionally cryopreserved. Cryopreservation can involve immersing the fish skin in a cryoprotectant solution before freezing. The cryoprotectant solution generally includes an appropriate buffer, one or more cryoprotectants, and optionally a solvent, such as an organic solvent that can be combined with water and undergo minimal expansion and contraction. Examples of cryoprotectants include sucrose, raffinose, dextran, trehalose, dimethylacetamide, eimethylsulfoxide, ethylene glycol, glycerol, propylene glycol, 2-methyl-2,4-pantandial, certain antifreeze proteins and peptides, and combinations thereof. Alternatively, if the decellularized fish skin is rapidly frozen (quickly frozen) before sublimation to minimize ice crystal formation during the freezing step, the fish skin can be optionally frozen in a cryoprotectant-free buffer. Cryopreservation is generally carried out at a pH of at least 5.5, such as 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0 or higher. In certain embodiments, the pH is between 7.0 and 9.0, e.g., between 7.5 and 8.5.

[0051] The decellularized fish skin can be packaged inside a sterile container such as a glass bottle or pouch. In one embodiment, a TYVEK® pouch is used. For example, the fish skin can be incubated in a cryoprotectant solution, packaged in a TYVEK® pouch, and then placed in a freeze-dryer to freeze at a rate compatible with the cryoprotectant.

[0052] Decellularized fish skin can be freeze-dried, i.e., frozen at low temperatures and under vacuum, resulting in the sequential removal of water from each ice crystal phase without recrystallization of the ice. During freeze-drying, water is generally removed first by sublimation, and then, if necessary, by desorption. Another method for removing remaining excess water after processing and before sterilization is vacuum pressing.

[0053] In certain embodiments, the decellularized fish skin is sterilized before and / or after being frozen. Sterilization methods are known in the art. For example, the decellularized fish skin can be placed in an ethylene oxide chamber and treated with an appropriate cycle of ethylene oxide. Other sterilization methods include sterilization by ozone, carbon dioxide, gaseous formaldehyde, or radiation (e.g., gamma rays, X-rays, electron beam treatment, and subatomic particles).

[0054] As an alternative to, or in addition to, water freezing, lyophilization, and / or vacuum compression, decellularized fish skin can be stored in a non-aqueous solution, such as alcohol.

[0055] The resulting product (scaffold material) is a sterilized collagen-based matrix with properties that can promote tissue regeneration, repair, and / or replacement (e.g., endogenous tissue repair, regeneration, and / or growth). The term "scaffold material" may refer in certain embodiments to a material comprising fish skin that has been decellularized as described above and optionally bleached, digested, freeze-dried, etc. In various aspects, the term "scaffold material" may refer to a particularized, i.e., shredded, graft product, such as decellularized fish skin.

[0056] The scaffold material is capable of providing an intact scaffold for endothelial and / or epithelial cell support, capable of being integrated by the host, biocompatible, not significantly calcifying, and capable of being stored and transported at ambient temperatures. Here, the phrase "integrated by the host" means that the patient's cells and tissues treated with the scaffold material are capable of growing within the scaffold material and that the scaffold material is actually integrated / absorbed into the patient's body. The term "biocompatible" refers to a material that is substantially non-toxic in the in vivo environment of its intended use and is not substantially rejected by the patient's physiological system (i.e., is non-antigenic).

[0057] This can be determined by a material's ability to pass biocompatibility tests as described in International Organization for Standardization (ISO) Standard No. 10993 and / or United States Pharmacopeia (USP) 23 and / or U.S. Food and Drug Administration (FDA) Blue Book Memorandum No. G95-1, entitled "Use of International Standard ISO-10993, Biological Evaluation of Medical Devices Part 1: Evaluation and Testing." Generally, these tests measure a substance's toxicity, infectivity, pyrogenicity, irritation potential, reactivity, hemolytic activity, carcinogenicity, and / or immunogenicity. A biocompatible structure or material, when introduced into the majority of patients, does not provoke a significant adverse, long-lasting, or escalating biological reaction or response, distinct from the mild, transient inflammation commonly associated with surgery or the implantation of a foreign substance into a living organism.

[0058] The scaffold material can contain proteins derived from the ECM of fish skin. ECM components in the scaffold material can include, for example, structural proteins, adhesive glycoproteins, proteoglycans, non-proteoglycan polysaccharides, and matricellular proteins. Examples of structural proteins include collagens (the most abundant proteins in the ECM), such as fibrillar collagens (types I, II, III, V, and XI), facit collagens (types IX, XII, and XIV), short-chain collagens (types VIII and X), basement membrane collagens (type IV), and other collagens (types VI, VII, and XIII), elastin, and laminin. Examples of adhesive glycoproteins include fibronectin, tenascin, and thrombospondin. Examples of proteoglycans include heparin sulfate, chondroitin sulfate, and keratan sulfate. An example of a non-proteoglycan polysaccharide is hyaluronic acid. Matricellular proteins are a structurally diverse group of extracellular proteins that regulate cellular function through interactions with cell surface receptors, cytokines, growth factors, proteases, and the ECM. Examples include thrombospondin (TSP) 1 and 2, tenascin, and SPARC (secreted protein, acidic and cysteine-rich).

[0059] In certain embodiments, decellularization (and any other processing steps) does not remove all of the naturally occurring lipids from the lipid layer of fish skin. Thus, the scaffold material may contain one or more lipids derived from fish skin, particularly from the lipid layer of fish skin. For example, the scaffold material may contain up to about 25% w / w lipid (of the dry weight of the total scaffold material after lyophilization), such as 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, or 24% w / w lipid. The presence of lipids in fish skin can be confirmed, for example, by organic solvent extraction followed by chromatography. Examples of suitable organic solvents include acetone and chloroform.

[0060] Lipids in fish skin may include, for example, fatty acyl (i.e., fatty acids, their conjugates and derivatives), glycerolipids, glycerophospholipids (i.e., phospholipids), sphingolipids, saccharolipids, polyketides, sterol lipids (i.e., sterols), certain fat-soluble vitamins, prenol lipids, and / or polyketides. Examples of fatty acyl include saturated fatty acids, such as polyunsaturated fatty acids, fatty esters, fatty amides, and eicosanoids. In certain embodiments, the fatty acids include omega-3 fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are found in high concentrations in fish oil. Other fatty acids found in fish oil include arachidic acid, gadoleic acid, arachidonic acid, butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, vaccenic acid, linoleic acid, α-linolenic acid, γ-linolenic acid, behenic acid, erucic acid, and lignoceric acid. Examples of glycerolipids include mono-, di-, and tri-substituted glycerols, such as monoacylglycerol, diacylglycerol, and triacylglycerol (i.e., monoglycerides, diglycerides, and triglycerides). Examples of glycerophospholipids include phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine. Examples of sphingolipids include sphingophospholipids and glycosphingolipids. Examples of sterol lipids include cholesterol, steroids, and secosteroids (various forms of vitamin D). Examples of prenol lipids include isoprenoids, carotenoids, and quinones and hydroquinones such as vitamin E and vitamin K.

[0061] The scaffold material can be used in a dry form. Alternatively, the scaffold material can be rehydrated before use. In certain embodiments, one or more scaffold materials are layered together to form a thicker scaffold material.

[0062] The scaffold material, in embodiments, can be provided in the form of particles, each configured to actively promote wound healing by serving as a physical scaffold for infiltrating cells involved in wound healing / repair, such as cell growth and neovascularization. The scaffold material particles of embodiments can be configured to advantageously preserve the three-dimensional ("3D") structure of the scaffold material, e.g., with an ECM discernible, e.g., by histological analysis. Furthermore, the particle dimensions can be configured to facilitate molding, packing, or otherwise applying the particles into the wound cavity with greater precision than existing approaches to wound treatment.

[0063] In an embodiment, the scaffold material particles have a maximum dimension within a predetermined maximum size threshold and a minimum size threshold effective for maintaining the matrix structure of the decellularized fish skin and promoting cell regeneration in-growth into the wound. That is, the maximum dimension, such as the maximum of the length, width, and / or thickness of the above particles, can be smaller than the maximum size, such as 1 mm, and larger than the minimum size, such as the size at which ECM is destroyed. In an embodiment, chopped decellularized fish skin particles for the scaffold material are obtained by providing a sheet of decellularized fish skin as described above, then chopping the sheet of decellularized fish skin, and optionally sieving the chopped particles until the chopped decellularized fish skin particles fall within the predetermined minimum and maximum size thresholds.

[0064] Biological skin substitutes may include, but are not limited to, autologous skin grafts, syngeneic skin grafts, allogeneic skin grafts, xenogeneic skin grafts such as porcine skin grafts, cadaver skin allografts, and skin grafts such as amniotic membrane tissue grafts. Certain biological skin substitutes can be made from biological materials containing intact or reconstituted collagen. Examples include brands such as Oasis, Matristem, Integra, and Puracol. These products are often referred to by clinicians as matrix products. The matrix product is inserted into the wound to attract cell growth. A secondary wet-dry wound dressing can then be applied over the wound dressing. The decellularized fish skin described in U.S. Pat. No. 8,613,957 is an example of a matrix product. The decellularized fish skin product described in U.S. Pat. No. 8,613,957 serves as a scaffold material, providing an intact scaffold to support the growth of endothelial and / or epithelial cells.

[0065] Other examples of biological skin substitutes include those described in U.S. Patent No. 6,541,023, which describes the use of a porous collagen gel derived from fish skin for use as a tissue engineering scaffold. The preparation of the collagen gel involves crushing the fish skin. Furthermore, Chinese Patent No. 1,068,703 describes a method for preparing fish skin for burn treatment, which involves separating the fish skin from the body of the fish and placing the skin in a preservative solution consisting of iodine tincture, ethanol, borneol, zinc sulfadiazine, and hydrochloric acid in an amount sufficient to establish a pH value of 2.5 to 3. However, because the product in U.S. Patent No. 6,541,023 is in a gel form, and the product in Chinese Patent No. 1,068,703 is stored in a solution, these products can be difficult to handle.

[0066] Furthermore, many extracellular matrix products for medical use are derived from human skin (ALLODERM® Regenerative Tissue Matrix (LifeCell)), fetal bovine dermis (PRIMATRIX TM Dermal Repair Scaffold (T.E.I. Biosciences), pig bladder (MATRISTEM TM Extracellular Matrix Wound Sheet (Medline Industries, Inc.), and porcine small intestinal submucosa (OASIS® Wound Matrix (HealthPoint Limited)).

[0067] Examples of commercially available synthetic skin substitutes include Biobrane®, Dermagraft®, Integra®, Apligraf®, MatriDerm®, OrCel®, Hyalomatrix®, and Renoskin®.

[0068] U.S. Patent Application Publication No. 2003 / 0059460 discloses a hybrid polymer skin substitute material containing a synthetic polymer and a natural polymer that can be used to regenerate living tissue. The hybrid contains a crosslinked natural polymer and a biodegradable, absorbable synthetic polymer. However, to produce the hybrid material, a series of complex process steps must be undertaken. Furthermore, the resulting hybrid material contains both natural and synthetic materials.

[0069] As noted above, the group of skin substitutes that can be used as examples of skin substitutes according to the present disclosure is large and diverse. The AHRQ Technology Assessment Program's Technology Overview, Project ID WNDT0818, entitled "Skin Substitutes for Treating Chronic Wounds," published February 2, 2020, and incorporated herein by reference, identified 76 commercially available products on pages 9-13, Table 2, but few studies have compared them internally. Each of these skin substitutes listed in the table may be an embodiment of a skin substitute according to the present disclosure.

[0070] In various examples, colorants may be used with skin graft materials, as described in U.S. Patent Application No. 17 / 703,650, filed March 24, 2022, the contents of which are incorporated herein by reference in their entirety. In its broadest sense, a colorant contemplated herein is a colorant, or coloring agent or combination of colorants, that imparts, changes color, or loses color to the skin substitute based on changes in conditions within the wound during the healing process or based on changes to the skin substitute. In preferred embodiments, the colorant can be degraded by attack by one or more proteases within the wound. For such colorants, the colorant loses its color due to degradation by one or more proteases. For example, the colorant can impart a blue or purple color to the skin substitute. However, after applying a wound treatment containing a skin substitute and a coloring agent, the color of the wound treatment skin substitute may also be degraded or lost due to attack by one or more proteases within the wound, thereby changing the color of the applied wound treatment to the original color of the skin substitute or a different color. However, the color change of the coloring agent is not limited thereto and may also include a color change in response to a change in conditions within the wound. For example, the color imparted by the coloring agent may be induced by application or addition of the coloring agent such that the original color of the skin substitute remains unchanged. However, a color change of the coloring agent may be induced or caused by a change in conditions within the wound, which may cause the wound treatment skin substitute to change to a new color different from the original color of the skin substitute.

[0071] For purposes of this application, the terms “adipose material,” “subcutaneous fat,” “dermal adipose material,” “adipose tissue,” or “fat graft” may include or otherwise be derived from the biological components of adipose tissue. In various examples, the adipose material may include, for example, aspirated adipose tissue material obtained from liposuction or similar procedures. The adipose material may include one or more of adipocytes, endothelial cells, fibroblasts, B and T lymphocytes, macrophages, myeloid cells, pericytes, preadipocytes, smooth muscle cells, collagen, fibronectin, laminin, and stromal cells. The adipose material may further include blood, lipids, debris from adipocyte rupture, and / or any surrounding biological components. It should be noted that visceral adipose material is not excluded from the disclosed embodiments, although the embodiments may be described with reference to subcutaneous adipose material for convenience and to reflect more common use.

[0072] As described above, the adipose material can include lipoaspirate obtained by a liposuction procedure. Generally, liposuction refers to a surgical procedure involving the use of suction, such as through a cannula, to remove adipose material from a body part. Liposuction procedures can include any known procedure for removing adipose material, such as tumescent liposuction, dry liposuction, or wet liposuction. In various embodiments, the adipose tissue can include apical fat, mantle fat, deep fat, camper's fascia, subscarpa's fascia, and / or other adipose tissue. The adipose material can be obtained from any part of the body that has adipose material and associated tissue.

[0073] In embodiments, the fat material may be recently harvested or may have been stored for an extended period of time prior to use. The fat material may be processed after extraction or provided directly to the skin graft material, according to various embodiments. Skin graft material can be prepared for use with a patient in wound treatment according to the present disclosure by applying fat material harvested from the same patient, from another patient, from lab-grown material, and / or from an animal. Advantageously, using fat material harvested from a patient with a wound for treatment allows for improved healing, allows for a ready source of fat material, and prevents allergic or similar complications.

[0074] Various embodiments and components used therewith 1 illustrates a system 100 for preparing a wound treatment according to an embodiment of the present disclosure. The system 100 can include a syringe 110 or similar device for delivering a fat material to a loading chamber 120. The loading chamber 120 can be connected by an extrusion die 130 to a preparation chamber 140, which is configured to hold a skin graft material 150 therein.

[0075] In one embodiment, system 100 can be provided as a kit or device for preparing a wound treatment for application to a patient's wound. In this regard, skin graft material 150 can be loaded into preparation chamber 140 via a closure element 142 in the form of a door, flap, or similarly accessible portion of preparation chamber 140. Skin graft material 150 can be held in place within preparation chamber 140 through the use of a stage, clips, closure element 142, clamps, or the like.

[0076] Fat material can be obtained from a patient or another source, such as by a liposuction procedure, via a syringe 110 or similar device. In the embodiment of FIG. 1 , the syringe 110 includes a syringe plunger 112 for generating positive or negative pressure within the syringe body 114 to load or remove fat material therein through the cannula 116 or needle. The syringe 110 can be configured to cooperate with an input channel 122 of the loading chamber 120 to load the fat material into the loading chamber 120. For example, the input channel 122 can have a length, diameter, or other dimension configured to match the length, diameter, or other dimension of the cannula 116. In certain embodiments, the input channel 122 can have an inner diameter of 6 mm or less, more particularly, an inner diameter of 4 mm or less. In embodiments, the input channel 122 can include an elastic material configured to elastically contract relative to the cannula 116 such that the inner diameter of the input channel 122 corresponds to the outer diameter of the cannula 116. Aligning the input channel 122 with the cannula 116 in accordance with embodiments of the present disclosure can advantageously prevent any fatty material from escaping the loading chamber 120 via the input channel 122 .

[0077] The loading chamber 120 can be formed into an extrusion device by using a plunger 124 or similar pressure-generating device configured to force the fat material from the loading chamber 120 through the extrusion die 130. In embodiments, the plunger 124 can be configured to apply and / or generate pressure within the loading chamber 120 by applying manual force, such that extrusion of the fat material can be triggered by hand. The extrusion die 130 can be configured to shape the fat material into smaller portions for application to the skin graft material 150 in the preparation chamber 140. In certain aspects of the disclosed embodiments, applying the fat material to the skin graft material 150 can include infusing, impregnating, mixing, or otherwise combining the fat material with the skin graft material 150. In various embodiments, the preparation chamber 140 may be configured to hold the skin graft material 150 directly against the extrusion die 130, or may be configured to hold the skin graft material 150 at another predetermined location to receive the extruded fat material.

[0078] Referring to FIG. 2 , an extrusion die 230 according to an embodiment of the present disclosure can include a surface 232 defining one or more openings 234 therethrough. The one or more openings 234 can be configured to have a circular, square, rectangular, or any other shape. The one or more openings can have a smallest dimension perpendicular to the direction of travel of the fat material of 2 mm or less, 1 mm or less, or 0.5 mm or less, thereby separating individual components of the fat material and / or reducing their size for application to the scaffold material. To facilitate separation of the individual components of the fat material, the one or more openings 234 can be provided with sharp edges and / or blades at the one or more openings 234, with the sharp edges or blades facing the plunger 124.

[0079] In another aspect, a system 300 for preparing a wound treatment can be provided according to FIG. 3 . In the system 300, a loading chamber 320 containing a fat material 326 can be connected to a preparation chamber 340 holding a skin graft material 350 therein. The preparation chamber 340 can include an input channel 344 configured to cooperate with the loading chamber 320. In certain examples, the loading chamber 320 can comprise a syringe or a syringe-like device. The input channel 344 can be configured to removably secure to the loading chamber 320 or otherwise have a shape corresponding to an end of the loading chamber 320, advantageously facilitating the movement of the fat material 326 from the loading chamber 320 to the preparation chamber 340. In embodiments, the input channel 340 can be configured to open and close, such as by applying a sealing element thereto or by actuating a valve therein.

[0080] A screen mesh 352 can be provided in the preparation chamber 340 between the input channel 344 and the output channel 346. The screen mesh 352 can be configured to hold the skin graft material 350 against positive or negative pressure in the preparation chamber 340 while allowing the fat material 326 to pass therethrough. In embodiments, a vacuum pump 360 can be connected to the output channel 346 to generate negative pressure within the preparation chamber 340. The vacuum pump 360 can draw the fat material 326 from the loading chamber 320 into the preparation chamber 340, passing through both the skin graft material 350 and the screen mesh 352. In this manner, the fat material 326 can be applied to the skin graft material 350 to prepare a wound treatment in accordance with the present disclosure.

[0081] In some embodiments, the vacuum pump 360 can comprise a peristaltic pump, a syringe, or similar device. The loading chamber 320 can be further configured to force the fat material 326 into the preparation chamber 340 under a compressive force, such as by actuating a plunger or similar element, as described with respect to the system 100 of FIG. 1. In various embodiments, the input channel 344 can be provided with an extrusion die, blades, or the like to separate individual components or portions of the fat material 326 as it is delivered to the preparation chamber 340.

[0082] In some embodiments, the screen mesh 352 may have a screen size of 3 mm or less, 2 mm or less, or 1 mm or less. The screen mesh 352 may be disposed between the skin graft material 350 and the output channel 346, such as to prevent the skin graft material 350 from contacting or entering the output channel 346. In certain embodiments, the screen mesh 352 may be secured to the preparation chamber 340 or may have a size and shape corresponding to the size and shape of the preparation chamber 340 such that the screen mesh 352 is substantially held in place therein. In one aspect, an additional screen mesh 352 may be provided between the skin graft material 350 and the input channel 344 such that the additional skin graft material 352 can separate individual components or portions of the fat material 326 as it passes through the preparation chamber 340.

[0083] The preparation chamber 340 can comprise a flexible bag or a rigid chamber. The preparation chamber 340 defines an interior volume accessible via a closure element, such as a door, flap, or similar accessible portion. Similar to the system 100 of FIG. 1, the skin graft material 350 can be held in place in the preparation chamber 340 using a stage, clips, closure elements, clamps, or the like.

[0084] FIG. 4 illustrates an embodiment of a preparation chamber 440 comprising a flexible bag including a sealable flap 448. The sealable flap 448 may include adhesive or similar connecting means for enclosing and / or sealing the preparation chamber 440. As shown, a screen mesh 452 and skin graft material 450 may be provided to the preparation chamber 440 and enclosed therein using the sealable flap 448. In certain aspects, the screen mesh 452 may be configured as a tray for receiving the skin graft material 450, such as having raised elements 454 defining its perimeter. An input channel 444 may be provided on a first side 441 of the preparation chamber 440 opposite an output channel 446 on a second side 443 of the preparation chamber 440.

[0085] The operation of the system 300 for preparing a wound treatment can be better understood with reference to the flow diagram of Figure 5. As shown in the embodiment of Figure 5, a loading chamber 520 can be provided in the form of a syringe loaded with fat material. In a first step 502, for example according to the embodiment of Figure 4, the loading chamber 520 can be connected to a preparation chamber 540 containing skin graft material and a screen mesh therein. A vacuum pump 560 in the form of a peristaltic pump can be connected to the preparation chamber 540 via an outlet channel 546 in the form of a tube, such as in the form of surgical tubing, which is connected to the preparation chamber 540.

[0086] The vacuum pump 560 can be activated in a second step 504 to generate vacuum pressure in the preparation chamber 540 via the outlet channel 546. The vacuum pressure can draw fat material from the loading chamber 520 into the preparation chamber 540, passing through both the skin graft material and the screen mesh so that the fat material can be applied to the skin graft material. In various embodiments, the outlet channel 546 can be configured to exhaust any fat material drawn therethrough or to collect the fat material, whether for another use or for disposal. Once a desired level of fat material application is achieved in the skin graft material 550, the preparation chamber 540 can be opened and the skin graft material with the fat material 551 applied can be removed in step 506. The prepared skin graft material 551 can then be used as a wound treatment, such as by applying it to a wound, or can be stored for a period of time for use on a wound.

[0087] In embodiments according to Figures 6A and 6B, systems 600A, 600B can be provided for preparing wound treatment using non-planar skin graft material, agglomerations of comminuted or particulate skin graft material, or skin graft material 650 provided in a rolled or folded configuration. Similar in some respects to the embodiment of Figure 3, in system 600A, a loading chamber 620A containing fat material 626 can be connected to a preparation chamber 640 that holds skin graft material 650 therein. Preparation chamber 640 can include an input channel 644 configured to cooperate with loading chamber 620A. In certain examples, loading chamber 620A can comprise a syringe or syringe-like device.

[0088] 1 , system 600B can include a syringe 610 or similar device for delivering fat material to loading chamber 620B. Loading chamber 620B, which contains fat material 626, can be connected to a preparation chamber 640, which holds skin graft material 650 therein. Preparation chamber 640 can include an input channel 655 configured to cooperate with loading chamber 620B. In certain examples, loading chamber 620B can include a sealable compartment that can receive the fat material from syringe 610.

[0089] A vacuum pump 660 or similar vacuum source can be connected to the output channel 646 of the preparation chamber 640 to generate a negative pressure within the preparation chamber 640. The vacuum pump 660 can draw the fat material 626 from the loading chambers 620A, 620B into the preparation chamber 640 and pass it through the non-planar, comminuted, or rolled skin graft material 650. In certain embodiments, the dimensions of the input channel 655 and / or the output channel 646 can be configured to prevent the skin graft material 650 from being pulled or pushed out of the preparation chamber 640 so that the fat material 626 can be applied to the skin graft material 650. In certain embodiments, the vacuum pump 660 can include a syringe, for example, the same syringe 610 used to load the loading chamber or a separate syringe.

[0090] The operation of systems 600A, 600B for preparing a wound treatment can be better understood with reference to the flow diagram of FIG. 7. As shown in the embodiment of FIG. 7, a syringe 710A loaded with adipose material 726 can be provided to a loading chamber 720 in a first step 702. In a second step 704, the adipose material 726 can be loaded from syringe 710A into loading chamber 720, such as by expelling the contents of syringe 710 through the plunger of syringe 710A. In a third step 706, syringe 710B, whether the same syringe or a separate syringe, can be connected to a preparation chamber 740 containing skin graft material 750, e.g., rolled skin graft material, therein. Syringe 710B can be connected to preparation chamber 740 via an outlet channel 746 opposite loading chamber 720 and can be used to generate negative pressure within preparation chamber 740 during third step 706.

[0091] Negative pressure can draw fat material 726 from the loading chamber 720 into the preparation chamber 740 and through the skin graft material 750 so that the fat material 726 can be applied to the skin graft material 750. Once a desired level of application of fat material is achieved on the scaffold material 750, the preparation chamber 740 can be opened and the skin graft material with the fat material 751 applied can be removed in step 708. The prepared skin graft material 751 can then be used as a wound treatment, such as by applying it to a wound, or can be stored for a period of time for use on a wound.

[0092] In various embodiments, system 800 can be configured to repeatedly move adipose material through skin graft material 850 contained in preparation chamber 840, such as by moving the adipose material back and forth between two syringes 810A, 810B or between two vacuum pumps as shown in FIG. 8. By repeatedly cycling through skin graft material 850, more complete application of the adipose material to skin graft material 850 can be achieved without increasing the amount of adipose material required, advantageously reducing the amount of aspirated liposuction that needs to be removed from the patient, for example.

[0093] 9A , system 900A can be configured to include two syringes 910A-1, 910A-2 or two vacuum pumps disposed on opposite sides of preparation chamber 940A to repeatedly move adipose material across skin graft material 950 contained within preparation chamber 940A, such as by moving the adipose material back and forth. A screen mesh 952A-1 can be provided between syringe 910A-1 and skin graft material 950 in preparation chamber 940A, and a screen mesh 952A-2 can be provided between syringe 910A-1 and skin graft material 950. Screen mesh structures 952A-1, 952A-2 can be configured to hold skin graft material 950 against positive or negative pressure within preparation chamber 940A while allowing adipose material 926 to pass through. In this manner, the fat material 926 can be impregnated into the skin graft material 950, preparing it for wound treatment according to the present disclosure. In some embodiments, the screen meshes 952A-1, 952A-2 can further separate individual components or portions of the fat material 926 as it passes through the preparation chamber 940A, which can improve the application of the fat material 926 to or within the skin graft material 950.

[0094] 9B , system 900B can be configured to include two syringes 910B-1, 910B-2 or two vacuum pumps arranged perpendicular to each other (or approximately perpendicular to each other) on a vertical surface of preparation chamber 940B to repeatedly move adipose material across skin graft material 950 contained within preparation chamber 940B, such as by moving the adipose material back and forth. A screen mesh 952B-1 can be provided between syringe 910B-1 and skin graft material 950 in preparation chamber 940B, and a screen mesh 952B-2 can be provided between syringe 910B-2 and skin graft material 950. Screen mesh structures 952B-1, 952B-2 can be configured to hold skin graft material 950 against positive or negative pressure within preparation chamber 940B while allowing adipose material 926 to pass through. In this manner, the fat material 926 can be impregnated into the skin graft material 950 to prepare a wound treatment according to the present disclosure. In some embodiments, the screen meshes 952B-1, 952B-2 can further separate individual components or portions of the fat material 926 as it passes through the preparation chamber 940B, which can improve the application of the fat material 926 to or within the skin graft material 950.

[0095] 9C , system 900C can be configured to include two syringes 910C-1, 910C-2 or two vacuum pumps positioned on the same side of preparation chamber 940C to repeatedly move adipose material across skin graft material 950 contained within preparation chamber 940C, such as by moving the adipose material back and forth. A screen mesh 952C-1 can be provided in preparation chamber 940C between syringes 910C-1, 910C-2 and skin graft material 950, and a screen mesh 952C-2 can be provided on the opposite side of skin graft material 950 from first screen mesh 952C-1. Screen mesh structures 952C-1, 952C-2 can be configured to hold skin graft material 950 against positive or negative pressure within preparation chamber 940C while allowing adipose material 926 to pass through. In this manner, by repeatedly operating syringes 910C-1, 910C-2, fat material 926 can be impregnated into skin graft material 950, thus preparing a wound treatment according to the present disclosure. In some embodiments, screen meshes 952C-1, 952C-2 can further separate individual components or portions of fat material 926 as it passes through preparation chamber 940C, which can improve application of fat material 926 to or within skin graft material 950.

[0096] 9D , a system 900D can be configured to include two syringes 910D-1, 910D-2 or two vacuum pumps positioned on the same side of the preparation chamber 940D to repeatedly move adipose material across a skin graft material 950 contained within the preparation chamber 940D, such as by moving the adipose material back and forth. A screen mesh 952D-1 can be provided between the syringe 910D-1 and the skin graft material 950 in the preparation chamber 940D, and a screen mesh 952D-2 can be provided on the opposite side of the skin graft material 950 from the first screen mesh 952D-1. The screen mesh structures 952D-1, 952D-2 can be configured to hold the skin graft material 950 against positive or negative pressure within the preparation chamber 940D while allowing the adipose material 926 to pass through. In this embodiment, a channel 970D can optionally be provided to facilitate the movement of fat material 926 injected by second syringe 910D-2 under the skin graft material 950 and through opening 975D, as shown by the arrow indicating the movement of fat material 926 under the skin graft material. In this manner, fat material 926 can impregnate skin graft material 950 from both sides, preparing the wound for treatment according to the present disclosure. In some embodiments, screen meshes 952D-1, 952D-2 can further separate individual components or portions of fat material 926 as it passes through preparation chamber 940D, improving the application of fat material 926 to or within skin graft material 950.

[0097] 9E , a system 900E can be configured to include two syringes 910E-1, 910E-2 or two vacuum pumps positioned on the same side of a preparation chamber 940E to repeatedly move adipose material across a skin graft material 950 contained within the preparation chamber 940E, such as by moving the adipose material back and forth. Syringe 910E-1 is positioned perpendicular or nearly perpendicular to the top surface of the preparation chamber 940E, while a second syringe 910E-2 is positioned offset from the perpendicular to the top surface of the preparation chamber 940E by an angle α. A screen mesh 952E-1 can be provided in the preparation chamber 940E between the syringes 910E-1, 910E-2 and the skin graft material 950, and a screen mesh 952E-2 can be provided on the opposite side of the skin graft material 950 from the first screen mesh 952E-1. The screen mesh structures 952E-1, 952E-2 can be configured to hold the skin graft material 950 against positive or negative pressure in the preparation chamber 940E while allowing the fat material 926 to pass through. In this manner, by repeatedly operating the syringes 910E-1, 910E-2, the fat material 926 can be impregnated into the skin graft material 950, thus preparing a wound treatment according to the present disclosure. In some embodiments, the screen meshes 952E-1, 952E-2 can further separate individual components or portions of the fat material 926 as it passes through the preparation chamber 940E, which can improve the application of the fat material 926 to or within the skin graft material 950.

[0098] 9F, a system 900F can be configured to include two syringes 910F-1, 910F-2 or two vacuum pumps positioned on the same side of the preparation chamber 940F to repeatedly move adipose material across skin graft material 950 contained within the preparation chamber 940F, such as by drawing the adipose material back and forth. Syringe 910F-1 is positioned perpendicular or generally perpendicular to the top surface of the preparation chamber 940F, while the second syringe 910F-2 is positioned offset from the perpendicular to the top surface of the preparation chamber 940F by an angle α. A screen mesh 952F-1 can be provided in the preparation chamber 940F between the syringes 910F-1, 910F-2 and the skin graft material 950, and a screen mesh 952F-2 can be provided on the side of the skin graft material 950 opposite the first screen mesh 952F-1. In this embodiment, a channel 970F can optionally be provided to facilitate the movement of fat material 926 injected by the second syringe 910F-2 underneath the skin graft material 950 and through opening 975F, as shown by the arrow indicating the movement of fat material 926 underneath the skin graft material. The screen mesh structures 952F-1, 952F-2 can be configured to hold the skin graft material 950 against positive or negative pressure in the preparation chamber 940F while allowing the fat material 926 to pass through. In this manner, by repeatedly operating the syringes 910F-1, 910F-2, the fat material 926 can impregnate the skin graft material 950, thus preparing a wound treatment in accordance with the present disclosure. In some embodiments, the screen meshes 952F-1, 952F-2 can further separate individual components or portions of the adipose material 926 as it passes through the preparation chamber 940F, which can improve the application of the adipose material 926 to or within the skin graft material 950.

[0099] 10 , a system 1000 for preparing a wound treatment can include a preparation chamber 1040, a support frame 1070, and a roller 1080. In this embodiment, a skin graft material 1050 and a fat material 1026 can be provided to the preparation chamber 1040, and the preparation chamber 1040 can be positioned against the frame 1070. The roller 1080 can be positioned against the frame 1070 such that the roller 1080 can roll along the preparation chamber 1040 and apply pressure to the skin graft material 1050 and the fat material 1026 therein to apply the fat material to the scaffold material 1050.

[0100] In some embodiments, the roller 1080 can have a smooth surface or a patterned surface. For example, the roller 1080 can have multiple protrusions on its surface to apply irregular pressure across the preparation chamber 1040 so that the fat material 1026 can be broken down into separate components and / or portions. The roller 1080 can include a handle or the like to allow a user to control the roller 1080 and manually apply pressure thereto. In embodiments, the interior of the preparation chamber 1040 can include a patterned surface that faces the skin graft material 1050. For example, the preparation chamber 1040 can have multiple protrusions on its inner surface to apply irregular pressure across the skin graft material 1050 so that the fat material 1026 can be broken down into separate components and / or portions during application to the skin graft material 1050.

[0101] In its simplest form, the frame 1070 may include a support surface 1072. In embodiments, the frame 1070 may include a retaining element 1074 in the form of a clip or clamp. In the embodiment shown in FIG. 10, the retaining element is movable between a first open configuration and a second closed configuration. In the closed configuration, the retaining element 1074 may be secured by a cooperating locking element, such as a protrusion 1076, or another securing means configured to interact with a corresponding portion of the retaining element 1074.

[0102] An embodiment of a method 1100 of using system 1000 is shown in FIG. 11. As shown, in a first step 1101, skin graft material can be provided to a preparation chamber. In a second step 1102, fat material can be added to the preparation chamber. The fat material can then be secured to the support frame (step 1103), for example, by fastening a clip connection between the retaining element and the support frame. A roller can then be run across the preparation chamber to apply the fat material to the skin graft material (step 1104). The preparation chamber can then be removed from the frame in step 1105, and the prepared skin graft material can be removed from the preparation chamber in step 1106.

[0103] In various embodiments, the holding element 1074 can include rollers such that the preparation chamber 1040 is secured to the frame 1070 such that the holding element 1074 can perform both a holding and a rolling action as it is moved along the length of the frame 1070. In this manner, the system 1000 can form a mill or roller mill for compressing the skin graft material 1050 and fat material 1026 within the preparation chamber 1040.

[0104] 12, a system 1200 for preparing a wound treatment can include a plurality of rollers 1280 and a preparation chamber 1240 enclosing skin graft material and fat material. The rollers 1280 can be configured to face each other at a predetermined distance, for example, by mounting them to a common or mountable frame 1270. The rollers 1280 can then be clipped or mounted to the preparation chamber 1240 between themselves so that the rollers 1280 can be moved back and forth across the preparation chamber 1240, similar to a rolling mill in metalworking.

[0105] In certain embodiments, the rollers 1280 and brewing chamber 1240 may be provided with corresponding protrusions 1282 and / or grooves to restrict movement of the brewing chamber 1240 to a predetermined path between the rollers 1280. The protrusions 1282 and / or grooves may be provided in or on the frame of the brewing chamber 1240, such as in the form of thickened sections that are more rigid than the rest of the brewing chamber 1240. In this way, the brewing chamber may be more fully secured between the rollers 1280, and movement of the rollers 1280 across the brewing chamber 1240 may be more easily accomplished while maintaining precise control over the movement of the rollers 1280 and / or brewing chamber 1240.

[0106] 13 , a system 1300 for preparing a wound treatment can be provided with additional configurations. In the system 1300, a loading chamber 1320 containing a fat material can be connected to a preparation chamber 1340 holding a skin graft material 1350 therein. The preparation chamber 1340 can include an input channel 1344 configured to cooperate with the loading chamber 1320. In certain examples, the loading chamber 1320 can include a syringe or syringe-like device, as shown. In embodiments, the preparation chamber 1340 can include cooperating elements that surround and connect the skin graft material 1350 and a screen mesh 1352.

[0107] 13 , multiple output channels 1346 can be provided in the preparation chamber 1340 opposite the input channel 1344. A screen mesh 1352 can be configured to hold the skin graft material 1350 in the preparation chamber 1340 against the positive pressure from the loading chamber 1320 while allowing the fat material to pass through. In an embodiment, the fat material can be impregnated into the skin graft material 1350, and excess fat material can be forced out of the preparation chamber 1340 through the multiple output channels 1346 and collected or disposed of. The preparation chamber 1340 can then be opened and the prepared scaffold material 1351 can be removed for use as a wound treatment.

[0108] In some embodiments, the preparation chamber 1340 can comprise a rigid material and / or be reusable after cleaning and sterilization. A handle 1390 can be provided for receiving the loading chamber 1320 to give the user more ergonometric support or leverage while using the loading chamber 1320. In certain embodiments, the handle 1390 can be configured to provide a predetermined resistance to reduce the pressure that can be applied to the fat material, or can be configured with auxiliary means, such as a spring, to increase the pressure that can be applied to the fat material.

[0109] In some embodiments, additional benefits can be realized through methods of preparing skin graft materials for application of adipose material. In particular, certain components of adipose material may be too large to infiltrate, impregnate, infuse, or bond with the structure of existing scaffold materials, such as the collagen structure of decellularized fish skin. As shown in the hematoxylin and eosin stained images of properly freeze-dried decellularized fish skin scaffold materials in accordance with FIGS. 14A and 14B, known methods for preparing skin graft materials 1450A, 1450B may result in tightly packed collagen structures with openings that are too small for adequate penetration with the adipose material according to the present disclosure. This may be due to the common belief that more complete preservation of ECM structure, including pore size, in skin graft materials may result in improved wound healing, whereas enlargement or destruction of pores is generally considered detrimental to wound healing.

[0110] For example, known scaffold materials can be frozen using liquid nitrogen or other specialized freezing equipment that can rapidly freeze skin to below −70° C. to preserve the collagen structure of the scaffold. In another common application, the scaffold material can be freeze-dried, i.e., frozen at low temperatures and vacuum conditions, where water is sequentially removed from each ice crystal phase without ice recrystallization, or cryopreserved, i.e., fish skin is immersed in a cryoprotectant solution before freezing, to preserve the collagen structure of the scaffold.

[0111] In contrast to known methods for preparing scaffold materials, the method for preparing a scaffold material for use in a skin graft material according to the present application allows for the intentional disruption of the collagen structure of the scaffold to facilitate impregnation of the scaffold material with adipose material, such as through annealing or fenestration of the scaffold material.

[0112] In method 1500 according to the embodiment of FIG. 15 , the scaffold material or skin graft material can be annealed so that the collagen structure of the scaffold material or skin graft material can be destroyed during freeze-drying. In one embodiment, a hydrated scaffold material can be provided (1502), whether during the initial processing of the fish skin or during subsequent reprocessing. The hydrated scaffold material can then be gradually frozen (1504). For example, the hydrated scaffold material can be cooled to 0°C and held at 0°C for 1-24 hours, or 2-12 hours, or 3-6 hours. In this way, the pores of the collagen structure can be enlarged, and the pore size can be balanced with the level of ECM destruction so that the scaffold material maintains a sufficiently intact ECM but has a pore size large enough to facilitate impregnation with the dermal fat material. The freezing process can then reach a temperature of −30°C or below, −40°C or below, or −70°C or below. During the freezing process, the pressure surrounding the scaffold material can be maintained at atmospheric pressure or a vacuum pressure below standard conditions for part or all of the freezing process. The resulting dehydrated scaffold material with annealed collagen structure can then receive adipose material 1506, such as using a system according to an embodiment of the present disclosure.

[0113] Surprisingly, as shown in the hematoxylin and eosin stained images of decellularized fish skin scaffold material annealed according to Figures 16A and 16B, the pore size of scaffold material 1650A, 1650B can be increased relative to scaffold material 1450A, 1450B while the ECM structure is largely maintained. The increased pore size can then provide new, unexpected, and surprising advantages of allowing for improved impregnation or combination with fat materials according to the present disclosure. While previously thought to reduce the effectiveness of skin graft materials, it has surprisingly been discovered by the inventors of the present application that increased pore size of skin graft material according to the described embodiments for use with fat materials improves wound healing, contrary to current expectations in the art.

[0114] The pore size of the annealed scaffold material can be configured to accommodate adipose material containing one or more of, for example, adipocytes, endothelial cells, fibroblasts, B and T lymphocytes, macrophages, myeloid cells, pericytes, preadipocytes, smooth muscle cells, collagen, fibronectin, laminin, stromal cells, blood, lipids, debris from adipocyte rupture, and / or any surrounding biological components. According to various embodiments of the present disclosure, skin graft materials can be annealed to include pores having sizes, dimensions, or diameters, for example, with average diameters ranging from 10 μm to 1000 μm, 20 μm to 600 μm, 40 μm to 300 μm, 80 μm to 300 μm, 100 μm to 500 μm, 20 μm to 100 μm, or 30 μm to 80 μm. In some embodiments, annealing a scaffold material can increase the pore size of the scaffold material by at least 50%, at least 100%, at least 150%, at least 200%, or at least 1000%.

[0115] In another method 1700 according to the embodiment of FIG. 17 , a scaffold material can be fenestrated to disrupt the collagen structure of the scaffold material. In an embodiment, a scaffold material can be provided (1702), whether during initial processing of fish skin or during subsequent reprocessing. The scaffold material can then be fenestrated (1704) or cut to form multiple new openings in the scaffold material. Cutting can be performed using a micro-fenestration procedure, such as using a computer-controlled cutting machine with a blade size of less than 2 mm or less than 1 mm. The perforated scaffold material can then receive a fat material (1706), such as using a system according to an embodiment of the present disclosure. The perforated scaffold material can advantageously have a larger pore size than the initial scaffold material, as well as an increased surface area for application of a fat material. Thus, improved application of fat material in scaffold material can be achieved according to embodiments of the present disclosure.

[0116] Fenestration according to the present disclosure may include abrasion of the surface of the scaffold or skin graft material to disrupt the surface of the scaffold or skin graft material for application of adipose material. In some embodiments, fenestration may include the use of laser light energy or similar means to cut or create openings in the scaffold or skin graft material.

[0117] In another embodiment, as shown in FIG. 18 , a system 1800 for preparing a wound treatment in the form of a skin graft material with adipose material is shown. The system includes a press 1801, which can be a handheld press, having opposing sections including a first section 1810 and an opposing section 1820. Handle portions 1815 and 1825 can be provided on the first section 1810 and the second section 1820, respectively, and the handle portions are configured to be held by a hand (or both hands) or by a physician, can include gripping surfaces, and more importantly, can provide leverage in the pivotal movement of the first and second sections 1810, 1820, respectively. Alternatively, the handles can be actuated by a robotic mechanism. The opposing sections 1810, 1820 can pivot about a hinged fulcrum 1811, which provides movement along a circular arc β. The upper section 1810 is provided with a convex pressing portion having a convex pressing surface 1830, while the lower section 1820 is provided with a concave pressing portion 1840 with a concave pressing surface 1845. However, in another embodiment, the upper section can have a concave portion with a concave pressing surface and the lower section can have a convex pressing portion with a convex pressing surface. Or, indeed, in another embodiment, the opposing pressing surfaces can actually be parallel and planar, or generally parallel and planar.

[0118] In the embodiment of FIG. 18 , the syringe 1810 has a supply of fat material 1826 dispensed therein. The upper section 1810 further includes a narrow passage 1831 through the convex pressing portion 1830. Skin graft material 1850 can be disposed within the pressing area, in this case on the concave pressing surface 1845, although the opposing surface can also be configured to receive the skin graft material 1850. Upon or prior to closing the press 1801, the syringe plunger 1812 is actuated to pass the fat material 1826 through the passage 1831 and contact the skin graft material 1850. The opposing sections 1810, 1820 can then apply force to compress the fat material 1826 to prepare a wound treatment according to the present disclosure. The opposing sections 1810 , 1820 can be closed and opened in a related manner to penetrate the fat material 1826 into the skin graft material 1850 .

[0119] In some embodiments, further advantages can be realized through the use of a kit including the system of the present disclosure along with corresponding skin graft material. The kit can be provided as a sterile, disposable kit for wound treatment. In use, the kit is opened, the skin graft material is cut or placed into the desired shape, fat material is obtained from the patient, the fat material is applied to the skin graft material using a system according to the present disclosure, and the resulting skin graft material, including the fat material, can then be applied to the wound.

[0120] Various changes and / or modifications of the inventive features described herein, and additional applications of the principles described herein, will occur to those skilled in the relevant art having access to this disclosure, and may be made to the illustrated embodiments without departing from the spirit and scope of the invention as defined by the claims, and are to be considered within the scope of this disclosure. Accordingly, while various aspects and embodiments have been disclosed herein, other aspects and embodiments are also contemplated. Although many methods and components similar or equivalent to those described herein can be used to practice embodiments of the present disclosure, only specific components and methods are described herein.

[0121] It is also recognized that systems, devices, products, kits, methods, and / or processes according to certain embodiments of the present disclosure can include, incorporate, or otherwise consist of properties, features (e.g., components, members, elements, parts, and / or portions) described in other embodiments disclosed and / or described herein. Thus, various features of particular embodiments can be compatible with, combined with, included in, and / or incorporated into other embodiments of the present disclosure. Thus, the disclosure of particular features with respect to a stated embodiment of the present disclosure should not be construed as limiting the application or inclusion of such features to the stated embodiment. Rather, it is recognized that other embodiments can also include such features, members, elements, parts, and / or portions without necessarily departing from the scope of the present disclosure.

[0122] Also, unless a feature is described as requiring another feature to be combined with it, any feature herein can be combined with any other feature of the same or different embodiments disclosed herein. Moreover, to avoid obscuring aspects of the example embodiments, various known aspects of the example systems, methods, devices, etc. are not described in particular detail herein. However, such aspects are also contemplated herein.

[0123] This disclosure provides various examples, embodiments, and features that improve the visual performance of a display and / or a camera that records images from the display. It should be understood that the various examples, embodiments, and features disclosed herein can be combined with other examples, embodiments, or features described herein, unless expressly stated otherwise or unless such examples, embodiments, and features are mutually exclusive.

[0124] In addition to the above, further embodiments and examples include the following enumerated embodiments of displays and methods, including the first group of enumerated embodiments of displays having acoustic and / or light trapping, the second group of enumerated embodiments of displays and methods having stacked pixels, the third group of enumerated embodiments of displays and methods having macroblocks with sync-banding (in 3D), and the fourth group of enumerated embodiments of displays and methods having multiplexing and staggered multiplexing. It is noted that the embodiments and examples listed in the tables of each of the following four groups can be combined with other embodiments and examples listed in the tables of any, all, or any or all combinations of embodiments and examples of the other groups.

[0125] 1. A system for preparing a wound for treatment, the system comprising an applicator configured to apply a fat material to a skin graft material.

[0126] 2. A system according to item 1 above, any one of items 3 to 36 below, or a combination thereof, further comprising a preparation chamber configured to hold skin graft material therein.

[0127] 3. A system according to 1 or 2 above, any one of 4 to 36 below, or a combination thereof, wherein the applicator comprises one or more of a syringe, a plunger, a roller, a screw, and a container.

[0128] 4. A system according to any one of 1 to 3 above or 5 to 36 below, or a combination thereof, wherein the applicator comprises an extrusion press.

[0129] 5. A system according to any one of items 1 to 4 above or 6 to 36 below, or a combination thereof, The above extrusion press is a loading chamber configured to receive the fat material via the input channel; a plunger at a first end of the loading chamber; an extrusion die at a second end of the loading chamber opposite the first end; Equipped with the extrusion die connects the loading chamber and the preparation chamber, and the plunger is configured to force the fat material from the loading chamber, through the extrusion die, and into the preparation chamber. system.

[0130] 6. A system according to any one of items 1 to 5 or 7 to 36 below, or a combination thereof, wherein the input channel has an inner diameter of 4 mm or less.

[0131] 7. A system according to any one of 1-6 above or 8-36 below, or a combination thereof, wherein the input channel comprises a resilient material configured to substantially seal the input channel around a needle or cannula.

[0132] 8. A system according to any one of items 1 to 7 or 9 to 36 below, or a combination thereof, wherein the plunger has a diameter approximately equal to the inner diameter of the loading chamber.

[0133] 9. A system according to any one of items 1 to 8 or 10 to 36 below, or a combination thereof, wherein the extrusion die comprises one or more openings connecting the loading chamber to the preparation chamber, each of the one or more openings having a diameter of 1 mm or less.

[0134] 10. A system according to any one of 1 to 9 above or 11 to 36 below, or a combination thereof, wherein the extrusion die has one or more blades facing the plunger.

[0135] 11. A system according to any one of items 1 to 10 or 12 to 36 below, or a combination thereof, wherein the one or more blades define the contours of the one or more openings in the extrusion die.

[0136] 12. A system according to any one of 1 to 11 above or 13 to 36 below, or a combination thereof, wherein the one or more blades are provided in the one or more openings of the extrusion die.

[0137] 13. A system according to any one of items 1 to 12 above or 14 to 36 below, or a combination thereof, wherein the preparation chamber comprises a holding element for fixing the skin graft material.

[0138] 14. A system according to any one of 1-13 above or 15-36 below, or a combination thereof, wherein the retaining element is configured to secure the skin graft material to the extrusion die.

[0139] 15. A system according to any one of items 1 to 14 above or items 16 to 36 below, or a combination thereof, The system is an output channel on a second side of the preparation chamber opposite the first side, the output channel configured to cooperate with a vacuum source; a mesh screen provided in the preparation chamber; Further equipped The system, wherein the vacuum source is configured to create a vacuum in the preparation chamber such that the fat material is drawn from the loading chamber through the skin graft material in the preparation chamber.

[0140] 16. A system according to any one of items 1 to 15 above or 17 to 36 below, or a combination thereof, wherein the vacuum source comprises a peristaltic pump.

[0141] 17. A system according to any one of items 1 to 16 above or items 18 to 36 below, or a combination thereof, wherein the mesh screen has a screen size of 1 mm or less.

[0142] 18. A system according to any one of items 1 to 17 above or 19 to 36 below, or a combination thereof, wherein the loading chamber comprises a syringe.

[0143] 19. A system according to any one of 1 to 18 above or 20 to 36 below, or a combination thereof, wherein the mesh screen is disposed between the skin graft material and the output channel.

[0144] 20. A system according to any one of 1-19 above or 21-36 below, or a combination thereof, comprising a further mesh screen disposed between the skin graft material and the input channel.

[0145] 21. A system according to any one of 1 to 20 above or 22 to 36 below, or a combination thereof, wherein the preparation chamber comprises a sealable bag.

[0146] 22. A system according to any one of 1 to 21 above or 23 to 36 below, or a combination thereof, wherein the output channel comprises tubing configured for use with a peristaltic pump.

[0147] 23. A system according to any one of 1 to 22 above or 24 to 36 below, or a combination thereof, wherein the mesh screen is larger than the skin graft material.

[0148] 24. A system according to any one of 1 to 23 above or 25 to 36 below, or a combination thereof, wherein the mesh screen has varying screen sizes.

[0149] 25. A system according to any one of items 1 to 24 above or items 26 to 36 below, or a combination thereof, A support frame; a roller configured to apply pressure to the brewing chamber opposite the support frame; The system further comprises:

[0150] 26. A system according to any one of 1 to 25 above or 27 to 36 below, or a combination thereof, wherein the roller includes a handle for operating the roller.

[0151] 27. A system according to any one of 1 to 26 above or 28 to 36 below, or a combination thereof, wherein the support frame includes a holding element for fixing the preparation chamber thereto.

[0152] 28. A system according to any one of 1 to 27 above or 29 to 36 below, or a combination thereof, wherein the holding element comprises a clamp arm movable between a first open position and a second closed position, the clamp arm extending from a surface of the support frame.

[0153] 29. A system according to any one of items 1 to 28 or 30 to 36 below, or a combination thereof, wherein the roller has a plurality of protrusions on its surface.

[0154] 30. A system according to any one of 1 to 29 above or 31 to 36 below, or a combination thereof, wherein the support frame is configured to fix the preparation chamber between the roller and a further opposing roller.

[0155] 31. A system according to any one of 1 to 30 above or 32 to 36 below, or a combination thereof, wherein the roller and the opposing roller are resiliently fixed to the frame such that the distance between the roller and the opposing roller can vary based on the resistance provided by the brewing chamber.

[0156] 32. A system according to any one of 1 to 31 above or 33 to 36 below, or a combination thereof, wherein the mesh screen divides the preparation chamber into a first portion and a second portion.

[0157] 33. A system according to any one of 1 to 32 above or 34 to 36 below, or a combination thereof, the system comprising an opposing vacuum source located opposite the vacuum source, the opposing vacuum source configured to generate an opposing vacuum in the preparation chamber such that the fat material is drawn back into the preparation chamber through the skin graft material.

[0158] 34. A system according to any one of 1 to 33 above or 35 to 36 below, or a combination thereof, wherein the frame has a plurality of protrusions on its surface.

[0159] 35. A system according to any one of 1 to 34 above, 36 below, or a combination thereof, wherein the preparation chamber comprises a sealable container.

[0160] 36. A system according to any one of 1 to 35 above, or a combination thereof, wherein the applicator comprises a heating element and / or a thermally conductive material.

[0161] 37. A wound care kit comprising: Skin graft material; an applicator configured to apply a fat material to the skin graft material; A wound treatment kit comprising:

[0162] 38. A kit according to item 37 above, any one of items 39-40 below, or a combination thereof, wherein the applicator comprises one or more of a syringe, an extrusion press, a loading chamber, a preparation chamber, and / or a pressure source.

[0163] 39. A kit according to any one of items 37-38 above, 40 below, or a combination thereof, wherein one or more of the syringe, the extrusion press, the loading chamber, the preparation chamber, and / or the pressure source comprise fluid connections for fluidly connecting together.

[0164] 40. A kit according to any one of 37 to 39 above, or a combination thereof, further comprising a container enclosing at least the skin graft material and the applicator in a sterile condition.

[0165] 41. A wound treatment composition comprising: Skin graft material; A fatty material; 10. A wound treatment composition comprising:

[0166] 42. The composition according to item 41 above, any one of items 43 to 60 below, or a combination thereof, wherein the skin graft material comprises a biological material.

[0167] 43. A composition according to any one of items 41-42 or 44-60 below, or a combination thereof, wherein the biological material comprises fish skin.

[0168] 44. A composition according to any one of items 41 to 43 or 45 to 60 below, or a combination thereof, wherein the biological material comprises pig skin.

[0169] 45. A composition according to any one of items 41 to 44 or 46 to 60 below, or a combination thereof, wherein the skin graft material comprises an extracellular matrix material.

[0170] 46. ​​A composition according to any one of items 41 to 45 or 47 to 60 below, or a combination thereof, wherein the skin graft material comprises decellularized skin.

[0171] 47. A composition according to any one of items 41-46 or 48-60 below, or a combination thereof, wherein the skin graft material comprises a synthetic scaffold material.

[0172] 48. A composition according to any one of items 41-47 or 49-60 below, or a combination thereof, wherein the skin graft material comprises a plurality of pulverized particles.

[0173] 49. A composition according to any one of items 41 to 48 or 50 to 60 below, or a combination thereof, wherein the skin graft material has a pore size larger than the fat cells.

[0174] 50. A composition according to any one of items 41 to 49 or 51 to 60 below, or a combination thereof, wherein the fatty material comprises subcutaneous fat and / or visceral fat.

[0175] 51. A composition according to any one of items 41 to 50 or 52 to 60 below, or a combination thereof, wherein the fatty material comprises dermal fat.

[0176] 52. A composition according to any one of items 41 to 51 or 53 to 60 below, or a combination thereof, wherein the fat material comprises aspirated fat tissue.

[0177] 53. A composition according to any one of items 41-52 or 54-60 below, or a combination thereof, wherein the fatty material comprises a plurality of comminuted particles.

[0178] 54. A composition according to any one of items 41 to 53 or 55 to 60 below, or a combination thereof, wherein the skin graft material comprises a plurality of pores.

[0179] 55. A composition according to any one of items 41 to 54 or items 56 to 60 below, or a combination thereof, wherein the pores of the skin graft material contain the fatty material.

[0180] 56. A composition according to any one of items 41 to 55 or 57 to 60 below, or a combination thereof, wherein the skin graft material is impregnated or injected with a fat material.

[0181] 57. A composition according to any one of items 41-56 or 58-60 below, or a combination thereof, wherein the fatty material constitutes at least 50% of the total weight of the composition.

[0182] 58. A composition according to any one of items 41-57 or 59-60 below, or a combination thereof, wherein the fatty material constitutes at least 50% of the total weight of the composition.

[0183] 59. A composition according to any one of items 41 to 58 above, item 60 below, or a combination thereof, wherein the skin graft material has a pore size larger than the fat cells.

[0184] 60. A composition according to any one of 41 to 59 above, or a combination thereof, wherein the fatty material comprises subcutaneous fat and / or visceral fat.

[0185] 61. A method for preparing a patient for wound treatment, comprising: providing a skin graft material; applying a fat material to the skin graft material; A method comprising:

[0186] 62. A method according to claim 61, any one of claims 63 to 80, or a combination thereof, wherein the step of providing the skin graft material includes providing the skin graft material in a preparation chamber.

[0187] 63. A method according to any one of items 61-62 or 64-80 below, or a combination thereof, further comprising the step of preparing the skin graft material to receive the fat material.

[0188] 64. A method according to any one of items 61 to 63 or 65 to 80 below, or a combination thereof, wherein the step of preparing the skin graft material includes increasing the pore size and / or surface area of ​​the skin graft material by annealing, fenestration, and / or comminution.

[0189] 65. A method according to any one of items 61-64 or 66-80 below, or a combination thereof, wherein the step of annealing the skin graft material includes freezing the skin graft material to form ice crystals therein.

[0190] 66. A method according to any one of items 61 to 65 or 67 to 80 below, or a combination thereof, wherein the freezing of the skin graft material is carried out at a temperature of 0°C for 1 to 24 hours.

[0191] 67. A method according to any one of items 61 to 66 or 68 to 80 below, or a combination thereof, wherein freezing the skin graft material increases the pore size of the skin graft material.

[0192] 68. A method according to any one of items 61 to 67 or 69 to 80 below, or a combination thereof, wherein the fenestration comprises cutting the skin graft material with a blade of 1 mm or less.

[0193] 69. A method according to any one of items 61-68 or 70-80 below, or a combination thereof, wherein the fenestration comprises applying laser energy to the skin graft material to form an opening therein.

[0194] 70. The method according to any one of 61-69 above or 71-80 below, or a combination thereof, wherein the fenestration comprises applying an abrasive to the skin graft material to form an opening therein.

[0195] 71. A method according to any one of items 61 to 70 or items 72 to 80 below, or a combination thereof, further comprising the step of obtaining said fatty material.

[0196] 72. A method according to any one of items 61-71 or 73-80 below, or a combination thereof, wherein the step of obtaining the adipose material includes performing a liposuction procedure.

[0197] 73. A method according to any one of items 61-72 or 74-80 below, or a combination thereof, wherein the step of obtaining the adipose material includes separating the adipose material from aspirated fat tissue from the liposuction procedure.

[0198] 74. A method according to any one of items 61 to 73 or items 75 to 80 below, or a combination thereof, wherein the step of obtaining the fatty material includes cutting the fatty material to a predetermined size.

[0199] 75. A method according to any one of items 61 to 74 or items 76 to 80 below, or a combination thereof, wherein the step of obtaining the fatty material includes grinding the fatty material into particles of a predetermined size.

[0200] 76. A method according to any one of items 61 to 75 or items 77 to 80 below, or a combination thereof, wherein the step of applying the fat material to the skin graft material includes applying pressure to the fat material and / or the skin graft material such that the fat material and the skin graft material are pressed or pulled together under pressure.

[0201] 77. A method according to any one of items 61-76 or 78-80 below, or a combination thereof, wherein the step of applying pressure to the fat material and / or the skin graft material includes applying pressure for a predetermined period of time.

[0202] 78. A method according to any one of items 61 to 77 or items 79 to 80 below, or a combination thereof, wherein the step of applying the fat material to the skin graft material includes impregnating and / or injecting the fat material into the skin graft material.

[0203] 79. A method according to any one of items 61 to 78 above, item 80 below, or a combination thereof, wherein the step of applying the fat material to the skin graft material includes infiltrating the fat material into the pores of the skin graft material.

[0204] 80. A method according to any one of 61 to 79 above, or a combination thereof, wherein the step of applying the fat material to the skin graft material includes applying thermal energy to the fat material and / or the skin graft material.

[0205] 81. A method for treating a wound, comprising: providing a skin graft material; applying fat material to the skin graft material to form a combined fat and skin graft material; applying the combined fat and skin graft material to the wound; A method comprising:

[0206] 82. The method according to claim 81, any one of claims 83 to 90, or a combination thereof, wherein the step of applying the combined fat and skin graft material to the wound is performed within 12 hours of the step of applying the fat material to the skin graft material.

[0207] 83. A method according to any one of items 81-82 above or 84-90 below, or a combination thereof, wherein the step of applying the combined fat and skin graft material to the wound is performed within 12 hours of removing the fat material from the patient.

[0208] 84. A method according to any one of items 81 to 83 or 85 to 90 below, or a combination thereof, wherein the fatty material is obtained from a patient having a wound.

[0209] 85. A method according to any one of items 81-84 or 86-00 below, or a combination thereof, further comprising the step of cutting the skin graft material to the shape of the wound.

[0210] 86. A method according to any one of items 81-85 or 87-90 below, or a combination thereof, wherein the skin graft material and / or the fat material comprises a plurality of comminuted particles, and the step of applying the combined fat and skin graft material to the wound further comprises shaping the combined fat and skin graft material to fit the wound.

[0211] 87. A method according to any one of items 81-86 above or 88-90 below, or a combination thereof, wherein the step of applying the combined fat and skin graft material to the wound includes sealing the wound.

[0212] 88. A method according to any one of items 81-87 above or 89-90 below, or a combination thereof, wherein the wound is sealed with a bandage and / or sutures at least partially covering the combined fat and skin graft material.

[0213] 89. A method according to any one of items 81-88 above, 90 below, or a combination thereof, wherein the step of applying the combined fat and skin graft material to the wound is performed within 2 hours of the step of applying the fat material to the skin graft material.

[0214] 90. A method according to any one of items 81 to 89 above, or a combination thereof, wherein the step of applying the combined fat and skin graft material to the wound is performed within 2 hours of removing the fat material from the patient.

[0215] 91. A wound treatment composition comprising: The wound treatment composition comprises a skin graft material having a plurality of pores; The pores have an average diameter of 20 μm to 1000 μm, Optionally, the skin graft material has been treated to increase the pore size and / or surface area of ​​the skin graft material by annealing, fenestration, and / or milling; or The skin graft material has been annealed to increase the pore size and / or surface area of ​​the skin graft material by freezing the skin graft material to form ice crystals therein. Wound treatment composition.

[0216] 92. The composition according to claim 91, any one of claims 93 to 100, or a combination thereof, wherein the skin graft material comprises a biological material.

[0217] 93. A composition according to any one of items 91-92 above or 94-100 below, or a combination thereof, wherein the biological material comprises fish skin.

[0218] 94. A composition according to any one of items 91-93 or 95-100 below, or a combination thereof, wherein the biological material comprises pig skin.

[0219] 95. A composition according to any one of items 91-95 or 96-100 below, or a combination thereof, wherein the skin graft material comprises an extracellular matrix material.

[0220] 96. A composition according to any one of items 91-95 or 97-100 below, or a combination thereof, wherein the skin graft material comprises decellularized skin.

[0221] 97. A composition according to any one of items 91-96 or 98-100 below, or a combination thereof, wherein the skin graft material comprises a synthetic scaffold material.

[0222] 98. A composition according to any one of items 91 to 97 above or 99 to 100 below, or a combination thereof, wherein the pores have an average diameter of 20 μm to 600 μm.

[0223] 99. The composition according to any one of items 91 to 98 above, item 100 below, or a combination thereof, wherein the pores have an average diameter of 20 μm to 300 μm.

[0224] 100. The composition according to any one of 91 to 99 above, or a combination thereof, wherein the pores have an average diameter of 20 μm to 100 μm.

[0225] 101. A method for preparing a patient for wound treatment, comprising: providing a skin graft material; treating the skin graft material to increase the pore size and / or surface area of ​​the skin graft material; A method comprising:

[0226] 102. The method according to claim 101, any one of claims 103 to 110, or a combination thereof, wherein the step of treating the skin graft material includes increasing the pore size and / or surface area of ​​the skin graft material by annealing, fenestration, and / or milling.

[0227] 103. A method according to any one of 101-102 above or 104-110 below, or a combination thereof, wherein the step of annealing the skin graft material comprises freezing the skin graft material to form ice crystals therein.

[0228] 104. A method according to any one of items 101 to 103 above or 105 to 110 below, or a combination thereof, wherein the freezing of the skin graft material is carried out at a temperature of 0°C for 1 to 24 hours.

[0229] 105. A method according to any one of 101-104 above or 106-110 below, or a combination thereof, wherein freezing the skin graft material increases the pore size of the skin graft material.

[0230] 106. A method according to any one of 101 to 105 above or 107 to 110 below, or a combination thereof, wherein the fenestration comprises cutting the skin graft material with a blade of 1 mm or less.

[0231] 107. The method according to any one of 101-106 or 108-110 below, or a combination thereof, wherein said fenestration comprises applying laser energy to said skin graft material to form an opening therein.

[0232] 108. The method according to any one of 101-107 above or 109-110 below, or a combination thereof, wherein said fenestration comprises applying an abrasive to skin graft material to form an opening therein.

[0233] 109. The method according to any one of 101-108 above, 110 below, or a combination thereof, wherein the skin graft material comprises a biological material.

[0234] 110. The method according to any one of 101 to 109 above, or a combination thereof, wherein treating the skin graft material increases the average diameter of the pores to within the range of 20 μm to 1000 μm.

[0235] A skilled artisan will recognize the interchangeability of the various disclosed features. In addition to the variations described herein, other known equivalents for each feature can be mixed and matched by those skilled in the art to prepare wound treatment and utilize methods for doing so based on the principles of the present disclosure. A skilled artisan will understand that the features described herein can be adapted for other types of wound treatment and health care applications generally.

[0236] Although a system or method for preparing a wound treatment including a skin graft material provided with a fat material has been disclosed in certain preferred embodiments and examples, it will be understood by those skilled in the art that the present disclosure extends beyond the disclosed embodiments to other alternative embodiments and / or uses of the system or method for preparing a wound treatment including a skin graft material provided with a fat material, as well as obvious modifications and equivalents. It is intended that the scope of the disclosed system or method for preparing a wound treatment including a skin graft material with a fat material should not be limited by the above-disclosed embodiments, but should be determined solely by a fair reading of the claims that follow.

Claims

1. A system for preparing a wound treatment product, the system comprising an applicator configured to apply a fat material to a skin graft material.

2. The system of claim 1 , further comprising a preparation chamber configured to hold the skin graft material therein.

3. The system of claim 1 or 2, wherein the applicator comprises one or more of a syringe, a plunger, a roller, a screw, and a container.

4. The system of any one of claims 1 to 3, wherein the applicator comprises an extrusion press.

5. The extrusion press comprises: a loading chamber configured to receive the fat material via an input channel; a plunger at a first end of the loading chamber; an extrusion die at a second end of the loading chamber opposite the first end; Equipped with the extrusion die connects the loading chamber and the preparation chamber, and the plunger is configured to force the fat material from the loading chamber, through the extrusion die, and into the preparation chamber. The system according to any one of claims 1 to 4.

6. 6. The system of claim 1, wherein the extrusion die comprises one or more openings connecting the loading chamber to the preparation chamber, each of the one or more openings having a diameter of 1 mm or less.

7. the extrusion die includes one or more blades facing the plunger; the one or more blades define the contours of the one or more openings in the extrusion die or are disposed in the one or more openings in the extrusion die; The system according to any one of claims 1 to 6.

8. the preparation chamber comprising a holding element for securing the skin graft material; the retaining element is configured to secure the skin graft material to the extrusion die; The system according to any one of claims 1 to 7.

9. The system comprises: an output channel on the second surface of the brewing chamber opposite the first surface, the output channel configured to cooperate with a vacuum source; a mesh screen provided in the preparation chamber; Furthermore, the vacuum source is configured to create a vacuum in the preparation chamber such that the fat material is drawn from the loading chamber through the skin graft material in the preparation chamber. The system according to any one of claims 1 to 7.

10. The system of any one of claims 1 to 9, wherein the loading chamber comprises a syringe.

11. The system of any one of claims 1 to 9, wherein the preparation chamber comprises a sealable bag.

12. A support frame; a roller configured to apply pressure to the brewing chamber opposite the support frame; The system of any one of claims 1 to 11, further comprising:

13. The system of claim 12 , wherein the roller includes a handle for manipulating the roller.

14. the system further comprising an opposing vacuum source located opposite the vacuum source; 14. The system of any one of claims 1 to 13, wherein the contralateral vacuum source is configured to create a contralateral vacuum in the preparation chamber such that the fat material is drawn back into the preparation chamber through the skin graft material.

15. the applicator comprising a plurality of syringes each configured to inject the fatty material into the preparation chamber; Optionally, each of said syringes is configured to draw said fatty material from said preparation chamber. The system of claim 2.

16. 3. The system of claim 2, wherein the applicator includes two opposing pressure sections, including a first pressure section and a second pressure section, configured to pivot relative to each other at a hinge point, each of the opposing pressure sections having respective opposing pressure surfaces that define a preparation chamber, the opposing pressure surfaces providing pressure to press the fat material into the skin graft material.

17. The system of claim 16 , wherein the opposing pressure surfaces include a concave pressure surface and a convex pressure surface.

18. the system includes a syringe disposed behind the first pressing section; the first pressing section includes a convex pressing section, a passageway through the convex pressing section being provided for providing a passageway for the fat material to be discharged from the syringe into the preparation chamber; 18. The system of claim 17.

19. 1. A method for preparing a skin graft material, comprising: providing a skin graft material; Providing a system according to any one of claims 1 to 18; applying a fat material to the skin graft material using the system; A method comprising:

20. 1. A wound treatment kit comprising: Skin graft material; An applicator according to any one of claims 1 to 18, configured to apply a fat material to the skin graft material; A wound treatment kit comprising:

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