Method for modulating endothelial glycocalyx structure

Abstract

To provide products, processes, and methods for treating (e.g., supporting and / or maintaining) the endothelial glycocalyx.SOLUTION: The present invention provides methods and devices for augmenting impaired glycocalyx barrier function in a subject in need thereof by contacting a sample (e.g., blood) obtained from the subject with a glycocalyx-mimetic adsorption media. The adsorption media include glycosaminoglycan structures and, optionally, proteoglycan core proteins, which are conducive to enhancing and / or restoring the impaired glycocalyx barrier function in a sample. The contacted sample is subsequently separated from the adsorption media, producing a treated sample that can be infused into the subject. Methods and devices for treating a patient suffering from a disease associated with glycocalyx barrier dysfunction are also provided herein.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Application No. 62 / 848,819, filed May 16, 2019, the contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] Impairment of the glycocalyx barrier through structural damage or depletion, dysfunction, or other mechanisms can contribute to microvascular endothelial dysfunction, including inflammatory and coagulative endothelial activation, vascular leakage of fluid, proteins, and other substances (e.g., cholesterol), failure to properly regulate perfused vascular density, and other adverse conditions. All of the foregoing leads to general and specific negative vascular health indicators. For example, an unhealthy endothelial glycocalyx is associated with a "leaky" endothelium, which can be evidenced by (1) the presence (or "leakage") of cholesterol (or other substances, such as fluids or proteins) in (or within) the subendothelial space and (2) a narrowed lumen, which can reduce blood flow or perfusion to distal capillaries, muscles, organs, etc., and potentially increase blood pressure. However, a healthy (thick and / or dense) endothelial glycocalyx is associated with a well-formed endothelium and the structural organization of healthy blood vessels.

[0003] U.S. Patent No. 2020 / 0023001 to Ebong teaches a composition comprising heparan sulfate and sphingosine-1-phosphate for regenerating the endothelial glycocalyx and treating vascular disease. Exogenous heparan sulfate is incorporated into the glycocalyx. Summary of the Invention [Problem to be solved by the invention]

[0004] Thus, there is a need for products, processes, and methods for treating (e.g., supporting and / or maintaining) the endothelial glycocalyx. The present disclosure addresses these and other needs. [Means for solving the problem]

[0005] Generally, provided herein are methods and devices for enhancing impaired glycocalyx barrier function in a subject in need thereof. In other words, the methods and devices described herein are for improving or restoring damaged or disrupted glycocalyx barrier function in a subject. Consequently, methods and devices for treating patients suffering from diseases associated with glycocalyx barrier dysfunction are also provided herein. A sample obtained from a subject with impaired glycocalyx barrier function is contacted with a glycocalyx-mimicking adsorption medium. The adsorption medium serves to enhance and / or restore impaired glycocalyx barrier function, acting as an artificial glycocalyx and comprising structures and materials that remove inflammatory mediators from the sample that would be regulated by an undamaged or intact glycocalyx barrier. The sample is then separated from the adsorption medium, resulting in enhanced impaired glycocalyx barrier function and / or a reduction in the amount of inflammatory mediators in the currently processed sample. The processed sample (e.g., blood) is then returned to the subject by reinfusion.

[0006] According to one aspect, the present disclosure provides a method for enhancing impaired glycocalyx barrier function in a subject in need thereof. The method comprises contacting a sample from the subject with a glycocalyx-mimetic adsorption medium to enhance and / or restore impaired glycocalyx barrier function, thereby treating the sample; and injecting the treated sample into the subject, wherein the glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof. The adsorbent does not leach from the solid substrate into the sample.

[0007] According to one aspect, the adsorbent is a glycosaminoglycan mixture comprising about 40% to about 96% w / w heparan sulfate and / or heparin, and optionally one or more of the following additional adsorbents: about 5% to about 30% w / w chondroitin sulfate, about 1% to about 25% w / w dermatan sulfate, about 0.01% to about 20% w / w keratan sulfate, and / or about 5% to about 50% w / w hyaluronic acid. According to some embodiments, the glycosaminoglycan optionally comprises at least one proteoglycan core protein selected from the group consisting of syndecan, glypican, perlecan, versican, decorin, biglycan, and mimecan.

[0008] According to some embodiments, the glycocalyx-mimicking adsorption medium aids in a member selected from the group consisting of vascular permeability, leukocyte adhesion, platelet adhesion, shear stress mediation, and inflammatory regulation. According to some embodiments, the adsorption medium functions as an endothelial surface layer to protect and / or maintain glycocalyx function. According to some embodiments, the adsorption medium reduces a member selected from the group consisting of capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulation, and loss of vascular reactivity.

[0009] According to some embodiments, the adsorption medium reduces glycocalyx shedding during reperfusion of tissue. According to some embodiments, the tissue is cardiac tissue during coronary artery bypass surgery. According to some embodiments, the tissue is perfused during organ transplantation.

[0010] According to some embodiments, the adsorption medium reduces glycocalyx shedding during sepsis. According to some embodiments, the adsorption medium removes tumor necrosis factor (TNF)-α and bacterial lipopolysaccharide (LPS). According to some embodiments, the adsorption medium reduces the risk of organ failure.

[0011] According to some embodiments, the method treats acute respiratory distress syndrome (ARDS) in a subject.

[0012] According to some embodiments, the method improves oxygen saturation in the subject.

[0013] According to some embodiments, the method improves hemodynamic stability in a subject.

[0014] According to some embodiments, the method treats Covid-19.

[0015] According to some embodiments, the adsorption medium reduces glycocalyx shedding due to atherosclerosis or diabetes. According to some embodiments, the adsorption medium removes low-density lipoproteins (LDL).

[0016] In some embodiments, the adsorption medium binds heparin-binding protein (HBP), and in some embodiments, the treated sample has an HBP content that is reduced by about 10% to about 100% compared to the HBP content of the sample before treatment.

[0017] According to some embodiments, the adsorption medium binds a member selected from the group consisting of exotoxins, endotoxins, ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, and cytokines.

[0018] In some embodiments, the sample is a member selected from the group consisting of whole blood, serum, and plasma. In some embodiments, the sample is whole blood.

[0019] According to some embodiments, the glycocalyx-mimetic adsorption medium is negatively charged.

[0020] According to some embodiments, the solid substrate comprises a non-toxic, non-leaching material. According to some embodiments, the solid substrate comprises a plurality of rigid polymer beads. According to some embodiments, the rigid polymer beads are selected from the group consisting of polyurethane, polymethyl methacrylate, polyethylene or copolymers of ethylene and other monomers, polyethyleneimine, polypropylene, and polyisobutylene. According to some embodiments, the solid substrate comprises one or more hollow fibers.

[0021] Another aspect of the present disclosure relates to an apparatus for enhancing impaired glycocalyx barrier function in a subject in need thereof, the apparatus comprising a cartridge having a first end plate and a second end plate with a glycocalyx-mimicking adsorption medium disposed thereon, wherein the glycocalyx-mimicking adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof. According to some embodiments, the adsorbent of the device is a glycosaminoglycan mixture comprising about 40% to about 96% w / w heparan sulfate and / or heparin, and optionally one or more of the following additional adsorbents: about 5% to about 30% w / w chondroitin sulfate, about 1% to about 25% w / w dermatan sulfate, about 0.01% to about 20% w / w keratan sulfate, and / or about 5% to about 50% w / w hyaluronic acid; the device further comprises a sample inlet port for allowing a sample to enter the device; and a sample outlet port for allowing a sample to exit the device, wherein the sample flows through the first end plate, through the adsorption medium, and out the outlet port. According to some embodiments, the glycosaminoglycan mixture comprises at least one proteoglycan core protein selected from the group consisting of syndecan, glypican, perlecan, versican, decorin, biglycan, and mimecan.

[0022] According to some embodiments, the glycocalyx-mimicking adsorption medium aids in a member selected from the group consisting of vascular permeability, leukocyte adhesion, platelet adhesion, shear stress mediation, and inflammatory regulation. According to some embodiments, the adsorption medium functions as an endothelial surface layer to protect and / or maintain glycocalyx function. According to some embodiments, the adsorption medium reduces a member selected from the group consisting of capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulation, and loss of vascular reactivity.

[0023] According to some embodiments, the glycocalyx-mimetic adsorption medium is negatively charged.

[0024] According to some embodiments, the solid substrate comprises a non-toxic, non-leaching material. According to some embodiments, the solid substrate comprises a plurality of rigid polymer beads. According to some embodiments, the rigid polymer beads are selected from the group consisting of polyurethane, polymethyl methacrylate, polyethylene or copolymers of ethylene and other monomers, polyethyleneimine, polypropylene, and polyisobutylene. According to some embodiments, the solid substrate comprises one or more hollow fibers.

[0025] According to yet another embodiment, the present disclosure provides a method for improving oxygen saturation in a subject in need thereof, comprising contacting a sample from the subject with a glycocalyx-mimetic adsorption medium to treat the sample thereby enhancing and / or restoring impaired glycocalyx barrier function; and injecting the treated sample into the subject, wherein the glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof.

[0026] These and other embodiments, aspects and objects will become more apparent when read in conjunction with the following detailed description taken in conjunction with the accompanying figures. [Brief explanation of the drawings]

[0027] [Figure 1] Figure 1A shows an electron micrograph representation of the glycocalyx. Figure 1B shows a magnified view of the electron micrograph representation of the glycocalyx. Figure 1C shows an electron micrograph representation of the glycocalyx.

[0028] [Figure 2] Figure 2A shows that heparin-binding protein (HBP) is adsorbed by heparin. Figure 2B shows that heparin-binding protein (HBP) is adsorbed by heparin.

[0029] [Figure 3] FIG. 3 shows that the methods and devices disclosed herein improve oxygen saturation. DETAILED DESCRIPTION OF THE INVENTION

[0030] The present disclosure relates in part to methods and devices for enhancing, improving, and / or restoring impaired (or disrupted) glycocalyx barrier function.

[0031] Disruption and damage to the glycocalyx is the cause of many disorders and diseases of the vascular system, such as sepsis and edema.

[0032] This method involves the use of a glycocalyx-mimicking adsorption medium that acts as an artificial glycocalyx, removing many mediators of inflammation in samples from subjects with impaired glycocalyx barrier function, thereby treating or "cleansing" the sample.

[0033] The treated or "washed" sample can then be re-injected into the subject, either continuously or intermittently.

[0034] A technical advantage of the embodiments described herein is that the use of glycocalyx-mimetic glycosaminoglycan adsorbents targets the cause of inflammatory and vascular disorders rather than treating the symptoms. Advantageously, the current methods safely and effectively restore the function of the damaged glycocalyx barrier. I. Definition

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. As used herein, the following terms have the following meanings:

[0036] As used herein, the terms "about" and "approximately equal to" are used to modify a numerical value and indicate a defined range surrounding that value. When "X" is a value, "about X" or "approximately equal to X" typically refers to a value between 0.90X and 1.10X. References to "about X" include at least the values ​​X, 0.90X, 0.91X, 0.92X, 0.93X, 0.94X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, 1.05X, 1.06X, 1.07X, 1.08X, 1.09X, and 1.10X. Thus, "about X" is intended to disclose, for example, "0.98X." When "about" is applied to the beginning of a numerical range, it applies to both ends of the range. Thus, "about 6 to 8.5" is equivalent to "about 6 to about 8.5." When "about" is applied to the first value in a set of values, it is applied to all values ​​in that set. Thus, "about 7, 9, or 11%" is equivalent to "about 7%, about 9%, or about 11%."

[0037] As used herein, the terms "comprising" or "comprises" are intended to mean that compositions, devices, and methods include the recited elements, but do not exclude others. "Consisting essentially of" refers to elements required for a given embodiment. This phrase allows for the presence of additional elements that do not materially affect the basic and novel or functional characteristics (e.g., compositions, devices, and methods) of a given embodiment. "Consisting of" refers to compositions, devices, methods, and their respective components described herein, excluding elements not recited in that description of the embodiment. Embodiments defined by each of these transition terms are within the scope of this disclosure.

[0038] As used herein, the term "enhancing impaired glycocalyx barrier function" refers to enhancing or improving the function of a disrupted, damaged, or impaired endothelial glycocalyx barrier. In a general sense, "enhancement" refers to the phenomenon in which a combination of two or more components provides an effect greater than the effect of the individual components or the sum of the components acting alone. Thus, in the context of the present disclosure, the combination of a glycocalyx-mimetic adsorption medium with a dysfunctional glycocalyx barrier results in an increase and improvement in the performance of the impaired glycocalyx barrier function that is greater than the performance of the impaired glycocalyx barrier function in the absence of the adsorption medium. For example, contacting a sample from a subject suffering from impaired glycocalyx barrier function with the glycocalyx-mimetic adsorption medium of the present disclosure enhances or ameliorates the impaired glycocalyx barrier function. In other words, contact with the adsorption medium improves and / or restores impaired glycocalyx barrier function to unimpaired or restored glycocalyx barrier function.

[0039] As used herein, the terms "glycocalyx barrier," "endothelial glycocalyx barrier," and "glycocalyx" are used interchangeably to refer to the carbohydrate-rich lining layer located at the interface between endothelial cells of vascular endothelium and circulating blood. The glycocalyx is connected to the endothelium primarily through proteoglycans and glycoproteins, incorporates soluble plasma molecules, and binds to each other directly or via soluble proteoglycans and / or glycosaminoglycans. Thus, the glycocalyx composition is a dynamic mesh of interactions between membrane-bound proteoglycans, glycoproteins, and glycosaminoglycans, and soluble glycosaminoglycans and plasma proteins. Endothelial function is regulated primarily through the glycocalyx to mediate platelet and leukocyte adhesion, hemostasis, and vascular barrier function, as described in detail herein.

[0040] As used herein, the term "impaired glycocalyx barrier function" refers to a decrease in the ability of the glycocalyx barrier to protect the vascular wall from direct bloodstream exposure. For example, impaired glycocalyx barrier function can be the inability of the glycocalyx to function as a vascular permeability barrier by losing its ability to regulate coagulation, prevent platelet adhesion to the vascular wall, prevent leukocyte adhesion to the vascular wall, regulate shear stress on endothelial cells, and regulate inflammatory processes. As used herein, "disrupting" or "disruption of" the glycocalyx refers to any process or condition that affects the glycocalyx such that it does not function normally (i.e., is dysfunctional). Disruption can be caused by inflammation or oxidation in the body. Disruption can result in thinning of the glycocalyx (enzymatic or shear-induced shedding) and loss of its component proteoglycans, resulting in impaired glycocalyx function. The following references describe various organ tissue models, animal models, clinical studies, and / or assays that demonstrate glycocalyx disruption and its dysfunction: Schott, U. et al. Scand J Trauma Resusc Emerg Med. 2016, 24(48), 1-8; Becker, BF, et al. Cardiovascular Research, 2010, 87, 300-310; Kolarova, H., et al. Mediators of Inflammation, 2014, 1-14; Vlahu, CA et al. J. Am. Soc. Nephrol. 2012, 23, 1900-1908; Yeo, TW, et al. Clinical Infectious Diseases, 2019, ciz038, https: / / doi.org / 10.1093 / cid / ciz038; and Mulivor, AW, et al. Am J Physiol Heart Circ Physiol. 2004, 286(5), H1672-80.

[0041] As used herein, the term "glycocalyx-mimetic adsorption medium" refers to a material having a surface modified, functionalized, coated, etc. with a composition (i.e., adsorbent) that may differ from the exact general composition and structure but function substantially similarly to a naturally occurring glycocalyx barrier. In the context of the present disclosure, a glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent that is a glycosaminoglycan mixture including heparin, heparan sulfate, and mixtures thereof. According to certain aspects, the glycosaminoglycan mixture includes heparin, heparan sulfate, and mixtures thereof, and optionally one or more additional glycosaminoglycans, such as, for example, chondroitin sulfate, dermatan sulfate, keratan sulfate, sialic acid / sialylated glycans, and / or hyaluronic acid. The glycosaminoglycan adsorbent mixture can optionally further include at least one proteoglycan core protein, such as, for example, syndecan, glypican, perlecan, versican, decorin, biglycan, mimecan, or a combination thereof. In other words, the glycocalyx-mimetic adsorption medium is a solid substrate containing glycosaminoglycan adsorbents that acts as a natural glycocalyx barrier capable of binding to analytes / adsorbates present in a sample, as described in detail herein. The glycocalyx-mimetic adsorption medium also acts as an endothelial surface layer to protect and / or maintain glycocalyx function that may have been impaired prior to contact with the glycocalyx-mimetic adsorption medium.

[0042] As used herein, the term "adsorbent" refers to a glycosaminoglycan mixture attached (e.g., linked, coupled, or bound) to the solid substrate of a glycocalyx-mimetic adsorption medium described herein. In certain instances, the adsorbent is the solid substrate itself. Thus, a "glycosaminoglycan adsorbent" is a solid substrate to which a glycosaminoglycan mixture is attached. For example, a glycosaminoglycan adsorbent is a polymeric resin to which a glycosaminoglycan mixture is attached. The glycosaminoglycan mixture includes heparin, heparan sulfate, and mixtures thereof, and optionally one or more additional glycosaminoglycans, such as, for example, chondroitin sulfate, dermatan sulfate, keratan sulfate, sialic acid / sialylated glycans, and / or hyaluronic acid. The glycosaminoglycan mixture can optionally further comprise at least one proteoglycan core protein, such as, for example, syndecan, glypican, perlecan, versican, decorin, biglycan, mimecan, or a combination thereof.

[0043] As used herein, the terms "analyte" and "adsorbate" are used interchangeably to refer to any molecule that disrupts glycocalyx barrier function and has affinity for the adsorbent. In the context of the present disclosure, a sample obtained from a subject suffering from impaired glycocalyx barrier function will contain adsorbates. Upon contact with the glycocalyx-mimetic adsorption medium of the present disclosure, the adsorbates bind to the surface of the adsorption medium and are therefore removed from the sample. By disrupting glycocalyx barrier function, these analytes promote or mediate inflammation and other conditions and diseases associated with impaired glycocalyx barrier function. Non-limiting examples of adsorbates include, but are not limited to, the following inflammatory mediators: lymphokines, interferons, chemokines, exotoxins, endotoxins, ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, cytokines, tumor necrosis factor (TNF)-α, bacterial lipopolysaccharide (LPS), low-density lipoprotein (LDL), and heparin-binding protein (HBP). For example, HPB (i.e., azurocidin or CAP37) is stored in neutrophil secretory vesicles and azurophilic granules and is released during inflammatory responses upon neutrophil adhesion and extravasation. Bacterial products induce the release of HBP, which acts on endothelial cells to increase vascular leakage. HBP binds to cell surface proteoglycans via heparan sulfate and chondroitin sulfate, thereby disrupting the glycocalyx and causing conditions associated with impaired glycocalyx barrier function (e.g., sepsis). See, e.g., Bentzer, P. et al. Intensive Care Medicine Experimental, 2016, 4(33), 1-16.

[0044] As used herein, the term "inflammation" refers to a protective response of tissues to injury or destruction to exclude or seal off damaging agents and damaged tissue and initiate tissue repair. Inflammation can cause pain, heat, redness, swelling, and loss of function. Inflammatory mediators (lymphokines, interferons, chemokines, exotoxins, endotoxins, LDL, HBP, etc.) can cause glycocalyx shedding. Inflammation can also cause leukocytes to degranulate enzymes that can degrade the glycocalyx.

[0045] As used herein, the phrase "condition associated with impaired glycocalyx barrier function" refers to a human disease or condition that is at least partially caused by or induces impaired glycocalyx barrier function. Thus, treating a condition associated with impaired glycocalyx barrier function refers to treating the condition, restoring impaired glycocalyx barrier function, or preventing or ameliorating conditions or symptoms resulting from impaired glycocalyx barrier function, such as inflammation, intimal proliferation, and thrombosis. In the context of the present disclosure, enhancing impaired glycocalyx barrier function using the glycocalyx-mimetic adsorption medium described herein can treat conditions associated with impaired glycocalyx barrier function. Non-limiting examples of conditions or diseases associated with impaired glycocalyx barrier function include capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, loss of vascular reactivity, sepsis, organ failure, atherosclerosis, and diabetes.

[0046] As used herein, the terms "glycosaminoglycan," "GAG," and "glycosaminoglycan chain" are used interchangeably to refer to the glycan components of glycosylated proteins (i.e., proteoglycans) present in the endothelial glycocalyx. Generally, proteoglycan structures comprise a core protein bearing multiple covalently bound and / or interacting GAG chains of variable length. Glycosaminoglycans of the glycocalyx are structurally diverse, unbranched carbohydrate polymers of about 5 to about 200,000 or more repeating disaccharide units (e.g., about 1 kDa to about 100,000 kDa or more), and negatively charged under physiological conditions. Each repeating disaccharide unit (i.e., "AB") contains a hexose or hexosamine (i.e., "A") glycosidically linked to a hexosamine (i.e., "B"), where the glycosidic bond configuration can be either α or β configuration, and each of the A and B components of every AB unit can be independently modified or unmodified, details of which are provided below. Non-limiting examples of GAGs that can be used in the embodiments described herein include chondroitin, chondroitin sulfate (CS), dermatan, dermatan sulfate (DS), heparan sulfate (HS), heparin, keratin, keratan sulfate, sialic acid / sialylated glycans, and haluronic acid (HA), details of which are provided herein. The "core protein" of endothelial proteoglycans may be directly bound to the endothelial cell membrane or may be present in the glycocalyx as a soluble plasma component. Endothelial proteoglycan core proteins vary in size (e.g., from about 20 kDa to about 500 kDa or more) and the number of attached GAG chains (e.g., from about to about 50 or more attached GAG chains). Thus, non-limiting examples of endothelial proteoglycan core proteins include syndecans, glypicans, perlecans, versicans, decorins, biglycans, and mimecans, each of which is described in detail herein.See, for example, Reitsma, S., et al. Pflugers Archiv: European Journal of Physiology, 2007, 454, 345-359; Kolarova, H., et al. Mediators of Inflammation, 2014, 1-14.

[0047] As used herein, the term "AB" refers to a single disaccharide unit of a glycosaminoglycan chain, where "A" represents either a hexose or hexolonic acid, and "B" represents a hexosamine. Generally, the disaccharide units of the GAG ​​chains described herein can include D-galactose (Gal), D-glucuronic acid (GlcA), or L-iduronic acid (IdoA) as component A and D-galactosamine (GalN) or glucosamine (GlcN) as component B. The glycosidic bond (i.e., COC bond) formed between the A and B components of an AB unit (and between two or more AB units, e.g., -[AB]1-[AB]2-[AB]3-) is a covalent bond formed from the hydroxyl groups of adjacent sugars. Bonds can occur between the 1st and 6th carbon atoms of adjacent sugars (i.e., a 1-6 bond, 1→6, or 1,6), between the 1st and 4th carbon atoms of adjacent sugars (i.e., a 1-4 bond, 1→4, or 1,4), between the 1st and 3rd carbon atoms of adjacent sugars (i.e., a 1-3 bond, 1→3, or 1,3), or between the 1st and 2nd carbon atoms of adjacent sugars (i.e., a 1-2 bond, 1→2, or 1,2), between the 2nd and 4th carbon atoms of adjacent sugars (i.e., a 2-4 bond, 2→4, or 2,4), or between the 2nd and 3rd carbon atoms of adjacent sugars (i.e., a 2-3 bond, 2→3, or 2,3). GAG chains can contain bonds between carbon atoms other than the 1st, 2nd, 3rd, 4th, and 6th carbon atoms.

[0048] Sugar moieties can be linked within GAGs such that the anomeric carbon is in either the α- or β-configuration. In this regard, the glycosidic bond formed between the A and B moieties of an AB unit can be referred to as either an α-linkage (or bond) or a β-linkage, relative to the anomeric carbon configuration of A. The glycosidic bond formed between each disaccharide unit of a GAG chain is also referred to as an α-linkage or a β-linkage (e.g., the bond between disaccharide [AB]1 and disaccharide [AB]2 in a -[AB]1-[AB]2- chain can be an α-linkage or a β-linkage, relative to the anomeric carbon configuration of B1). For example, a GAG chain can include repeating disaccharide units of A1α(1→3)B1, in which carbon 1 of A1 is in the α-configuration and is glycosidically linked to carbon 3 of B1. The disaccharide sequence 4A1α(1→3)B1β1 indicates that the 4-carbon of A1 is glycosidically linked to an unspecified carbon of the preceding adjacent sugar in the sequence, and the 1-carbon of B1 is in the β configuration and glycosidically linked to an unspecified carbon of the next adjacent sugar in the sequence.

[0049] Each A and / or B component of the AB unit can be unmodified or can include one or more modifications, such as O-sulfation and / or N-sulfation or N-acetylation, depending on the GAG ​​polymer. In this regard, modifications of hexose and hexuronic acid residues can include substituting one or more hydroxyl groups at positions 2, 3, 4, and 6 with O-sulfate. Modifications of hexosamine residues include substituting the amino group at position 2 with either N-acetyl or N-sulfate, and / or substituting one or more hydroxyl groups at positions 3, 4, and 6 with O-sulfate.

[0050] Examples of modified A components (i.e., hexose / hexuronic acid) include, but are not limited to, Gal2S, which contains an O-sulfate at the 2-position of D-galactose (Gal); Gal3S, which contains an O-sulfate at the 3-position of D-galactose (Gal); Gal4S, which contains an O-sulfate at the 4-position of D-galactose (Gal); Gal6S, which contains an O-sulfate at the 6-position of D-galactose (Gal); GlcA2S, which contains an O-sulfate at the 2-position of D-glucuronic acid (GlcA); GlcA3S, which contains an O-sulfate at the 3-position of D-glucuronic acid (GlcA); GlcA4S, which contains an O-sulfate at the 4th position of D-glucuronic acid (GlcA); GlcA6S, which contains an O-sulfate at the 6th position of D-glucuronic acid (GlcA); IdoA2S, which contains an O-sulfate at the 2nd position of L-iduronic acid (IdoA); IdoA3S, which contains an O-sulfate at the 3rd position of L-iduronic acid (IdoA); IdoA4S, which contains an O-sulfate at the 4th position of L-iduronic acid (IdoA); and IdoA6S, which contains an O-sulfate at the 6th position of L-iduronic acid (IdoA).

[0051] Examples of modified B components (i.e., hexosamines), specifically modifications to D-galactosamine (GalN), include, but are not limited to, the following: GalN3S, which contains an O-sulfate at position 3 of D-galactosamine (GalN); GalN4S, which contains an O-sulfate at position 4 of D-galactosamine (GalN); GalN6S, which contains an O-sulfate at position 6 of D-galactosamine (GalN); GalN3S6S, which contains an O-sulfate at position 3 and an O-sulfate at position 6 of D-galactosamine (GalN); GalN3S4S, which contains an O-sulfate at position 3 and an O-sulfate at position 4 of D-galactosamine (GalN); GalN4S6S, which contains an O-sulfate at position 4 and an O-sulfate at position 6 of D-galactosamine (GalN); GalNAc containing an N-acetyl at position 2 of D-galactosamine (GalN) (DN-acetylgalactosamine); GalNAc3S containing an O-sulfate at position 3 of DN-acetylgalactosamine (GalNAc); GalNAc4S containing an O-sulfate at position 4 of DN-acetylgalactosamine (GalNAc); GalNAc6S containing an O-sulfate at position 6 of DN-acetylgalactosamine (GalNAc); GalNAc3S6S containing an O-sulfate at position 3 and an O-sulfate at position 6 of DN-acetylgalactosamine (GalNAc); GalNAc3S4S containing an O-sulfate at position 3 and an O-sulfate at position 4 of DN-acetylgalactosamine (GalNAc); GalNAc4S6S containing an O-sulfate at position 4 and an O-sulfate at position 6 of DN-acetylgalactosamine (GalNAc); GalNS containing an N-sulfate at the 2nd position of D-galactosamine (GalN) (DN-sulfogalactosamine); GalNS3S containing an O-sulfate at the 3rd position of DN-sulfogalactosamine (GalNS); GalNS4S containing an O-sulfate at the 4th position of DN-sulfogalactosamine (GalNS); GalNS6S containing an O-sulfate at the 6th position of DN-sulfogalactosamine (GalNS); GalNS3S6S containing an O-sulfate at the 3rd position and an O-sulfate at the 6th position of DN-sulfogalactosamine (GalNS); GalNS3S4S containing an O-sulfate at the 3rd position and an O-sulfate at the 4th position of DN-sulfogalactosamine (GalNS);GalNS4S6S contains O-sulfate at position 4 and O-sulfate at position 6 of DN-sulfogalactosamine (GalNS);

[0052] Examples of modified B components (i.e., hexosamines), specifically modifications to D-glucosamine (GlcN), include, but are not limited to, the following: GlcN3S, which contains an O-sulfate at position 3 of D-glucosamine (GlcN); GlcN4S, which contains an O-sulfate at position 4 of D-glucosamine (GlcN); GlcN6S, which contains an O-sulfate at position 6 of D-glucosamine (GlcN); GlcN3S6S, which contains an O-sulfate at position 3 and an O-sulfate at position 6 of D-glucosamine (GlcN); GlcN3S4S, which contains an O-sulfate at position 3 and an O-sulfate at position 4 of D-glucosamine (GlcN); GlcN4S6S, which contains an O-sulfate at position 4 and an O-sulfate at position 6 of D-glucosamine (GlcN); GlcNAc containing an N-acetyl group at the 2nd position of D-glucosamine (GlcN) (DN-acetylglucosamine); GlcNAc3S containing an O-sulfate group at the 3rd position of DN-acetylglucosamine (GlcNAc); GlcNAc4S containing an O-sulfate group at the 4th position of DN-acetylglucosamine (GlcNAc); GlcNAc6S containing an O-sulfate group at the 6th position of DN-acetylglucosamine (GlcNAc); GlcNAc3S6S containing an O-sulfate group at the 3rd position and an O-sulfate group at the 6th position of DN-acetylglucosamine (GlcNAc); GlcNAc3S4S containing an O-sulfate group at the 3rd position and an O-sulfate group at the 4th position of DN-acetylglucosamine (GlcNAc); GlcNAc4S6S containing an O-sulfate group at the 4th position and an O-sulfate group at the 6th position of DN-acetylglucosamine (GlcNAc); GlcNS containing an N-sulfate at position 2 of D-glucosamine (GlcN) (DN-sulfoglucosamine); GlcNS3S containing an O-sulfate at position 3 of DN-sulfoglucosamine (GlcNS); GlcNS4S containing an O-sulfate at position 4 of DN-sulfoglucosamine (GlcNS); GlcNS6S containing an O-sulfate at position 6 of DN-sulfoglucosamine (GlcNS); GlcNS3S6S containing an O-sulfate at position 3 and an O-sulfate at position 6 of DN-sulfoglucosamine (GlcNS); GlcNS3S4S containing an O-sulfate at position 3 and an O-sulfate at position 4 of DN-sulfoglucosamine (GlcNS); and GlcNS4S6S containing an O-sulfate at position 4 and an O-sulfate at position 6 of DN-sulfoglucosamine (GlcNS).Thus, the GAG ​​chains of the glycocalyx may contain a variety of different disaccharide units, where each hexose / hexosic acid residue and each hexosamine residue of every hexose / hexosic acid-hexosamine pair (i.e., disaccharide unit) can optionally be independently mono- or polysubstituted (i.e., unmodified or modified).

[0053] As used herein, the terms "heparan sulfate" and "HS" are used interchangeably to refer to glycosaminoglycan polymers of repeating disaccharide A-B units, which can include GlcA, IdoA, and IdoA2S residues as possible A components, and GlcN, GlcNAc, GlcNAc6S, GlcNS, GlcNS6S, and GlcNS3S6S residues as possible B components. Generally, heparan sulfate contains about 40-70% unsulfated disaccharide sequences, about 30-65% various monosulfated disaccharide sequences, and about 1-10% disulfated and / or trisulfated disaccharide sequences. Heparan sulfate polymers can have about 65 N-sulfates per 100 disaccharides, an N-sulfation to O-sulfation ratio of about 2:3 to about 3:4, and an average of about 0.8 to about 1.8 sulfate groups per disaccharide unit. The most common disaccharide sequence in heparan sulfate is 4GlcAβ(1→4)GlcNAcα1, which may comprise up to approximately 50% or more of the HS chains. While GlcA is the more common hexuronic acid, IdoA may comprise approximately 30% to approximately 50% of heparan sulfate and may occur as the unsulfated unit 4IdoAα(1→4)GlcNAcα1 and / or any of the monosulfated, disulfated, and / or trisulfated disaccharide units, such as 4IdoAα(1→4)GlcNSα1, 4IdoAα(1→4)GlcNAc6Sα1, and 4IdoAα(1→4)GlcNS6Sα1. Heparan sulfate polymers can have from about 20 to about 200 or more repeating disaccharide units and can have molecular weights of about 10 kDa to about 100 kDa.For example, see the following: Shriver, S. et al. Handb. Exp. Pharmacol. 2012, 207, 159-176; Zhang, F. et al. Chapter 3 - Glycosaminoglycans. 2010. In: Richard D. Cummings, J. Michael Pierce, et al., editors. Handbook of Glycomics, Academic Press, 2010, 59-80; Gandhi, N. et al. Chem. Biol. Drug. Des. 2008, 72, 455-482; Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17。

[0054] As used herein, the terms "heparin" and "Hep" are used interchangeably to refer to glycosaminoglycan polymers of repeating disaccharide A-B units, which can include GlcA, IdoA, and IdoA2S residues as possible A components, and GlcN, GlcNAc, GlcNAc6S, GlcNS, GlcNS6S, and GlcNS3S6S residues as possible B components. Generally, heparin contains approximately 70-90% trisulfated disaccharide sequences, approximately 10-30% various nonsulfated, monosulfated, and disulfated disaccharide sequences, and an average of approximately 1.8 to 2.8 sulfate groups per disaccharide unit. The most common disaccharide sequence in heparin is 4IdoA2Sα(1→4)GlcNS6Sα1, which may comprise up to approximately 70% or more of Hep chains. Heparin polymers can have from about 15 to about 60 or more repeating disaccharide units and can have a molecular weight of from about 10 kDa to about 35 kDa. See, for example: Shriver, S. et al. Handb. Exp. Pharmacol. 2012, 207, 159-176; Zhang, F. et al. Chapter 3 - Glycosaminoglycans. 2010. In: Richard D. Cummings, J. Michael Pierce, et al., editors. Handbook of Glycomics, Academic Press, 2010, 59-80; Gandhi, N. et al. Chem. Biol. Drug. Des. 2008, 72, 455-482; Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0055] As used herein, the terms "chondroitin sulfate" and "CS" are used interchangeably to refer to glycosaminoglycan polymers of repeating disaccharide A-B units that can contain GlcA, GlcA2S, and GlcA3S residues as possible A components, and GalNAc, GalNAc4S, GalNAc6S, and GalNAc4S6S residues as possible B components. Various types of chondroitin sulfate exist, including SA, CS-C, CS-D, and CS-D. The CS-B subtype is known as dermatan sulfate and is described in detail herein. Generally, chondroitin sulfate type A contains repeating disaccharide units of 4GlcAβ(1→3)GalNAc4Sβ1, chondroitin sulfate type C contains repeating disaccharide units of 4GlcAβ(1→3)GalNAc6Sβ1, chondroitin sulfate type D contains repeating disaccharide units of 4GlcA2Sβ(1→3)GalNAc6Sβ1, and chondroitin sulfate type E contains repeating disaccharide units of 4GlcAβ(1→3)GalNAc4S6Sβ1. Chondroitin sulfate chains can be hybrid structures containing more than one type of chondroitin disaccharide unit. In other words, the hybrid CS polymer can contain the repeating disaccharide sequences 4GlcAβ(1→3)GalNAc4Sβ1 (chondroitin sulfate A), 4GlcAβ(1→3)GalNAc6Sβ1 (chondroitin sulfate C), 4GlcA2Sβ(1→3)GalNAc6Sβ1 (chondroitin sulfate D), and / or 4GlcAβ(1→3)GalNAc4S6Sβ1 (chondroitin sulfate E) in any amounts and combinations thereof. As a non-limiting example, a hybrid chondroitin sulfate chain may comprise about 40-60% 4GlcAβ(1→3)GalNAc4Sβ1, about 40-60% 4GlcAβ(1→3)GalNAc6Sβ1, and, optionally, about 1-20% of various unsulfated, disulfated, and / or trisulfated disaccharide sequences, e.g., 4GlcAβ(1→3)GalNAcβ1, 4GlcA2Sβ(1→3)GalNAc6Sβ1, and / or 4GlcAβ(1→3)GalNAc4S6Sβ1.Chondroitin sulfate polymers (non-hybrid or hybrid) can have from about 4 to about 155 or more repeating disaccharide units and can have a molecular weight of from about 2 kDa to about 70 kDa or more. See, for example: Zhang, F. et al. Chapter 3 - Glycosaminoglycans. 2010. In: Richard D. Cummings, J. Michael Pierce, et al., editors. Handbook of Glycomics, Academic Press, 2010, 59-80; Gandhi, N. et al. Chem. Biol. Drug. Des. 2008, 72, 455-482; Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0056] As used herein, the terms "dermatan sulfate" and "DS" are used interchangeably to refer to glycosaminoglycan polymers of repeating disaccharide A-B units, which can contain GlcA, IdoA, and IdoA2S residues as possible A components, and GalNAc, GalNAc4S, and GalNAc6S residues as possible B components. Dermatan sulfate generally contains approximately 70-95% monosulfated disaccharide sequences, approximately 5-20% disulfated disaccharide sequences, and approximately 1-10% nonsulfated disaccharide sequences. The most common disaccharide sequence in DS polymers is 4IdoAα(1→3)GalNAc4Sβ1, which can constitute up to 80% or more of the DS chains. Dermatan sulfate polymers can have from about 20 to about 155 or more repeating disaccharide units and can have molecular weights of from about 10 kDa to about 70 kDa or more. See, for example: Zhang, F. et al. Chapter 3 - Glycosaminoglycans. 2010. In: Richard D. Cummings, J. Michael Pierce, et al., editors. Handbook of Glycomics, Academic Press, 2010, 59-80; Gandhi, N. et al. Chem. Biol. Drug. Des. 2008, 72, 455-482; Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0057] As used herein, the terms "keratan sulfate" and "KS" are used interchangeably to refer to a glycosaminoglycan polymer of repeating disaccharide AB units that may contain Gal and Gal6S residues as possible A components and GlcNAc and GlcNAc6S residues as possible B components. Keratan sulfate contains a mixture of mostly unsulfated disaccharide sequences 3Galβ(1→4)GlcNAcβ1, monosulfated disaccharide sequence 3Galβ(1→4)GlcNAc6Sβ1, and disulfated disaccharide sequence 3Gal6Sβ(1→4)GlcNAc6Sβ1. Disaccharides within the repeating regions of keratan sulfate may be fucosylated, with N-acetylneuraminic acid capping the chain ends. See, for example, Stanley, P. et al. Structures Common to Different Glycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 14. KS polymers can have about 10 = about 70 or more repeating disaccharide units and can have a molecular weight of about 5 kDa to about 30 kDa or more.For example, see the following: Zhang, F. et al. Chapter 3 - Glycosaminoglycans. 2010. In: Richard D. Cummings, J. Michael Pierce, et al., editors. Handbook of Glycomics, Academic Press, 2010, 59-80; Gandhi, N. et al. Chem. Biol. Drug. Des. 2008, 72, 455-482; Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17。

[0058] As used herein, the terms "hyaluronic acid," "hyaluronic acid," "HA," and "hyaluronan" are used interchangeably to refer to a glycosaminoglycan polymer of repeating disaccharide AB units that may contain a GlcA residue as a possible A component and GlcNAc as a possible B component. HA is the only non-sulfated GAG polymer and contains the repeating disaccharide sequence 4GlcAβ(1→3)GalNAcβ1. Hyaluronic acid can be purified from animal and non-animal sources. HA polymers can have from about 10 to about 100,000 repeating disaccharide units and can have a molecular weight of from about 4 kDa to about 20,000 kDa. See, for example: Zhang, F. et al. Chapter 3 - Glycosaminoglycans. 2010. In: Richard D. Cummings, J. Michael Pierce, et al., editors. Handbook of Glycomics, Academic Press, 2010, 59-80; Gandhi, N. et al. Chem. Biol. Drug. Des. 2008, 72, 455-482; Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0059] As used herein, the term "syndecan" refers to the membrane-bound core protein of the glycocalyx, which is connected to the membrane via a transmembrane domain. There are four subtypes of syndecan core proteins, ranging from about 20 kDa to about 45 kDa. Syndecan proteoglycan core proteins can contain about five or more HS and CS glycosaminoglycan chains, such as a mixture of two to three HS chains or three to four HS and one to two CS chains. Syndecans can function in regulating cell adhesion, migration, and actin cytoskeleton organization, as well as controlling ligand clearance from the cell surface. See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0060] As used herein, the term "glypican" refers to the membrane-bound core protein of the glycocalyx, which is connected to the membrane via a glycosylphosphatidylinositol anchor. There are six subtypes of glypican core proteins, ranging from approximately 55 kDa to approximately 70 kDa. Glypican proteoglycan core proteins can contain approximately three or more HS glycosaminoglycan chains. Glypicans can function as coreceptors, regulating signal transduction through associated receptors (e.g., tyrosine kinase receptors). See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0061] As used herein, the term "perlecan" refers to a secreted core protein present in the glycocalyx as a soluble plasma component. The average core mass of the perlecan proteoglycan core protein is approximately 400 kDa and can contain approximately three or more HS glycosaminoglycan chains, and optionally one, two, or more CS chains (e.g., one to four HS chains or one to three HS and zero to two CS chains). Perlecan can exhibit the following functions: extracellular matrix (ECM) assembly, regulated cell migration through integrin interactions, and sequestration of growth factors (e.g., FGF). See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0062] As used herein, the term "versican" refers to a secreted core protein present in the glycocalyx as a soluble plasma component. The average core mass of the versican proteoglycan core protein is approximately 370 kDa and can contain approximately 10-30 or more CS and DS glycosaminoglycan chains (e.g., a mixture of 5-15 CS chains and 10-20 DS chains). Versican is involved in multiple ECM interactions, regulation of inflammation, and cell adhesion and migration. See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0063] As used herein, the term "decorin" refers to a secreted core protein present in the glycocalyx as a soluble plasma component and is a member of the small leucine-rich proteoglycan (SLRP) family. As an SLRP, decorin contains leucine-rich repeats flanked by cysteines in the central domain. The average core mass of the decorin proteoglycan core protein is approximately 35-40 kDa and can contain at least one CS chain and / or at least one DS chain. Decorin regulates interstitial collagen fibrillogenesis and is involved in the inhibition of TGF-β signaling. See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0064] As used herein, the term "biglycan" refers to the secreted core protein of the SLRP family, which is present in the glycocalyx as a soluble plasma component. The biglycan proteoglycan core protein has an average core mass of approximately 36-40 kDa and can contain approximately two or more CS and DS chains. Biglycan is involved in collagen matrix assembly and activation of the innate immune system. See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0065] As used herein, the term "mimecan" refers to the secreted core protein of the SLRP family, which is present in the glycocalyx as a soluble plasma component. The mimecan proteoglycan core protein has an average core mass of approximately 25-35 kDa and can contain approximately two or more KS chains. Mimecan is involved in collagen matrix assembly, bone formation, and corneal transparency. See, for example, Lindahl, U, et al. Proteoglycans and Sulfated Glycosaminoglycans. 2017. In: Varki A, Cummings RD, Esko JD, et al., editors. Essentials of Glycobiology [Internet]. 3rd edition. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 2015-2017. Chapter 17.

[0066] As used herein, the term "contacting" refers to the process of contacting at least two different species such that they touch or are in direct or local proximity. In the context of the present disclosure, a sample from a subject suffering from impaired glycocalyx barrier function is contacted with a glycocalyx-mimetic adsorption medium. Thus, the analytes / adsorbates within the sample contact the glycocalyx-mimetic adsorption medium. Upon contact with the glycocalyx-mimetic adsorption medium, the analytes / adsorbates remain attached to the glycocalyx-mimetic adsorption medium, thereby removing the analytes / adsorbates from the sample.

[0067] As used herein, the term "sample" refers to any biological sample that may contain analytes / adsorbates obtained from a subject suffering from impaired glycocalyx barrier function. Typically, a sample is in liquid form or can be converted into a liquid form. Non-limiting examples of samples include whole blood, serum, and plasma. In the context of this disclosure, a sample that has not been contacted with a glycocalyx-mimicking adsorption medium is considered an "untreated sample." Thus, an untreated sample contains molecules (i.e., analytes / adsorbates) that disrupt glycocalyx barrier function. A "treated sample" is a sample that has been contacted with a glycocalyx-mimicking adsorption medium. Treated samples contain a reduced amount of analytes / adsorbates and can therefore be considered "washed" or "clean."

[0068] As used herein, the term "perfusion" refers to the passage of fluid (i.e., blood) over and / or through an organ, while the term "reperfusion" refers to the passage of fluid over and / or through an organ that was previously not perfused (e.g., an artery that has been clamped during surgery to prevent the passage of blood). Stated differently, reperfusion refers to the act of restoring blood flow to an organ or tissue (heart, kidney, etc.).

[0069] As used herein, the term "hard polymer beads" refers to beads, granules, pellets, spheres, particles, microcapsules, spheres, microspheres, nanospheres, microbeads, nanobeads, microparticles, nanoparticles, and the like made from polymer resins or other biocompatible matrix materials.

[0070] The term "percent weight," unless otherwise specified, refers to % or the percentage of a component measured by weight per total weight of a particular composition. Weight percent is expressed as "%" or "% w / w." In the context of the present disclosure, a glycosaminoglycan adsorbent (e.g., a glycosaminoglycan mixture) attached to a solid substrate of a glycocalyx-mimetic adsorption medium described herein will contain a specific amount of GAG chains and, in some cases, a specific amount of proteoglycan core protein based on the total weight of the glycosaminoglycan mixture. Thus, the % w / w of each GAG chain (and optional one or more core proteins) in the adsorption medium is based on the total weight of the glycosaminoglycan mixture used to coat or functionalize the surface of the solid substrate. II. Definitions and Devices

[0071] Disclosed herein are materials, compounds, compositions, and components that can be used in, can be used in combination with, can be used in preparation for, or are products of the disclosed methods, compositions, and devices. These and other materials are disclosed herein, and when combinations, subsets, interactions, groups, etc. of these materials are disclosed, it is understood that specific reference to each of the various individual and collective combinations and permutations of these materials is not expressly disclosed, and each is specifically contemplated and may be described herein. For example, when a method, composition, or device is disclosed, or several modifications that can be made to several components of the method, composition, or device are discussed, all possible combinations and permutations are specifically contemplated unless specifically indicated to the contrary.

[0072] Thus, if a class of components or elements A, B, and C and a class of components or elements D, E, and F are disclosed, and an example of a method, composition, or device AD ​​is disclosed, each is individually and collectively contemplated, even though each is not individually recited. Thus, in this example, each of the combinations AE, AF, BD, BE, BF, CD, CE, and CF is specifically contemplated and should be considered disclosed from the disclosure of A, B, and C; D, E, and F; and example combination AD. Likewise, subsets or combinations of these are also specifically contemplated and disclosed. Thus, for example, the subgroup of AE, BF, and CE is specifically contemplated and should be considered disclosed from the disclosure of A, B, and C; D, E, and F; and example combination AD. This concept applies to all aspects of the present disclosure, including, but not limited to, method steps for making and using the disclosed compositions and devices. Thus, if there are various additional steps that can be performed, it is understood that each of these additional steps can be performed with any particular aspect or combination of aspects of the disclosed methods, and that each such combination is to be considered specifically contemplated and disclosed.

[0073] Figures 1A-C show electron micrographs of the glycocalyx. As shown in Figure 1A and the inset, the glycocalyx 101 is a slippery, gel-like network of negatively charged molecules present on the luminal side of the vascular endothelium. The lumen of the vessel is shown as 102. Figure 1B is a magnified view and shows that the negatively charged glycocalyx consists of a web of membrane-bound glycoproteins 126, 131, 140 associated with various glycosaminoglycans and proteoglycans 125. It is cooperatively connected to the vascular endothelium by a host of scaffolding molecules (mainly the aforementioned proteoglycans and glycoproteins) that form a network into which various water-soluble molecules are incorporated. Glycosaminoglycans can bind up to 10,000 times their own weight in water and therefore contribute significantly to the total volume of the endothelial glycocalyx.

[0074] Figure 1C shows that the glycocalyx 110 is a carbohydrate-rich layer connected to the endothelium 115 via skeletal proteoglycans and glycoproteins. A complex network of plasma- and epithelial-derived soluble molecules is continuously incorporated into the glycocalyx. A dynamic equilibrium is formed between blood components and the glycocalyx (see C. Biddle, AANA Journal, 81, (6) (2013)).

[0075] The present disclosure provides a method for enhancing impaired glycocalyx barrier function in a subject in need thereof, comprising: contacting a sample from the subject with a glycocalyx-mimicking adsorption medium to enhance and / or restore impaired glycocalyx barrier function, thereby treating the sample; and injecting the treated sample into the subject, wherein the glycocalyx-mimicking adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan mixture comprising at least one glycosaminoglycan (GAG) chain and, optionally, one or more proteoglycan core proteins.

[0076] Impaired glycocalyx barrier function can result from alterations, disruption, or damage to the structure or composition of the glycocalyx barrier. Located on the apical surface of vascular endothelial cells lining the lumen, the glycocalyx is a negatively charged, dynamic network of proteoglycans, glycoproteins, and glycolipids, containing a variety of enzymes and proteins that regulate cell and molecule adhesion and transport. The primary role of the glycocalyx in the vascular system is to maintain plasma and vessel wall homeostasis. Enzymes, proteins, and other molecular entities serve to reinforce the glycocalyx barrier against vascular and other diseases. Another function of the glycocalyx within the vascular endothelium is to protect the vessel wall from direct exposure to the blood flow, acting as a vascular permeability barrier. Thus, shear generated by blood flow regulates the balance between biosynthesis and shedding of various glycocalyx components. Its protective function is universal throughout the vascular system, and its relative importance varies depending on its precise location within the vascular system. The glycocalyx is involved in filtration of fluid from plasma to the interstitial space, protecting the endothelium from blood cell adhesion, and mediating the signal for nitric oxide (NO) production by endothelial cells. As a result, the glycocalyx serves as a protective barrier for vital vasculature, including the brain, spinal cord, organs, lungs, and lymphatic system. Therefore, alterations or damage to this complex and dynamic glycocalyx barrier structure ultimately leads to impaired glycocalyx barrier function, rendering the vasculature more susceptible to injury and disease.

[0077] The subject suffering from impaired glycocalyx barrier function may be an animal. According to some embodiments, the subject is a mammal. According to some embodiments, the subject is a human. The subject may be of any gender or age. According to some embodiments, the subject suffering from impaired glycocalyx barrier function has one or more of the following conditions: capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, loss of vascular reactivity, sepsis, organ failure, atherosclerosis, and diabetes.

[0078] The sample from the subject can be any bodily fluid collected from the subject. According to some embodiments, the sample includes whole blood. According to some embodiments, the sample includes serum. According to some embodiments, the sample includes plasma. According to some embodiments, the sample includes cerebrospinal fluid. The sample can be collected from the subject in the form of a separate sample to be processed in this manner. The sample can be collected from the subject in the form of a continuous or semi-continuous stream. The amount of sample that can be used in the claimed method is not intended to be limiting. It can range from less than 1 mL to more than 1 L, including the subject's entire blood volume if the sample includes blood and continuous recirculation into the subject is employed. If desired, one or more "passes" through the sorbent bed can be used. The term "sorbent bed" refers to a container, chamber, column, etc., that holds a glycocalyx-mimetic sorbent medium. The sorbent bed can be part of a dialysis or extracorporeal circuit. In another aspect, it can be part of a blood bag.

[0079] According to some embodiments, samples are drawn from the subject at the following rates: about 5 mL / min, about 10 mL / min, about 15 mL / min, about 20 mL / min, about 25 mL / min, about 30 mL / min, about 35 mL / min, about 40 mL / min, about 45 mL / min, about 50 mL / min, about 60 mL / min, about 70 mL / min, about 80 mL / min, about 90 mL / min, about 100 mL / min, about 150 mL / min, about 200 mL / min, about 250 mL / min, about 300 mL / min, about 350 mL / min, about 400 mL / min, about 450 mL / min, about 500 mL / min, about 550 mL / min, about 600 mL / min, about 700 mL / min, about 800 mL / min, about 900 mL / min, about 1000 mL / min, or even about 2000-6000 mL / min. / min.

[0080] The glycocalyx-mimetic adsorption medium of the present disclosure is designed to function similarly to an intact, naturally occurring glycocalyx barrier. In this manner, the glycocalyx-mimetic adsorption medium enhances, strengthens, improves, and / or enhances the ability of a damaged / dysfunctional endothelial glycocalyx barrier to function properly. In other words, contacting a sample from a subject suffering from impaired glycocalyx barrier function with the glycocalyx-mimetic adsorption medium enhances the impaired glycocalyx barrier function. According to some embodiments, enhancing impaired glycocalyx barrier function by contact with the glycocalyx-mimetic adsorption medium enhances or restores the impaired glycocalyx's ability to regulate coagulation, prevent platelet adhesion to vascular walls, prevent leukocyte adhesion to vascular walls, regulate shear stress on endothelial cells, and regulate inflammatory processes.

[0081] A sample from a subject suffering from impaired glycocalyx barrier function can be contacted with the glycocalyx-mimetic adsorption medium for a time sufficient to enhance, strengthen, improve, and / or reverse impaired glycocalyx barrier function in the sample. According to some embodiments, the sample is contacted with the glycocalyx-mimetic adsorption medium for a period of 1 minute to 12 hours, or longer. According to some embodiments, the sample is contacted with the glycocalyx-mimetic adsorption medium for a duration of 1 minute, 5 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 45 minutes, 60 minutes, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, or 12 hours. According to some embodiments, the sample is contacted with the glycocalyx-mimetic adsorption medium in an uninterrupted or continuous flow, where the sample flows continuously over, on, or through the glycocalyx-mimetic adsorption medium. Continuous flow of the sample includes a constant or variable fluid flow at a set rate or rate at which the sample contacts the glycocalyx-mimetic adsorption medium. According to some embodiments, the sample is contacted with the glycocalyx-mimicking adsorption medium at the following rates: about 5 mL / min, about 10 mL / min, about 15 mL / min, about 20 mL / min, about 25 mL / min, about 30 mL / min, about 35 mL / min, about 40 mL / min, about 45 mL / min, about 50 mL / min, about 60 mL / min, about 70 mL / min, about 80 mL / min, about 90 mL / min, about 100 mL / min, about 150 mL / min, about 200 mL / min, about 250 mL / min, about 300 mL / min, about 350 mL / min, about 400 mL / min, about 450 mL / min, about 500 mL / min, about 550 mL / min, about 600 mL / min, about 700 mL / min, about 800 mL / min, about 900 mL / min, or about 1000 mL / min.

[0082] A sample from a subject suffering from impaired glycocalyx barrier function can be contacted with a glycocalyx-mimetic adsorption medium one or more times to enhance, strengthen, improve, and / or restore impaired glycocalyx barrier function in the sample. According to some embodiments, the sample is contacted with the glycocalyx-mimetic adsorption medium at least once. According to some embodiments, the sample is contacted with the glycocalyx-mimetic adsorption medium 1 to 20 times or more. According to some embodiments, the sample is contacted with the glycocalyx-mimetic adsorption medium 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 times.

[0083] The glycocalyx-mimicking adsorption medium for enhancing impaired glycocalyx barrier function of a sample and processing the sample can be a microporous medium such as activated carbon or size-exclusion chromatography resin made hemocompatible. According to some embodiments, the glycocalyx-mimicking adsorption medium does not include activated carbon. The glycocalyx-mimicking adsorption medium can be in a container such as a column, cartridge, tube, centrifuge tube, bottle, flexible bag, etc., or any container that allows removal of the processed sample without disturbing the glycocalyx-mimicking adsorption medium.

[0084] In the disclosed methods and devices, a variety of materials, in terms of shape and composition, can be used as solid substrates for glycocalyx-mimetic adsorption media. All suitable solid substrates provide a high surface area, thereby enhancing impaired glycocalyx barrier function, while facilitating the transport of adsorbates (primarily) to their binding adsorbent sites by forced convection or diffusive transport. Useful solid substrates for creating glycocalyx-mimetic adsorption media include: nonporous rigid beads, particles, or packings; reticulated foams; rigid monolithic beds (e.g., formed from sintered beads or particles); columns packed with woven or nonwoven fabrics; columns packed with yarns or solid or hollow mesoporous or microporous monofilament fibers; spiral-wound cartridges formed from flat films or dense membranes; or media combinations such as mixed bead / fabric cartridges. According to some embodiments, the solid substrate is initially mesoporous or microporous but becomes essentially nonporous when the surface is treated before, during, or after the creation of adsorption sites. According to some embodiments, the substrate comprises polymers or rigid polymer beads. The matrix may also be a metal, ceramic, glass, natural mineral, silica, etc. Typically, the matrix does not leach impurities that would cause clinically significant problems if introduced into the patient's blood.

[0085] According to some embodiments, the total surface area of ​​the solid substrate can be in the range of 0.1 to 10,000 square meters, preferably in the range of 0.5 to 50 square meters, for example, 0.5, 1, 1, 2, 2, 5, 10, 25, 40, 50 square meters, and values ​​therebetween. According to some embodiments, the material of the solid substrate is selected from the group consisting of glass, silica, latex, cellulose, cellulose acetate, chitin, chitosan, cross-linked dextran, cross-linked agarose, cross-linked alginate, polyethylene, polypropylene, polystyrene, polycarbonate, polysulfone, polyacrylonitrile, silicone, fluoropolymers (such as polytetrafluoroethylene), polyurethane, and other synthetic polymers. Other materials commonly used in medical applications can also be used. According to some embodiments, the solid substrate comprises a cross-linked polysaccharide. The solid substrate can comprise multiple adsorbent monolayers, filters, membranes, solid fibers, hollow fibers, particles, or beads. Optionally, the solid substrate can exist in other forms or shapes that provide a large surface area.

[0086] According to some embodiments, the solid substrate is a mixed-media solid substrate created by layering different glycocalyx-mimicking adsorption media on the solid substrate in a parfait-type arrangement so that the sample contacts the different media in serial or parallel flow. A specific mixed-media embodiment is disclosed in U.S. Patent No. 8,758,286, incorporated herein by reference. The different media arrangement involves placing unmixed glycocalyx-mimicking adsorption media (i.e., anionic media) in the fluid inlet and / or fluid outlet regions of the solid substrate, and optionally, the mixed region includes other materials interposed between the inlet and outlet regions, such as cationic media. For media in fiber form, mixed woven, knitted, or nonwoven structures can be prepared by methods well known in the textile industry for forming fabrics from mixed fibers. According to some embodiments, yarns are prepared from thinner multifilament yarns or monofilaments made from two or more fibers with different surface chemistries, with one fiber type containing a surface that actively prevents blood clotting upon contact. This mixed-fiber yarn can then be used to prepare a fabric suitable for contact with a sample (such as blood).

[0087] The solid substrate of the glycocalyx-mimetic adsorption medium is modified, functionalized, coated, etc. with an adsorbent, wherein the adsorbent is a glycosaminoglycan mixture comprising at least one glycosaminoglycan (GAG) chain. According to some embodiments, the solid substrate of the glycocalyx-mimetic adsorption medium is modified, functionalized, coated, etc. with a glycosaminoglycan adsorbent, wherein the adsorbent is a glycosaminoglycan mixture comprising at least one glycosaminoglycan (GAG) chain and, optionally, one or more proteoglycan core proteins. According to some embodiments, the solid substrate of the glycocalyx-mimetic adsorption medium is modified, functionalized, coated, etc. with a glycosaminoglycan adsorbent, wherein the adsorbent is a glycosaminoglycan mixture comprising heparin, heparan sulfate, and mixtures thereof. According to some embodiments, the glycosaminoglycan mixture adsorbent optionally further comprises one or more of the following: chondroitin sulfate, dermatan sulfate, keratan sulfate, sialic acid / sialylated glycans, and / or hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture comprises heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, sialic acid, sialylated glycans, and hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture comprises heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, sialic acid, and hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture comprises heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, sialic acid, and hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture comprises heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, sialylated glycans, and hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture comprises heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, and hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture consists essentially of heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate, and hyaluronic acid.According to some embodiments, the solid substrate of the glycocalyx-mimetic adsorption medium is modified, functionalized, coated, etc. with an adsorbent, wherein the adsorbent is a glycosaminoglycan mixture comprising about 30% to about 98% w / w heparan sulfate and, optionally, one or more of the following: about 0.1% to about 50% w / w chondroitin sulfate, about 0.1% to about 50% w / w dermatan sulfate, about 0.001% to about 40% w / w keratan sulfate, and / or about 0.1% to about 60% w / w hyaluronic acid. According to some embodiments, the glycosaminoglycan mixture comprises about 30% to about 98% w / w heparan sulfate, about 0.1% to about 50% w / w chondroitin sulfate, about 0.1% to about 50% w / w dermatan sulfate, about 0.001% to about 40% w / w keratan sulfate, and about 0.1% to about 60% w / w hyaluronic acid.

[0088] According to some embodiments, the adsorbent is 100% heparin. According to particular aspects, the adsorbent is 100% heparan sulfate. According to particular aspects, the adsorbent is 99% to 1% heparin and 1% to 99% heparan sulfate.

[0089] According to some embodiments, glycosaminoglycan adsorbents containing about 30% to about 98% w / w heparan sulfate may also contain heparin. In certain cases, heparin can be used in place of or substituted for heparan sulfate at any percentage. For example, if a glycosaminoglycan mixture (i.e., a glycosaminoglycan adsorbent of a glycocalyx-mimetic adsorption medium) contains 50% w / w heparan sulfate, any amount of HS (e.g., about 1% to 50%, or 25% to 50%, or 50%, etc.) can be substituted with heparin. Thus, in certain cases, the adsorption medium contains about 30% to about 98% w / w heparin instead of heparan sulfate. According to some embodiments, the adsorption medium contains about 30% to about 98% w / w heparin-heparan sulfate mixture. According to some embodiments, the heparin polymer comprises about 15 to about 60 repeating disaccharide units, such as, for example, about 16, 17, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 44, 48, 50, 52, 56, or about 60 repeating disaccharide units. According to some embodiments, the heparin polymer has an average molecular weight in the range of about 10 kDa to about 35 kDa, e.g., about 11 kDa, or about 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, or about 34 kDa.

[0090] According to some embodiments, the glycosaminoglycan adsorbent can comprise one or more proteoglycan core proteins to which GAG chains of the glycosaminoglycan mixture are covalently attached. Alternatively, or in addition, the glycosaminoglycan adsorbent can comprise one or more proteoglycan core proteins and the GAG ​​chains of the glycosaminoglycan mixture, where the GAG ​​chains are not covalently attached to the one or more proteins. The one or more core proteins can be syndecan, glypican, perlecan, versican, decorin, biglycan, mimecan, or a combination thereof. The glycosaminoglycan adsorbent can have from about 0.001% to about 50% w / w of the one or more core proteins. According to some embodiments, the amount of one or more core proteins in the glycosaminoglycan mixture ranges from about 0.001% to about 30% w / w, or from about 0.001% to about 5% w / w, from about 0.05% to about 10% w / w, from about 1% to about 15% w / w, or from about 5% to about 20% w / w. According to some embodiments, the amount of one or more core proteins included in the mixture of glycosaminoglycans is about 0.0015% w / w, or about 0.002%, 0.0025%, 0.005%, 0.0075%, 0.01%, 0.025%, 0.05%, 0.075%, 0.1%, 0.25%, 0.5%, 0.75%, 1.0%, 1.25%, 1.5%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 12.0%, 14.0%, 15.0%, 18.0%, 20.0%, 22.0%, 24.0%, or about 25.0% w / w. Thus, according to some embodiments, the adsorbent of the glycocalyx-mimetic adsorption medium solid substrate optionally comprises one or more core proteins selected from the group consisting of syndecan, glypican, perlecan, versican, decorin, biglycan, mimecan, or combinations thereof, in an amount ranging from about 0.001% to about 50% w / w, in addition to about 30% to about 98% w / w heparan sulfate and one or more of the following: about 0.1% to about 50% w / w chondroitin sulfate; about 0.1% to about 50% w / w dermatan sulfate; about 0.001% to about 40% w / w keratan sulfate; and about 0.1% to about 60% w / w hyaluronic acid.According to some embodiments, the glycosaminoglycan adsorbent does not include one or more core proteins.

[0091] According to some embodiments, the heparan sulfate polymer of the adsorbent comprises about 20 to about 250 repeating disaccharide units. According to some embodiments, the heparan sulfate polymer comprises about 20 repeating disaccharide units, or about 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or about 250 repeating disaccharide units. According to some embodiments, the heparan sulfate polymer of the glycosaminoglycan adsorbent comprises about 22 to about 240 repeating disaccharide units. According to some embodiments, the heparan sulfate polymer of the glycosaminoglycan adsorbent comprises about 24 repeating disaccharide units, about 60 repeating disaccharide units, about 80 repeating disaccharide units, about 120 repeating disaccharide units, or about 240 repeating disaccharide units.

[0092] According to some embodiments, the heparan sulfate polymers of the adsorbent have an average molecular weight in the range of about 10 kDa to about 100 kDa. According to some embodiments, the average molecular weight of the heparan sulfate polymers is about 10 kDa, or about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or about 100 kDa. According to some embodiments, the heparan sulfate polymers of the glycosaminoglycan adsorbent have an average molecular weight of about 10 kDa to about 98 kDa. According to some embodiments, the heparan sulfate polymers of the adsorbent have an average molecular weight of about 12 kDa, about 35 kDa, about 50 kDa, or about 85 kDa.

[0093] In some embodiments, the amount of heparan sulfate contained in the solid adsorbent matrix of the glycocalyx-mimetic adsorption medium ranges from about 40% to about 96% w / w. In some embodiments, the amount of heparan sulfate in the adsorbent is about 40% w / w, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 96% w / w. In some embodiments, the amount of heparan sulfate in the adsorbent ranges from about 50% to about 90% w / w. In some embodiments, the amount of heparan sulfate in the adsorbent ranges from about 55% to about 85% w / w.

[0094] According to some embodiments, the chondroitin sulfate polymer of the adsorbent comprises from about 4 to about 155 repeating disaccharide units. According to some embodiments, the chondroitin sulfate polymer comprises about 6 repeating disaccharide units, or about 8, 10, 12, 15, 18, 20, 22, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, or about 155 repeating disaccharide units. According to some embodiments, the chondroitin sulfate polymer of the glycosaminoglycan adsorbent comprises from about 4 to about 152 repeating disaccharide units. According to some embodiments, the chondroitin sulfate polymer of the glycosaminoglycan adsorbent comprises about 4 repeating disaccharide units, about 50 repeating disaccharide units, about 54 repeating disaccharide units, about 110 repeating disaccharide units, or about 148 repeating disaccharide units.

[0095] According to some embodiments, the chondroitin sulfate polymers of the adsorbent have an average molecular weight in the range of about 2 kDa to about 70 kDa. According to some embodiments, the average molecular weight of the chondroitin sulfate polymers is about 4 kDa, or about 6, 8, 10, 12, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or about 70 kDa. According to some embodiments, the chondroitin sulfate polymers of the glycosaminoglycan adsorbent have an average molecular weight of about 2 kDa to about 68 kDa. According to some embodiments, the chondroitin sulfate polymers of the adsorbent have an average molecular weight of about 2 kDa, about 25 kDa, about 50 kDa, or about 70 kDa.

[0096] According to some embodiments, the amount of chondroitin sulfate contained in the solid adsorbent matrix of the glycocalyx-mimetic adsorption medium ranges from about 5% to about 30% w / w. According to some embodiments, the amount of chondroitin sulfate in the adsorbent is about 5% w / w, about 7% w / w, about 10% w / w, about 12% w / w, about 15% w / w, about 18% w / w, about 20% w / w, about 22% w / w, about 25% w / w, about 27% w / w, or about 30% w / w. According to some embodiments, the amount of chondroitin sulfate in the adsorbent ranges from about 10% to about 24% w / w. According to some embodiments, the amount of chondroitin sulfate in the adsorbent ranges from about 12% to about 22% w / w.

[0097] According to some embodiments, the ratio of heparan sulfate to chondroitin sulfate in the adsorbent solid substrate of the glycocalyx-mimetic adsorption medium can range from about 10:1 to about 1:10. According to some embodiments, the ratio of heparan sulfate to chondroitin sulfate can range from about 8:1 to about 1:8, about 6:1 to about 1:6, or about 4:1 to about 1:4. According to some embodiments, the ratio of heparan sulfate to chondroitin sulfate can be about 5:1, 5:2, 5:3, 5:4, 5:5, 4:5, 3:5, 2:5, or about 1:5. According to some embodiments, the ratio of heparan sulfate to chondroitin sulfate can be about 4:1, 2:1, 4:3, 1:1, 3:4, 1:2, or about 1:4. According to some embodiments, the ratio of heparan sulfate to chondroitin sulfate can be about 6:1, 5:1, 4:1, 3:1, 2:1, or about 1: 1. According to some embodiments, the ratio of heparan sulfate to chondroitin sulfate is about 4:1.

[0098] In some embodiments, the dermatan sulfate polymer of the adsorbent comprises about 20 to about 155 repeating disaccharide units. In some embodiments, the dermatan sulfate polymer comprises about 20 repeating disaccharide units, or about 22, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, or about 155 repeating disaccharide units. In some embodiments, the dermatan sulfate polymer of the glycosaminoglycan adsorbent comprises about 22 to about 152 repeating disaccharide units. According to some embodiments, the dermatan sulfate polymer of the glycosaminoglycan adsorbent comprises about 22 repeating disaccharide units, about 36 repeating disaccharide units, about 60 repeating disaccharide units, about 65 repeating disaccharide units, about 110 repeating disaccharide units, or about 148 repeating disaccharide units.

[0099] In some embodiments, the dermatan sulfate polymer of the adsorbent has an average molecular weight in the range of about 10 kDa to about 70 kDa. In some embodiments, the average molecular weight of the dermatan sulfate polymer is about 10 kDa, or about 12, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or about 70 kDa. In some embodiments, the dermatan sulfate polymer of the glycosaminoglycan adsorbent has an average molecular weight of about 10 kDa to about 68 kDa. In some embodiments, the dermatan sulfate polymer of the adsorbent has an average molecular weight of about 10 kDa, about 30 kDa, about 52 kDa, or about 70 kDa.

[0100] According to some embodiments, the amount of dermatan sulfate contained in the adsorbent solid matrix of the glycocalyx-mimetic adsorption medium ranges from about 1% to about 25% w / w. According to some embodiments, the amount of dermatan sulfate in the adsorbent is about 1%, about 3%, about 5%, about 7%, about 10%, about 12%, about 15%, about 18%, about 20%, about 22%, or about 25% w / w. According to some embodiments, the amount of dermatan sulfate in the adsorbent ranges from about 4% to about 22% w / w. According to some embodiments, the amount of dermatan sulfate in the adsorbent ranges from about 7% to about 18% w / w.

[0101] According to some embodiments, the keratan sulfate polymer of the adsorbent comprises about 10 to about 70 repeating disaccharide units. According to some embodiments, the keratan sulfate polymer comprises about 10 repeating disaccharide units, or about 11, 12, 13, 14, 15, 18, 20, 22, 25, 30, 35, 40, 45, 50, 55, 60, 65, or about 70 repeating disaccharide units. According to some embodiments, the keratan sulfate polymer of the glycosaminoglycan adsorbent comprises about 10 to about 68 repeating disaccharide units. According to some embodiments, the keratan sulfate polymer of the glycosaminoglycan adsorbent comprises about 11 repeating disaccharide units, about 35 repeating disaccharide units, about 40 repeating disaccharide units, about 56 repeating disaccharide units, or about 67 repeating disaccharide units.

[0102] According to some embodiments, the keratan sulfate polymers of the adsorbent have an average molecular weight in the range of about 5 kDa to about 30 kDa. According to some embodiments, the average molecular weight of the keratan sulfate polymers is about 5 kDa, or about 6, 8, 10, 12, 14, 15, 16, 18, 20, 22, 24, 25, 26, 28, or about 30 kDa. According to some embodiments, the keratan sulfate polymers of the glycosaminoglycan adsorbent have an average molecular weight of about 5 kDa to about 28 kDa. According to some embodiments, the keratan sulfate polymers of the adsorbent have an average molecular weight of about 5 kDa, about 25 kDa, about 50 kDa, or about 70 kDa.

[0103] According to some embodiments, the amount of keratan sulfate contained in the adsorbent solid matrix of the glycocalyx-mimetic adsorption medium ranges from about 0.01% to about 20% w / w. According to some embodiments, the amount of keratan sulfate in the adsorbent is about 0.01%, about 0.05%, about 0.1%, about 0.5%, about 1%, about 2%, about 3%, about 5%, about 6%, about 8%, about 10%, about 12%, about 14%, about 15%, about 16%, about 18%, or about 20% w / w. According to some embodiments, the amount of keratan sulfate in the adsorbent ranges from about 0.5% to about 15% w / w. According to some embodiments, the amount of keratan sulfate in the adsorbent ranges from about 1% to about 5% w / w.

[0104] According to some embodiments, the hyaluronic acid polymer of the adsorbent comprises from about 10 to about 100,000 repeating disaccharide units. According to some embodiments, the hyaluronic acid polymer comprises from about 10 repeating disaccharide units, or from about 25, 50, 75, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 10000, 11000, 12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000, 21000, 22000, 23000, 24 ... 000, 10,000, 12,000, 15,000, 20,000, 25,000, 30,000, 35,000, 40,000, 45,000, 50,000, 55,000, 60,000, 65,000, 70,000, 75,000, 80,000, 85,000, 90,000, 95,000, or about 100,000 repeating disaccharide units. According to some embodiments, the hyaluronic acid polymer of the glycosaminoglycan adsorbent has from about 10 to about 90,000 repeating disaccharide units, or from about 40 to about 80,000 repeating disaccharide units, from about 60 to about 70,000, from about 80 to about 60,000, from 100 to about 50,000, from about 150 to about 40,000, from about 200 to about 30,000, from about 250 to about 20,000, from about 300 to about 10,000, from about 350 to about 9,000, from about 400 to about 8,000, from about 450 to about 50,000, or from about 500 to about 60,000. The hyaluronic acid polymer of the glycosaminoglycan adsorbent contains about 7,000 to about 500 to about 6,000, about 550 to about 5,000, about 600 to about 4,000, about 650 to about 3,000, about 700 to about 2,000, or about 750 to about 1,000 repeating disaccharide units. According to some embodiments, the hyaluronic acid polymer of the glycosaminoglycan adsorbent contains about 11 repeating disaccharide units, about 1,200 repeating disaccharide units, about 2,200 repeating disaccharide units, about 4,100 repeating disaccharide units, about 7,900 repeating disaccharide units, about 12,300 repeating disaccharide units, 21,100 repeating disaccharide units, about 55,400 repeating disaccharide units, or about 98,700 repeating disaccharide units.

[0105] According to some embodiments, the hyaluronic acid polymers of the adsorbent have an average molecular weight in the range of about 4 kDa to about 40,000 kDa. According to some embodiments, the average molecular weight of the hyaluronic acid polymers is about 5 kDa, or about 15, 20, 25, 50, 75, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 10,000, 12,000, 15,000, 20,000, 25,000, 30,000, 35,000, or about 40,000 kDa. According to some embodiments, the hyaluronic acid polymer of the glycosaminoglycan adsorbent has a molecular weight of about 5 kDa to about 38,000 kDa, or about 10 to about 36,000 kDa, about 12 to about 35,000, about 18 to about 32,000, about 28 to about 30,000, about 35 to about 28,000, about 42 to about 25,000, about 46 to about 22,000, about 52 to about 18,000, about 64 to about 14,000, about 78 to about 11,000, The hyaluronic acid polymers of the adsorbent have an average molecular weight of about 120 to about 9500, about 180 to about 8000, about 320 to about 5500, about 460 to about 3500, about 570 to about 2200, about 630 to about 1800, or about 800 to about 1000 kDa. According to some embodiments, the hyaluronic acid polymers of the adsorbent have an average molecular weight of about 4 kDa, about 890 kDa, about 1340 kDa, about 3000 kDa, about 5900 kDa, about 8000 kDa, about 11,900 kDa, or about 34,000 kDa.

[0106] According to some embodiments, the amount of hyaluronic acid contained in the adsorbent solid matrix of the glycocalyx-mimetic adsorption medium ranges from about 5% to about 50% w / w. According to some embodiments, the amount of hyaluronic acid in the adsorbent is about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% w / w. According to some embodiments, the amount of hyaluronic acid in the adsorbent ranges from about 8% to about 50% w / w. According to some embodiments, the amount of hyaluronic acid in the adsorbent ranges from about 10% to about 50% w / w.

[0107] In some embodiments, the adsorbent of the glycomimetic adsorption medium solid substrate comprises about 40% to about 96% w / w heparan sulfate, about 5% to about 30% w / w chondroitin sulfate, about 1% to about 25% w / w dermatan sulfate, about 0.01% to about 20% w / w keratan sulfate, and about 5% to about 50% w / w hyaluronic acid. In some embodiments, the adsorbent comprises about 50% to about 90% w / w heparan sulfate, about 10% to about 24% w / w chondroitin sulfate, about 4% to about 22% w / w dermatan sulfate, about 0.5% to about 15% w / w keratan sulfate, and about 8% to about 50% w / w hyaluronic acid.

[0108] In certain aspects, the adsorbent of the glycocalyx-mimetic adsorption medium solid substrate comprises one or more core proteins selected from the group consisting of syndecan, glypican, perlecan, versican, decorin, biglycan, mimecan, or combinations thereof, in an amount ranging from about 0.001% to about 30% w / w; about 30% to about 96% w / w heparan sulfate; about 0.1% to about 30% w / w chondroitin sulfate; about 0.1% to about 25% w / w dermatan sulfate; about 0.001% to about 20% w / w keratan sulfate; and about 0.1% to about 50% w / w hyaluronic acid.

[0109] According to some embodiments, the solid substrate of the glycocalyx-mimetic adsorption medium has the contents of any one of Formulations A-FF in Tables 1-8 below: [Table 1] [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7] [Table 8]

[0110] The glycosaminoglycan / core protein of proteoglycans can be attached to the surface of the adsorption medium by a single covalent end-point bond (e.g., via the terminal residues of HS (and / or Hep), CS, DS, KS, and HA molecules). Compared to non-covalent or multipoint bonds, a single covalent bond at the end group of the attached molecule advantageously provides better control over the orientation of the immobilized molecules while maximizing their surface density. According to some embodiments, the end-point attachment of these long-chain carbohydrates provides a brush-type molecular surface structure that results in a higher concentration of accessible positions on the GAG ​​chains available for sample contact and / or analyte binding. According to some embodiments, full-length GAG chains are used in glycosaminoglycan adsorbents to coat substrate surfaces. According to some embodiments, fragmented GAG chains are used in glycosaminoglycan adsorbents to coat substrate surfaces.

[0111] Covalent attachment of GAG / core protein to the solid substrate, compared to non-covalent attachment, provides control over parameters such as surface density and orientation of the immobilized molecules, providing sample contact and adsorbate binding to the immobilized molecules. According to some embodiments, the surface concentration of adsorbent on the solid substrate can be between 0.01 and about 0.5 μg / cm. 2 range, e.g., 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19 or 0.2 μg / cm 2 According to another embodiment, the surface concentration of the absorbent on the solid substrate is 0.001 to 2.0 μg / cm 2According to another embodiment, the surface concentration of the absorbent on the solid substrate is in the range of 0.005 to 0.5 μg / cm 2 is in the range.

[0112] According to some embodiments, the surface concentration of the adsorbent on the solid substrate is greater than 1 μg / cm 2 ~20μg / cm 2 in the range of, for example, 1 μg / cm 2 , 2 μg / cm 2 , 3 μg / cm 2 , 4 μg / cm 2 , 5 μg / cm 2 , 6 μg / cm 2 , 7 μg / cm 2 , 8 μg / cm 2 , 9 μg / cm 2 , 10 μg / cm 2 , 11 μg / cm 2 , 12 μg / cm 2 , 13 μg / cm 2 , 14 μg / cm 2 , 15 μg / cm 2 , 16 μg / cm 2 , 17 μg / cm 2 , 18 μg / cm 2 , 19 μg / cm 2 , and 20 μg / cm 2 According to another embodiment, the surface concentration of the adsorbent on the solid substrate is 5 μg / cm 2 ~15μg / cm 2 range, e.g., 5 μg / cm 2 , 6 μg / cm 2 , 7 μg / cm 2 , 8 μg / cm 2 , 9 μg / cm 2 , 10 μg / cm 2 , 11 μg / cm 2 , 12 μg / cm 2 , 13 μg / cm 2 , 14 μg / cm 2 , and 15 μg / cm 2 is.

[0113] According to some embodiments, GAGs and / or core proteins are reductively conjugated to primary amines on aminated substrates, such as aminated beads, by reductive amination. Coupling the open aldehyde form of the reducing end of a GAG chain to a bead yields a stable secondary amine. The non-reducing end of a GAG chain bearing a reactive amine can be attached to beads with an intermediate bearing an aldehyde functionality. For example, an adsorbent can be attached to an amine-containing substrate by (a) contacting the aminated substrate with an aqueous solution containing mannose to form a Schiff base intermediate, and (b) contacting the Schiff base with a reducing agent to attach the GAG. The GAG ​​can be dissolved in an aqueous solution, such as an acidic aqueous solution. The aqueous GAG solution is contacted with an aminated substrate, such as aminated beads. A Schiff base is generated. The Schiff base is then reduced with a reducing agent. The reducing agent can be, for example, sodium cyanoborohydride or sodium borohydride. In certain cases, the solid substrate can be aldehyde-activated beads that can be reacted with proteoglycan core proteins that have reactive primary amines.

[0114] According to some embodiments, covalent attachment of full-length GAG molecules to a surface can be achieved by reaction of the aldehyde group of the GAG ​​molecule with a primary amino group present on the surface of the adsorption medium. A unique property of all carbohydrates is that they possess a hemiacetal at their reducing end. This acetal is in equilibrium with the aldehyde form and can form a Schiff base with a primary amine. These Schiff bases can be reduced to stable secondary amines. According to some embodiments, full-length GAG molecules are surface-immobilized on a solid substrate by covalent bonding. According to other embodiments, full-length GAGs are covalently attached to the adsorption medium via a stable secondary amino group.

[0115] In certain instances, various methods of coating solid substrates with adsorbents are disclosed in U.S. Pat. Nos. 8,663,148 and 8,758,286; and U.S. Patent Application Publication Nos. 2009 / 0136586, 2012 / 0305482, and U.S. Pat. No. 2014 / 231357, the disclosures of which are incorporated herein by reference for all purposes.

[0116] According to some embodiments, the solid substrate of the glycocalyx-mimetic adsorption medium can be in the form of multiple solid beads or particles. The beads can be made of a material that is sufficiently rigid to resist deformation or compression under the flow rates and pressures encountered. According to some embodiments, sufficient substrate rigidity is defined as a rigidity that does not result in a significant increase in pressure drop across the adsorption bed during approximately one hour of water or saline flow at a typical clinical flow rate. For example, adequate substrate rigidity will result in an increase in pressure drop of less than 10-50% compared to the initial pressure drop (e.g., measured within the first minute of flow) when measured at a similar flow rate of, for example, saline.

[0117] The size of the solid substrate can be selected depending on the amount of sample to be processed and other parameters. According to some embodiments, each bead of the plurality of rigid polymer beads has an average outer diameter of about 1 μm to about 1 mm, e.g., 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 45 μm, 55 μm, 60 μm, 65 μm, 70 μm, 75 μm, 80 μm, 85 μm, 90 μm, 95 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, or 1 mm. For example, polyethylene beads from DSM Biomedical (Berkeley, Calif.) having an average diameter of 300 μm are suitable for the methods and devices disclosed herein. According to other embodiments, each bead of the plurality of rigid polymer beads has an average diameter of about 10 μm to about 200 μm, e.g., 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 45 μm, 55 μm, 60 μm, 65 μm, 70 μm, 75 μm, 80 μm, 85 μm, 90 μm, 95 μm, 100 μm, 105 μm, 110 μm, 115 μm, 120 μm, 125 μm, 130 μm, 135 μm, 140 μm, 145 μm, 150 μm, 155 μm, 160 μm, 165 μm, 170 μm, 175 μm, 180 μm, 185 μm, 190 μm, 195 μm, 200 μm, or more. Generally, particle sizes in the range of 20-200 μm, such as 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 μm, are useful, although larger particles may be required for high flow applications. In certain cases, particles sized 120 μm or less are preferred for use with plasma and serum.

[0118] Methods for making such beads are known in the art. For example, suitable polyethylene and other polyolefin beads are produced directly during certain synthesis processes. In some cases, the beads are processed to the required size and shape. Other polymers may need to be milled or spray dried and classified, or otherwise processed to produce beads of the desired size distribution and shape.

[0119] Beads can be sintered into a monolithic porous structure by either chemical or physical means. Polyethylene beads can be sintered by heating the beads above their melting temperature in a cartridge and applying pressure. The resulting interstitial pore size is slightly reduced from that of a packed bed of unsintered beads of the same size. This reduction can be empirically determined and used to produce the desired final interstitial pore size.

[0120] The solid substrate may comprise one or more hollow or solid fibers. According to an embodiment of the device of the present invention in which the solid substrate comprises hollow fibers, the hollow fibers may preferably comprise a material selected from the group consisting of polysulfone, polyfluorocarbon, polyamide, polynitrile, polypropylene, cross-linked alginate, and cellulose. Other materials commonly used in hollow fibers for medical applications may also be used. The hollow fibers may preferably comprise polysulfone.

[0121] The size and porosity of the solid substrate must be selected for each application or treatment to allow adequate blood flow through the device with an acceptable pressure drop across the device. Certain applications requiring high blood flow rates and low pressure drop require larger diameter particles, pores, hollow fibers, or other solid substrates. Other applications not requiring high blood flow rates and low pressure drop can use smaller diameter particles, pores, hollow fibers, or other solid substrates. According to one embodiment of the present disclosure, in which the solid substrate is in the form of a hollow fiber, the inner diameter of the fiber can range from 1 μm to 1000 μm, e.g., 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 μm. Generally, inner diameters in the range of 20 to 200 μm are useful, although larger or smaller diameter fibers can be used in certain applications.

[0122] As described above, the glycocalyx-mimetic adsorption medium of the present disclosure is designed to function similarly to a naturally occurring, fully functional glycocalyx barrier. Thus, the glycocalyx-mimetic adsorption medium acts as an endothelial surface layer to protect and / or maintain the glycocalyx function of an impaired glycocalyx barrier. In other words, enhancing impaired glycocalyx barrier function with the glycocalyx-mimetic adsorption medium not only enhances and / or restores impaired glycocalyx barrier function, but the glycocalyx-mimetic adsorption medium also protects and / or maintains the enhanced and / or restored glycocalyx barrier function. In addition to enhancing, strengthening, restoring, protecting, and / or maintaining glycocalyx barrier function, the glycocalyx-mimetic adsorption medium can bind to adsorbates that cause glycocalyx barrier dysfunction. Thus, a sample obtained from a subject suffering from impaired glycocalyx barrier function contains adsorbates that bind to the glycocalyx-mimetic adsorption medium upon contact and are therefore removed from the sample, thereby forming a processed sample. According to some embodiments, the glycocalyx-mimetic adsorption medium removes adsorbates from a sample, thereby processing the sample, wherein the adsorbates are selected from the group including lymphokines, interferons, chemokines, exotoxins, endotoxins, ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, cytokines, tumor necrosis factor (TNF)-α, bacterial lipopolysaccharide (LPS), low-density lipoprotein (LDL), and heparin-binding protein (HBP).

[0123] HBP is an early marker of organ dysfunction. Heparin-binding protein (HBP), released by activated neutrophils, is a potent inducer of vascular leakage. Plasma levels of HBP can be used as an early diagnostic marker for severe sepsis. In other words, high plasma levels of HBP help identify patients at imminent risk of developing sepsis with circulatory collapse. (See Linder, A et al., ClinInfectDis. 2009 Oct 1; 49(7):1044-50.)

[0124] According to some embodiments, the glycocalyx-mimicking adsorption medium reduces the content (e.g., amount, level, etc.) of at least one adsorbate by about 5% to about 100% compared to the adsorbate content (e.g., amount, level, etc.) of the sample before processing. According to some embodiments, the glycocalyx-mimicking adsorption medium reduces the content of at least one adsorbate by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more compared to the adsorbate content of the sample before processing. In some embodiments, the glycocalyx-mimetic adsorption medium reduces the content of at least one adsorbate by about 10% to about 100% compared to the adsorbate content of the sample before treatment. The reduction in adsorbate levels can be detected using methods known in the art for detecting transcripts (e.g., quantitative polymerase chain reaction) or adsorbates (proteins) directly (e.g., immunoblot, enzyme-linked immunosorbent assay) or indirectly by measuring the activity of the adsorbate (protein) (e.g., degradation of extracellular matrix components).

[0125] According to some embodiments, the glycocalyx-mimicking adsorption medium reduces the content of at least one adsorbate selected from the group consisting of lymphokines, interferons, chemokines, exotoxins, endotoxins, ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, cytokines, tumor necrosis factor (TNF)-α, bacterial lipopolysaccharide (LPS), low-density lipoprotein (LDL), and heparin-binding protein (HBP) by about 10% to about 100% compared to the adsorbate content of the sample before treatment. According to some embodiments, the glycocalyx-mimicking adsorption medium reduces the content of ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, cytokines, tumor necrosis factor (TNF)-α, bacterial lipopolysaccharide (LPS), low-density lipoprotein (LDL), heparin-binding protein (HBP), or a combination thereof, by about 10% to about 100% compared to the adsorbate content of the sample before treatment. According to some embodiments, the glycocalyx-mimetic adsorption medium reduces the content of tumor necrosis factor (TNF)-α, bacterial lipopolysaccharide (LPS), low-density lipoprotein (LDL), heparin-binding protein (HBP), or a combination thereof by about 10% to about 100% compared to the adsorbate content of the sample before treatment. According to some embodiments, the glycocalyx-mimetic adsorption medium reduces the content of tumor necrosis factor (TNF)-alpha by about 10% to about 100% compared to the content of tumor necrosis factor (TNF)-α in the sample before treatment. According to some embodiments, the glycocalyx-mimetic adsorption medium reduces the content of low-density lipoprotein (LDL) by about 10% to about 100% compared to the content of low-density lipoprotein (LDL) in the sample before treatment. According to some embodiments, the glycocalyx-mimetic adsorption medium reduces the content of heparin-binding protein (HBP) by about 10% to about 100% compared to the content of heparin-binding protein (HBP) in the sample before treatment.

[0126] Also provided herein are methods for treating a condition associated with impaired glycocalyx barrier function in a subject in need thereof, the method comprising contacting a sample from the subject with a glycocalyx-mimetic adsorption medium to (a) enhance and / or restore impaired glycocalyx barrier function and / or (b) remove at least one adsorbate; and infusing the treated sample into the subject, thereby treating the condition associated with impaired glycocalyx barrier function. According to some embodiments, the condition associated with impaired glycocalyx barrier function is selected from the group consisting of capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, loss of vascular reactivity, sepsis, organ failure, atherosclerosis, and diabetes.

[0127] One specific example of a condition that may be treated according to the methods described herein is sepsis. Sepsis can be defined as a severe endothelial dysfunction syndrome in response to intravascular and extravascular infections that causes reversible or irreversible damage to the microcirculation, resulting in multiple organ failure. Sepsis is a potentially life-threatening complication of infection that can result from sepsis (i.e., organisms, their metabolic end products, or toxins in the bloodstream), including bacteremia (i.e., bacteria in the blood), and toxemia (i.e., bacteria in the blood), including endotoxemia (i.e., toxins in the blood). Sepsis can occur when endothelial and neutrophil cells become activated and release chemicals (i.e., inflammatory mediators) into the bloodstream to fight infection and trigger a systemic inflammatory response. Such chemicals or inflammatory mediators (e.g., lymphokines, interferons, chemokines, exotoxins, endotoxins, ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, cytokines, tumor necrosis factor (TNF)-α, bacterial lipopolysaccharide (LPS), low-density lipoprotein (LDL), and heparin-binding protein (HBP)) can trigger a series of changes that cause the body's immune system to attack healthy tissues, damaging multiple organ systems and leading to multiple organ failure in highly perfused organs. Sepsis can progress to septic shock, in which blood pressure drops dramatically and can lead to death. Septic shock occurs as a complication of infection, in which toxins can trigger a systemic inflammatory response. Septic shock can occur when blood pressure drops to dangerously low levels after infection. Reducing HBP and other inflammatory mediators modulates the immune response and may function as an organ-preserving strategy.

[0128] Von Willebrand factor (VWF) is a multimeric protein that mediates platelet adhesion and aggregation at sites of vascular injury. VWF, stored in Weibel-Palade bodies within alpha granules of endothelial cells and platelets, is released from stimulated endothelium as highly active ultra-large VWF multimers (ULVWF). ULVWF has a high affinity for platelets, favoring platelet aggregation, leading to platelet activation and microvascular thrombosis. Coactivation of coagulation factors and platelets can lead to disseminated intravascular coagulation (DIC) and may contribute to organ dysfunction. Hyperactive ULVWF is cleaved into a less active form by an enzyme known as ADAMTS-13 (a disintegrin and thrombospondin type 1 motif-containing metalloprotease, member 13). However, insufficient proteolysis of ULVWF due to reduced ADAMTS-13 activity leads to thrombotic microangiopathy (thrombosis) and disseminated platelet-rich thrombi in the microcirculation, as seen in organ failure. Decreased levels of ADAMTS-13 are particularly seen in thrombotic thrombocytopenic purpura, liver disease, malignant tumors, systemic lupus erythematosus, disseminated intravascular coagulation, and severe sepsis. ADAMTS-13 deficiency and elevated plasma levels of ULVWF are associated with sepsis, resulting in the net accumulation of thrombogenic ULVWF on the surface of endothelial cells with disrupted glycocalyx barriers, propagating pathological platelet-endothelial interactions and combining with other prothrombotic changes. In sepsis, this may contribute to microvascular thrombosis, platelet consumption, disseminated intravascular coagulation, edema, and ultimately multiple organ failure. (See, e.g., Blockmeyer, CL, et al. Haematologica, 2008, 93, 137-140; Karim, F. et al. BMC Pediatrics, 2013, 13(44), 1-5.) Therefore, removal of ULVWF and other inflammatory mediators from patient blood and / or plasma samples can treat conditions associated with glycocalyx barrier dysfunction, such as sepsis, capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, loss of vascular reactivity, organ failure, atherosclerosis, and diabetes.

[0129] After processing the sample, a processed sample can be formed by separating the sample from the glycocalyx-mimicking adsorption medium. The resulting separated sample is washed because the adsorbates present in the unprocessed sample remain as part of the glycocalyx-mimicking adsorption medium. Separation can be achieved, for example, by movement of the sample across the glycocalyx-mimicking adsorption medium. Movement can be, for example, by diffusion or forced convection. According to some embodiments, the flow of the sample across the glycocalyx-mimicking adsorption medium is driven by a pump, such as a positive displacement pump, an impulse pump, a velocity pump, a gravity pump, or a valveless pump. According to some embodiments, the pump is a centrifugal pump. According to some embodiments, the flow of the sample across the glycocalyx-mimicking adsorption medium is driven by cardiac activity of the subject.

[0130] In therapeutic applications, the washed sample can be reinjected back into the subject. The washed sample can be injected into the subject immediately after formation. The washed sample can be held for any period of time before injection into the subject. One or more components can be added to the washed sample after formation and before injection. According to some embodiments, a liquid is added to the washed sample to adjust its volume after formation and before injection. Injection can be performed in the form of discrete amounts of sample washed in this manner. Injection can be performed in the form of a continuous or semi-continuous flow. The amount of washed sample that can be injected in the claimed method is not intended to be limiting. If the sample contains blood and continuous recirculation into the subject is employed, it can range from less than 1 mL to more than 1 L, up to the patient's entire blood volume. If desired, one or more "passes" through the sorbent bed can be used. According to some embodiments, the washed sample is infused into the subject at the following rates: about 5 mL / min, about 10 mL / min, about 15 mL / min, about 20 mL / min, about 25 mL / min, about 30 mL / min, about 35 mL / min, about 40 mL / min, about 45 mL / min, about 50 mL / min, about 60 mL / min, about 70 mL / min, about 80 mL / min, about 90 mL / min, about 100 mL / min, about 150 mL / min, about 200 mL / min, about 250 mL / min, about 300 mL / min, about 350 mL / min, about 400 mL / min, about 450 mL / min, about 500 mL / min, about 550 mL / min, about 600 mL / min, about 700 mL / min, about 800 mL / min, about 900 mL / min, or about 1000 mL / min.

[0131] According to yet another embodiment, the present disclosure provides a method for improving oxygen saturation in a subject in need thereof, comprising contacting a sample from the subject with a glycocalyx-mimetic adsorption medium to treat the sample thereby enhancing and / or restoring impaired glycocalyx barrier function; and injecting the treated sample into the subject, wherein the glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof.

[0132] Normal arterial blood oxygen is approximately 75 to 100 millimeters of mercury (mmHg). A value below 60 mmHg typically indicates the need for supplemental oxygen. Normal pulse oximeter readings are typically in the 95 to 100 percent range. A value below 90% is considered low. Current methods improve low blood oxygen levels, bringing them back into the normal range. For example, a pulse oximeter reading in the low 80 to 90% range is restored to 95 to 100%. Similarly, a reading of 50 to 60 mmHg is restored to 75 to 100 mmHg.

[0133] Also provided is a device for enhancing impaired glycocalyx barrier function in a subject in need thereof. The device is characterized by a cartridge including an adsorption medium, inlet and outlet ports that allow a sample to enter and exit the device, and an end plate that prevents substantially all of the adsorption medium from exiting the cartridge. One such device is disclosed in U.S. Patent Application No. 14 / 860,589, filed September 21, 2015. [Example]

[0134] The following examples are provided to illustrate, but not limit, the disclosure. Example 1

[0135] This example demonstrates the use of a glycocalyx-mimetic adsorption medium to remove ultra-large VWF (ULVWF) multimers in plasma samples from individuals suspected of having sepsis.

[0136] A 45-year-old female patient presented to the emergency department with evidence of sepsis-like symptoms, including chills, dizziness, fatigue, flushing, hypothermia, and shivering. The patient's medical history revealed a recent diagnosis of a bladder infection and administration of oral antibiotics, including levofloxacin, approximately 48 hours prior. On examination, she was febrile with a temperature of 101.5°F and had low blood pressure (systolic pressure of approximately 90 mmHg). Physical examination confirmed the patient's pain near her kidneys, lower back, and a rash on her legs. A sepsis screen to diagnose possible bacterial sepsis or systemic inflammatory response syndrome confirmed that the female patient indeed had sepsis, with an ADAMTS-13 level of 26% (less than the lower limit of normal, 40%) and the presence of ULVWF. Sepsis in the patient's plasma sample was confirmed using gel electrophoresis. See, for example, Blockmeyer, CL, et al. Haematologica, 2008, 93, 137-140.

[0137] A patient is treated for sepsis by first obtaining whole blood from the patient through a catheter, which is then transfused into a treatment cartridge constructed with beads coated with the disclosed glycosaminoglycan adsorbent. The treatment cartridge consists of a sealed 300 mL adsorption column filled with glycosaminoglycan adsorbent-coated beads and mounted on a vertical stand. Plasma is then repeatedly transfused over the adsorption column multiple times. Each time the plasma passes through the column, ULVWF and other inflammatory mediators in the plasma adhere to the adsorption medium, reducing the concentration of ULVWF and other inflammatory mediators in the plasma. The plasma is then returned to the patient. The glycosaminoglycan mixture removes ULVWF from the plasma sample, effectively treating individuals suffering from sepsis. Example 2

[0138] This example demonstrates the removal of heparin-binding proteins (HBPs) using the methods of the present disclosure.

[0139] Figures 2A-B show the removal of HBP using heparinized beads. Figures 2A-B show that approximately 99.9% (A) and 95% (B) of HBP, respectively, were removed using heparinized beads. As shown, the negatively charged control hydrophilic beads (Figure 2A) removed more HBP than the positively charged hydrophilic beads (Figure 2B). Example 3

[0140] This example demonstrates that the methods of the present disclosure improve oxygen saturation in humans with various medical conditions.

[0141] The Seraph® 100 Microbind® Affinity Blood Filter device is a sterile, single-use, disposable column packed with ultra-high molecular weight polyethylene (UHMWPE) beads, which are surface-modified with non-leaching, end-point-attached heparin, and is composed of the following components and materials: [Table 9]

[0142] The functional mechanism of the Seraph® 100 Microbind® Affinity Blood Filter is as follows: The Seraph® 100 Microbind® Affinity Blood Filter device is an extracorporeal broad-spectrum adsorbent hemoperfusion device designed to reduce bacteria, viruses, toxins, cytokines, and other inflammatory mediators from whole blood. The Seraph® 100 Microbind® Affinity Blood Filter device is designed to share a very similar form factor to other blood filters, such as hemodialyzers and hemoperfusion filters, and is therefore compatible with hemodialysis systems that use industry-standard blood line connectors for ease of operation, training, and utility.

[0143] The Seraph® 100 Microbind® Affinity Blood Filter device achieves its intended performance by relying on the natural affinity that many adsorbates (pathogens, toxins, inflammatory mediators, etc.) have for surface-bound heparin. To efficiently remove pathogens and other adsorbates, the Seraph® 100 Microbind® Affinity Blood Filter device requires a large surface area of ​​surface-bound heparin. This is achieved by filling the device with heparin-coated microparticles.

[0144] Next, whole blood circulates through the Seraph® 100 Microbind® Affinity Blood Filter device, exposing pathogens and adsorbates to heparin. The Seraph 100 Microbind Affinity Blood Filter device interacts with pathogens and various inflammatory mediators via affinity adsorption. Heparin is composed of many different potential specific binding sites that match the chemical sequences of many inflammatory mediators, cytokines, and pathogens. Many microorganisms, including bacteria, viruses, and parasites, attach to heparan sulfate receptors (glycosaminoglycans) on the surface of mammalian cells.

[0145] The following study uses a Seraph® 100 Microbind® Affinity Blood Filter, but each of the formulations in Tables 1 to 8 (i.e., formulations A to FF) can be used.

[0146] In an exploratory medical device study, 15 subjects underwent a successful procedure using the Seraph® 100 Microbind® Affinity Blood Filter (Seraph® 100). As the patient's blood flowed through the Seraph® 100 Microbind® Affinity Blood Filter, it passed through polyethylene microbeads containing heparin. No serious adverse events (SAEs) were reported while the subjects were connected to the study device. No device complications or unexpected serious adverse effects were reported. No subjects died during the procedure. The study consisted of 15 dialysis patients with a mean age of 72.2 years. During the procedure, patients received treatment with the Seraph® for approximately 4 hours. Unexpectedly, a significant increase in oxygen saturation was observed, as measured by pulse oximetry during the procedure. The data are shown in Figure 3. The data clearly demonstrate that the oxygen saturation in the patients' blood increased to normal values.

[0147] Patient Case 1: Persistent Drug-Resistant S. aureus A 70-year-old female patient had been suffering from persistent drug-resistant S. aureus for at least five days before being treated with Seraph® for five hours. During treatment, her bloodstream infection resolved and her oxygen saturation increased from 96% to 100%. Patient Case 2. Severe S. aureus pneumonia An 86-year-old female patient suffered from severe S. aureus pneumonia with positive blood cultures. She was treated with Seraph® for 6 hours. During treatment, the device removed recent blood flow, and her oxygen saturation increased from 90% to 96%. Example 4

[0148] This example demonstrates that the disclosed method improves hemodynamic stability. Case 1 of a COVID-19 patient.

[0149] Hemodynamic stability can be described as stable blood flow. If a person is hemodynamically stable, it means that he / she has a stable cardiac pump and good blood circulation. Hemodynamic instability is defined as blood pressure instability that can lead to insufficient arterial blood flow to organs. It is also a state in which physiological and mechanical support is required to ensure adequate cardiac input / output or blood pressure.

[0150] While most COVID-19 patients experience mild or uncomplicated illness, approximately 14% develop severe illness requiring hospitalization and oxygen support, and 5% require admission to an intensive care unit. In severe cases, COVID-19 can be complicated by acute respiratory distress syndrome (ARDS), sepsis and septic shock, and multiple organ failure, including acute kidney injury and cardiac failure. Elderly age and comorbidities have been reported as risk factors for death, with older age, higher Sequential Organ Failure Assessment (SOFA) scores, and d-dimer levels above 1 μg / L at admission associated with higher mortality rates.

[0151] A 67-year-old patient was suffering from COVID-19-associated acute respiratory distress syndrome (ARDS) and was in hemodynamic shock. During treatment with Seraph®, his norepinephrine dose was 0.3 mcg / kg / min, maintaining his mean arterial pressure above 60 mmHg. During the first 3 hours of treatment, his norepinephrine dose was reduced to less than 0.1 mcg / kg / min, and his mean arterial pressure (MAP) rose above 80. By the end of 24 hours of treatment, norepinephrine was discontinued, and his MAP remained stable. Case 2 of a COVID-19 patient.

[0152] A 59-year-old patient suffered from COVID-19-associated acute respiratory distress syndrome (ARDS) and went into shock with increased vasopressor and norepinephrine requirements. The patient was treated for 8 hours, during which the norepinephrine dose was reduced from 0.14 mcg / kg / min when MAP was 60 to 0.07 mcg / kg / min when MAP was greater than 80 mmHg. His fraction of inspired oxygen (FiO2) level also decreased from 70% to 60%, indicating improved pulmonary function. Discussion

[0153] Rapid hemodynamic stabilization is important because myocardial injury has been associated with mortality in COVID-19 patients. Because it is still too early to use Seraph® to treat COVID-19 patients, the specific mechanism of hemodynamic stabilization is still under investigation. However, mycordial cell damage may be caused by direct interaction with the virus, a systemic inflammatory response, unstable coronary plaque, and exacerbated hypoxia. Increased troponin T (TnT) levels have been shown to correlate with outcomes. Increased TnT levels have also been found to be associated with increases in C-reactive protein and NT-proBNP. N-terminal pro-b-type natriuretic peptide (NT-proBNP) is an inflammatory marker and is produced when the heart is under stress.

[0154] NT-proBNT or proBNP levels have been monitored in at least two patients treated with Seraph®. The data are summarized in the chart below. Given that systemic heparin does not interfere with the analysis of NT-proBNP, it is possible that heparin does not bind to NT-proBNP. Therefore, the mechanism of NT-proBNP is unclear. [Table 10]

[0155] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, those skilled in the art will appreciate that certain changes and modifications may be practiced within the scope of the appended claims. Furthermore, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference were individually incorporated by reference.

Claims

1. 1. A method for enhancing impaired glycocalyx barrier function in a subject in need thereof, comprising: contacting a sample from said subject with a glycocalyx-mimicking adsorption medium to enhance and / or restore impaired glycocalyx barrier function, thereby treating the sample; and injecting the processed sample into a subject; wherein the glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof.

2. 2. The method of claim 1, wherein the adsorbent is a glycosaminoglycan mixture comprising about 40% to about 96% w / w heparan sulfate and / or heparin, and optionally one or more of the following additional adsorbents: about 5% to about 30% w / w chondroitin sulfate, about 1% to about 25% w / w dermatan sulfate, about 0.01% to about 20% w / w keratan sulfate, and / or about 5% to about 50% w / w hyaluronic acid.

3. 3. The method of claim 1 or 2, wherein the glycocalyx-mimetic adsorption medium aids in a member selected from the group consisting of vascular permeability, leukocyte adhesion, platelet adhesion, mediation of shear stress, and regulation of the inflammatory machinery.

4. The method according to any one of claims 1 to 3, wherein the adsorption medium acts as an endothelial surface layer to protect and / or maintain glycocalyx function.

5. 5. The method of any one of claims 1 to 4, wherein the adsorption medium reduces a member selected from the group consisting of capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, and loss of vascular reactivity.

6. The method of any one of claims 1 to 5, wherein the adsorption medium reduces glycocalyx shedding during tissue reperfusion.

7. The method of claim 6 , wherein the tissue is cardiac tissue undergoing coronary artery bypass surgery.

8. 7. The method of claim 6, wherein the tissue is perfused during organ transplantation.

9. 9. The method of any one of claims 1 to 8, for treating acute respiratory distress syndrome (ARDS) in a subject.

10. The method of any one of claims 1 to 9, wherein the method improves oxygen saturation in a subject.

11. The method of any one of claims 1 to 10, wherein the method improves hemodynamic stability in a subject.

12. The method of any one of claims 1 to 11, wherein the adsorption medium reduces glycocalyx shedding during sepsis.

13. The method of any one of claims 1 to 12, wherein the adsorption medium removes tumor necrosis factor (TNF)-α and bacterial lipopolysaccharide (LPS).

14. The method of any one of claims 1 to 13, wherein the adsorption medium reduces the risk of organ failure.

15. 15. The method of any one of claims 1 to 14, wherein the adsorption medium reduces glycocalyx shedding due to atherosclerosis or diabetes.

16. 16. The method of any one of claims 1 to 15, wherein the adsorption medium removes low density lipoproteins (LDL).

17. The method of any one of claims 1 to 16, wherein the adsorption medium binds heparin binding protein (HBP).

18. 18. The method of any one of claims 1 to 17, wherein the treated sample has an HBP content that is reduced by about 10% to about 100% compared to the HBP content of the sample before treatment.

19. 19. The method of any one of claims 1 to 18, wherein the adsorption medium binds a member selected from the group consisting of exotoxins, endotoxins, ultra-large von Willebrand factor (ULVWF), histones, exosomes, microvesicles, and cytokines.

20. The method of any one of claims 1 to 19, wherein the sample is a member selected from the group consisting of whole blood, serum, and plasma.

21. The method of any one of claims 1 to 20, wherein the sample is whole blood.

22. The method according to any one of claims 1 to 21, wherein the glycocalyx-mimetic adsorption medium is negatively charged.

23. The method of any one of claims 1 to 22, wherein the solid substrate comprises a non-toxic, non-leaching material.

24. The method of any one of claims 1 to 23, wherein the solid substrate comprises a plurality of rigid polymer beads.

25. 25. The method of any one of claims 1 to 24, wherein the hard polymer beads are selected from the group consisting of polyurethane, polymethyl methacrylate, polyethylene or copolymers of ethylene and other monomers, polyethyleneimine, polypropylene, and polyisobutylene.

26. The method of any one of claims 1 to 25, wherein the solid substrate comprises one or more hollow fibers.

27. The method of any one of claims 1 to 26, wherein the glycosaminoglycan mixture comprises at least one proteoglycan core protein selected from the group consisting of syndecan, glypican, perlecan, versican, decorin, biglycan, and mimecan.

28. 1. A device for enhancing impaired glycocalyx barrier function in a subject in need thereof, comprising:

1. A device comprising: a cartridge having a first end plate and a second end plate with a glycocalyx-mimicking adsorption medium disposed therein, wherein the glycocalyx-mimicking adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof.

29. the adsorbent is a glycosaminoglycan mixture comprising about 40% to about 96% w / w heparan sulfate and / or heparin, and optionally one or more of the following additional adsorbents: about 5% to about 30% w / w chondroitin sulfate, about 1% to about 25% w / w dermatan sulfate, about 0.01% to about 20% w / w keratan sulfate, and / or about 5% to about 50% w / w hyaluronic acid; a sample injection port for allowing the sample to enter the device; and 30. The device of claim 28, comprising a sample outlet port for allowing sample to flow into the device, wherein the sample flows through the first end plate, through the sorbent medium, and out the outlet port.

30. 30. The device of claim 28 or 29, wherein the glycocalyx-mimetic adsorption medium aids in a member selected from the group consisting of vascular permeability, leukocyte adhesion, platelet adhesion, mediation of shear stress, and modulation of the inflammatory machinery.

31. A device according to any one of claims 28 to 30, wherein the adsorption medium acts as an endothelial surface layer to protect and / or maintain glycocalyx function.

32. 32. The method of any one of claims 28-31, wherein the adsorption medium reduces a member selected from the group consisting of capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, and loss of vasoreactivity.

33. The device according to any one of claims 28 to 32, wherein the glycocalyx-mimetic adsorption medium is negatively charged.

34. The device of any one of claims 28 to 33, wherein the solid substrate comprises a non-toxic, non-leaching material.

35. The device of any one of claims 28 to 34, wherein the solid substrate comprises a plurality of rigid polymer beads.

36. 36. The device of any one of claims 28 to 35, wherein the hard polymer beads are selected from the group consisting of polyurethane, polymethyl methacrylate, polyethylene or copolymers of ethylene and other monomers, polyethyleneimine, polypropylene, and polyisobutylene.

37. The device of any one of claims 28 to 36, wherein the solid substrate comprises one or more hollow fibers.

38. The device of any one of claims 28 to 37, wherein the glycosaminoglycan mixture comprises at least one proteoglycan core protein selected from the group consisting of syndecan, glypican, perlecan, versican, decorin, biglycan, and mimecan.

39. 1. A method for improving oxygen saturation in a subject in need thereof, comprising: contacting a sample from said subject with a glycocalyx-mimicking adsorption medium to enhance and / or restore impaired glycocalyx barrier function, thereby treating the sample; and injecting the processed sample into a subject; wherein the glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof.

40. 40. The method of claim 39, wherein the adsorbent is a glycosaminoglycan mixture comprising about 40% to about 96% w / w heparan sulfate and / or heparin, and optionally one or more of the following additional adsorbents: about 5% to about 30% w / w chondroitin sulfate, about 1% to about 25% w / w dermatan sulfate, about 0.01% to about 20% w / w keratan sulfate, and / or about 5% to about 50% w / w hyaluronic acid.

41. 41. The method of claim 39 or 40, wherein the glycocalyx-mimetic adsorption medium aids in a member selected from the group consisting of vascular permeability, leukocyte adhesion, platelet adhesion, mediation of shear stress, and regulation of the inflammatory machinery.

42. 42. The method according to any one of claims 39 to 41, wherein the adsorption medium acts as an endothelial surface layer to protect and / or maintain glycocalyx function.

43. 43. The method of any one of claims 39-42, wherein the adsorption medium reduces a member selected from the group consisting of capillary leak syndrome, edema formation, inflammation, platelet hyperaggregation, hypercoagulability, and loss of vasoreactivity.

44. 44. The method of any one of claims 39 to 43, wherein oxygen saturation is returned to normal.

45. 1. A method for treating Covid-19 in a subject in need thereof, comprising: contacting a sample from said subject with a glycocalyx-mimicking adsorption medium to enhance and / or restore impaired glycocalyx barrier function, thereby treating the sample; and injecting the processed sample into a subject; wherein the glycocalyx-mimetic adsorption medium is a solid substrate having an adsorbent, the adsorbent being a glycosaminoglycan, including heparin, heparan sulfate, or a mixture thereof.

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