GELMA polymer composition containing cells

Abstract

The present disclosure provides improved polymer compositions, such as GelMA polymer compositions. In certain embodiments, the improved polymer compositions can be used to deliver one or more therapeutic agents, such as cells, to a target treatment area, such as the eye of a subject. In certain embodiments, the improved polymer composition is a hydrogel comprising gelatin methacryloyl (i.e., GelMA) or a polymer cross-linked derivative thereof.

Description

Technical Field

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 332,963, filed on April 20, 2022, and U.S. Provisional Patent Application No. 63 / 410,041, filed on September 26, 2022. The entire contents of these applications are incorporated herein by reference.

[0002] The present disclosure provides improved polymer compositions, such as GelMA polymer compositions. In certain embodiments, the improved polymer composition can be used to deliver one or more therapeutic agents, such as cells, to a target treatment area, such as an eye of a subject. In certain embodiments, the improved polymer composition is a hydrogel comprising gelatin methacryloyl (i.e., GelMA) or a polymer cross-linked derivative thereof.

Background Art

[0003] GelMA polymer compositions have emerged as effective materials for use in sealing, repairing, and / or treating injuries, defects, or diseases in the soft tissues of a subject. The design and generation of improved GelMA polymer compositions for this purpose is an active area of research.

[0004] There remains a need for improved GelMA polymer compositions, methods for generating GelMA polymer compositions, and therapeutic uses for GelMA polymer compositions.

Summary of the Invention

[0005] Details of various embodiments of the present disclosure are set forth in the following description.

[0006] In certain embodiments, the present disclosure provides a polymer composition comprising at least one chemically modified gelatin. In certain embodiments, the polymer composition may comprise at least one acrylated gelatin. In certain embodiments, the polymer composition comprises at least one methacrylated gelatin (GelMA). In certain embodiments, the polymer composition comprises from about 0.5% to about 5.0% w / v of the chemically modified gelatin. In certain embodiments, the polymer composition comprises from about 0.5% to about 5.0% w / v of the acrylated gelatin. In certain embodiments, the polymer composition comprises from about 0.5% to about 5.0% w / v of GelMA.

[0007] In certain embodiments, the polymer composition comprises at least one chemically modified gelatin (optionally an acrylated gelatin such as GelMA) and at least one polymer crosslinking initiator (e.g., a photoinitiator). In certain embodiments, the polymer composition comprises (i) at least one chemically modified gelatin (optionally an acrylated gelatin), (ii) at least one polymer crosslinking initiator, and (iii) at least one therapeutic agent (e.g., a cell). In certain embodiments, the polymer composition comprises (i) at least one chemically modified gelatin (optionally a methacrylated gelatin such as GelMA), (ii) at least one polymer crosslinking initiator, and (iii) at least one cell. In certain embodiments, the polymer composition is a precursor polymer composition. In certain embodiments, the polymer composition is a gel polymer composition.

[0008] In certain embodiments, the polymer composition comprises at least one crosslinking initiator. In certain embodiments, the crosslinking initiator comprises one or more photoactivated photoinitiators and optionally one or more photoinitiators activated by visible light. In certain embodiments, the polymer crosslinking initiator comprises eosin Y, N-vinylcaprolactam, triethanolamine, or any combination thereof. In certain embodiments, the polymer crosslinking initiator comprises eosin Y disodium salt (EYDS), N-vinylcaprolactam (NVC), triethanolamine, or any combination thereof. In certain embodiments, the polymer crosslinking initiator comprises eosin Y disodium salt (EYDS), N-vinylpyrrolidone (NVP), triethanolamine, or any combination thereof. In certain embodiments, the polymer crosslinking initiator comprises (i) about 50 μM of eosin Y or eosin Y disodium salt (EYDS), (ii) about 3.5 to about 5.0 μL / mL of N-vinylcaprolactam (NVC) or N-vinylpyrrolidone (NVP), and (iii) triethanolamine. In certain embodiments, the polymer crosslinking initiator comprises (i) about 50 μM of eosin Y disodium salt (EYDS), (ii) about 5.0 μL / mL of N-vinylpyrrolidone (NVP), and (iii) about 1.5% v / v of triethanolamine.

[0009] In certain embodiments, the chemically modified gelatin is acrylated gelatin. In certain embodiments, the acrylated gelatin has a degree of acylation of about 5 to 40%. In certain embodiments, the acrylated gelatin has a degree of acylation of 5 to 20%. In certain embodiments, the acrylated gelatin has a degree of acylation of about 5%, about 10%, or about 15%.

[0010] In certain embodiments, the chemically modified gelatin is methacrylated gelatin (GelMA). In certain embodiments, GelMA has a degree of methacrylation of about 5 to 40%. In certain embodiments, GelMA has a degree of methacrylation of about 5 to 20%. In certain embodiments, GelMA has a degree of methacrylation of about 5%, about 10%, or about 15%.

[0011] In certain embodiments, the polymer composition comprises from about 2% to about 5% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises from about 3% to about 5% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises from about 3% to about 4% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises about 3.5% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises from about 3% to about 4% w / v of GelMA. In certain embodiments, the polymer composition comprises about 3.5% w / v of GelMA.

[0012] In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture. In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture, and the total GelMA in the polymer composition is from about 0.5% to about 5% w / v. In certain embodiments, the total GelMA in the polymer composition is from about 2% to about 5% w / v. In certain embodiments, the total GelMA in the polymer composition is from about 2% to about 4% w / v. In certain embodiments, the total GelMA in the polymer composition is from about 3% to about 5% w / v. In certain embodiments, the total GelMA in the polymer composition is from about 3% to about 4% w / v. In certain embodiments, the total GelMA in the polymer composition is about 3.5% w / v.

[0013] In certain embodiments, the polymer composition comprises from about 0.5% to about 3% of a first GelMA mixture and from about 0.5% to about 3% of a second GelMA mixture. In certain embodiments, the polymer composition comprises from about 0.5% to about 1.5% of a first GelMA mixture and from about 2% to about 3% of a second GelMA mixture. In certain embodiments, the polymer composition comprises about 1% of a first GelMA mixture and about 2.5% of a second GelMA mixture.

[0014] In certain embodiments, the first GelMA mixture comprises GelMA having a high average molecular weight and a low degree of methacrylation (DOM). In certain embodiments, the second GelMA mixture comprises GelMA having a low average molecular weight and a high degree of methacrylation (DOM). In certain embodiments, the first GelMA mixture comprises GelMA having a high average molecular weight and a low DOM, and the second GelMA mixture comprises GelMA having a low average molecular weight and a high DOM. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of 140-180 kDa and a DOM of 5%-40%, and the second GelMA mixture comprises GelMA having an average molecular weight of 75-115 kDa and a DOM of 50%-80%. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of 140-180 kDa and a DOM of 5%-20%, and the second GelMA mixture comprises GelMA having an average molecular weight of 80-100 kDa and a DOM of 50%-70%. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of about 160 kDa and a DOM of about 10%, and the second GelMA mixture comprises GelMA having an average molecular weight of about 90 kDa and a DOM of about 60%.

[0015] In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 3% w / v comprising GelMA having a high average molecular weight and low DOM, and a second GelMA mixture of about 0.5% to about 3% comprising GelMA having a low average molecular weight and high DOM. In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 3% w / v comprising GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 40%, and a second GelMA mixture of about 0.5% to about 3% w / v comprising GelMA having an average molecular weight of 75 - 115 kDa and a DOM of 50% - 80%. In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 1.5% w / v comprising GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 40%, and a second GelMA mixture of about 2% to about 3% w / v comprising GelMA having an average molecular weight of 75 - 115 kDa and a DOM of 50% - 80%. In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 1.5% w / v comprising GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 20%, and a second GelMA mixture of about 2% to about 3% w / v comprising GelMA having an average molecular weight of 85 - 100 kDa and a DOM of 50% - 70%.

[0016] In certain embodiments, the polymer composition comprises a first GelMA mixture of about 1% w / v comprising GelMA having an average molecular weight of about 160 kDa and a DOM of about 10%, and a second GelMA mixture of about 2.5% w / v comprising GelMA having an average molecular weight of about 90 kDa and a DOM of about 60%.

[0017] In certain embodiments, at least one cell comprises an endothelial cell. In certain embodiments, at least one cell comprises a human umbilical vein endothelial cell (HUVEC). In certain embodiments, at least one cell comprises an epithelial cell. In certain embodiments, at least one cell comprises a human retinal pigment epithelial cell (HRPEC). In certain embodiments, at least one cell comprises a human retinal pigment epithelial cell (HRPEC) derived from induced pluripotent stem cells (iPSCs). In certain embodiments, at least one cell comprises an eye cell, or an eye cell derived from pluripotent or embryonic stem cells.

[0018] In certain embodiments, the polymer composition further comprises at least 0.1% (w / v) of a hydrophilic nonionic surfactant. In certain embodiments, the hydrophilic nonionic surfactant comprises at least one poloxamer surfactant such as poloxamer 407. In certain embodiments, the composition comprises about 0.2% (w / v) of a poloxamer surfactant such as poloxamer 407.

[0019] In certain embodiments, the present disclosure describes a precursor polymer composition comprising the polymer composition of the present disclosure. In certain embodiments, the present disclosure describes a gel polymer composition formed by photocrosslinking the precursor polymer composition of the present disclosure. In certain embodiments, the present disclosure describes a hydrogel polymer composition formed by photocrosslinking the precursor polymer composition of the present disclosure.

[0020] In certain embodiments, the present disclosure provides a method for treating and / or repairing a defect, injury, and / or disease in a target soft tissue of a subject. In certain embodiments, the present disclosure provides a method for treating and / or repairing a defect, injury, and / or disease in a target soft tissue of a subject, the method comprising providing a precursor polymer composition of the present disclosure, administering the precursor polymer composition onto or under the surface of the target soft tissue of the subject, optionally at the location of the soft tissue defect, injury, and / or disease, and crosslinking the precursor polymer composition by exposing a polymer crosslinking initiator in the polymer composition to crosslinking conditions, wherein crosslinking of the precursor polymer composition produces a gel polymer composition.

[0021] In certain embodiments, the present disclosure provides a method for treating a defect, injury, and / or disease in a target soft tissue of a subject, the method comprising providing a gel polymer composition of the present disclosure and administering the gel polymer composition onto, under, or near the surface of the target soft tissue of the subject. In certain embodiments, the gel polymer composition is administered at the location of the soft tissue defect, injury, and / or disease.

[0022] In certain embodiments, the target soft tissue is eye tissue. In certain embodiments, the target soft tissue is subconjunctival eye tissue or retinal eye tissue. In certain embodiments, the polymer composition is applied onto or under the surface of the eye tissue by subconjunctival injection, subretinal injection, or suprachoroidal injection.

[0023] In certain embodiments, the defect, injury, and / or disease of the target soft tissue includes an eye defect, injury, and / or disease and optionally an eye ulcer such as a corneal ulcer from an infection, injury, perforation, or other defect. In certain embodiments, the eye defect, injury, and / or disease includes a retinal degenerative disease. In certain embodiments, the eye defect, injury, and / or disease includes age-related macular degeneration (AMD). In certain embodiments, the eye defect, injury, and / or disease includes retinitis pigmentosa. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The foregoing and other objects, features, and advantages will become apparent from the following description of particular embodiments of the present disclosure, as illustrated in the accompanying drawings. The drawings are not necessarily to scale and are not inclusive, and instead focus on explaining the principles of the various embodiments of the present disclosure.

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DETAILED DESCRIPTION OF THE INVENTION

[0046] I. POLYMER COMPOSITION Overview The present disclosure provides a polymer composition (e.g., a GelMA polymer composition) having one or more advantages over compositions currently in commercial use or known in the art. In certain embodiments, the polymer composition has one or more of the following advantages compared to one or more compositions currently in commercial use or known in the art: (i) lower cost, (ii) easier production, (iii) improved biocompatibility, (iv) faster and / or stronger crosslinking and stability, (v) easier and / or more stable application, (vi) stronger adhesion and / or retention to a target surface, (vii) designable and adjustable degradation characteristics, (viii) a smooth surface when applied, and / or (ix) higher cell survival or delivery for encapsulated cells. In certain embodiments, the polymer compositions of the present disclosure enable the controlled sustained release of one or more therapeutic agents or cells over a period of time. Thus, the polymer compositions of the present disclosure present a clear improvement over compositions currently in commercial use and currently known in the art.

[0047] As used herein, the term "polymer composition" can refer to a precursor polymer composition (e.g., a polymer composition prior to crosslinking polymerization) and / or a gel polymer composition (e.g., a polymer composition after crosslinking polymerization) provided by the corresponding context of the present disclosure. Examples of gel polymer compositions include hydrogels and polymer compositions having an increase in viscosity resulting from crosslinking polymerization in the polymer composition (e.g., soft gels).

[0048] Generally, references to polymer components (e.g., GelMA, MeHA, PEGDA) in this disclosure can refer to polymer precursor components (e.g., monomers or precursor oligomers), cross-linked forms of polymer components in oligomers (e.g., cross-linked oligomers), and / or polymerized forms of polymer components in gel polymer compositions (e.g., hydrogel polymers), according to the context within this disclosure.

[0049] In certain embodiments, the polymer composition includes a chemically modified gelatin such as gelatin methacryloyl (i.e., GelMA). In certain embodiments, the polymer composition includes a chemically modified gelatin (e.g., GelMA) and one or more cross-linking agents. In certain embodiments, the polymer composition includes a chemically modified gelatin (e.g., GelMA) and one or more polymer cross-linking initiators such as a photoactivatable photoinitiator element. In certain embodiments, the polymer composition can include a chemically modified gelatin (e.g., GelMA), one or more cross-linking agents, and one or more polymer cross-linking initiators such as a photoactivatable photoinitiator element.

[0050] In certain embodiments, the polymer composition includes a chemically modified gelatin (e.g., GelMA) and a chemically modified hyaluronic acid (e.g., MeHA). In certain embodiments, the polymer composition includes a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), and one or more cross-linking agents. In certain embodiments, the polymer composition includes a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), and one or more polymer cross-linking initiators such as a photoactivatable photoinitiator element. In certain embodiments, the polymer composition includes a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), one or more cross-linking agents, and one or more polymer cross-linking initiators such as a photoactivatable photoinitiator element. In certain embodiments, the polymer composition includes unmodified HA. In certain embodiments, the polymer composition includes unmodified HA and chemically modified HA (e.g., MeHA).

[0051] In certain embodiments, the polymer composition comprises a chemically modified gelatin (e.g., GelMA) and a chemically modified poly(ethylene glycol) (PEG) (e.g., PEGDA). In certain embodiments, the polymer composition comprises a chemically modified gelatin (e.g., GelMA), a chemically modified PEG (e.g., PEGDA), and one or more crosslinking agents. In certain embodiments, the polymer composition comprises one or more polymer crosslinking initiators, such as a chemically modified gelatin (e.g., GelMA), a chemically modified PEG (e.g., PEGDA), and a photoactivatable photoinitiator element. In certain embodiments, the polymer composition comprises a chemically modified gelatin (e.g., GelMA), a chemically modified PEG (e.g., PEGDA), one or more crosslinking agents, and one or more polymer crosslinking initiators, such as a photoactivatable photoinitiator element. In certain embodiments, the polymer composition comprises unmodified PEG. In certain embodiments, the polymer composition comprises unmodified PEG and a chemically modified PEG (e.g., PEGDA).

[0052] In certain embodiments, the polymer composition comprises a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), and a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), a chemically modified PEG (e.g., PEGDA), and one or more crosslinking agents. In certain embodiments, the polymer composition comprises one or more polymer crosslinking initiators, such as a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), a chemically modified PEG (e.g., PEGDA), and a photoactivatable photoinitiator element. In certain embodiments, the polymer composition comprises a chemically modified gelatin (e.g., GelMA), a chemically modified hyaluronic acid (e.g., MeHA), a chemically modified PEG (e.g., PEGDA), one or more crosslinking agents, and one or more polymer crosslinking initiators, such as a photoactivatable photoinitiator element. In certain embodiments, the polymer composition comprises unmodified HA and / or unmodified PEG.

[0053] In certain embodiments, the polymer composition does not contain a hydrolase enzyme. In certain embodiments, the polymer composition does not contain a glycosidase hydrolase enzyme.

[0054] In certain embodiments, the gel polymer composition is a hydrogel. A hydrogel generally comprises a cross-linked polymer framework that encompasses a network of pores filled with an interstitial solvent (e.g., a fluid) containing water. In certain embodiments, the hydrogel polymer composition has a water content of about 80% or more. In certain embodiments, the hydrogel polymer composition has a water content of greater than about 80%, greater than about 81%, greater than about 82%, greater than about 83%, greater than about 84%, greater than about 85%, greater than about 86%, greater than about 87%, greater than about 88%, greater than about 89%, greater than about 90%, greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, or greater than about 99%.

[0055] In certain embodiments, the polymer composition (e.g., a hydrogel or hydrogel precursor) of the present disclosure (e.g., a GelMA polymer composition) is a primary polymer composition and can include or be combined with one or more secondary hydrogel-forming polymer components (i.e., a polymer or its precursor). In certain embodiments, the polymer composition (e.g., a hydrogel or hydrogel precursor) of the present disclosure (e.g., a GelMA polymer composition) is a primary polymer composition and is selected from acrylamide, acrylic acid, alginate, methacrylated alginate, cellulose, chitosan, methacrylated chitosan, dimethacrylamide, gelatin, methacrylated gelatin, glycol chitosan, methacrylated glycol chitosan, hexyl methacrylate, hyaluronic acid, methacrylated hyaluronic acid, hydroxyethyl methacrylate, hydroxyethyl acrylate, isopropylacrylamide, isopropylmethacrylamide, methacrylamide, methacrylic acid, polyamide, polycaprolactone, polyethylene-glycol (PEG), polyethylene terephthalate, polylactic acid, polyurethane, polyvinyl alcohol, polyethylene oxide dimethacrylate, siloxane, polysiloxane, or an oligomer, polymer, and / or combinations thereof, and can include or be combined with one or more secondary hydrogel-forming polymer components. In certain embodiments, the secondary hydrogel polymer (formed from a second hydrogel-forming polymer precursor) is covalently crosslinked to the primary gel polymer composition (e.g., a GelMA polymer composition). In certain embodiments, the secondary hydrogel polymer (formed from a secondary logel-forming polymer precursor) is not covalently crosslinked to the primary gel polymer composition (e.g., a GelMA polymer composition); for example, the secondary hydrogel polymer forms a polymer network that interweaves with the polymer network of the primary gel polymer composition.

[0056] In certain embodiments, the polymer compositions of the present disclosure include one or more biocompatible polymer components or polysaccharides. In certain embodiments, the polymer compositions of the present disclosure include one or more biocompatible polymer components or polysaccharides selected from agarose, alginate, amylopectin, amylose, carrageenan, cellulose, chitin, chitosan, chondroitin sulfate, collagen, dermatan sulfate, dextran, elastin, elastin-like polypeptide (ELP), tropoelastin, fibrin, fibrinogen, fibronectin, gelatin, glycogen, heparan, heparan sulfate, heparin, heparin sulfate, hyaluronan, hyaluronic acid, keratan sulfate, laminin, pectin, polyglycerol sebacate (PGS), polyethylene glycol (PEG), polylactic acid (PLA), polylysine, starch, thrombin, derivatives thereof, or combinations thereof.

[0057] In certain embodiments, the polymer compositions of the present disclosure include one or more cell adhesives selected from fibronectin, laminin, vitronectin, RGD, vixapatin, derivatives thereof, or combinations thereof.

[0058] In certain embodiments, the polymer compositions of the present disclosure include one or more synthetic polymer components, such as biocompatible synthetic polymer components. In certain embodiments, the polymer composition includes one or more synthetic polymer components selected from polyurethane, polysiloxane, silicone, polyethylene, polyvinylpyrrolidone, polyhydroxyethyl methacrylate (poly-HEMA), polymethyl methacrylate, polyvinyl alcohol, polyacrylic acid, polyacrylamide, polyethylene-co-vinyl acetate, polyethylene glycol, polymethacrylic acid, polylactic acid, polyglycolic acid, polylactide-co-glycolide, nylon, polyamide, polyanhydride, polyethylene-co-vinyl alcohol, polycaprolactone, polyvinyl acetate, polyvinyl hydroxide, polyethylene oxide, polyorthoester, polyallylamine, polyethyleneimine, polylysine, polyarginine, derivatives thereof, or combinations and / or copolymers thereof.

[0059] In certain embodiments, the polymer compositions of the present disclosure include one or more polymer components (e.g., monomers, precursors, polymers) that contain crosslinkable groups. In certain embodiments, the polymer compositions of the present disclosure include one or more polymer components that contain crosslinkable groups selected from (or formed from reactions with) anhydrides, acid halides, carboxylic acids, diols, acrylic anhydrides, methacrylic anhydrides, acryloyl chloride, acryloyl bromide, methacryloyl chloride, methacryloyl bromide, acrylic acid, glycidyl methacrylate, methacrylic acid, dopamine, their derivatives, or combinations thereof.

[0060] In certain embodiments, hydroxyethyl methacrylate (HEMA) or its polymer may be present in the polymer composition at a concentration of about 1% to about 60% weight / volume (w / v).

[0061] In certain embodiments, the polymer compositions of the present disclosure include one or more stabilizers and / or enhancers. In certain embodiments, the polymer compositions of the present disclosure include one or more stabilizers and / or enhancers selected from polar amino acids (e.g., tyrosine, cysteine, serine, threonine, asparagine, glutamine, aspartic acid, glutamic acid, arginine, lysine, and histidine), amino acid analogs, amino acid derivatives, collagen, divalent cation chelating agents (e.g., ethylenediaminetetraacetic acid (EDTA) or its salts), or combinations thereof.

[0062] In certain embodiments, the polymer compositions of the present disclosure can be transparent and / or translucent. In certain embodiments, the polymer composition can be partially translucent or partially opaque. In certain embodiments, the polymer composition can be opaque.

[0063] In certain embodiments, the polymer compositions of the present disclosure can include, can be produced, can be analyzed, or can be used with a polymer component or a therapeutic component as disclosed in U.S. Patent Application Publication No. 20140377326, U.S. Patent Application Publication No. 20150274805, U.S. Patent Application Publication No. 20160175488, U.S. Patent Application Publication No. 20170232138, U.S. Patent Application Publication No. 20190022280(A1), International Publication No. 2020051133, and International Publication No. 2020081673. Each of these documents is hereby incorporated by reference in its entirety to the extent it describes the composition, production, analysis, and use of acrylated gelatin polymer compositions such as GelMA hydrogels.

[0064] Formulation In certain embodiments, the polymer compositions of the present disclosure include chemically modified gelatin (e.g., GelMA), chemically modified hyaluronic acid (e.g., MeHA), chemically modified PEG (e.g., PEGDA), or any combination thereof. In certain embodiments, the polymer composition includes chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition includes chemically modified hyaluronic acid (e.g., MeHA). In certain embodiments, the polymer composition includes chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition includes chemically modified gelatin (e.g., GelMA) and chemically modified hyaluronic acid (e.g., MeHA). In certain embodiments, the polymer composition includes chemically modified gelatin (e.g., GelMA) and chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition includes chemically modified hyaluronic acid (e.g., MeHA) and chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition includes chemically modified gelatin (e.g., GelMA), chemically modified hyaluronic acid (e.g., MeHA), and chemically modified PEG (e.g., PEGDA).

[0065] In certain embodiments, the polymer compositions of the present disclosure include combinations of precursor polymer components according to Table 1 (percentages are w / v concentrations in the total precursor polymer formulation). Unless otherwise specified, the GelMA materials in Table 1 are 160 / 80 (i.e., having a molecular weight (MW) of 160 kDa and a degree of methacrylation (DoM) of 80%). Unless otherwise specified, the HAMA materials in Table 1 are 500 / 30 (i.e., having a molecular weight (MW) of 500 kDa and a degree of methacrylation (DoM) of 30%). Unless otherwise stated, the PEGDA materials in Table 1 are formed from 35 kDa PEG materials. Pluronic F127 = Px407.

Table 1-1

Table 1-2

[0066] In certain embodiments, the polymer composition includes from about 4 to 20% w / v of a chemically modified gelatin (e.g., GelMA), from about 0 to 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and from about 0 to 5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition includes from about 4 to 10% w / v of a chemically modified gelatin (e.g., GelMA), from about 1 to 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and from about 0.1 to 5% w / v of a chemically modified PEG (e.g., PEGDA).

[0067] In certain embodiments, the polymer composition comprises GelMA having a molecular weight (MW) of about 160 kDa. In certain embodiments, the polymer composition comprises GelMA having a molecular weight (MW) of about 160 kDa. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 80% to about 90%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 85%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 80%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 75% to about 85%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 70% to about 80%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 75%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 70%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 65% to about 75%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 60% to about 70%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 65%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 60%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 55% to about 65%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 50% to about 60%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 55%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 50%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 45% to about 55%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 40% to about 50%.In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 45%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 40%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of from about 35% to about 45%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of from about 30% to about 40%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 35%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 30%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of from about 25% to about 35%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of from about 20% to about 30%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of about 25%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 20%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of from about 15% to about 25%. In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylation (DoM) of from about 10% to about 20%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 15%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 10%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 5%. In certain embodiments, the polymer composition comprises GelMA having a DoM of from about 10 to 40%. In certain embodiments, the polymer composition comprises GelMA having a DoM of from about 50 to 80%. In certain embodiments, the polymer composition comprises GelMA having a DoM of from about 10 to 20%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 7%. In certain embodiments, the polymer composition comprises GelMA having a DoM of about 5%. In certain embodiments, the polymer composition comprises GelMA having a DoM of from about 5 to 40%.In certain embodiments, the polymer composition comprises GelMA having about 5-20% DoM. In certain embodiments, the polymer composition comprises GelMA having about 1-10% DoM.

[0068] In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture. In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture, and the total GelMA in the polymer composition is from about 0.5% to about 5%. In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture, and the total GelMA in the polymer composition is from about 2% to about 5%. In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture, and the total GelMA in the polymer composition is from about 2% to about 4%. In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture, and the total GelMA in the polymer composition is from about 3% to about 5%. In certain embodiments, the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture, and the total GelMA in the polymer composition is from about 3% to about 4%.

[0069] In certain embodiments, the polymer composition comprises about 0.5% to about 3% of the first GelMA mixture. In certain embodiments, the polymer composition comprises about 0.5% to about 3% of the second GelMA mixture. In certain embodiments, the polymer composition comprises about 0.5% to about 3% of the first GelMA mixture and about 0.5% to about 3% of the second GelMA mixture. In certain embodiments, the polymer composition comprises about 0.5% to about 1.5% of the first GelMA mixture and about 2% to about 3% of the second GelMA mixture. In certain embodiments, the polymer composition comprises about 2% to about 3% of the first GelMA mixture and about 0.5% to about 1.5% of the second GelMA mixture. In certain embodiments, the polymer composition comprises about 1% of the first GelMA mixture and about 2.5% of the second GelMA mixture.

[0070] In certain embodiments, the first GelMA mixture comprises GelMA having a high average molecular weight (e.g., 140 - 180 kDa, or about 160 kDa). In certain embodiments, the first GelMA mixture comprises GelMA having a low degree of methacrylation (DOM) (e.g., 5% - 40%, or about 10%). In certain embodiments, the first GelMA mixture comprises GelMA having a high average molecular weight (e.g., 140 - 180 kDa, or about 160 kDa) and a low (DOM) (e.g., 5% - 40%, or about 10%). In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 40%. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 20%. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of about 160 kDa and a DOM of about 10%.

[0071] In certain embodiments, the second GelMA mixture comprises GelMA having a low average molecular weight (e.g., 75 - 115 kDa, or about 90 kDa). In certain embodiments, the second GelMA mixture comprises GelMA having a high degree of methacrylation (DOM) (e.g., 50% - 80%, or about 60%). In certain embodiments, the second GelMA mixture comprises GelMA having a low average molecular weight (e.g., 75 - 115 kDa, or about 90 kDa) and a high (DOM) (e.g., 50% - 80%, or about 60%). In certain embodiments, the second GelMA mixture comprises GelMA having an average molecular weight of 75 - 115 kDa and a DOM of 50% - 80%. In certain embodiments, the second GelMA mixture comprises GelMA having an average molecular weight of 80 - 115 kDa and a DOM of 50% - 70%. In certain embodiments, the second GelMA mixture comprises GelMA having an average molecular weight of about 90 kDa and a DOM of about 60%.

[0072] In certain embodiments, the first GelMA mixture comprises GelMA having a high average molecular weight (e.g., 140 - 180 kDa, or about 160 kDa) and a low degree of methacrylation (DOM) (e.g., 5% - 40%, or about 10%), and the second GelMA mixture comprises GelMA having a low average molecular weight (e.g., 75 - 115 kDa, or about 90 kDa) and a high DOM (e.g., 50% - 80%, or about 60%). In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 40%, and the second GelMA mixture comprises GelMA having an average molecular weight of 75 - 115 kDa and a DOM of 50% - 80%. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of 140 - 180 kDa and a DOM of 5% - 20%, and the second GelMA mixture comprises GelMA having an average molecular weight of 80 - 100 kDa and a DOM of 50% - 70%. In certain embodiments, the first GelMA mixture comprises GelMA having an average molecular weight of about 160 kDa and a DOM of about 10%, and the second GelMA mixture comprises GelMA having an average molecular weight of about 90 kDa and a DOM of about 60%.

[0073] In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 3% w / v, which comprises GelMA having a high average molecular weight (e.g., 140 to 180 kDa, or about 160 kDa) and a low degree of methacrylation (DOM) (e.g., 5% to 40%, or about 10%). In certain embodiments, the polymer composition comprises a second GelMA mixture of about 0.5% to about 3% w / v, which comprises GelMA having a low average molecular weight (e.g., 75 to 115 kDa, or about 90 kDa) and a high DOM (e.g., 50% to 80%, or about 60%). In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 3% w / v, which comprises GelMA having a high average molecular weight (e.g., 140 to 180 kDa, or about 160 kDa) and a low DOM (e.g., 5% to 40%, or about 10%), and a second GelMA mixture of about 0.5% to about 3% w / v, which comprises GelMA having a low average molecular weight (e.g., 75 to 115 kDa, or about 90 kDa) and a high DOM (e.g., 50% to 80%, or about 60%).

[0074] In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 1.5% w / v, which comprises GelMA having an average molecular weight of 140 to 180 kDa and a DOM of 5% to 40%, and a second GelMA mixture of about 2% to about 3% w / v, which comprises GelMA having an average molecular weight of 75 to 115 kDa and a DOM of 50% to 80%. In certain embodiments, the polymer composition comprises a first GelMA mixture of about 0.5% to about 1.5% w / v, which comprises GelMA having an average molecular weight of 140 to 180 kDa and a DOM of 5% to 20%, and a second GelMA mixture of about 2% to about 3% w / v, which comprises GelMA having an average molecular weight of 85 to 100 kDa and a DOM of 50% to 70%.

[0075] In certain embodiments, the polymer composition comprises a first GelMA mixture of about 1% w / v, which comprises GelMA having an average molecular weight of about 160 kDa and a DOM of about 10%, and a second GelMA mixture of about 2.5% w / v, which comprises GelMA having an average molecular weight of about 90 kDa and a DOM of about 60%.

[0076] In certain embodiments, the polymer composition comprises a combination of GelMA having about 10% DoM (e.g., GelMA 160 / 10) and GelMA having about 80% DoM (e.g., GelMA 160 / 80). In certain embodiments, the polymer composition comprises a combination of GelMA having about 10% DoM and GelMA having about 80% DoM, and the polymer composition comprises about 4 - 10% w / v GelMA. In certain embodiments, the polymer composition comprises a combination of GelMA having about 10% DoM (e.g., GelMA 160 / 10) and GelMA having about 80% DoM (e.g., GelMA 160 / 80), and the ratio of 10% DoM GelMA to 80% DoM GelMA is from about 1:9 to about 9:1. In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 1:9. In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 2:8 (i.e., about 1:4). In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 3:7. In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 4:6 (i.e., about 2:3). In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 5:5 (i.e., about 1:1). In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 4:6 (i.e., about 2:3). In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 3:7. In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 2:8 (i.e., about 1:4). In certain embodiments, the ratio of 10% DoM GelMA to 80% DoM GelMA is about 1:9.

[0077] In certain embodiments, the polymer composition comprises glycidyl methacrylate-functionalized GelMA having about 45% DoM (e.g., GelMA 160 / 45). In certain embodiments, the polymer composition comprises glycidyl methacrylate-functionalized GelMA (GelMA 160 / 45) having about 45% DoM and 160 kDa. In certain embodiments, the polymer composition comprises glycidyl methacrylate-functionalized GelMA (GelMA 90 / 45) having about 45% DoM and 90 kDa.

[0078] In certain embodiments, the polymer composition comprises MeHA having a molecular weight (MW) of about 500 kDa. In certain embodiments, the polymer composition comprises MeHA having a degree of methacrylation (DoM) of about 30%. In certain embodiments, the polymer composition comprises PEGDA formed from a PEG material of about 35 kDa. In certain embodiments, the polymer composition comprises PEGDA formed from a PEG material of about 2 kDa.

[0079] In certain embodiments, the polymer composition comprises a poloxamer surfactant (e.g., poloxamer 407). In certain embodiments, the polymer composition comprises about 0.1 - 0.5% w / v (e.g., about 0.2% w / v) of a poloxamer surfactant (e.g., poloxamer 407). In certain embodiments, the polymer composition comprises a tyloxapol surfactant. In certain embodiments, the polymer composition comprises about 0.1 - 0.5% w / v (e.g., about 0.1% w / v) of a tyloxapol surfactant.

[0080] In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1 - 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1 - 5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1 - 5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1 - 5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA).In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA).

[0081] In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1-1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA).In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA).

[0082] In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1 - 1.5% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.1 - 5% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.1 - 5% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1.5% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.1 - 5% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1.5% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of chemically modified PEG (e.g., PEGDA).In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1.5% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1.5% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of chemically modified gelatin (e.g., GelMA), about 1.5% w / v of chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of chemically modified PEG (e.g., PEGDA).

[0083] In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1-1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA).In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), about 1.5% w / v of a chemically modified hyaluronic acid (e.g., MeHA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA).

[0084] In certain embodiments, the polymer composition comprises about 4-20% w / v of a chemically modified gelatin (e.g., GelMA), and about 0-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4-10% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1-5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 4% w / v of a chemically modified gelatin (e.g., GelMA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 5% w / v of a chemically modified gelatin (e.g., GelMA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 7% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA).In certain embodiments, the polymer composition comprises about 7% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 7% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 7% w / v of a chemically modified gelatin (e.g., GelMA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 10% w / v of a chemically modified gelatin (e.g., GelMA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.1% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.5% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), and about 0.67% w / v of a chemically modified PEG (e.g., PEGDA). In certain embodiments, the polymer composition comprises about 20% w / v of a chemically modified gelatin (e.g., GelMA), and about 1.0% w / v of a chemically modified PEG (e.g., PEGDA).

[0085] In certain embodiments, the polymer composition comprises about 4% GelMA (10 - 40% DoM) and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 2% GelMA (10 - 40% DoM), about 2% gelatin, and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 4% gelatin acrylate (10 - 40% DoM) and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 2% gelatin acrylate (10 - 40% DoM), about 2% gelatin, and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 4% GelMA (10 - 40% DoM), about 1% PEGDA (35 kDa), and about 1 - 20% PEG methyl ether acrylate (35 kDa). In certain embodiments, the polymer composition comprises about 4% GelMA (10 - 40% DoM), about 1% HAMA (500 kDa, 5 - 40% DoM), and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 2% GelMA (10 - 40% DoM), about 2% gelatin, about 1% HAMA (500 kDa, 5 - 40% DoM), and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 4% gelatin acrylate (10 - 40% DoM), about 1% HAMA (500 kDa, 5 - 40% DoM), and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 2% gelatin acrylate (10 - 40% DoM), about 2% gelatin, about 1% HAMA (500 kDa, 5 - 40% DoM), and about 1% PEGDA (35 kDa). In certain embodiments, the polymer composition comprises about 4% GelMA (10 - 40% DoM), about 1% HAMA (500 kDa, 5 - 40% DoM), about 1% PEGDA (35 kDa), and about 1 - 20% PEG methyl ether acrylate (35 kDa). In certain embodiments, the polymer composition comprises about 5 - 20% GelMA (10 - 40% DoM).In certain embodiments, the polymer composition comprises about 5-20% GelMA (10-40% DoM), and about 1% HAMA (500 kDa, 5-40% DoM).

[0086] In certain embodiments, the polymer composition comprises about 4% GelMA (80% DoM), about 1% PEGDA (2 kDa), and about 0.2% (w / v) of a poloxamer surfactant (e.g., poloxamer 407), and optionally an active agent (e.g., a corticosteroid). In certain embodiments, the polymer composition comprises about 4% GelMA (40% DoM), about 1% PEGDA (35 kDa), and about 0.2% (w / v) of a poloxamer surfactant (e.g., poloxamer 407), and optionally an active agent (e.g., a corticosteroid). In certain embodiments, the polymer composition comprises about 4% GelMA (10% DoM), about 1% PEGDA (35 kDa), and about 0.2% (w / v) of a poloxamer surfactant (e.g., poloxamer 407), and optionally an active agent (e.g., a corticosteroid). In certain embodiments, the polymer composition comprises about 20% GelMA (40% DoM), and about 0.2% (w / v) of a poloxamer surfactant (e.g., poloxamer 407), and optionally an active agent (e.g., a corticosteroid).

[0087] Chemically modified gelatin Gelatin is a naturally occurring biocompatible mixture of peptides and proteins derived from collagen, which is a major structural component of animal tissues including eye tissue, bone, and skin. The native substrate peptides and proteins (e.g., denatured collagen) that can be used in the production of the gelatin materials of the present disclosure can include gelatin components derived from animals including, but not limited to, pigs, cows, horses, chickens, and fish. In certain embodiments, the gelatin material can be derived from connective tissue proteins such as collagen. In certain embodiments, the gelatin material can be derived from bone, skin, or eye tissue. In certain embodiments, the gelatin material can be prepared by acid hydrolysis and / or base hydrolysis of connective tissue proteins (e.g., collagen).

[0088] In certain embodiments, the polymer composition of the present disclosure comprises a chemically modified gelatin. In certain embodiments, the polymer composition comprises acrylated gelatin. In certain embodiments, the polymer composition comprises gelatin methacryloyl (i.e., GelMA). In certain embodiments, the chemically modified gelatin can be included in the precursor polymer composition of the present disclosure. In certain embodiments, the chemically modified gelatin comprises a photocrosslinkable derivative of gelatin. In certain embodiments, the chemically modified gelatin can be modified with acrylic anhydride or acryloyl chloride (substituted or unsubstituted) to form acryloyl-substituted gelatin. In certain embodiments, the chemically modified gelatin can be modified with one or more crosslinkable groups selected from methyl acrylate, ethyl acrylate, propyl acrylate, methyl methacrylate, ethyl methacrylate, methacryloyl, catechol, ethylene oxide, or propylene oxide. In certain embodiments, the chemically modified gelatin can be modified with methacrylic anhydride (MA) (also known as methacryloyl anhydride) to form methacryloyl-substituted gelatin (commonly referred to as gelatin methacryloyl, or GelMA). FIG. 1A provides an example of a reaction in which gelatin is modified with methacrylic anhydride to form methacryloyl-substituted gelatin (GelMA).

[0089] In certain embodiments, the acryloyl modification of gelatin can be carried out by a synthetic reaction of a functionalized compound containing an acrylate group with gelatin. In certain embodiments, the methacryloyl modification of gelatin can be carried out by a synthetic reaction of gelatin with methacrylic anhydride, methacryloyl chloride, 2-isocyanatoethyl methacrylate, 2-hydroxyethyl methacrylate, glycidyl methacrylate, N-hydroxysuccinimide ester of methacrylic acid, allyl methacrylate, vinyl methacrylate, bis(2-methacryloyl)oxyethyl disulfide, 2-hydroxy-5-N-methacrylamidobenzoic acid, or a mixture thereof.

[0090] As used herein, the terms "acryloyl-substituted gelatin" and "acrylated gelatin" can describe gelatin having free amines (e.g., lysine, arginine, asparagine, or glutamine side chains) and / or free hydroxyls (e.g., serine, threonine, aspartic acid or glutamic acid side chains) substituted with at least one acryloyl group. Generally, the acryloyl group is of the formula H 2 C=CR’-C(=O)-R, an a,b-unsaturated carbonyl compound, where R’ is, without limitation, hydrogen, halogen, hydroxyl, C 1 ~C 5 alkoxy, C 1 ~C 5 alkyl, C 3 ~C 8 cycloalkyl, C 1 ~C 5 heteroalkyl, C 3 ~C 8 heterocycloalkyl, aryl, heteroaryl or an amino group, each optionally substituted with halogen, C 1 ~C 5 alkoxy, C 1 ~C 5 alkyl, C 3 ~C 8 cycloalkyl, C 1 ~C 5 heteroalkyl, C 3 ~C 8 heterocycloalkyl, aryl, heteroaryl or an amino group. For the acryloyl-substituted gelatin of the present disclosure, the R group represents the terminal amine and / or hydroxyl group on the gelatin that is the subject of acryloyl functionalization.

[0091] In certain embodiments, the R’ group of the acryloyl moiety is methyl, and is commonly referred to as a methacryloyl group. As used herein, the terms “methacryloyl-substituted gelatin,” “gelatin methacryloyl,” and “GelMA” can describe gelatin having at least one methacryloyl group, such as a methacrylamide group (derived from a free amine on the gelatin) and / or a methacrylate group (derived from a free hydroxyl on the gelatin), substituting a free amine (e.g., lysine, arginine, asparagine, or glutamine side chain) and / or a free hydroxyl (e.g., serine, threonine, aspartic acid, or glutamic acid side chain).

[0092] In certain embodiments, the chemically modified gelatin (e.g., GelMA) can be present in the polymer composition at a concentration of about 1% to about 60% weight / volume (w / v). In certain embodiments, the chemically modified gelatin (e.g., GelMA) can be present in the polymer composition at a weight / volume concentration (w / v) of about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, or about 60%. In certain embodiments, the chemically modified gelatin (e.g., GelMA) can be present in the polymer composition at a weight / volume concentration (w / v) of about 1 - 3%, about 3 - 6%, about 6 - 10%, about 1 - 5%, about 1 - 10%, about 5 - 10%, about 11 - 13%, about 13 - 16%, about 16 - 20%, about 10 - 20%, about 10 - 15%, about 15 - 20%, about 21 - 23%, about 23 - 26%, about 26 - 30%, about 20 - 30%, about 20 - 25%, about 25 - 30%, about 31 - 33%, about 33 - 36%, about 36 - 40%, about 30 - 40%, about 30 - 35%, about 35 - 40%, about 41 - 43%, about 43 - 46%, about 46 - 50%, about 40 - 50%, about 40 - 45%, about 45 - 50%, about 51 - 53%, about 53 - 56%, about 56 - 60%, about 50 - 60%, about 50 - 55%, or about 55 - 60%.

[0093] In certain embodiments, the polymer composition comprises acrylated gelatin (i.e., GelMA) having an acryloyl substitution degree (i.e., methacryloyl functionalization). As used herein, the term "acryloyl substitution degree" can describe the proportion of free amines and hydroxyls in gelatin substituted with acryloyl groups. As used herein, the term "methacryloyl substitution degree" can describe the proportion of free amines and hydroxyls in gelatin substituted with methacryloyl groups. In certain embodiments, the polymer composition comprises acrylated gelatin having an acryloyl substitution degree of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90%. In certain embodiments, the polymer composition comprises acrylated gelatin having an acryloyl substitution degree of about 10 - 99%. In certain embodiments, the acryloyl substitution degree is about 1 - 5%, about 5 - 10%, about 10 - 15%, about 15 - 20%, about 20 - 25%, about 25 - 30%, about 30 - 35%, about 35 - 40%, about 40 - 45%, about 45 - 50%, about 50 - 55%, about 55 - 60%, about 60 - 65%, about 65 - 70%, about 70 - 75%, about 75 - 80%, about 80 - 85%, about 85 - 90%, about 90 - 95%, or about 95 - 99%. In certain embodiments, the polymer composition comprises GelMA having a methacryloyl substitution degree of about 1 - 5%, about 5 - 10%, about 10 - 15%, about 15 - 20%, about 20 - 25%, about 25 - 30%, about 30 - 35%, about 35 - 40%, about 40 - 45%, about 45 - 50%, about 50 - 55%, about 55 - 60%, about 60 - 65%, about 65 - 70%, about 70 - 75%, about 75 - 80%, about 80 - 85%, about 85 - 90%, about 90 - 95%, or about 95% - 99%.

[0094] In certain embodiments, the polymer composition comprises GelMA having a methacrylamide substitution degree (i.e., methacrylamide functionalization). In certain embodiments, the polymer composition comprises GelMA having a methacrylamide substitution degree of at least about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or at least about 90%. In certain embodiments, the polymer composition comprises GelMA having a methacrylamide substitution degree of about 20 - 90%. In certain embodiments, the methacrylamide substitution degree is about 1 - 5%, about 5 - 10%, about 10 - 15%, about 15 - 20%, about 20 - 25%, about 25 - 30%, about 30 - 35%, about 35 - 40%, about 40 - 45%, about 45 - 50%, about 50 - 55%, about 55 - 60%, about 60 - 65%, about 65 - 70%, about 70 - 75%, about 75 - 80%, about 80 - 85%, or about 85 - 90%. In certain embodiments, the methacrylamide substitution degree can be measured using proton nuclear magnetic resonance. In certain embodiments, the methacrylamide substitution degree can be measured using a fluoraldehyde assay.

[0095] In certain embodiments, the polymer composition can include GelMA having a methacrylate substitution degree (i.e., methacrylate functionalization). In certain embodiments, the polymer composition includes GelMA having a methacrylate substitution degree of at least about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, or at least about 90%. In certain embodiments, the polymer composition includes GelMA having a methacrylate substitution degree of about 20 - 90%. In certain embodiments, the methacrylate substitution degree is about 1 - 5%, about 5 - 10%, about 10 - 15%, about 15 - 20%, about 20 - 25%, about 25 - 30%, about 30 - 35%, about 35 - 40%, about 40 - 45%, about 45 - 50%, about 50 - 55%, about 55 - 60%, about 60 - 65%, about 65 - 70%, about 70 - 75%, about 75 - 80%, about 80 - 85%, or about 85 - 90%. In certain embodiments, the methacrylate substitution degree can be measured using proton nuclear magnetic resonance. In certain embodiments, the methacrylate substitution degree can be measured using an Fe(III)-hydroxamate-based assay. In certain embodiments, the measurement of the methacrylate substitution degree can include an aminolysis reaction that converts the methacrylate group to an N-hydroxymethacrylamide group (e.g., by exposure to a hydroxylamine solution).

[0096] In certain embodiments, the polymer composition comprises GelMA having a degree of methacrylamide substitution and a degree of methacrylate substitution. In certain embodiments, the ratio of methacrylamide substitution to methacrylate substitution in GelMA is from about 1:1 to 99:1. In some embodiments, the ratio of methacrylamide substitution to methacrylate substitution is about 1:1 to 2:1, about 2:1 to 3:1, about 3:1 to 4:1, about 4:1 to 5:1, about 1:1 to 5:1, about 5:1 to 10:1, about 10:1 to 15:1, about 15:1 to 20:1, about 20:1 to 25:1, about 25:1 to 30:1, about 30:1 to 35:1, about 35:1 to 40:1, about 40:1 to 45:1, about 45:1 to 50:1, about 50:1 to 55:1, about 55:1 to 60:1, about 60:1 to 65:1, about 65:1 to 70:1, about 70:1 to 75:1, about 75:1 to 80:1, about 80:1 to 85:1, about 85:1 to 90:1, about 90:1 to 95:1, or about 95:1 to 99:1. In certain embodiments, the ratio of methacrylate substitution to methacrylamide substitution in GelMA is from about 1:1 to 99:1. In some embodiments, the ratio of methacrylate substitution to methacrylamide substitution is about 1:1 to 2:1, about 2:1 to 3:1, about 3:1 to 4:1, about 4:1 to 5:1, about 1:1 to 5:1, about 5:1 to 10:1, about 10:1 to 15:1, about 15:1 to 20:1, about 20:1 to 25:1, about 25:1 to 30:1, about 30:1 to 35:1, about 35:1 to 40:1, about 40:1 to 45:1, about 45:1 to 50:1, about 50:1 to 55:1, about 55:1 to 60:1, about 60:1 to 65:1, about 65:1 to 70:1, about 70:1 to 75:1, about 75:1 to 80:1, about 80:1 to 85:1, about 85:1 to 90:1, about 90:1 to 95:1, or about 95:1 to 99:1.

[0097] In certain embodiments, the polymer composition comprises GelMA having a methacryloyl modification of gelatin effected by reaction of gelatin with methacrylic anhydride. In certain embodiments, the polymer composition comprises GelMA having a methacryloyl modification of gelatin effected by reaction of gelatin with glycidyl methacrylate.

[0098] In certain embodiments, gelatin can be functionalized with an anchor integrin and / or a protein (e.g., a protein that binds to a surface protein of a target surface). Such functionalization can occur with a poly(ethylene glycol) (PEG) or other polymeric linker between the gelatin and the integrin and / or protein.

[0099] Chemically modified hyaluronic acid Hyaluronic acid (HA) is a viscoelastic and biocompatible glycosaminoglycan that occurs naturally in the cornea and other tissues. In certain embodiments, the polymeric compositions of the present disclosure include chemically modified hyaluronic acid (HA). In certain embodiments, the polymeric composition includes acryloyl-substituted hyaluronic acid. In certain embodiments, the polymeric composition includes methacrylated hyaluronic acid (MeHA). In certain embodiments, the chemically modified HA can be included in the precursor polymeric compositions of the present disclosure. In certain embodiments, the chemically modified HA includes a photocrosslinkable derivative of HA. In certain embodiments, the chemically modified HA includes methacrylated hyaluronic acid (MeHA). In certain embodiments, the chemically modified HA includes methacrylated hyaluronic acid (MeHA) containing methacrylic anhydride-hyaluronic acid (HAMA), i.e., MeHA formed by the reaction of methacrylic anhydride and hyaluronic acid. In certain embodiments, the chemically modified HA includes methacrylated hyaluronic acid (MeHA) containing glycidyl methacrylate-hyaluronic acid (HAGM), i.e., MeHA formed by the reaction of glycidyl methacrylate and hyaluronic acid. In certain embodiments, methacrylation of HA can be carried out by a combination of a ring-opening reaction of the HA backbone and a reversible transesterification reaction. Figure 1B provides an example of a reaction in which hyaluronic acid is modified with glycidyl methacrylate to form the HAGM type of methacrylated hyaluronic acid (MeHA).

[0100] In certain embodiments, the chemically modified HA (e.g., MeHA) can be present in the polymer composition at a concentration of about 1% to about 60% weight / volume (w / v). In certain embodiments, the chemically modified HA (e.g., MeHA) can be present in the polymer composition at a weight / volume concentration (w / v) of about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, or about 60%. In certain embodiments, the chemically modified HA (e.g., MeHA) is present in the polymer composition at a weight / volume concentration (w / v) of about 1-3%, about 3-6%, about 6-10%, about 1-5%, about 1-10%, about 5-10%, about 11-13%, about 13-16%, about 16-20%, about 10-20%, about 10-15%, about 15-20%, about 21-23%, about 23-26%, about 26-30%, about 20-30%, about 20-25%, about 25-30%, about 31-33%, about 33-36%, about 36-40%, about 30-40%, about 30-35%, about 35-40%, about 41-43%, about 43-46%, about 46-50%, about 40-50%, about 40-45%, about 45-50%, about 51-53%, about 53-56%, about 56-60%, about 50-60%, about 50-55%, or about 55-60%.

[0101] In certain embodiments, the polymer compositions of the present disclosure comprise acryloyl-substituted gelatin (e.g., GelMA) and acryloyl-substituted hyaluronic acid (e.g., MeHA) in a ratio of from about 30:1 to about 1:30 w / w. In certain embodiments, the polymer compositions of the present disclosure comprise acryloyl-substituted gelatin and acryloyl-substituted hyaluronic acid in a ratio (w / w) of about 30:1, about 29:1, about 28:1, about 27:1, about 26:1, about 25:1, about 24:1, about 23:1, about 22:1, about 21:1, about 20:1, about 19:1, about 18:1, about 17:1, about 16:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, or about 1:30.

[0102] In certain embodiments, acryloyl-substituted hyaluronic acid (e.g., MeHA) can be synthesized as taught in Bencherif et al., Biomaterials 29, 1739-1749 (2008), or Prata et al., Biomacromolecules 11, 769-775 (2010), each of which is incorporated herein by reference in its entirety to the extent that each describes the composition, production, analysis, and use of acryloyl-substituted hyaluronic acid polymer compositions such as MeHA.

[0103] Chemically modified poly(ethylene glycol) Poly(ethylene glycol) (PEG) is a synthetic linear polymer known to have high biocompatibility and immunotolerance in the human body and is soluble in many aqueous and organic solvents. In certain embodiments, the polymer compositions of the present disclosure include chemically modified PEG. In certain embodiments, the polymer compositions include acryloyl-substituted PEG. In certain embodiments, the polymer compositions include one or more acryloyl-substituted PEGs selected from the following: PEG diacrylate (PEGDA), PEG monoacrylate, PEG dimethacrylate, PEG monomethacrylate, methoxy PEG acrylate, methoxy PEG methacrylate, ethoxy PEG acrylate, ethoxy PEG methacrylate, propoxy PEG acrylate, or propoxy PEG methacrylate.

[0104] In certain embodiments, the polymer compositions include poly(ethylene glycol) diacrylate (PEGDA). In certain embodiments, the chemically modified PEG can be included in the precursor polymer compositions of the present disclosure. In certain embodiments, the chemically modified PEG includes a photocrosslinkable derivative of PEG. In certain embodiments, the chemically modified PEG includes poly(ethylene glycol) diacrylate (PEGDA). In certain embodiments, the chemical modification of PEG can be carried out by reacting PEG with acryloyl chloride or a functionally similar acrylated compound. Figure 1C provides an example of a reaction in which poly(ethylene glycol) (PEG) is modified with acryloyl chloride to form poly(ethylene glycol) diacrylate (PEGDA).

[0105] In certain embodiments, the chemically modified PEG has a molecular weight of from about 5 kDa to about 200 kDa. In certain embodiments, the chemically modified PEG can have a molecular weight of about 5 - 10 kDa, about 10 - 15 kDa, about 15 - 20 kDa, about 20 - 25 kDa, about 25 - 30 kDa, about 30 - 35 kDa, about 35 - 40 kDa, about 40 - 45 kDa, about 45 - 50 kDa, about 50 - 55 kDa, about 55 - 60 kDa, about 60 - 65 kDa, about 65 - 70 kDa, about 70 - 75 kDa, about 75 - 80 kDa, about 80 - 85 kDa, about 85 - 90 kDa, about 90 - 95 kDa, about 95 - 100 kDa, about 100 - 105 kDa, about 105 - 110 kDa, about 110 - 115 kDa, about 115 - 120 kDa, about 120 - 125 kDa, about 125 - 130 kDa, about 130 - 135 kDa, about 135 - 140 kDa, about 140 - 145 kDa, about 145 - 150 kDa, about 150 - 155 kDa, about 155 - 160 kDa, about 160 - 165 kDa, about 165 - 170 kDa, about 170 - 175 kDa, about 175 - 180 kDa, about 180 - 185 kDa, about 185 - 190 kDa, or about 195 - 200 kDa.

[0106] In certain embodiments, the chemically modified PEG (e.g., PEGDA) can be present in the polymer composition at a concentration of about 1 wt% to about 60% weight / volume (w / v). In certain embodiments, the chemically modified PEG (e.g., PEGDA) can be present in the polymer composition at a weight / volume concentration (w / v) of about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, or about 60%. In certain embodiments, the chemically modified PEG (e.g., PEGDA) can be present in the polymer composition at a weight / volume concentration (w / v) of about 1-3%, about 3-6%, about 6-10%, about 1-5%, about 1-10%, about 5-10%, about 11-13%, about 13-16%, about 16-20%, about 10-20%, about 10-15%, about 15-20%, about 21-23%, about 23-26%, about 26-30%, about 20-30%, about 20-25%, about 25-30%, about 31-33%, about 33-36%, about 36-40%, about 30-40%, about 30-35%, about 35-40%, about 41-43%, about 43-46%, about 46-50%, about 40-50%, about 40-45%, about 45-50%, about 51-53%, about 53-56%, about 56-60%, about 50-60%, about 50-55%, or about 55-60%.

[0107] In certain embodiments, the polymer compositions of the present disclosure comprise acryloyl-substituted gelatin (e.g., GelMA) and acryloyl-substituted PEG (e.g., PEGDA) in a ratio of about 30:1 to about 1:30 w / w. In certain embodiments, the polymer compositions of the present disclosure comprise acryloyl-substituted gelatin and acryloyl-substituted PEG in a ratio of about 30:1, about 29:1, about 28:1, about 27:1, about 26:1, about 25:1, about 24:1, about 23:1, about 22:1, about 21:1, about 20:1, about 19:1, about 18:1, about 17:1, about 16:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, or about 1:30 (w / w).

[0108] In certain embodiments, the polymer compositions of the present disclosure comprise acryloyl-substituted PEG (e.g., PEGDA) and acryloyl-substituted hyaluronic acid (e.g., MeHA) in a ratio of about 30:1 to about 1:30 w / w. In certain embodiments, the polymer compositions of the present disclosure comprise acryloyl-substituted PEG and acryloyl-substituted hyaluronic acid in a ratio of about 30:1, about 29:1, about 28:1, about 27:1, about 26:1, about 25:1, about 24:1, about 23:1, about 22:1, about 21:1, about 20:1, about 19:1, about 18:1, about 17:1, about 16:1, about 15:1, about 14:1, about 13:1, about 12:1, about 11:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, or about 1:30 (w / w).

[0109] In certain embodiments, the polymer compositions of the present disclosure comprise one or more synthetic polymer components (i.e., polymers or precursors) selected from methacrylate-oligolactide-PEO-oligolactide-methacrylate, polyethylene glycol (PEG), polyglycerol sebacate (PGS), polylactic acid (PLA), polypropylene glycol (PPO), PEG-PPO-PEG copolymers (e.g., pluronics), polyphosphazenes, polymethacrylic acid, poly(N-vinylpyrrolidone), and polyethyleneimine.

[0110] Crosslinking agent In certain embodiments, the polymer compositions of the present disclosure include a crosslinking agent. As used herein, the phrase "crosslinking agent" can describe a substance that forms, facilitates, or modulates intermolecular bonds (covalent, ionic, hydrogen bonds) between polymer units or chains to create a network of polymer chains. A crosslinking agent typically exhibits one or more, optionally two or more, bonding functional groups capable of forming chemical bonds between two or more polymer chains. The crosslinking agent can include, for example, two vinyl bonds (tetrafunctional) or three amines (trifunctional).

[0111] In certain embodiments, the polymer composition includes a crosslinking agent that can be used to activate or facilitate the polymerization, gelation, and solidification of the polymer composition from a precursor polymer composition to a gel polymer composition. In certain embodiments, when the polymer composition of the present disclosure (e.g., a precursor polymer composition) is exposed to crosslinking conditions (e.g., exposure to visible light in the presence of a photoinitiator), one or more acryloyl groups in the polymer composition (e.g., acryloyl-substituted gelatin, acryloyl-substituted HA, acryloyl-substituted PEG, acryloyl-substituted tropoelastin, and other acryloyl-based crosslinking agents) can react with other acryloyl groups to crosslink the polymer composition and form a gel polymer composition (e.g., a GelMA hydrogel).

[0112] In certain embodiments, the polymer compositions of the present disclosure (e.g., precursor polymer compositions) may include one or more crosslinking agents at about 1% to about 50% (w / v). In certain embodiments, the polymer composition may include one or more crosslinking agents at a concentration (w / v) of at least about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, or about 40%. In certain embodiments, the polymer composition may include one or more crosslinking agents at a concentration (w / v) of about 50%, about 45%, about 40%, about 35%, or about 30% or less. In certain embodiments, the polymer composition includes one or more crosslinking agents at a concentration (w / v) of about 1-3%, about 3-6%, about 6-10%, about 1-5%, about 1-10%, about 5-10%, about 11-13%, about 13-16%, about 16-20%, about 10-20%, about 10-15%, about 15-20%, about 21-23%, about 23-26%, about 26-30%, about 20-30%, about 20-25%, about 25-30%, about 31-33%, about 33-36%, about 36-40%, about 30-40%, about 30-35%, about 35-40%, about 41-43%, about 43-46%, about 46-50%, about 40-50%, about 40-45%, or about 45-50%.

[0113] In certain embodiments, the polymer compositions of the present disclosure may include one or more crosslinking agents selected from glutaraldehyde, epoxides (e.g., bis-oxirane), oxidized dextran, p-azidobenzoyl hydrazide, N-(α-maleimidopropionyloxy) succinimide ester, p-azidophenylglyoxal monohydrate, bis((4-azidosalicylamido)ethyl) disulfide, suberic acid bis(sulfo-succinimidyl), dithiobis(propionic acid succinimidyl), suberic acid disuccinimidyl, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), ethoxylated trimethylolpropane triacrylate, N-hydroxysuccinimide (NHS), derivatives thereof, or combinations thereof.

[0114] In certain embodiments, the polymer composition of the present disclosure comprises one or more crosslinking agents selected from polyethylene oxide dimethacrylate, methylene bisacrylamide, methylene bis(2-methylacrylamide), methylene diacrylate, methylene bis(2-methylacrylate), diethylene glycol diacrylate, hexamethylene diacrylate, hexamethylene diisocyanate, oxybis(methylene)bis(2-methylacrylate), oxybis(ethane-2,1-diyl)bis(2-methylacrylate), trimethylolpropane triacrylate, pentaerythritol triacrylate, tris(2-hydroxyethyl)isocyanurate triacrylate, isocyanuric acid tris(2-acryloyloxyethyl)ester, ethoxylated trimethylolpropane triacrylate, pentaerythrityl triacrylate, and glycerol triacrylate, phosphinylidene tris(oxyethylene)triacrylate, derivatives thereof, or combinations thereof.

[0115] Polymer Crosslinking Initiator / Photoinitiator In certain embodiments, the polymer composition comprises one or more polymer crosslinking initiators such as a photoinitiator element. In certain embodiments, the polymer crosslinking initiator forms free radicals when exposed to certain polymer crosslinking conditions (e.g., acidic conditions, basic conditions, high salt conditions, low salt conditions, high temperature, stirring, dissolution conditions, light exposure), and the free radicals can result in the formation of bonds between reactive groups in the composition, such as vinyl bond crosslinking between methacrylate groups in the GelMA polymer composition.

[0116] In certain embodiments, the polymer composition comprises one or more photoinitiator elements (i.e., crosslinking initiators that are initiated or activated by absorbing light of a specific wavelength). In certain embodiments, the precursor polymer composition of the present disclosure comprises one or more photoinitiator elements. In certain embodiments, the photoinitiator element can be activated by exposure to light. In certain embodiments, light exposure can activate the photoinitiator to form free radicals, which can result in the formation of bonds between reactive groups in the composition, such as vinyl bond crosslinking between methacrylate groups in the GelMA polymer composition.

[0117] In certain embodiments, the photoinitiator element can be activated by exposure to one or more light sources selected from visible light sources (e.g., white or blue light), ultraviolet (UV) light sources, near-infrared (NIR) light sources, and fluorescent light sources. In certain embodiments, the photoinitiator element comprises a visible light-activated photoinitiator, such as a visible light-activated photoinitiator that is activated upon exposure to light having a wavelength of from about 380 nm to about 740 nm. In certain embodiments, the visible light-activated photoinitiator can be activated upon exposure to light having a wavelength of about 380 - 435 nm (i.e., violet light), about 435 - 500 nm (i.e., blue light), about 500 - 565 nm (i.e., green light), about 565 - 600 nm (i.e., yellow light), about 600 - 650 nm (i.e., orange light), or about 650 - 740 nm (i.e., red light). In certain embodiments, the photoinitiator element comprises an ultraviolet-activated photoinitiator. In certain embodiments, the photoinitiator element comprises a near-infrared light (NIR)-activated photoinitiator. In certain embodiments, the photoinitiator element comprises a white light-activated photoinitiator. In certain embodiments, the photoinitiator element comprises a blue light-activated photoinitiator.

[0118] In certain embodiments, the polymer composition comprises triethanolamine; 1-vinyl-2-pyrrolidone (NVP); N-vinylcaprolactam (NVC); ethylene glycol diacrylate (EGDA); riboflavin; azobisisobutyronitrile; benzoyl peroxide; 1-benzoylcyclohexanol; di-tert-butyl peroxide; eosin Y (e.g., disodium salt), (2-(2,4,5,7-tetrabromo-6-oxide-3-oxo-3H-xanthen-9-yl)benzoate); 4,6-trimethylbenzoylphosphinate; triethanolamine; 2,3-diketo-1,7,7-trimethylnorbornane; 1-phenyl-1,2-propadione; 2,4,6-trimethylbenzoyl-diphenylphosphine oxide; bis(2,6-dichlorobenzoyl)-(4-propylphenyl)phosphine oxide; 4,4'-bis(dimethylamino)benzophenone; 4,4'-bis(diethylamino)benzophenone; 2-chlorothioxanthone-9-one, 4-(dimethylamino)benzophenone; phenanthrenequinone; ferrocene; 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone; 2-hydroxy-2-methylpropiophenone; diphenyl(2,4,6trimethylbenzoyl)phosphine oxide / 2-hydroxy-2-methylpropiophenone (blend); benzoin methyl ether; benzoin isopropyl ether; 2,2-diethoxyacetophenone; dibutoxyacetophenone; 2,2-dimethoxy-2-phenyl-1-phenylethanone; 2,2-dimethoxy-2-phenylacetophenone; dibenzosuberone; (benzene)tricarbonylchromium; resazurin; resorufin; benzoyltrimethylgermanium; lithium phenyl-2,4,6-trimethyl-benzoylphosphinate; camphorquinone; 2-methyl-1-(4-(methylthio)phenyl)-2-morpholinopropan-2-one; 2-benzyl-2-(dimethylamino)-4'-morpholinobutyrophenone; 2-benzyl-2-dimethylamino-1-(4-morpholinophenyl)butan-1-one; methybenzoylformate;Bis(2,4,6-trimethylbenzoyl)-phenylphosphine oxide, bis(2,4-cyclopentadien-1-yl)-bis(2,6-difluoro-3-(1H-pyrrol-1-yl)-phenyl)titanium; 5,7-diiodo-3-butoxy-6-fluorone; 2,4,5,7-tetraiodo-3-hydroxy-6-fluorone; 2,4,5,7-tetraiodo-3-hydroxy-9-cyano-6-fluorone; dimethoxyhydroxy-acetophenone; 2-naphthalenesulfonyl chloride; 1-phenyl-1,2-propanedione-2-(O-ethoxy-carbonyl)oxime; 2-ethylthioxanthone; 2-isopropylthioxanthone; 2,4-diethylthioxanthone; 2-tert-butylthioxanthone; 2-chlorothioxanthone; 2-propoxythioxanthone; methylphenylglycoxylate; phenyl 2-hydroxy-2-propyl ketone; 4-isopropylphenyl 2-hydroxy-2-propyl ketone; 4-n-dodecylphenyl 2-hydroxy-2-propyl ketone; 4-(2-hydroxyethoxy)phenyl 2-hydroxy-2-propyl ketone; 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propan-1-one); 4-(2-acryloyloxyethoxy)phenyl 2-hydroxy-2-propyl ketone; one or more photoinitiator elements selected from derivatives thereof, or combinations thereof. In certain embodiments, the polymer composition comprises a combination of eosin Y, triethanolamine, and / or vinylcaprolactam. In certain embodiments, the polymer crosslinking initiator comprises eosin Y disodium salt (EYDS), N-vinylcaprolactam (NVC), triethanolamine, or any combination thereof. In certain embodiments, the polymer crosslinking initiator comprises eosin Y disodium salt (EYDS), N-vinylpyrrolidone (NVP), triethanolamine, or any combination thereof.;

[0119] In certain embodiments, the polymer crosslinking initiator comprises (i) about 50 μM eosin Y or eosin Y disodium salt (EYDS), optionally about 50 μM eosin Y disodium salt (EYDS), (ii) about 3.5 to about 5.0 μL / mL N-vinylcaprolactam (NVC) or N-vinylpyrrolidone (NVP), optionally about 3.5 to about 5.0 μL / mL NVP, optionally about 5.0 μL / mL NVP, and (iii) triethanolamine, optionally about 1.5% v / v triethanolamine.

[0120] In certain embodiments, the polymer composition is acetophenone, anisoin, anthraquinone, anthraquinone-2-sulfonic acid, sodium salt monohydrate, (benzene)tricarbonylchromium, 4-(boc-aminomethyl)phenyl isothiocyanate, benzene, benzoin, benzoin ethyl ether, benzoin isobutyl ether, benzoin methyl ether, benzoic acid, benzoyl-phenyl-hydroxycyclohexyl phenyl ketone, 3,3’,4,4’-benzophenone tetracarboxylic dianhydride, 4-benzoylbiphenyl, 2-benzyl-2-(dimethylamino)-4’-morpholinobutyrophenone, 4,4’-bis(diethylamino)benzophenone, 4,4’-bis(dimethylamino)benzophenone, Michler's ketone, camphorquinone, 2-chlorothioxanthen-9-one, 5-dibenzosuberone, (cumene)cyclopentadienyliron(II) hexafluorophosphate, dibenzosuberone, 2,2-diethoxyacetophenone, 4,4’-dihydroxybenzophenone, 2,2-dimethoxy-2-phenylacetophenone, 4-(dimethylamino)benzophenone, 4,4’-dimethylbenzyl, 2,5-dimethylbenzophenone, 3,4-dimethylbenzophenone, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide, 2-hydroxy-2-methylpropiophenone, 4’-ethoxyacetophenone, 2-ethylanthraquinone, ferrocene, 3’-hydroxyacetophenone, 4’-hydroxyacetophenone, 3-hydroxybenzophenone, 4-hydroxybenzophenone, 1-hydroxycyclohexyl phenyl ketone, 2-hydroxy-2-methylpropiophenone, 2-methylbenzophenone, 3-methylbenzophenone, methybenzoylformate, 2-methyl-4’-(methylthio)-2-morpholinopropiophenone, 9,10 - phenanthrenequinone, 4'-phenoxyacetophenone, thioxanthen-9-one, triarylsulfonium hexafluoroantimonate, triallylsulfonium hexafluorophosphate, 3-mercapto-1-propanol, mercapto-1-undecanol, 1-mercapto-2-propanol, 3-mercapto-2-butanol, hydrogen peroxide, benzoyl peroxide, 4,4'-dimethoxybenzoin, 2,2-dimethoxy-2-phenylacetophenone, dibenzoyldisulfide, diphenyldithiocarbonate, 2,2'-azobisisobutyronitrile, 2,2'-azobis[2-methyl-N-(2-hydroxyethyl)propionamide], camphorquinone, eosin, dimethylaminobenzoate, dimethoxy-2-phenyl-acetophenone; Quanta-cure ITX X-ray sensitizer, Irgacure (e.g., 907, 2959, 651), Darocur 2959, ethyl-4-N,N-dimethylaminobenzoate, 1-[- (4-benzoylphenylsulfanyl)phenyl]-2-methyl-2-(4-methylphenylsulfonyl)propan-1-one, 1-hydroxy-cyclohexyl-phenyl-ketone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide, diphenyl(2,4,6-trimethylbenzoyl)phosphine, 2-ethylhexyl-4-dimethylaminobenzoate, 2-hydroxy-2-methyl-1-phenyl-1-propanone, oligo[2-hydroxy-2-methyl-1-[4-(methylvinyl)phenyl]propanone] and propoxylated glyceryl triacrylate, benzii dimethyl ketal, benzophenone, blend of benzophenone and α-hydroxy-cyclohexyl-phenylketone, blend of Esacure KIP 150 and Esacure TZT, blend of Esacure KIP150 and Esacure TZT, blend of Esacure KIP150 and TPGDA; phosphine oxide, blend of Esacure KIP 150 and Esacure TZT; difunctional α-hydroxyketone, ethyl 4-dimethylaminobenzoate, isopropylthioxanthone, 2-hydroxy-2methyl-phenylpropanone, 2,4,6,- One or more photoinitiator elements selected from blends of trimethylbenzoyldiphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 4-methylbenzophenone and benzophenone, oligo(2-hydroxy-2-methyl-1-(4(1-methylvinyl)phenyl)propanone, oligo(2-hydroxy-2-methyl-1-4(1-methylvinyl)phenylpropanone and 2-hydroxy-2-methyl-1-phenyl-1-propanone, 4-methylbenzophenone, trimethylbenzophenone and methylbenzophenone; and aqueous emulsions of 2,4,6-trimethylbenzoylphosphine oxide, alpha-hydroxyketone, trimethylbenzophenone, and 4-methylbenzophenone.,

[0121] In certain embodiments, the polymer composition comprises one or more cationic and / or anionic photoinitiator elements selected from titanium tetrachloride, vanadium tetrachloride, bis(cyclopentadienyl)titanium dichloride, ferrocene, cyclopentadienylmanganetricarbonyl, manganese decacarbonyl, diazonium salts, diaryliodonium salts (e.g., 3,3'-dinitrodiphenyliodonium hexafluoroarsenate, diphenyliodonium fluoroborate, 4-methoxydiphenyliodonium fluoroborate) and triarylsulfonium salts.,

[0122] For photoinitiation and photoinitiators, see Rabek, Mechanisms of Photophysical Processes and Photochemical Reactions in Polymers, New York: Wiley & Sons, 1987, and Fouassier, Photoinitiation, Photopolymerization, Photocuring, Cincinnati, Ohio: Hanser / Gardner, Fisher et al., 2001, Annu. Rev. Mter. Res., 31:171, each of which is incorporated herein by reference in its entirety to the extent each describes the polymerization and use of photoinitiators in the production of polymer compositions containing acryloyl gelatin such as GelMA hydrogels.

[0123] In certain embodiments, the polymer composition comprises one or more metal 2+ ions and / or metal 3+ ion-containing crosslinkers or initiators. In certain embodiments, the polymer composition comprises Fe 2+ 、Fe 3+ 、Ni 2+ 、Zn 2+ 、Cu 2+ 、Ag 2+ 、Au 3+ 、Co 2+ 、Co 3+ 、Cr 2+ 、Cr 3+ 、Cd 2+ 、Mn 2+ 、Mg 2+ 、Pd 2+ 、Pt 2+ 、Al 3+ 、or one or more metal 2+ ions and / or metal 3+ ion-containing crosslinkers selected from combinations thereof. In certain embodiments, the precursor polymer composition of the present disclosure comprises both one or more photoinitiator elements and one or more metal 2+ / 3+ ions.

[0124] In certain embodiments, the polymer composition includes a crosslinking agent or initiator that uses click bioconjugation chemistry for polymer crosslinking. In certain embodiments, the polymer composition includes a crosslinking agent or initiator that uses click bioconjugation chemistry selected from azide-alkyne cycloaddition reactions with metal catalysts, strain-promoted azide-alkyne cycloadditions, strain-promoted alkyne-nitrone cycloadditions (e.g., alkene / azide [3+2] cycloadditions, alkene / tetrazine inverse electron demand Diels-Alder reactions, alkene / tetrazole photoclick reactions).

[0125] II. Physical, Mechanical, and Structural Characteristics Viscosity, Shear Strength, and Shear Resistance The viscosity of a material is a measure of the resistance of the material to deformation at a given rate. The viscosity of a fluid material often correlates with the thickness and / or density of that material.

[0126] In certain embodiments, the polymer compositions of the present disclosure can have a therapeutically effective viscosity. In certain embodiments, the polymer composition can have a viscosity that provides strong adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the precursor polymer compositions of the present disclosure can have a viscosity that provides strong adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the precursor polymer composition can have a viscosity greater than water. In certain embodiments, the precursor polymer composition can have a viscosity equivalent to that of a paste. In certain embodiments, the gel polymer compositions of the present disclosure can have a viscosity that provides strong adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the precursor polymer composition can have a viscosity equivalent to that of water. In certain embodiments, the gel polymer composition can retain its shape and / or consistency on the surface of the target tissue for one hour or more, one day or more, or one week or more.

[0127] In certain embodiments, the polymeric composition can have a viscosity of about 0.5 Pascal-seconds (Pa·s) to about 300 Pa·s at a low shear rate (e.g., a shear rate of about 0.001 s -1 to about 1 s -1 . In certain embodiments, the polymeric composition can have a viscosity of about 0.5 to 100 Pa·s at a low shear rate. In certain embodiments, the polymeric composition can have a viscosity of about 0.5 to 5 Pa·s, about 5 to 10 Pa·s, about 10 to 15 Pa·s, about 15 to 20 Pa·s, about 20 to 25 Pa·s, about 25 to 30 Pa·s, about 30 to 35 Pa·s, about 35 to 40 Pa·s, about 40 to 45 Pa·s, about 45 to 50 Pa·s, about 50 to 55 Pa·s, about 55 to 60 Pa·s, about 60 to 65 Pa·s, about 65 to 70 Pa·s, about 70 to 75 Pa·s, about 75 to 80 Pa·s, about 80 to 85 Pa·s, about 85 to 90 Pa·s, about 90 to 95 Pa·s, about 95 to 100 Pa·s, about 100 to 125 Pa·s, about 125 to 150 Pa·s, about 150 to 175 Pa·s, about 175 to 200 Pa·s, about 200 to 225 Pa·s, about 225 to 250 Pa·s, about 250 to 275 Pa·s, or about 275 to 300 Pa·s at a low shear rate.

[0128] Shear strength and / or resistance is a measure of the ability of a material to resist an external shear stress (i.e., shear load) without breaking (i.e., loss of adhesion or integrity). In certain embodiments, the polymeric compositions of the present disclosure can have a therapeutically effective shear strength. In certain embodiments, the polymeric composition can have a shear strength that provides durable adhesion and high retention of the polymeric composition on a target tissue of a subject. In certain embodiments, the gel-polymeric compositions of the present disclosure can have a shear strength that provides durable adhesion and high retention of the polymeric composition on a target tissue of a subject. In certain embodiments, the gel-polymeric composition can have a shear strength that enables the polymeric composition to maintain its shape, adhesion, connectivity, and / or consistency on the surface of the target tissue for more than one hour, more than one day, or more than one week.

[0129] In certain embodiments, the polymer composition can have a shear strength of from about 1 to about 360 kPa. In certain embodiments, the polymer composition can have a shear strength of from about 100 to 360 kPa. In certain embodiments, the polymer composition can have a shear strength of from about 200 to 360 kPa. In certain embodiments, the polymer composition can have a shear strength of from about 1 to 20 kPa, from about 20 to 40 kPa, from about 40 to 60 kPa, from about 60 to 80 kPa, from about 80 to 100 kPa, 100 to 120 kPa, from about 120 to 140 kPa, from about 140 to 160 kPa, from about 160 to 180 kPa, from about 180 to 200 kPa, 200 to 220 kPa, from about 220 to 240 kPa, from about 240 to 260 kPa, from about 260 to 280 kPa, from about 280 to 300 kPa, and from about 300 to 320 kPa, from about 320 to 340 kPa, and from about 340 to 360 kPa.

[0130] In certain embodiments, the shear strength of the polymer composition can be measured using the deformation of ASTM F2255-05, or a modified lap shear test thereof.

[0131] Swelling and water content In certain embodiments, the polymer composition includes a gel. A gel generally includes a cross-linked polymer framework that encompasses a network of pores filled with an interstitial solvent (e.g., a fluid). In certain embodiments, the polymer composition includes a hydrogel and the interstitial fluid includes water. In certain embodiments, the polymer composition includes an alcogel and the interstitial fluid includes an alcohol (e.g., methanol, ethanol).

[0132] When a gel material absorbs and holds additional interstitial fluid within the pore network of the gel, swelling (i.e., an increase in volume) may occur within the gel. Similarly, when the gel material expels interstitial fluid from the pore network of the gel, shrinkage (i.e., a decrease in volume) may occur within the gel. The ability and / or tendency of a gel material to swell and / or shrink in a particular solvent environment depends on the chemical properties of the polymer and the solvent (e.g., solubility, hydrophobicity, pore structure, affinity) and the elasticity of the polymer network of the gel. The swelling ratio of a gel is a measure of the slight increase in the weight of the gel due to fluid absorption (e.g., the weight increase of a hydrogel due to water absorption).

[0133] In certain embodiments, the polymer compositions of the present disclosure may have a therapeutically effective swelling ratio and / or water content. In certain embodiments, the polymer composition may have a swelling ratio and / or water content that provides strong adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the gel polymer compositions of the present disclosure may have a swelling ratio and / or water content that provides strong adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the gel polymer composition may have a swelling ratio and / or water content that enables the polymer composition to maintain its shape, adhesiveness, connectivity, and / or consistency on the surface of the target tissue for one hour or more, one day or more, or one week or more.

[0134] In certain embodiments, the polymer composition can have a swelling ratio of about 5% to about 50%. In certain embodiments, the polymer composition can have a swelling ratio of at least about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, or about 40%. In certain embodiments, the polymer composition can have a swelling ratio of about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, or about 10% or less. In certain embodiments, the polymer composition has a swelling ratio of about 25% or less, about 20% or less, about 15% or less, or about 10% or less. In certain embodiments, the polymer composition can have a swelling ratio of about 1-3%, about 3-6%, about 6-10%, about 1-5%, about 1-10%, about 5-10%, about 11-13%, about 13-16%, about 16-20%, about 10-20%, about 10-15%, about 15-20%, about 21-23%, about 23-26%, about 26-30%, about 20-30%, about 20-25%, about 25-30%, about 31-33%, about 33-36%, about 36%-40%, about 30-40%, about 30%-35%, about 35%-40%, about 41-43%, about 43-46%, about 46-50%, about 40-50%, about 40%-45%, or about 45%-50%. In certain embodiments, the polymer composition can have a swelling ratio of about 1-3%, about 3-6%, about 6-10%, about 1-5%, about 1-10%, about 5-10%, about 11-13%, about 13-16%, about 16-20%, about 10-20%, about 10-15%, about 15-20%, about 21-23%, about 23-26%, about 26-30%, about 20-30%, about 20-25%, about 25-30%, about 31-33%, about 33-36%, about 36%-40%, about 30-40%, about 30%-35%, about 35%-40%, about 41-43%, about 43-46%, about 46-50%, about 40-50%, about 40%-45%, or about 45%-50% for the short-term swelling ratio (i.e., the swelling ratio measured for about 1-24 hours).In certain embodiments, the polymer composition can have a medium swelling rate (i.e., the swelling rate measured over about 1 - 7 days) of about 1 - 3%, about 3 - 6%, about 6 - 10%, about 1 - 5%, about 1 - 10%, about 5 - 10%, about 11 - 13%, about 13 - 16%, about 16 - 20%, about 10 - 20%, about 10 - 15%, about 15 - 20%, about 21 - 23%, about 23 - 26%, about 26 - 30%, about 20 - 30%, about 20 - 25%, about 25 - 30%, about 31 - 33%, about 33 - 36%, about 36 - 40%, about 30 - 40%, about 30 - 35%, about 35 - 40%, about 41 - 43%, about 43 - 46%, about 46 - 50%, about 40 - 50%, about 40 - 45%, or about 45 - 50%. In certain embodiments, the polymer composition can have a long - term swelling rate (i.e., the swelling rate measured over about 1 - 4 weeks) of about 1 - 3%, about 3 - 6%, about 6 - 10%, about 1 - 5%, about 1 - 10%, about 5 - 10%, about 11 - 13%, about 13 - 16%, about 16 - 20%, about 10 - 20%, about 10 - 15%, about 15 - 20%, about 21 - 23%, about 23 - 26%, about 26 - 30%, about 20 - 30%, about 20 - 25%, about 25 - 30%, about 31 - 33%, about 33 - 36%, about 36 - 40%, about 30 - 40%, about 30 - 35%, about 35 - 40%, about 41 - 43%, about 43 - 46%, about 46 - 50%, about 40 - 50%, about 40 - 45%, or about 45 - 50%.

[0135] In certain embodiments, the hydrogel polymer composition can have a water content of from about 5% to about 99%. In certain embodiments, the hydrogel polymer composition can have a water content of from about 50% to about 99%. In certain embodiments, the hydrogel polymer composition can have a water content of from about 65% to about 85%. In certain embodiments, the polymer composition can have a water content of at least about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or about 80%. In certain embodiments, the polymer composition can have a swelling ratio of about 99% or less, about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, about 40% or less, about 35% or less, or about 30% or less. In certain embodiments, the polymer composition can have a water content of from about 1 - 3%, about 3 - 6%, about 6 - 10%, about 1 - 5%, about 5 - 10%, about 1 - 10%, about 11 - 13%, about 13 - 16%, about 16 - 20%, about 10 - 15%, about 15 - 20%, about 10 - 20%, about 21 - 23%, about 23 - 26%, about 26 - 30%, about 20 - 25%, about 25 - 30%, about 20 - 30%, about 31 - 33%, about 33 - 36%, about 36 - 40%, about 30 - 35%, about 35 - 40%, about 30 - 40%, about 41 - 43%, about 43 - 46%, about 46 - 50%, about 40 - 45%, about 45 - 50%, about 40 - 50%, about 51 - 53%, about 53 - 56%, about 56 - 60%, about 50 - 55%, about 55 - 60%, about 50 - 60%, about 61 - 63%, about 63 - 66%, about 66 - 70%, about 60 - 65%, about 65 - 70%, about 60 - 70%, about 71 - 73%, about 73 - 76%, about 76 - 80%, about 70 - 75%, about 75 - 80%, about 70 - 80%, about 81 - 83%, about 83 - 86%, about 86 - 90%, about 80 - 85%, about 85 - 90%, about 80 - 90%, about 91 - 93%, about 93 - 96%, about 96 - 99%, about 90 - 95%, about 95 - 99%, or about 90 - 99%.

[0136] In certain embodiments, the hydrogel polymer compositions of the present disclosure enable the controlled sustained release of one or more therapeutic agents over a period of time. In certain embodiments, the hydrogel polymer compositions enable the release of a therapeutic agent at least 1 μg / day, at least 2 μg / day, at least 3 μg / day, at least 4 μg / day, at least 5 μg / day, at least 6 μg / day, at least 7 μg / day, at least 8 μg / day, at least 9 μg / day, at least 10 μg / day, at least 11 μg / day, at least 12 μg / day, at least 13 μg / day, at least 14 μg / day, at least 15 μg / day, at least 16 μg / day, at least 17 μg / day, at least 18 μg / day, at least 19 μg / day, at least 20 μg / day, at least 25 μg / day, at least 30 μg / day, at least 35 μg / day, at least 40 μg / day, at least 45 μg / day, at least 50 μg / day, at least 60 μg / day, at least 70 μg / day, at least 80 μg / day, at least 90 μg / day, at least 100 μg / day, at least 150 μg / day, at least 200 μg / day, at least 250 μg / day, at least 300 μg / day, at least 350 μg / day, at least 400 μg / day, at least 450 μg / day, at least 500 μg / day, at least 600 μg / day, at least 700 μg / day, at least 800 μg / day, at least 900 μg / day, or at least 1000 μg / day. In certain embodiments, the hydrogel polymer compositions enable the release of a therapeutic agent at least 10 μg / day.

[0137] Durability and degradation In certain embodiments, the polymer compositions of the present disclosure may have a therapeutically effective polymer degradation rate (i.e., degradation rate). In certain embodiments, the polymer composition may have a degradation rate that provides for sustained adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the gel polymer compositions of the present disclosure may have a degradation rate that provides for sustained adhesion and high retention of the polymer composition on the target tissue of the subject. In certain embodiments, the gel polymer composition may have a degradation rate that allows the polymer composition to maintain its shape, adhesiveness, connectivity, and / or consistency on the surface of the target tissue for one hour or more, one day or more, or one week or more.

[0138] In certain embodiments, the polymer composition may have a degradation rate of 1 to 50 days. In certain embodiments, the polymer composition may have a degradation rate of about 1 to 3 days, about 3 to 6 days, about 6 to 10 days, about 1 to 5 days, about 1 to 10 days, about 5 to 10 days, about 11 to 13 days, about 13 to 16 days, about 16 to 20 days, about 10 to 20 days, about 10 to 15 days, about 15 to 20 days, about 21 to 23 days, about 23 to 26 days, about 26 to 30 days, about 20 to 30 days, about 20 to 25 days, about 25 to 30 days, about 31 to 33 days, about 33 to 36 days, about 36 to 40 days, about 30 to 40 days, about 30 to 35 days, about 35 to 40 days, about 41 to 43 days, about 43 to 46 days, about 46 to 50 days, about 40 to 50 days, about 40 to 45 days, or about 45 to 50 days.

[0139] Biocompatibility In certain embodiments, the polymer compositions of the present disclosure are biocompatible with the target tissue of the subject. In certain embodiments, the biomechanical properties of the polymer composition are similar and / or biocompatible to the biomechanical properties of the target tissue of the subject (e.g., the cornea of the subject).

[0140] In certain embodiments, the biocompatibility of the polymer composition may be demonstrated by the low level of inflammatory response in the target tissue or subject. In certain embodiments, the biocompatibility of the polymer composition may be demonstrated by the survival rate of cells from the target tissue transplanted or incorporated into a portion of the polymer composition.

[0141] Shape In certain embodiments, the polymer compositions of the present disclosure can be formed as molded, extruded, or shaped gel compositions. Molded, extruded, or shaped hydrogels can be prepared, for example, using the methods described in U.S. Patent Application Publication No. 20050008675 or U.S. Patent Application Publication No. 20040258729, each of which is incorporated herein by reference in its entirety to the extent that each describes the composition, production (including molding), analysis, and use of hydrogels comprising acrylated gelatin polymer compositions such as GelMA hydrogels.

[0142] In certain embodiments, the polymer compositions of the present disclosure (e.g., hydrogel polymer compositions) can be formed into cylinders, each cylinder having a length and a diameter. In certain embodiments, the polymer compositions of the present disclosure (e.g., hydrogel polymer compositions) can conform to the shape of the target surface. In certain embodiments, the polymer composition conforms to the convex, concave, or curved shape of the target surface.

[0143] In certain embodiments, the polymer composition can be formed into a cylindrical rod. As used herein, "cylindrical rod" or "rod" describes a cylinder having a cylinder length that is at least three times (3×) the cylinder diameter. By way of non-limiting example, the cylindrical rod can have a length of about 3 mm and a diameter of about 0.75 mm, or a length of about 2.5 mm and a diameter of about 0.75 mm. In certain embodiments, the hydrogel rod of the present disclosure can be about 3 mm in length and about 0.75 mm in diameter. In certain embodiments, the hydrogel rod of the present disclosure can be about 6 mm in length and about 0.75 mm in diameter.

[0144] In certain embodiments, the polymer composition can be formed into a cylindrical disk. As used herein, "cylindrical disk" or "disk" describes a cylinder having a cylinder diameter that is at least 2 times (2×) the cylinder length. By way of non-limiting example, the cylindrical disk can have a length of about 2.5 mm and a diameter of about 6 mm, or a length of about 2 mm and a diameter of about 6 mm.

[0145] III. Gel Preparation In certain embodiments, the polymer compositions of the present disclosure (e.g., GelMA polymer compositions) can be produced as described in the art, including Nichol et al., Biomaterials, 2010 Jul, 31(21):5536 - 44, Assmann et al., Biomaterials, 2017, 140:115 - 127, Noshadi et al., Biomater. Sci., 2017, 5:2093 - 2105, each of which is hereby incorporated by reference in its entirety to the extent each describes the production of polymer compositions, including acryloyl gelatin polymer compositions such as GelMA hydrogels.

[0146] In certain embodiments, the polymer compositions of the present disclosure can be formed by crosslinking two or more chemically modified gelatin components in a precursor polymer composition to form a gel polymer composition. In certain embodiments, the polymer compositions of the present disclosure can crosslink, polymerize, and / or gelate under wet, aqueous, and / or biological conditions to form a gel polymer composition. In certain embodiments, the crosslinking of two or more chemically modified gelatin components is initiated, facilitated, or enabled when exposed to certain crosslinking conditions (e.g., acidic conditions, basic conditions, high salt conditions, low salt conditions, high temperature, agitation, dissolution conditions). In certain embodiments, the crosslinking of two or more chemically modified gelatin components is initiated, facilitated, or enabled by a crosslinking agent. In certain embodiments, the crosslinking of two or more chemically modified gelatin components is initiated, facilitated, or enabled by a crosslinking agent under certain crosslinking conditions.

[0147] In certain embodiments, the present disclosure provides a method for generating a gel polymer composition, such as a hydrogel polymer composition. In certain embodiments, the present disclosure provides a method for generating a GelMA hydrogel polymer composition. FIG. 2 provides a method 100 for generating a gel polymer composition. In step 110, a precursor polymer composition is provided that includes a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA). In optional step 115, one or more additional chemically modified polymer precursors having crosslinkable groups (e.g., MeHA, PEGDA) are added to the precursor polymer composition. In certain embodiments, the polymer composition includes unmodified HA and / or unmodified PEG and / or unmodified tropoelastin. In step 120, a solution including one or more crosslinking agents and / or photoinitiators is added to the precursor polymer composition. In optional step 125, a therapeutic agent, cells, and / or particles (i.e., microparticles or nanoparticles) are added to the precursor polymer composition. In step 130, the precursor polymer composition is polymerized / crosslinked to generate a gel polymer composition.

[0148] In certain embodiments, the method for generating a gel polymer composition can include providing a precursor polymer composition that includes a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA). In certain embodiments, the chemically modified gelatin includes acrylated gelatin. In certain embodiments, the chemically modified gelatin includes gelatin methacryloyl (i.e., GelMA).

[0149] In certain embodiments, the precursor polymer composition includes one or more solvents or liquid vehicles, diluents, dispersion media, dispersants, granulating agents, binders, disintegrants, suspending agents, surface active agents, emulsifiers (emulsifler or emulsifying agent), isotonic agents, thickening agents, preservatives, solid binders, buffering agents, lubricants, coloring agents, coating agents, sweeteners, flavoring agents, fragrances, or combinations thereof.

[0150] In certain embodiments, the precursor polymer composition can include one or more solvents. In certain embodiments, the solvent includes an aqueous solvent. Examples of aqueous solvents include, but are not limited to, distilled water, deionized water, physiological saline, Dulbecco's phosphate buffered saline (DPBS), and Ringer's solution. In certain embodiments, the solvent includes DPBS. In certain embodiments, the solvent includes an organic solvent. Examples of organic solvents include, but are not limited to, hexane, benzene, toluene, acetone, diethyl ether, chloroform, dichloromethane, isopropanol, methanol, ethanol, n-propanol, and n-butanol.

[0151] In certain embodiments, the precursor polymer composition may be in a sprayable form. In certain embodiments, the precursor polymer composition may be in a high-viscosity form (e.g., paste-like viscosity). In certain embodiments, the precursor polymer composition may be in a low-viscosity form (e.g., liquid-like viscosity).

[0152] In certain embodiments, a method for generating a gel polymer composition can include adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition. In certain embodiments, a method for generating a gel polymer composition can include (i) providing a precursor polymer composition that includes a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA), and (ii) adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition. In certain embodiments, the one or more additional chemically modified polymer precursors include a chemically modified hyaluronic acid such as acryloyl-substituted hyaluronic acid. In certain embodiments, the chemically modified hyaluronic acid includes methacrylated hyaluronic acid (MeHA). In certain embodiments, the one or more additional chemically modified polymer precursors include a chemically modified poly(ethylene glycol) (PEG) such as acryloyl-substituted PEG. In certain embodiments, the chemically modified hyaluronic acid includes poly(ethylene glycol) diacrylate (PEGDA).

[0153] In certain embodiments, a method for generating a gel polymer composition can include adding one or more crosslinking agents and / or a polymer crosslinking initiator (e.g., a photoinitiator) to a precursor polymer composition. In certain embodiments, a method for generating a gel polymer composition can include (i) providing a precursor polymer composition comprising a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA), and (ii) adding one or more crosslinking agents and / or a polymer crosslinking initiator (e.g., a photoinitiator) to the precursor polymer composition. In certain embodiments, a method for generating a gel polymer composition can include (i) providing a precursor polymer composition comprising a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA), (ii) adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition, and (iii) adding one or more crosslinking agents and / or a polymer crosslinking initiator (e.g., a photoinitiator) to the precursor polymer.

[0154] In certain embodiments, one or more crosslinking agents and / or a polymer crosslinking initiator (e.g., a photoinitiator) can be added to the precursor polymer before adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition. In certain embodiments, a method for generating a gel polymer composition can include (i) providing a precursor polymer composition comprising a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA), (ii) adding one or more crosslinking agents and / or a polymer crosslinking initiator (e.g., a photoinitiator) to the precursor polymer composition, and (iii) adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition.

[0155] In certain embodiments, the polymer composition comprises one or more polymer crosslinking initiators (e.g., crosslinking initiators that form free radicals when exposed to certain polymer crosslinking conditions such as acidic conditions, basic conditions, high salt conditions, low salt conditions, high temperature, agitation, dissolution conditions, and light exposure). In certain embodiments, the polymer composition comprises one or more photoinitiator elements (i.e., crosslinking initiators that are initiated or activated by absorbing light of a specific wavelength). In certain embodiments, the precursor polymer composition of the present disclosure comprises one or more photoinitiator elements (i.e., crosslinking initiators that are initiated or activated by visible light). In certain embodiments, the photoinitiator element can be activated by exposure to light. In certain embodiments, light exposure can activate the photoinitiator to form free radicals, which can lead to the formation of bonds between reactive groups in the composition, such as vinyl bond crosslinking between methacrylate groups in the GelMA polymer composition. Figure 3 provides an example of a series of reactions for generating a GelMA hydrogel polymer composition, where (i) the photoinitiator element is activated by light energy (hv) to form free radicals (R*), which then initiate the formation of bonds between reactive groups on separate gelatin methacryloyl polymer precursors, thereby forming a crosslinked GelMA polymer network. The continuous reaction between reactive groups on the gelatin methacryloyl component results in the formation of a more extensive GelMA hydrogel polymer composition.

[0156] In certain embodiments, the photoinitiator element can be activated by exposure to one or more light sources selected from visible light sources (e.g., white or blue light), ultraviolet (UV) light sources, near-infrared (NIR) light sources, and fluorescent light sources. In certain embodiments, the photoinitiator element includes a visible light-activated photoinitiator, such as a visible light-activated photoinitiator that is activated upon exposure to light having a wavelength of from about 380 nm to about 740 nm. In certain embodiments, the visible light-activated photoinitiator can be activated upon exposure to light having a wavelength of about 380 - 435 nm (i.e., violet light), about 435 - 500 nm (i.e., blue light), about 500 - 565 nm (i.e., green light), about 565 - 600 nm (i.e., yellow light), about 600 - 650 nm (i.e., orange light), or about 650 - 740 nm (i.e., red light). In certain embodiments, the photoinitiator element includes an ultraviolet-activated photoinitiator. In certain embodiments, the photoinitiator element includes a near-infrared light (NIR)-activated photoinitiator. In certain embodiments, the photoinitiator element includes a white light-activated photoinitiator. In certain embodiments, the photoinitiator element includes a blue light-activated photoinitiator.

[0157] In certain embodiments, a method for generating a gel polymer composition can include adding one or more therapeutic agents and / or particles (i.e., microparticles or nanoparticles) to a precursor polymer composition. In certain embodiments, one or more therapeutic agents and / or particles can be added to the precursor polymer before adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition. In certain embodiments, one or more therapeutic agents and / or particles can be added to the precursor polymer before adding one or more crosslinking agents and / or polymer crosslinking initiators (e.g., photoinitiators) to the precursor polymer composition. In certain embodiments, a method for generating a gel polymer composition can include: (i) providing a precursor polymer composition comprising a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA); (ii) optionally adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition; (iii) adding one or more crosslinking agents and / or polymer crosslinking initiators (e.g., photoinitiators) to the precursor polymer; and (iv) optionally adding one or more therapeutic agents and / or particles.

[0158] In certain embodiments, the precursor polymer composition can be clarified, purified, or processed for quality and / or purity prior to any polymerization / crosslinking steps. In certain embodiments, the precursor polymer composition can be filtered. In certain embodiments, the precursor polymer composition can be lyophilized. In certain embodiments, the precursor polymer composition can be frozen for storage.

[0159] In certain embodiments, a method for generating a gel polymer composition can include the step of polymerizing / crosslinking a precursor polymer composition to generate a gel polymer composition. In certain embodiments, a method of making a gel polymer composition can include: (i) providing a precursor polymer composition comprising a chemically modified gelatin having crosslinkable groups (e.g., acryloyl-substituted gelatin, GelMA); (ii) optionally adding one or more additional chemically modified polymer precursors having crosslinkable groups to the precursor polymer composition; (iii) adding one or more crosslinking agents and / or polymer crosslinking initiators (e.g., photoinitiators) to the precursor polymer; (iv) optionally adding one or more therapeutic agents and / or particles; and (v) polymerizing / crosslinking the precursor polymer composition to generate a gel polymer composition.

[0160] In certain embodiments, crosslinking of the chemically modified gelatin component and any additional chemically modified polymer precursors (e.g., MeHA, PEGDA) is initiated, facilitated, or enabled by exposure to UV or visible light in the presence of a photoinitiator component. In certain embodiments, exposure to UV or visible light in the presence of a photoinitiator causes acryloyl groups on one chemically modified gelatin molecule to react with acryloyl groups on other chemically modified gelatin molecules to crosslink the acryloyl-substituted gelatin component and generate a gel (e.g., hydrogel). In certain embodiments, exposure to visible light in the presence of a photoinitiator causes methacryloyl groups on one methacryloyl gelatin molecule to react with methacryloyl groups on other methacryloyl gelatin molecules to crosslink the methacryloyl-substituted gelatin component and generate a gelatin methacryloyl (GelMA) hydrogel.

[0161] In certain embodiments, the polymer composition is exposed to a light source for 1 to 60 minutes. In certain embodiments, the polymer composition is exposed to a light source for a time of 1 minute or more, 5 minutes or more, 10 minutes or more, 15 minutes or more, 20 minutes or more, 25 minutes or more, or 30 minutes or more. In certain embodiments, the polymer composition is exposed to a light source for a time of within 1 minute, within 5 minutes, within 10 minutes, within 15 minutes, within 20 minutes, within 25 minutes, or within 30 minutes, within 35 minutes, or within 40 minutes. In certain embodiments, the polymer composition is exposed to a light source for about 5 seconds, about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 60 seconds, about 65 seconds, about 70 seconds, about 75 seconds, about 80 seconds, about 85 seconds, about 90 seconds, about 95 seconds, about 100 seconds, about 105 seconds, about 110 seconds, about 115 seconds, about 120 seconds, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, about 20 minutes, about 21 minutes, about 22 minutes, about 23 minutes, about 24 minutes, about 25 minutes, about 26 minutes, about 27 minutes, about 28 minutes, about 29 minutes, about 30 minutes, about 31 minutes, about 32 minutes, about 33 minutes, about 34 minutes, about 35 minutes, about 36 minutes, about 37 minutes, about 38 minutes, about 39 minutes, about 40 minutes, about 41 minutes, about 42 minutes, about 43 minutes, about 44 minutes, about 45 minutes, about 46 minutes, about 47 minutes, about 48 minutes, about 49 minutes, about 50 minutes, about 51 minutes, about 52 minutes, about 53 minutes, about 54 minutes, about 55 minutes, about 56 minutes, about 57 minutes, about 58 minutes, about 59 minutes, or about 60 minutes.In certain embodiments, the polymer composition is exposed to a light source for about 1 to 3 minutes, about 3 to 6 minutes, about 6 to 10 minutes, about 1 to 5 minutes, about 1 to 10 minutes, about 5 to 10 minutes, about 11 to 13 minutes, about 13 to 16 minutes, about 16 to 20 minutes, about 10 to 20 minutes, about 10 to 15 minutes, about 15 to 20 minutes, about 21 to 23 minutes, about 23 to 26 minutes, about 26 to 30 minutes, about 20 to 30 minutes, about 20 to 25 minutes, about 25 to 30 minutes, about 31 to 33 minutes, about 33 to 36 minutes, about 36 to 40 minutes, about 30 to 40 minutes, about 30 to 35 minutes, about 35 to 40 minutes, about 41 to 43 minutes, about 43 to 46 minutes, about 46 to 50 minutes, about 40 to 50 minutes, about 40 to 45 minutes, about 45 to 50 minutes, about 51 to 53 minutes, about 53 to 56 minutes, about 56 to 60 minutes, about 50 to 60 minutes, about 50 to 55 minutes, or about 55 to 60 minutes.

[0162] In certain embodiments, the polymer composition can have a thickness of about 1 μm to about 10,000 μm. In certain embodiments, the polymer composition can have a thickness of about 1 to 50 μm, about 50 to 100 μm, about 100 to 150 μm, about 150 to 200 μm, about 200 to 250 μm, about 250 to 300 μm, about 300 to 350 μm, about 350 to 400 μm, about 400 to 450 μm, about 450 to 500 μm, about 500 to 550 μm, about 550 to 600 μm, about 600 to 650 μm, about 650 to 700 μm, about 700 to 750 μm, about 750 to 800 μm, about 800 to 850 μm, about 850 to 900 μm, about 900 to 950 μm, about 950 to 1000 μm, about 1000 to 1500 μm, about 1500 to 2000 μm, about 2000 to 2500 μm, about 2500 to 3000 μm, about 3000 to 3500 μm, about 3500 to 4000 μm, about 4000 to 4500 μm, about 4500 to 5000 μm, about 5000 to 5500 μm, about 5500 to 6000 μm, about 6000 to 6500 μm, about 6500 to 7000 μm, about 7000 to 7500 μm, about 7500 to 8000 μm, about 8000 to 8500 μm, about 8500 to 9000 μm, about 9000 to 9500 μm, or about 9500 to 10,000 μm.

[0163] In certain embodiments, the precursor polymer composition can be cooled before or during the crosslinking reaction. In certain embodiments, the precursor polymer composition can be cooled to a temperature of about 0 °C to about 30 °C before or during the crosslinking reaction. In certain embodiments, the precursor polymer composition can be cooled to a temperature of about 0 to 5 °C, about 5 to 10 °C, about 0 to 10 °C, about 10 to 15 °C, about 15 to 20 °C, about 10 to 20 °C, about 20 to 25 °C, about 25 to 30 °C, or about 20 to 30 °C. In certain embodiments, the precursor polymer composition can be heated before or during the crosslinking reaction. In certain embodiments, the precursor polymer composition can be heated to a temperature of about 30 °C to about 150 °C before or during the crosslinking reaction. In certain embodiments, the precursor polymer composition can be heated to a temperature of about 30 to 35 °C, about 35 to 40 °C, about 30 to 40 °C, about 40 to 45 °C, about 45 to 50 °C, about 40 to 50 °C, about 50 to 55 °C, about 55 to 60 °C, about 50 to 60 °C, about 60 to 65 °C, about 65 to 70 °C, about 60 to 70 °C, about 70 to 75 °C, about 75 to 80 °C, about 70 to 80 °C, about 80 to 85 °C, about 85 to 90 °C, about 80 to 90 °C, about 90 to 95 °C, about 95 to 100 °C, about 90 to 100 °C, about 100 to 105 °C, about 105 to 110 °C, about 100 to 110 °C, about 110 to 115 °C, about 115 to 120 °C, about 110 to 120 °C, about 130 to 135 °C, about 135 to 140 °C, about 130 to 140 °C, about 140 to 145 °C, about 145 to 150 °C, or about 140 to 150 °C.

[0164] Once the crosslinking reaction is complete or stopped, the resulting gel polymer material can be clarified, purified, or processed for quality, purity, and / or therapeutic viability. In certain embodiments, the gel polymer composition can be dialyzed to remove any unreacted compounds from the gel mixture or structure. In certain embodiments, the gel polymer composition can be dialyzed against a dialysis buffer containing deionized water. In certain embodiments, the gel polymer composition can be filtered. In certain embodiments, the gel polymer composition can be dried. In certain embodiments, the gel polymer composition can be lyophilized. In certain embodiments, the gel polymer composition can be frozen for storage.

[0165] In certain embodiments, the polymer compositions of the present disclosure can be manufactured or processed by forming, molding, extrusion weaving, or other methods into fibers, films, disks, fabrics, tubes, conduits, rods, rings, meshes, or any other form or shape of polymer or gel materials known in the art. In certain embodiments, the polymer compositions of the present disclosure can be manufactured or processed by forming, molding, extrusion weaving, or other methods into a single-layer structure or a multi-layer structure (e.g., two-layer, three-layer, four-layer, etc.).

[0166] In certain embodiments, the polymer compositions of the present disclosure include polymeric polymers and / or fiber elements that are woven or intertwined within the interstitial porous network of the polymer composition but are not chemically connected to the core cross-linked polymer network. Non-limiting examples of such polymers include polycaprolactone, gelatin, gelatin methacrylate, alginate, alginate methacrylate, chitosan, chitosan methacrylate, glycol chitosan, glycol chitosan methacrylate, hyaluronic acid, hyaluronic acid methacrylate, and other non-crosslinked natural or synthetic polymer chains. Gel materials containing woven polymer structures can be referred to as composite structures or composite gels. Examples of hydrogel / fiber composites are described, for example, in Moutos et al. Nat. Mater., 2007, 6(2), p. 162-7, and the document is incorporated herein by reference in its entirety to the extent it describes the composition, production, analysis, and use of the composite gel material. In certain embodiments, the precursor polymer composition may be in a high-viscosity form (e.g., paste-like viscosity) and may be incorporated into a polymeric polymer matrix (e.g., a fibrous mat or tissue matrix). In certain embodiments, the precursor polymer composition may be in a low-viscosity form (e.g., liquid-like viscosity) and may be incorporated into a polymeric polymer matrix (e.g., a fibrous mat or tissue matrix).

[0167] In certain embodiments, the crosslinked polymer composition may have a substantially covalent matrix form. In certain embodiments, the crosslinked polymer composition may have an amorphous matrix form (i.e., a matrix formed primarily via ionic and / or hydrogen bonds).

[0168] In certain embodiments, the polymer compositions of the present disclosure can be formed as a patterned gel composition (e.g., a micropatterned hydrogel). Micropatterned hydrogels can be prepared, for example, using the methods described in U.S. Patent No. 6,423,252, which is hereby incorporated by reference in its entirety to the extent it describes the composition, generation (including micropatterning), analysis, and use of hydrogels, including acrylated gelatin polymer compositions such as GelMA hydrogels. For example, the method includes (i) contacting a precursor polymer composition with a mold or surface that includes a three-dimensional negative structure of a micropattern (i.e., a template), and (ii) crosslinking and / or polymerizing the precursor polymer composition to produce a crosslinked gel polymer composition (e.g., a GelMA hydrogel) that includes a micropattern on at least the surface of the hydrogel.

[0169] In certain embodiments, the polymer compositions of the present disclosure can be formed as a molded, cast, or shaped gel composition. Molded, cast, or shaped hydrogels can be prepared, for example, using the methods described in U.S. Patent Application Publication No. 20050008675 or U.S. Patent Application Publication No. 20040258729, each of which is hereby incorporated by reference in its entirety to the extent it describes the composition, generation (including molding), analysis, and use of hydrogels, including acrylated gelatin polymer compositions such as GelMA hydrogels.

[0170] IV. Administration and Treatment Overview The use of sutures, tissue grafts, and tissue adhesives are common treatments for soft tissue defects and / or traumatic injuries (such as corneal or scleral tissue). However, each treatment is associated with risks and complications: (i) sutures require advanced surgical skills and early intervention, often cause irregular stigmatism, and frequently lead to microbial entrapment and infection; (ii) tissue grafts and transplantation require donor tissue (associated with high costs), advanced surgical skills, and carry a high risk of immune response or complete rejection of the transplanted tissue; (iii) tissue adhesives (cyanoacrylate adhesives, fibrin adhesives, or polyethylene glycol (PEG)-based sealants) have limited effectiveness and adhesiveness (especially in aqueous and physiological environments), limited durability, may be difficult to apply and control the texture, have a high potential for leakage, lack biocompatibility (e.g., inflammation), may be toxic, lack translucency / transparency, have a high risk of infection (including risks associated with high porosity), and generally do not have FDA safety approval for the reduction of corneal defects or the repair of corneal incisions, perforations, or trauma.

[0171] There remains a need for improved polymer compositions effective for the treatment and / or sealing of injuries, defects, and diseases of a subject's soft tissue (i.e., body tissue excluding bone).

[0172] In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant composition for treating or repairing soft tissue in a subject. In certain embodiments, the polymer compositions of the present disclosure can be used as a delivery vehicle for administering a therapeutic agent for treating or repairing soft tissue in a subject. In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant composition for treating or repairing soft tissue in a subject and as a delivery vehicle for administering a therapeutic agent for treating or repairing the subject's soft tissue.

[0173] In certain embodiments, the methods and compositions of the present disclosure can be used to adhere to, seal, or treat a target soft tissue of interest. In certain embodiments, the methods and compositions of the present disclosure can be used to adhere to, seal, or treat one or more target soft tissues selected from adipose tissue, bladder tissue, bone marrow, cardiovascular tissue (e.g., the heart), dura mater, endocrine glands, gastrointestinal tissue, hair follicles, kidney tissue, liver tissue, lung tissue, lymph nodes, muscle tissue, neural / nerve tissue (e.g., the peripheral nervous system), ocular tissue (e.g., the cornea), oral tissue (e.g., craniofacial, teeth, periodontal), pancreatic tissue, renal tissue, skin tissue (e.g., for the treatment of local ulcers such as diabetic ulcers), urethral tissue, vascular tissue. In certain embodiments, the methods and compositions of the present disclosure can be used to adhere to, seal, or treat one or more target soft tissues in a stress environment and / or a physiological environment, or in similar applications that require an elastic and / or adhesive composition.

[0174] The polymer compositions of the present disclosure (e.g., GelMA polymer compositions) can be administered by any route that provides a therapeutically effective result.

[0175] In certain embodiments, the method includes applying a pre-gelled polymer composition to an applicator, placing the applicator containing the pre-gelled polymer composition on the surface of the target tissue of interest, and crosslinking (e.g., photocrosslinking) the polymer composition by exposing the pre-gelled polymer composition to crosslinking conditions (e.g., visible light with a photoinitiator). In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target tissue without using an applicator. In certain embodiments, application to the surface of the target tissue includes application to the outer surface of the target tissue (e.g., topical application). In certain embodiments, application to the surface of the target tissue includes application / injection into the space immediately beneath the surface of the target tissue (e.g., subconjunctival application to ocular tissue, subretinal application to ocular tissue).

[0176] In certain embodiments, the target soft tissue can be treated or sealed by applying a first layer comprising a first polymer composition of the present disclosure designed to have certain physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity), and then applying a second layer comprising a second polymer composition of the present disclosure designed to have different physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity). In certain embodiments, the method may include applying one or more additional layers (e.g., a third layer, a fourth layer, etc.) each comprising a polymer composition of the present disclosure designed to have certain physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity).

[0177] In certain embodiments, the target soft tissue can be treated by (i) forming a polymer composition pre-formed by polymerizing a polymer composition of the present disclosure, and (ii) applying the pre-formed polymer composition onto or under the surface of the target tissue of the subject (e.g., subconjunctival, subretinal). In certain embodiments, the application to the surface of the target tissue (e.g., onto or under the surface) includes application / injection into the space immediately under the surface of the target tissue (e.g., subconjunctival application to eye tissue, subretinal application to eye tissue). In certain embodiments, the pre-formed polymer composition can be designed to have certain physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity).

[0178] In certain embodiments, the target soft tissue can be treated by: (i) forming a pre-formed hydrogel polymer composition by polymerizing the polymer composition of the present disclosure; (ii) drying the hydrogel polymer by removing a substantial portion of the interstitial fluid from the hydrogel (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the interstitial fluid); (iii) applying the pre-formed polymer composition onto or beneath the surface of the target tissue of interest (e.g., subconjunctival, subretinal); and (iv) optionally rehydrating the dried hydrogel polymer to a substantially hydrated form (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the interstitial fluid volume). In certain embodiments, the application to the surface of the target tissue (e.g., onto or beneath the surface) includes application / injection into the space immediately beneath the surface of the target tissue (e.g., subconjunctival application to ocular tissue, subretinal application to ocular tissue). In certain embodiments, the pre-formed polymer composition can be designed to have certain physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity).

[0179] Therapeutic composition In certain embodiments, the polymer composition of the present disclosure may be formulated as, or included in, a therapeutic composition. In certain embodiments, the hydrogel polymer composition of the present disclosure may be formulated as, or included in, a therapeutic composition. In certain embodiments, the GelMA hydrogel polymer composition of the present disclosure may be formulated as, or included in, a therapeutic composition. Such compositions include one or more polymer compositions of the present disclosure (optionally including one or more therapeutic agents or active ingredients) and one or more pharmaceutically acceptable excipients (e.g., carriers, solvents, or delivery vehicles).

[0180] The relative amounts of the polymer composition (e.g., GelMA hydrogel polymer composition), the therapeutically acceptable excipient, and / or any additional components in the therapeutic composition of the present disclosure can vary depending on the nature, size, and / or condition of the subject or tissue being treated, and further depending on the route by which the composition is administered or applied. In certain embodiments, the therapeutic composition comprises from 0.1% to 99% (w / v) of the polymer composition of the present disclosure in the volume of the therapeutic composition. In certain embodiments, the therapeutic composition comprises the polymer composition of the present disclosure at a weight / volume concentration (w / v) of about 0.5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 88%, about 79%, about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%.In certain embodiments, the therapeutic composition comprises the polymeric composition of the present disclosure at a weight / volume concentration (w / v) of about 1-3%, about 3-6%, about 6-10%, about 1-5%, about 5-10%, about 1-10%, about 11-13%, about 13-16%, about 16-20%, about 10-15%, about 15-20%, about 10-20%, about 21-23%, about 23-26%, about 26-30%, about 20-25%, about 25-30%, about 20-30%, about 31-33%, about 33-36%, about 36-40%, about 30-35%, about 35-40%, about 30-40%, about 41-43%, about 43-46%, about 46-50%, about 40-45%, about 45-50%, about 40-50%, about 51-53%, about 53-56%, about 56-60%, about 50-55%, about 55-60%, about 50-60%, about 61-63%, about 63-66%, about 66-70%, about 60-65%, about 65-70%, about 60-70%, about 71-73%, about 73-76%, about 76-80%, about 70-75%, about 75-80%, about 70-80%, about 81-83%, about 83-86%, about 86-90%, about 80-85%, about 85-90%, about 80-90%, about 91-93%, about 93-96%, about 96-99%, about 90-95%, about 95-99%, or about 90-99%.

[0181] In certain embodiments, the therapeutic compositions and formulations of the present disclosure include, but are not limited to, saline, liposomes (e.g., unilamellar vesicles, multilamellar vesicles), lipid particles (including microparticles and nanoparticles), and / or polymer particles (including microparticles and nanoparticles). In certain embodiments, the therapeutic compositions and formulations of the present disclosure include the polymeric composition of the present disclosure that incorporates, but is not limited to, saline, liposomes, lipid particles (including microparticles and nanoparticles), polymer particles (including microparticles and nanoparticles), or combinations thereof.

[0182] In certain embodiments, the therapeutic compositions and formulations of the present disclosure are aqueous formulations (i.e., formulations containing water). In certain embodiments, the therapeutic compositions and formulations of the present disclosure include water, purified water, or water for injection (WFI).

[0183] In certain embodiments, the therapeutic compositions and formulations of the present disclosure include one or more of the following: pH buffer solutions (e.g., phosphate buffered saline (PBS), HEPES, TES, MOPS), isotonic saline, Ringer's solution, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol), alginic acid, ethyl alcohol, and therapeutically acceptable mixtures thereof. In certain embodiments, the therapeutic compositions and formulations of the present disclosure include phosphate buffered saline (PBS).

[0184] The formulations of the present disclosure can be used in any step of manufacturing, processing, preparing, storing, expanding, or administering the polymer compositions of the present disclosure.

[0185] In certain embodiments, the therapeutic compositions of the present disclosure may include one or more pharmaceutically acceptable excipients (e.g., a vehicle capable of suspending or dissolving the polymeric compound. Excipients may include, for example, antiadhesives, antioxidants, binders, coating agents, compression aids, disintegrants, dyes (pigments), emollients, emulsifiers, fillers (diluents), film-forming or coating agents, flavoring agents, fragrances, lubricants (glidants), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, and water of hydration. Exemplary excipients include, but are not limited to, acetic acid, aluminum stearate, butylated hydroxytoluene (BHT), calcium carbonate, calcium chloride, calcium phosphate (dibasic), calcium stearate, carboxymethyl cellulose, croscarmellose, crospovidone, cysteine, ethyl cellulose, gelatin, glucose, glucuronic acid, gluconic acid, hydroxypropyl cellulose, hydroxypropyl methylcellulose, hydroxyl-butanedioic acid, inositol, lactose, magnesium chloride, magnesium stearate, maltitol, mannitol, methionine, methyl cellulose, methyl paraben, microcrystalline cellulose, phosphoric acid, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, sucrose, shellac, silicon dioxide, sodium acetate, sodium carbonate, sodium bicarbonate, sodium carboxymethyl cellulose, sodium chloride, sodium citrate, sodium hydroxide, sodium phosphate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, xylitol, zinc stearate, and combinations thereof.

[0186] Therapeutic agent In certain embodiments, the polymeric compositions of the present disclosure may include a therapeutic agent. In certain embodiments, the polymeric compositions of the present disclosure may include a therapeutic agent as a delivery payload.

[0187] In certain embodiments, the polymer compositions of the present disclosure can contain a therapeutic agent at a concentration of about 0% to about 40% (w / v). In certain embodiments, the precursor polymer compositions of the present disclosure can contain a therapeutic agent at a concentration of about 0% to about 40% (w / v). In certain embodiments, the gel polymer compositions of the present disclosure can contain a therapeutic agent at a concentration of about 0% to about 40% (w / v). In certain embodiments, the polymer compositions of the present disclosure can contain a therapeutic agent at a concentration of about 1-2%, about 2-4%, about 4-6%, about 6-8%, about 8-10%, about 1-5%, about 5-10%, about 1-10%, 10-12%, about 12-14%, about 14-16%, about 16-18%, about 18-20%, about 10-15%, about 15-20%, about 10-20%, about 20-22%, about 22-24%, about 24-26%, about 26-28%, about 28-30%, about 20-25%, about 25-30%, about 20-30%, about 30-32%, about 32-34%, about 34-36%, about 36-38%, about 38-40%, about 30-35%, about 35-40%, or about 30-40% (w / v).

[0188] In certain embodiments, the precursor polymer compositions of the present disclosure can contain a therapeutic agent at a concentration of about 0.1 mg / mL to about 500 mg / mL. In certain embodiments, the polymer compositions of the present disclosure can contain a therapeutic agent at a concentration of about 0.1-0.5 mg / mL, about 0.5-1.0 mg / mL, about 1.0-2.5 mg / mL, about 2.5-5.0 mg / mL, about 5.0-10.0 mg / mL, about 10.0-25.0 mg / mL, about 25.0-50.0 mg / mL, about 50.0-100.0 mg / mL, about 100-150 mg / mL, about 150-200 mg / mL, about 200-250 mg / mL, about 250-300 mg / mL, about 300-350 mg / mL, about 350-400 mg / mL, about 400-450 mg / mL, about 450-500 mg / mL, about 500-550 mg / mL, about 550-600 mg / mL, about 600-650 mg / mL, about 650-700 mg / mL, about 700-750 mg / mL, about 750-800 mg / mL, about 800-850 mg / mL, about 850-900 mg / mL, about 900-950 mg / mL, or about 950-1000 mg / mL.

[0189] In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in less than 1 hour. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in less than 1 day. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in about 0 - 2 hours, about 2 - 4 hours, about 4 - 6 hours, about 6 - 8 hours, about 8 - 10 hours, about 10 - 12 hours, about 12 - 16 hours, about 16 - 20 hours, about 20 - 24 hours, about 24 - 30 hours, about 30 - 36 hours, about 36 - 42 hours, or about 42 - 48 hours. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in less than 1 week. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in about 0 - 2 days, about 2 - 4 days, about 4 - 6 days, about 6 - 8 days, about 8 - 10 days, about 10 - 12 days, about 12 - 16 days, about 16 - 20 days, about 20 - 24 days, about 24 - 30 days, about 30 - 35 days, about 35 - 40 days, about 40 - 45 days, about 45 - 50 days, about 50 - 55 days, about 55 - 60 days. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in less than 1 month. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in less than 12 months. In certain embodiments, the polymeric composition can deliver the therapeutic agent to peak concentration in about 0 - 1 month, about 1 - 2 months, about 2 - 3 months, about 3 - 4 months, about 4 - 5 months, about 5 - 6 months, about 6 - 7 months, about 7 - 8 months, about 8 - 9 months, about 9 - 10 months, about 10 - 11 months, or about 11 - 12 months.

[0190] In certain embodiments, the therapeutic agent is one or more of a growth factor, hemostatic agent, analgesic, anesthetic, antifungal agent, antibiotic, antibacterial agent, anti-inflammatory agent, antimicrobial agent, anthelmintic, antidote, antiemetic, antihistamine, antihypertensive agent, antimalarial agent, antipsychotic agent, antipyretic, antiseptic, anti-arthritis agent, antituberculosis agent, antitussive, antiviral agent, cardiac agent, cathartic, chemotherapeutic agent, coloring or fluorescent contrast agent, corticosteroid (such as steroid), antidepressant, inhibitor, diagnostic aid, diuretic, enzyme, expectorant, hormone, sleeping pill, immunosuppressant, mineral, nutritional supplement, parasympathomimetic, potassium supplement, radiosensitizer, radioisotope, sedative, sulfonamide, stimulant, sympathomimetic, psychotropic agent, urinary anti-infective agent, vasoconstrictor, vasodilator, vitamin, xanthine derivative, organic molecule, small molecule inhibitor, glycosaminoglycan, organometallic drug, chelated metal or metal salt, peptide-based drug, vitamin, nutritional supplement, glycoprotein (such as collagen), extracellular matrix protein or fragment thereof, fibronectin, peptide and / or protein, polysaccharide, carbohydrate (both simple and / or complex), proteoglycan, antigen, oligonucleotide (sense and / or antisense DNA and / or RNA), antibody, nucleic acid sequence, gene therapy agent, triamcinolone acetonide, ovalbumin, or a combination thereof.

[0191] In certain embodiments, the therapeutic agent comprises one or more anti - acanthamoebal agents, antiviral agents, and / or antibacterial agents. In certain embodiments, the therapeutic agent is selected from acyclovir, valacyclovir, famciclovir, penciclovir, trifluridine, vidarabine, hydroxychloroquine, gatifloxacin, daptomycin, tigecycline, telavancin, chloramphenicol, fusidic acid, chlorhexidine, polyhexamethylene biguanide, propamidine, hexamidine, bacitracin, metronidazole, rifampin, ethambutol, streptomycin, isoniazid, silver nanoparticles, copper oxide nanoparticles, glycopeptides (e.g., teicoplanin, vancomycin), aminoglycosides (e.g., gentamicin, tobramycin, amikacin, netimicin), cephalosporins (e.g., cefazolin, cefoxitin, cefotaxime, cefuroxime, moxalactam), macrolides (e.g., erythromycin), oxazolidinones (e.g., linezolid), quinolones, polymyxins, sulfonamides, tetracyclines, penems, carbapenems, monobactams, lincosides, spectinomycin, clindamycin, ansamycins, daptomycin, nitrofurans, trimethoprim - sulfamethoxazole combination, chitosan, penicillin, ciprofloxacin, or combinations thereof.

[0192] In certain embodiments, the therapeutic agent comprises one or more antifungal agents. In certain embodiments, the therapeutic agent is selected from amphotericin B, natamycin, candicin, filipin, hamycin, nystatin, rimocidin, voriconazole, imidazoles, triazoles, thiazoles, allylamines, echinocandins, benzoic acid, ciclopirox, flucytosine, griseofulvin, andexin, haloprogin, tolnaftate, undecylenic acid, and povidone iodine, or combinations thereof.

[0193] In certain embodiments, the therapeutic agent comprises one or more antimicrobial agents. In certain embodiments, the therapeutic agent is polymyxin B, vancomycin, cholera toxin, diphtheria toxin, lysostaphin, hemolysin, bacitracin, besifloxacin, albavancin, daptomycin, enfuvirtide, oritavancin, teicoplanin, telavancin, teravancin, guavanin 2, Maximin H5, dermcidicin, cecropin, andropin, moricin, ceratotoxin, melittin, magainin, delumecin, brevinin-1, esculentin, buforin II, CAP18, LL37, baecin, apidaecin, prophenin, indolicidin, antimicrobial peptide (AMP) (e.g., Tet213), chlorhexidine, chlorhexidine salts, triclosan, polymyxin, tetracycline, aminoglycoside (e.g., gentamicin, tobramycin), rifampicin, erythromycin, neomycin, chloramphenicol, miconazole, quinolone, penicillin, fusidic acid, cephalosporin, mupirocin, metronidazole, secropin, protegrin, bacteriolcin, defensin, nitrofurazone, mafenide, aracylovir, clindamycin, lincomycin, sulfonamide, norfloxacin, pefloxacin, nalidizic acid, cinnamycin, anti-DEFA5, duramycin, nisin, pediocin, Abaecin, Ct-AMP1, Apidaecin IA, Apidaecin IB, Bactenecin, Bactenecin 5, Bactenecin 7, Bactericidin B-2, Aurein family, SMAP-29, Temporin B, Pleurocidin, tachyplesin III, LL-37, citropin 1.1. It contains one or more antimicrobial agents selected from BMAP-27, BMAP-28, Agelaia-MP, Tempol 1Ola, NA-CATH, histatin, Latarcin, Halocidin, bombinin, cathelicidin, Malacidin, MP196, MS100a7a15, murepavadin, Myticin, Mytilin, Paenibacterin, pardaxin, peptibol, SAAP-148, sarcotoxin, Stomoxyn, tachyplesin, thioester-containing protein 1, cionin, alamethicin, arenicin, dermorphin, deltorphin, dermaseptin, pseudin, bombesin, maculatin, LEAP2, eflapeptin, Arylomycin, capreomycin, gramicidin B, Antiamoebin, Bacillomycin, teixobactin, tyrothricin, viomycin, oxalic acid, or combinations thereof.

[0194] In certain embodiments, the therapeutic agent comprises one or more anti-inflammatory agents. In certain embodiments, the therapeutic agent is a steroidal anti-inflammatory agent (e.g., prednisolone), a corticosteroid (e.g., loteprednol etabonate), a salicylate, a non-steroidal anti-inflammatory drug (e.g., bromfenac), an mTOR inhibitor, a calcineurin inhibitor, a synthetic or natural anti-inflammatory protein, dexamethasone, 5-fluorouracil, daunomycin, paclitaxel, curcumin, resveratrol, mitomycin, methylprednisolone, prednisolone, hydrocortisone, fludrocortisone, prednisone, celecoxib, ketorolac, piroxicam, diclofenac, ibuprofen, and ketoprofen, rapamycin, cyclosporine, tacrolimus / FK-506, or combinations thereof, comprising one or more anti-inflammatory agents selected from the group consisting of.

[0195] In certain embodiments, the therapeutic agent comprises one or more growth factors. In certain embodiments, the therapeutic agent comprises growth factors including recombinant hepatocyte growth factor or recombinant nerve growth factor. In certain embodiments, the therapeutic agent is activin (e.g., activin A, activin B, activin AB), adrenomedullin (AM), albumin, alpha-2 macroglobulin, annexin, angiopoietin (Ang), artemin, autocrine motility factor, bone morphogenetic protein (BMP) (e.g., BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP-9), brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor family, ciliary neurotrophic factor (CNTF), connective tissue activating peptide (CTAP), epidermal growth factor (EGF), ephrin (e.g., ephrin A1, ephrin A2, ephrin A3, ephrin A4, ephrin A5, ephrin B1, ephrin B2, ephrin B3), erythropoietin (EPO), fibroblast growth factor (FGF) (e.g., FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGF10, FGF11, FGF12, FGF13, FGF14, FGF15, FGF16, FGF17, FGF18, FGF19, FGF20, FGF21, FGF22, FGF23), basic fibroblast growth factor (bFGF), acidic fibroblast growth factor (aFGF), fetal bovine serum (FBS), glial cell line-derived neurotrophic factor (GDNF), granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), growth differentiation factor (GDF) (e.g., GDF1, GDF9), heparin-binding growth factor, hepatocyte growth factor (HGF), hepatocyte growth factor-like protein (HGFLP), hepatoma-derived growth factor (HDGF), inhibin (e.g., inhibin A, inhibin B), insulin, insulin-like growth factor (IGF) (e.g., IGF-1, IGF-2), interleukin (IL) (e.g., IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-11, and IL-13), keratinocyte growth factor (KGF), leukemia inhibitory factor (LIF), macrophage colony-stimulating factor (M-CSF), macrophage stimulating protein (MSP),One or more growth factors selected from migratory stimulating factor (MSF), myostatin, neuregulin (NRG) (e.g., NRG1, NRG2, NRG3, NRG4), neurotrophin (NT) (e.g., NT-1, NT-2, NT-3, NT-4), neuritin, nerve growth factor (NGF), bone morphogenetic factor, persephin, placental growth factor (PGF), platelet-derived growth factor (PDGF), lenalase (RNLS), stromal cell-derived factor-1, T cell growth factor (TCGF), thrombopoietin (TPO), transforming growth factor alpha (TGF-α), transforming growth factor beta (TGF-β), tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), anti-vascular endothelial growth factor (anti-VEGF) (e.g., bevacizumab, ranibizumab, aflibercept), and bioactive analogs, fragments, derivatives, or combinations thereof of such growth factors are included.

[0196] In certain embodiments, the therapeutic agent comprises one or more hormones. In certain embodiments, the therapeutic agent is anti-Müllerian hormone, Müllerian-inhibiting factor or hormone), adiponectin, adrenocorticotropic hormone, corticotropin, angiotensinogen, angiotensin, antidiuretic hormone, vasopressin, arginine vasopressin, atrial natriuretic peptide, atriopeptin, calcitonin, cholecystokinin, corticotropin-releasing hormone, erythropoietin, follicle-stimulating hormone, gastrin, ghrelin, glucagon, gonadotropin-releasing hormone, growth hormone-releasing hormone, human chorionic gonadotropin, human placental lactogen, growth hormone, somatomedin, leptin, luteinizing hormone, melanocyte-stimulating hormone, orexin, oxytocin, parathyroid hormone, prolactin, relaxin, secretin, somatostatin, thrombopoietin, thyroid-stimulating hormone, thyrotropin, thyrotropin-releasing hormone, or one or more hormones selected from combinations thereof.

[0197] In certain embodiments, the polymer composition of the present disclosure may include one or more growth factors at a concentration (w / v) of from about 0.001 μg / mL to about 2 g / mL. In certain embodiments, the polymer composition of the present disclosure may include one or more growth factors at a concentration (w / v) of from about 0.001 μg / mL to about 1000 μg / mL. In certain embodiments, the polymer composition may include one or more growth factors at a concentration (w / v) of from about 0.01 μg / mL to about 500 μg / mL. In certain embodiments, the polymer composition may include one or more growth factors at a concentration (w / v) of from about 0.1 μg / mL to about 200 μg / mL.In certain embodiments, the polymer composition may comprise one or more growth factors at a concentration (w / v) of about 0.1 - 0.5 μg / mL, about 0.5 - 1.0 μg / mL, about 1 - 2 μg / mL, about 2 - 4 μg / mL, about 4 - 6 μg / mL, about 6 - 8 μg / mL, about 8 - 10 μg / mL, about 10 - 12 μg / mL, about 12 - 14 μg / mL, about 14 - 16 μg / mL, about 16 - 18 μg / mL, about 18 - 20 μg / mL, about 20 - 22 μg / mL, about 22 - 24 μg / mL, about 24 - 26 μg / mL, about 26 - 28 μg / mL, about 28 - 30 μg / mL, about 30 - 35 μg / mL, about 35 - 40 μg / mL, about 40 - 45 μg / mL, about 45 - 50 μg / mL, about 50 - 55 μg / mL, about 55 - 60 μg / mL, about 60 - 65 μg / mL, about 65 - 70 μg / mL, about 70 - 75 μg / mL, about 75 - 80 μg / mL, about 80 - 85 μg / mL, about 85 - 90 μg / mL, about 90 - 95 μg / mL, about 95 - 100 μg / mL, about 100 - 125 μg / mL, about 125 - 150 μg / mL, about 150 - 175 μg / mL, about 175 - 200 μg / mL, about 200 - 225 μg / mL, about 225 - 250 μg / mL, about 250 - 275 μg / mL, about 275 - 300 μg / mL, about 300 - 325 μg / mL, about 325 - 350 μg / mL, about 350 - 375 μg / mL, about 375 - 400 μg / mL, about 400 - 425 μg / mL, about 425 - 450 μg / mL, about 450 - 475 μg / mL, about 475 - 500 μg / mL, about 500 - 550 μg / mL, about 550 - 600 μg / mL, about 600 - 650 μg / mL, about 650 - 700 μg / mL, about 700 - 750 μg / mL, about 750 - 800 μg / mL, about 800 - 850 μg / mL, about 850 - 900 μg / mL, about 900 - 950 μg / mL, about 950 - 1000 μg / mL, about 1000 - 1100 μg / mL, about 1100 - 1200 μg / mL, about 1200 - 1300 μg / mL, about 1300 - 1400 μg / mL, about 1400 - 1500 μg / mL, about 1500 - 1600 μg / mL, about 1600 - 1700 μg / mL, about 1700 - 1800 μg / mL, about 1800 - 1900 μg / mL, or about 1900 - 2000 μg / mL.

[0198] In certain embodiments, the therapeutic agent comprises one or more hemostatic agents (i.e., materials that promote hemostasis) and / or immunosuppressive agents. In certain embodiments, the therapeutic agent comprises one or more agents selected from platelets, platelet-like nanoparticles (e.g., silicate nanoparticles), blood coagulation factors (e.g., thrombin, prothrombin), alkylating agents, antimetabolites, mycophenolic acid, cyclosporine, tacrolimus, rapamycin, or combinations thereof. In certain embodiments, the therapeutic agent comprises an anticoagulant or blood thinner (e.g., heparin).

[0199] In certain embodiments, the polymer compositions of the present disclosure can incorporate or coat cells or cell precursors of a target tissue. In certain embodiments, the polymer composition can incorporate or coat one or more cells or cell precursors of a target tissue selected from nerve cells, muscle cells, myocytes, cardiomyocytes, hepatocytes, keratinocytes, melanocytes, ameloblasts, fibroblasts, osteoblasts, osteoblasts, osteoclasts, endothelial cells, epithelial cells, mesenchymal stem cells, nerve sheath cells (i.e., Schwann cells), embryonic stem cells, adult stem cells, pluripotent stem cells, multipotent stem cells, hematopoietic stem cells, adipose-derived stem cells, bone marrow-derived stem cells, bone cells, nerve cells, or combinations thereof. In certain embodiments, the polymer composition can incorporate or coat endothelial cells (e.g., corneal endothelial cells). In certain embodiments, the polymer composition can incorporate or coat eye cells. In certain embodiments, the polymer composition can incorporate or coat adherent cell types (i.e., cells that form an intercellular network, a 3D vascular system). In certain embodiments, the polymer composition can incorporate or coat monolayer cell types (i.e., 2D). In certain embodiments, the polymer composition can incorporate or coat epithelial cells, endothelial cells, corneal stromal cells, and combinations thereof. In certain embodiments, the polymer composition can incorporate or coat human umbilical vein endothelial cells (HUVECs) or vascular endothelial cells. In certain embodiments, the polymer composition can incorporate or coat human retinal pigment epithelial cells (HRPECs), human neuroepithelial cells, human photoreceptor cells, human corneal endothelial cells, human neural crest cells, human retinal ganglion cells, human ciliary body cells, human cardiomyocytes, human hepatocytes, human dermal cells, human gastrointestinal epithelial cells, human neurons, and human pancreatic islet cells, human immune cells, and other therapeutic human cells.

[0200] In certain embodiments, cells or cell precursors can be incorporated into or onto the polymer Gel matrix by placing the polymer gel composition in the cell culture mixture for a period of time. The culture time can vary depending on the cells used, but can generally be from 1 to 21 days. In certain embodiments, the exposure of the polymer gel composition to the cell culture is repeated to increase the cell density in or on the GelMA matrix.

[0201] In certain embodiments, the polymer compositions of the present disclosure can incorporate cells or cell precursors according to the procedures disclosed in International Publication No. WO 2013 / 040559, or Loessner et al., Nature protocols. 2016 Apr;11(4):727.A1, each of which is incorporated herein by reference in its entirety to the extent each describes the incorporation of cells or cell precursors onto or into a gel matrix such as a GelMA hydrogel.

[0202] Therapeutic use In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant and / or therapeutic composition for treating and / or repairing soft tissue in a subject. In certain embodiments, the polymer compositions of the present disclosure can be used as a delivery vehicle for administering a therapeutic agent for treating and / or repairing soft tissue in a subject. In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant and / or therapeutic composition for treating and / or repairing soft tissue in a subject and / or as a delivery vehicle for administering a therapeutic agent for treating and / or repairing the soft tissue of a subject.

[0203] In certain embodiments, the methods and compositions of the present disclosure can be used to adhere, seal, or treat one or more target soft tissues selected from eye tissue (i.e., the eye), lung, cardiovascular system, skin, kidney, bladder, urethra, dura mater, liver, gastrointestinal, or oral (i.e., mouth) tissue. In certain embodiments, the methods and compositions of the present disclosure can be used to adhere, seal, or treat one or more target soft tissues in a stress environment and / or a physiological environment, or in similar applications that require an elastic and / or adhesive composition.

[0204] In certain embodiments, the present disclosure provides a method for treating and / or repairing soft tissue in a subject using the polymer compositions of the present disclosure. In certain embodiments, the present disclosure provides a method for treating and / or repairing a defect, injury, and / or disease in a subject's soft tissue using the polymer compositions of the present disclosure. In certain embodiments, the method includes applying a pre-gelled polymer composition of the present disclosure (e.g., a polymer composition comprising acryloyl-substituted gelatin) to an applicator, disposing the applicator containing the pre-gelled polymer composition on the surface of a target soft tissue of the subject (e.g., the site of a soft tissue defect, injury, and / or disease), and cross-linking (e.g., photo-cross-linking) the polymer composition by exposing the pre-gelled polymer composition to a cross-linking initiator (e.g., a photoinitiator and visible light). In certain embodiments, the method includes removing the applicator from the gel polymer composition and / or the soft tissue surface after the polymer cross-linking and / or gelation of the polymer composition is complete. In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target soft tissue without using an applicator. In certain embodiments, the pre-gelled polymer composition is applied on or near the target soft tissue (e.g., on or under the same tissue). In certain embodiments, the pre-gelled polymer composition may have strong and persistent adhesion and high retention force on the target soft tissue of the subject. In certain embodiments, the gel polymer composition may have strong and persistent adhesion and high retention force on the target soft tissue of the subject. In certain embodiments, the polymer composition is designed to exhibit physical, mechanical, structural, chemical, and / or biological properties (elasticity, water content) that match or are similar to the target soft tissue. In certain embodiments, the polymer composition is designed to distribute a therapeutic agent to the target soft tissue.

[0205] Eye injuries and diseases In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant and / or therapeutic composition for treating and / or repairing the ocular soft tissues in the eye of a subject. In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant and / or therapeutic composition for treating and / or repairing an ocular defect, an ocular surface injury, or an eye disease in the eye of a subject. In certain embodiments, the ocular defect, injury, or disease is a defect, injury, or disease of the cornea or sclera. In certain embodiments, the injury to the cornea or sclera is a laceration (partial or full thickness), a perforation, an incision (e.g., a surgical incision), or a similar surface trauma (e.g., trauma from a foreign body or projectile). In certain embodiments, the ocular defect, injury, or disease is an eye ulcer such as a corneal ulcer from a severe infection, injury, perforation, or other defect. In certain embodiments, the target soft tissue is an ocular tissue, optionally a subconjunctival ocular tissue or a retinal ocular tissue.

[0206] In certain embodiments, the present disclosure provides a method of treating an eye defect, an eye surface injury, or an eye disease in a subject using the polymer compositions of the present disclosure. In certain embodiments, the method comprises applying a pre-gelled polymer composition of the present disclosure (e.g., a polymer composition comprising acryloyl-substituted gelatin) to an applicator, placing the applicator containing the pre-gelled polymer composition on the surface of the subject's eye, and cross-linking (e.g., photocross-linking) the polymer composition by exposing the pre-gelled polymer composition to a cross-linking initiator (e.g., visible light). In certain embodiments, the method comprises removing the applicator from the gelled polymer composition and / or the eye surface after the polymer cross-linking and / or gelation of the polymer composition is complete. In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target eye tissue without using an applicator. In certain embodiments, the pre-gelled polymer composition can have strong and persistent adhesion and high retention on the subject's eye tissue. In certain embodiments, the gelled polymer composition can have strong and persistent adhesion and high retention on the subject's eye tissue. In certain embodiments, the polymer composition is designed to exhibit physical, mechanical, structural, chemical, and / or biological properties (elasticity, water content) that match or are similar to those of the target eye tissue (e.g., corneal tissue).

[0207] In certain embodiments, the applicator is a curved concave surface. In certain embodiments, the applicator is a curved lens (e.g., a contact lens). In certain embodiments, the curvature of the applicator is similar to the curvature of the target eye surface.

[0208] In certain embodiments, an eye defect, eye surface injury, or eye disease in a target eye tissue can be treated by: (i) forming a pre-formed polymer composition by polymerizing the polymer composition of the present disclosure; and (ii) applying the pre-formed polymer composition onto or under the surface of the target tissue of the subject (e.g., subconjunctival, subretinal). In certain embodiments, application to the surface of the target tissue includes application / injection into the space immediately beneath the surface of the target tissue (e.g., subconjunctival application to the eye tissue, subretinal application to the eye tissue). In certain embodiments, the pre-formed polymer composition can be designed to have certain physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity).

[0209] In certain embodiments, an eye defect, eye surface injury, or eye disease in a target eye tissue can be treated by: (i) forming a pre-formed hydrogel polymer composition by polymerizing the polymer composition of the present disclosure; (ii) drying the hydrogel polymer by removing a substantial portion of the interstitial fluid from the hydrogel (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the interstitial fluid); (iii) applying the pre-formed polymer composition onto or under the surface of the target tissue of the subject (e.g., subconjunctival, subretinal); and (iv) optionally rehydrating the dried hydrogel polymer to a substantially hydrated form (e.g., at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the interstitial fluid volume). In certain embodiments, application to the surface of the target tissue includes application / injection into the space immediately beneath the surface of the target tissue (e.g., subconjunctival application to the eye tissue, subretinal application to the eye tissue). In certain embodiments, the pre-formed polymer composition can be designed to have certain physical, mechanical, structural, chemical, and / or biological properties (e.g., elasticity, biodegradability, porosity).

[0210] Oral injuries and oral diseases In certain embodiments, the polymer compositions of the present disclosure can be used as sealants and / or therapeutic compositions for treating and / or repairing soft tissues in a subject's oral cavity. In certain embodiments, the polymer compositions can be used for the treatment and / or repair of oral tissues associated with periodontal disease, injury, or illness. In certain embodiments, the periodontal disease, injury, or illness can include those associated with periodontal implants, including peri-implant diseases (PID) such as peri-implant mucositis (PIM) and peri-implantitis (PI). These conditions are often associated with inflammation of the soft tissues around the periodontal implant (due to bacterial accumulation and biofilm formation), which can result in hemorrhagic suppuration, erythema, swelling, and infection of the oral tissues, as well as the potential for progressive bone loss leading to implant failure.

[0211] In certain embodiments, the polymer compositions of the present disclosure can be used to seal areas of soft tissue around periodontal implants. In certain embodiments, the polymer compositions of the present disclosure can be used to deliver therapeutic agents (e.g., antimicrobial or anti-inflammatory agents) to areas of soft tissue around periodontal implants. In certain embodiments, the polymer composition includes an osteoinductive agent. In certain embodiments, the polymer composition includes one or more osteoinductive agents selected from silicate nanoparticles (SN), calcium salts, bioglass, hydroxyapatite, demineralized bone matrix (DBM), or combinations thereof. In certain embodiments, the polymer composition includes one or more silicate nanoparticles including SN containing one or more metals such as calcium, aluminum, silver, gold, platinum, palladium, lithium, magnesium, sodium, titanium, vanadium, chromium, manganese, iron, cobalt, nickel, copper, zinc, iridium, or combinations thereof. In certain embodiments, the silicate nanoparticles include laponite nanoparticles. In certain embodiments, the polymer composition includes one or more calcium salts such as calcium phosphate, calcium sulfate, calcium hydroxide, calcium bromide, calcium fluoride, calcium iodide, calcium hydride, or combinations thereof.

[0212] In certain embodiments, the present disclosure provides a method of treating a defect, injury, or disease in a target oral soft tissue using the polymer compositions of the present disclosure. In certain embodiments, the method comprises applying a pre-gelled polymer composition of the present disclosure (e.g., a polymer composition comprising acryloyl-substituted gelatin) to an applicator, placing the applicator containing the pre-gelled polymer composition on the surface of a target oral soft tissue (e.g., soft tissue around a periodontal implant), and cross-linking (e.g., photo-cross-linking) the polymer composition by exposing the pre-gelled polymer composition to a cross-linking initiator (e.g., visible light). In certain embodiments, the method comprises removing the applicator from the gel polymer composition and / or the oral soft tissue surface after the polymer cross-linking and / or gelation of the polymer composition is complete. In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target oral soft tissue without using an applicator. In certain embodiments, the pre-gelled polymer composition can have strong and persistent adhesion and high retention on the target oral soft tissue. In certain embodiments, the gel polymer composition can have strong and persistent adhesion and high retention on the target oral soft tissue. In certain embodiments, the polymer composition is designed to exhibit physical, mechanical, structural, chemical, and / or biological properties (elasticity, water content) that match or are similar to those of the target oral soft tissue (e.g., soft tissue surrounding a periodontal implant).

[0213] Nerve injuries and diseases In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant and / or therapeutic composition for treating and / or repairing soft tissue in a target nervous system (e.g., central nervous system (CNS), peripheral nervous system (PNS)) of a subject. In certain embodiments, the polymer composition can be used for the treatment and / or repair of nerve tissue associated with traumatic or surgical injuries, including peripheral nerve injury (PNI). Typical surgical interventions for these conditions (including suturing and / or commercially available adhesives) are often associated with inflammation, enhanced foreign body reaction (FBR), scarring, slow nerve regeneration, or loss of nerve function (partial or complete).

[0214] In certain embodiments, nerve tissue can be treated or sealed by applying the polymer compositions of the present disclosure to the target nerve tissue. In certain embodiments, nerve tissue can be treated or sealed by applying the polymer compositions of the present disclosure to the lumen of a nerve conduit at the site of a nerve injury.

[0215] In certain embodiments, the present disclosure provides methods of treating a deficit, injury, or disease in a subject's nerve or CNS tissue using the polymer compositions of the present disclosure. In certain embodiments, the method includes applying a pre-gelled polymer composition of the present disclosure (e.g., a polymer composition containing acryloyl-substituted gelatin) to an applicator, positioning the applicator containing the pre-gelled polymer composition on the surface of the subject's nerve or CNS tissue (e.g., a nerve of the peripheral nervous system), and crosslinking (e.g., photocrosslinking) the polymer composition by exposing the pre-gelled polymer composition to a crosslinking initiator (e.g., visible light). In certain embodiments, the method includes removing the applicator from the gel polymer composition and / or the nerve / CNS tissue surface after the polymer crosslinking and / or gelation of the polymer composition is complete. In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target nerve or CNS tissue without using an applicator. In certain embodiments, the pre-gelled polymer composition can have strong and persistent adhesion and high retention on the target nerve or CNS tissue of the subject. In certain embodiments, the gel polymer composition can have strong and persistent adhesion and high retention on the target nerve or CNS tissue of the subject. In certain embodiments, the polymer composition is designed to exhibit physical, mechanical, structural, chemical, and / or biological properties (elasticity, water content) that match or are similar to those of the target nerve or CNS tissue (e.g., a nerve of the peripheral nervous system).

[0216] In certain embodiments, the polymer compositions of the present disclosure can include a polymer or a therapeutic component, or can be generated, analyzed, or used by the methods disclosed in U.S. Patent Application Publication No. 20190070338 (including the treatment of nerve injury), and the literature is incorporated herein by reference in its entirety to the extent that it describes the composition, generation, analysis, and use of acrylated gelatin polymer compositions such as GelMA hydrogels.

[0217] Cardiovascular injuries and diseases In certain embodiments, the polymer compositions of the present disclosure can be used as a sealant and / or therapeutic composition for treating and / or repairing soft tissue in a subject's cardiovascular system (e.g., the heart). In certain embodiments, the polymer composition can be used for the treatment and / or repair of cardiovascular tissue associated with traumatic or surgical injury involving heart tissue. Typical surgical interventions for these diseases (including suturing and / or commercially available adhesives) are often associated with inflammation and infection, scarring, slow tissue regeneration, or loss of function (partial or complete).

[0218] In certain embodiments, the vascular / cardiovascular tissue can be treated or sealed by applying the polymer compositions of the present disclosure to the target vascular / cardiovascular tissue. In certain embodiments, the vascular / cardiovascular tissue can be treated or sealed by applying the cell-laden hydrogel compositions of the present disclosure to the target vascular / cardiovascular tissue. In certain embodiments, the cell-laden hydrogel composition includes cells or cell precursors that promote or facilitate the repair, recovery, replacement, or regeneration of vascular / cardiovascular tissue (e.g., heart tissue). In certain embodiments, the cell-laden hydrogel composition includes one or more cells or cell precursors selected from smooth muscle cells, cardiomyocytes, fibroblasts, mesenchymal stem cells, bone marrow stem cells, or combinations thereof. In certain embodiments, the cell-laden hydrogel composition is in a form suitable for use as a mat, fabric, mesh, or other shape of coating or implant.

[0219] In certain embodiments, the present disclosure provides a method of treating a defect, injury, or disease in a target cardiovascular tissue using the polymer compositions of the present disclosure. In certain embodiments, the method comprises applying a pre-gelled polymer composition of the present disclosure (e.g., a polymer composition comprising acryloyl-substituted gelatin) to an applicator, disposing the applicator containing the pre-gelled polymer composition on the surface of a target cardiovascular tissue (e.g., heart tissue), and crosslinking (e.g., photocrosslinking) the polymer composition by exposing the pre-gelled polymer composition to a crosslinking initiator (e.g., visible light). In certain embodiments, the method comprises removing the applicator from the gel polymer composition and / or the cardiovascular tissue surface after the polymer crosslinking and / or gelation of the polymer composition is complete. In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target cardiovascular tissue without using an applicator. In certain embodiments, the pre-gelled polymer composition can have strong and persistent adhesion and high retention force on the target cardiovascular tissue. In certain embodiments, the gel polymer composition can have strong and persistent adhesion and high retention force on the target cardiovascular tissue. In certain embodiments, the polymer composition is designed to exhibit physical, mechanical, structural, chemical, and / or biological properties (elasticity, water content) that match or are similar to those of the target cardiovascular tissue (e.g., heart tissue).

[0220] In certain embodiments, the polymer compositions of the present disclosure can include a polymer or a therapeutic component, or can be produced, analyzed, or used by the methods disclosed in International Publication No. WO 2014 / 063194 (including the treatment of cardiovascular injury), the disclosure of which is incorporated herein by reference in its entirety to the extent it describes the composition, production, analysis, and use of acrylated gelatin polymer compositions such as GelMA hydrogels.

[0221] Lung injury and disease In certain embodiments, the polymer compositions of the present disclosure can be used as sealants and / or therapeutic compositions for treating and / or repairing soft tissue in the lungs of a subject. In certain embodiments, the polymer compositions can be used for treating and / or repairing lung tissue associated with traumatic or surgical injury. Typical surgical interventions for these diseases (including suturing and / or commercially available adhesives) are often associated with inflammation and infection, scarring, slow tissue regeneration, or loss of function (partial or complete).

[0222] In certain embodiments, lung tissue can be treated or sealed by applying the polymer compositions of the present disclosure to the target vascular / cardiovascular tissue. In certain embodiments, lung tissue can be treated or sealed by applying the cell-laden hydrogel compositions of the present disclosure to the target lung tissue. In certain embodiments, the cell-laden hydrogel compositions include cells or cell precursors that promote or facilitate the repair, recovery, replacement, or regeneration of lung tissue. In certain embodiments, the cell-laden hydrogel compositions are in the form of a mat, fabric, mesh, or other shape suitable for use as a coating or implant.

[0223] In certain embodiments, the polymer composition includes acryloyl-substituted gelatin (e.g., GelMA) and acryloyl-substituted PEG (e.g., PEGDA) in a ratio of about 30:1 to about 1:30 w / w. In certain embodiments, the polymer composition includes acryloyl-substituted gelatin (e.g., GelMA) and acryloyl-substituted hyaluronic acid (e.g., MeHA) in a ratio of about 30:1 to about 1:30 w / w. In certain embodiments, the polymer composition includes acryloyl-substituted gelatin (e.g., GelMA), acryloyl-substituted PEG (e.g., PEGDA), and acryloyl-substituted hyaluronic acid (e.g., MeHA).

[0224] In certain embodiments, the present disclosure provides a method of treating a defect, injury, or disease in a subject's lung tissue using the polymer compositions of the present disclosure. In certain embodiments, the method comprises applying a pre-gelled polymer composition of the present disclosure (e.g., a polymer composition comprising acryloyl-substituted gelatin) to an applicator, placing the applicator containing the pre-gelled polymer composition on the surface of the subject's lung tissue, and cross-linking (e.g., photocross-linking) the polymer composition by exposing the pre-gelled polymer composition to a cross-linking initiator (e.g., visible light). In certain embodiments, the method comprises removing the applicator from the gel polymer composition and / or the lung tissue surface after the polymer cross-linking and / or gelation of the polymer composition is complete. In certain embodiments, the pre-gelled polymer composition is applied directly to the surface of the target lung tissue without using an applicator. In certain embodiments, the pre-gelled polymer composition can have strong and persistent adhesion and high retention on the subject's lung tissue. In certain embodiments, the gel polymer composition can have strong and persistent adhesion and high retention on the subject's lung tissue. In certain embodiments, the polymer composition is designed to exhibit physical, mechanical, structural, chemical, and / or biological properties (elasticity, water content) that match or are similar to those of the target lung tissue.

[0225] V. Definitions Throughout the present disclosure, substituents or properties of the compounds of the present disclosure are disclosed within groups or ranges. The present disclosure is specifically intended to include all individual or sub-combinations of the members of such groups and ranges.

[0226] Unless otherwise defined, the following terms and phrases have the meanings set forth below. The definitions are not intended to be limiting in nature but rather to aid in a clearer understanding of certain aspects of the present disclosure.

[0227] GelMA polymer composition: As used herein, the term "GelMA polymer composition" refers to.

[0228] Administer: As used herein, the term "administer" refers to providing a composition to a subject.

[0229] Improve: As used herein, the term "improve" or "improving" refers to alleviating the severity of at least one indicator of a condition or disease.

[0230] Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In certain embodiments, "animal" refers to a human at any stage of development. In certain embodiments, "animal" refers to a non-human animal at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, or pig). In certain embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, and insects. In certain embodiments, the animal is a transgenic animal, a genetically engineered animal, or a clone.

[0231] Approximately: As used herein, the term "approximately" or "about", when applied to one or more values of interest, refers to a value similar to the referenced value. The term can refer to + / - 10% of the recited value. In certain embodiments, the term refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the recited reference value in either direction (greater than or less than) the recited reference value, unless otherwise stated or apparent from the context (except where such a numerical value exceeds 100% of the possible values).

[0232] Associated with: As used herein, when used with respect to two or more moieties, the terms “associated with,” “conjugated to,” “linked to,” “coupled to,” and “tethered to” mean that the moieties are physically associated or bonded to each other, either directly or through one or more additional moieties that function as a linker, so as to form a sufficiently stable structure such that the moieties remain physically associated, for example, under the conditions in which the structure is used, such as physiological conditions. “Association” does not necessarily mean a chemical bond by direct covalent bonding strictly speaking. Also, it can be suggested that ionic bonds or hydrogen bonds, or hybridization-based connectivity are sufficiently stable such that the “associated” entities remain physically associated.

[0233] Biocompatible: As used herein, the term “biocompatible” refers to a substance that results in minimal or zero toxicity, harmfulness, or immune response in living tissue.

[0234] Biodegradable: As used herein, the term “biodegradable” refers to a material that can be partially or completely degraded under physiological conditions into biologically treatable by-products. For example, a material can be considered biodegradable if at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the material degrades within a desired period (e.g., minutes, hours, days, weeks, or months, depending on the nature of the material and the physiological application) under physiological conditions. The term “biodegradable” can encompass the term “bioabsorbable,” which describes a substance that degrades under physiological conditions into products that are bioabsorbed by the host subject (e.g., as metabolites of biochemical systems).

[0235] Biologically active: As used herein, the term “biologically active” refers to the characteristic of any substance or material that has activity in a biological system and / or organism. For example, a material that has a biological effect on an organism when administered to the organism is considered to be biologically active.

[0236] Compounds: The compounds of the present disclosure include all isotopes of atoms occurring in the intermediates or final compounds. "Isotope" refers to an atom having the same atomic number but a different mass number due to a different number of neutrons in the atomic nucleus. For example, isotopes of hydrogen include tritium and deuterium. The compounds and salts of the present disclosure can be prepared to form solvates and hydrates in combination with solvents or water molecules by conventional methods.

[0237] Crosslinking: As used herein, the term "crosslinking" or "crosslink" refers to the formation of a bond (e.g., a covalent bond) that links one polymer unit to another polymer unit.

[0238] Encapsulation: As used herein, the term "encapsulation" means to surround, enclose, or envelop.

[0239] Designed: As used herein, embodiments of the present disclosure are "designed" if they are designed to have features or characteristics that are different from the starting point or natural molecules, whether structural or chemical.

[0240] Effective amount: As used herein, the term "effective amount" of an agent is an amount sufficient to produce a beneficial or desired result, e.g., a clinical result, and thus the effective amount depends on the circumstances in which it is applied. For example, in the context of administering an agent to treat an ocular trauma or disorder, the effective amount of the agent is an amount sufficient to achieve treatment of the ocular trauma or disorder, e.g., compared to the response obtained without administration of the agent.

[0241] Feature: As used herein, "feature" refers to a characteristic, trait, or distinguishing element.

[0242] In vitro: As used herein, the term "in vitro" refers to events that occur not within a living organism (e.g., an animal, plant, or microorganism), but in an artificial environment, e.g., a test tube or reaction vessel, cell culture, Petri dish, etc.

[0243] In vivo: As used herein, the term "in vivo" refers to an event that occurs within an organism (e.g., an animal, a plant, or a microorganism or its cells or tissues).

[0244] Modification: As used herein, "modification" refers to an altered state or structure of a molecule of the present disclosure. A molecule can be modified in many chemical, structural, and functional ways. As used herein, embodiments of the present disclosure are modified when they have or possess a feature or property different from the starting or native molecule, whether structural or chemical.

[0245] Non-human animal: As used herein, the term "non-human animal" includes all animals (e.g., vertebrates) other than Homo sapiens, including wild and domestic animal species. Examples of non-human vertebrates include, but are not limited to, mammals such as alpaca, badger, bison, camel, cat, cow, deer, dog, donkey, gayal, goat, guinea pig, horse, llama, yak, pig, rabbit, reindeer, water buffalo, and yak. Non-human animals include non-human primates.

[0246] Pharmaceutically acceptable: The term "pharmaceutically acceptable" or "therapeutically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0247] Pharmaceutically acceptable excipient: As used herein, the term "pharmaceutically acceptable excipient" or "therapeutically acceptable excipient" refers to components other than the polymeric compositions described herein (e.g., a vehicle capable of suspending or dissolving a polymeric compound) and having substantially non-toxic and non-inflammatory properties in a subject.

[0248] Pharmaceutically acceptable salts: The present disclosure also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds in which the existing acid or base moiety is modified by converting it to its salt form (e.g., by reacting a free base moiety with an appropriate organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, alkali or organic salts of acidic residues such as carboxylic acids, and the like. Representative acid addition salts include acetate, acetic acid, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzenesulfonic acid, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, 4-(2-hydroxyethyl)-1-piperazineethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, maleate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. The pharmaceutically acceptable salts of the present disclosure include, for example, conventional non-toxic salts of the parent compounds formed from non-toxic inorganic or organic acids.The pharmaceutically acceptable salts of the present disclosure can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or free base forms of these compounds with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or a mixture of the two, and generally, a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile can be used.

[0249] Subject: As used herein, the term "subject" refers to any organism to which a composition according to the present disclosure can be administered for, for example, experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (such as mammals such as mice, rats, rabbits, non-human primates, and humans) and / or plants. A subject or patient can be seeking, in need of, receiving, about to receive, or being treated by a trained professional for a particular disease or condition.

[0250] Substantially: As used herein, the term "substantially" refers to a qualitative state that exhibits the entire or almost entire range or degree of a characteristic or property of a subject. One of ordinary skill in the art will understand that biological and chemical phenomena rarely complete and / or proceed to a complete state, or achieve or avoid a result absolutely. Thus, the term "substantially" is used herein to explicitly capture the potential lack of completeness inherent in many biological and chemical phenomena. Similarly, the exclusion of the term "substantially" does not exclude a similar potential lack of completeness inherent in many biological and chemical phenomena.

[0251] Synthesis: The term "synthesis" means being produced, prepared, and / or manufactured by human hands. The synthesis of a polynucleotide or polypeptide, or other molecule of the present disclosure can be chemical or enzymatic.

[0252] Therapeutic agent: The term "therapeutic agent" refers to any agent that, when administered to a subject, has a therapeutic, diagnostic, and / or prophylactic effect and / or induces a desirable biological and / or pharmacological effect. Examples of therapeutic agents include, but are not limited to, oligonucleotides (e.g., sense and / or antisense DNA and / or RNA), proteins and polypeptides (e.g., hormones, growth factors), small molecules and pharmaceuticals, and cells (e.g., stem cells, epithelial cells).

[0253] Treat: As used herein, the term "treat" refers to the partial or complete alleviation, amelioration, recovery, reduction, prevention, delay in the onset, inhibition of progression, decrease in severity, and / or decrease in incidence of one or more symptoms or features of a particular infectious disease, disorder, impairment, and / or condition. Treatment can be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition associated with the medical condition, and / or to a subject who exhibits only early signs of a disease, disorder, and / or condition, for the purpose of reducing the risk of developing the medical condition associated with the disease, disorder, and / or condition.

[0254] One of ordinary skill in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described herein. The scope of the disclosure is not intended to be limited to the above description.

[0255] In the claims, unless the contrary indication appears or is otherwise clear from the context, articles such as "a," "an," and "the" may mean one or more. A claim or description that includes "or" between one or more members of a group is considered satisfied, unless the contrary indication appears or is otherwise clear from the context, when one, two or more, or all of the group members are present in, used in, or related to a given product or process. The disclosure may include embodiments where exactly one member of the group is present in, used in, or related to a given product or process. The disclosure may include embodiments where two or more, or all group members are present in, used in, or related to a given product or process.

[0256] It should also be noted that the term "comprising" is intended to be open-ended and allows for the inclusion of additional elements or steps, but is not necessarily required. Thus, when the term "comprising" is used herein, the term "consisting of" is also included and disclosed.

[0257] The abbreviation "e.g." (exempli gratia in Latin) is used herein to indicate non-limiting examples. Thus, the abbreviation "e.g." is synonymous with the term "for example".

[0258] The abbreviation "i.e." (id est in Latin) is used herein to indicate non-limiting rephrases or explanations. Thus, the abbreviation "i.e." is synonymous with the term "that is".

[0259] When ranges are given, endpoints are included. Further, unless otherwise indicated by the context of the disclosure or clear to one of ordinary skill in the art in another manner, values expressed as ranges are to be construed to include any specific value or sub-range within the recited range that is within the unit of one tenth of the lower limit of the range, to the extent that the context of the disclosure does not clearly indicate otherwise.

[0260] Furthermore, it should be understood that any particular embodiment of the disclosure belonging to the prior art may be explicitly excluded from any one or more of the claims. Such embodiments are considered to be known to one of ordinary skill in the art and thus may be excluded even if not explicitly recited herein. Any particular embodiment of the compositions of the disclosure (e.g., any antibiotic, therapeutic agent, or active ingredient, any manufacturing method, any method of use, etc.) may be excluded from any one or more of the claims for any reason, whether or not related to the existence of prior art.

[0261] The terms used are explanatory rather than limiting, and it should be understood that modifications may be made within the scope of the appended claims without departing from the true scope and spirit of the disclosure in its broader aspects.

[0262] Although this disclosure has been described with some length and particularity with respect to several described embodiments, it is not intended to be limited to such details or embodiments, or any particular embodiment, but rather, the appended claims are to be construed with the broadest possible interpretation in view of the prior art and, accordingly, to effectively encompass the intended scope of this disclosure.

[0263] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. Further, the section headings, materials, methods, and examples are illustrative only and not intended to be limiting.

Examples

[0264] Example 1. Preparation of a precursor polymer composition (a) Preparation of a gelatin methacryloyl (GelMA) precursor polymer composition The GelMA precursor polymer composition can be synthesized as described in the art. For example, GelMA is synthesized by dissolving 10% (w / v) gelatin (e.g., porcine gelatin) in phosphate buffered saline (PBS) and then heating at 60 °C for 20 minutes. After heating, 8% (v / v) methacrylic anhydride is added dropwise at 50 °C for 3 hours (with continuous stirring), followed by dilution with PBS and dialysis at 40 - 50 °C for about 7 days (using deionized water). The resulting mixture is filtered and lyophilized for 4 days. The resulting GelMA precursor polymer composition can be stored at -80 °C until further use.

[0265] In one alternative method, GelMA is synthesized by dissolving 10 grams of fish skin-derived gelatin in 100 ml of DPBS at 60 °C for 30 minutes. Next, 8% (v / v) methacrylic anhydride is added dropwise to the solution at 60 °C with stirring for 3 hours. An additional 300 ml of DPBS is added to stop the reaction. The resulting mixture is dialyzed at 50 °C for about 5 days using a deionized water bath to remove unreacted methacrylic anhydride. The resulting solution is filtered and lyophilized for about 4 days.

[0266] (b) Preparation of methacrylated hyaluronic acid (MeHA) precursor polymer composition The MeHA precursor polymer composition can be synthesized as described in the art, such as those presented in Bencherif et al., Biomaterials 29, 1739 - 1749 (2008), Prata et al., Biomacromolecules 11, 769 - 775 (2010). For example, MeHA is synthesized by dissolving approximately 2 grams of sodium hyaluronate salt in 200 ml of deionized water, followed by sequentially adding 8.0 mL of triethylamine, 8.0 mL of glycidyl methacrylate, and 4.0 g of tetrabutylammonium bromide (with 1 hour of stirring between each sequential addition). The resulting mixture is incubated at 55 °C for 1 hour, then cooled (ice bath) and precipitated in acetone (4 L) to form a white solid precipitate. The precipitate is rinsed with fresh acetone, dissolved in pure water, dialyzed for 2 days, then frozen and lyophilized for storage.

[0267] (c) Preparation of polyethylene glycol diacrylate (PEGDA) precursor polymer composition The PEGDA precursor polymer composition can be synthesized as described in the art. For example, PEGDA is synthesized by reacting 10 grams (10% w / v) of PEG in dichloromethane with triethylamine and acryloyl chloride (1:4:4 molar ratio) at 4 °C under inert conditions (stirring overnight). The resulting mixture is filtered and then precipitated using ice-cold ether. The resulting precipitate product is filtered and dried in a vacuum desiccator overnight to remove the residual material.

[0268] In one alternative method, PEGDA is synthesized by dissolving PEG diol in benzene and then azeotropically distilling with toluene using a Dean-Stark trap to remove water to ensure dry acrylation conditions. The acrylation of PEG is carried out by dissolving PEG in a dichloromethane solution (under argon), followed by adding acryloyl chloride and triethylamine in a molar ratio of 2:3:3 of the OH groups of PEG:acryloyl chloride:triethylamine. The resulting mixture is stirred overnight at room temperature (in a darkroom condition). The resulting product is then precipitated using diethyl ether, cooled to 4 °C, followed by filtration recovery and vacuum oven drying.

[0269] Example 2: Preparation of Hydrogel Polymer Composition The hydrogel polymer composition can be synthesized as described in the art. For example, the lyophilized GelMA precursor polymer composition produced according to Example 1(a) is dissolved in PBS or (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffered saline at a concentration of 5-25% (w / v). Either 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone or eosin Y or eosin Y disodium salt is added as a photoinitiator, and the mixture is dissolved at 20-80 °C. The resulting precursor polymer composition is photocrosslinked by visible light irradiation (e.g., blue or white light) to form a GelMA hydrogel polymer composition. In one alternative method, the MeHA precursor polymer composition [Example 1(b)] and / or the PEGDA precursor polymer composition [Example 1(c)] can be added to the precursor polymer solution at the target concentration, and the amounts of each component are added based on the desired physical, mechanical, structural, chemical, and / or biological properties of the hydrogel polymer composition.

[0270] In one alternative method, the GelMA hydrogel polymer composition is first synthesized by dissolving 7-15% w / v of the gelatin methacryloyl of Example 1 in a solution containing at least one photoinitiator element such as a mixture of triethanolamine (about 2% w / v) and N-vinylcaprolactam (about 1.25% w / v) in distilled water at room temperature. Then, a solution of eosin Y disodium salt (0.5 mM) is added to the gelatin methacryloyl solution, and the resulting precursor polymer composition is photocrosslinked for 120 seconds under exposure to visible light (420-480 nm). In one alternative method, the MeHA precursor polymer composition [Example 1(b)] and / or the PEGDA precursor polymer composition [Example 1(c)] can be added to the precursor polymer solution at the target concentration, and the amounts of each component are added based on the desired physical, mechanical, structural, chemical, and / or biological properties of the hydrogel polymer composition.

[0271] In one alternative, microparticles (e.g., micelles) containing a therapeutic agent (e.g., an ophthalmic antibiotic such as ciprofloxacin) are incorporated into the GelMA precursor polymer composition prior to photocrosslinking.

[0272] Porosity can be measured and analyzed by creating a freeze-dried gold sputter-coated hydrogel sample, which can then be imaged using a scanning electron microscope (SEM).

[0273] Samples can also be subjected to various mechanical tests, including elasticity, swelling, compression testing, texture, and tensile testing.

[0274] In one alternative, the GelMA hydrogel polymer composition is formed on the surface of the target tissue. The resulting samples can be subjected to various mechanical and therapeutic tests, including adhesion, burst pressure, wound closure strength, shear strength, and durability / degradation rate.

[0275] Example 3: Preparation of Hydrogel Polymer Composition A hydrogel polymer composition was prepared according to the following steps.

[0276] A photoinitiator mixture containing 0.35 mg / mL eosin Y (20% v / v), 12.5 mg / mL N-vinylcaprolactam, and 18.75 mg / mL triethanolamine (80% v / v) in phosphate buffered saline (PBS; pH 7) was prepared, and the pH was adjusted using concentrated HCl as needed.

[0277] The polymer precursors were obtained from the following suppliers: (1) GelMA - Rousselot Biosciences (160P80 or GelMA 160P40 or GelMA 160P10), (2) HAMA - HTL Biotechnology (BLo - RD029 - 008), (3) HAGM - synthesized in - house according to methods known in the art (see, for example, Example 1(b)), and (4) PEGDA - Jen Kem (ACLT - PEG35K - ACLT). The polymer precursors were allowed to reach room temperature (RT) before being incorporated into the hydrogel polymer precursor composition.

[0278] The PEGDA precursor material (when applicable to the target formulation) was first added to the photoinitiator mixture at the desired concentration (e.g., 0.1 - 20% w / v) and dissolved at 37 °C for about 5 minutes.

[0279] Next, the GelMA precursor material (when applicable to the target formulation) was added to the hydrogel precursor mixture at the desired concentration (e.g., 4 - 20% w / v) and dissolved at 60 °C for about 2 hours while vortexing occasionally.

[0280] Next, the MeHA (i.e., HAMA or HAGM) precursor material (when applicable to the target formulation) was added to the hydrogel precursor mixture at the desired concentration (e.g., 1 - 3% w / v) and dissolved at 60 °C overnight with stirring (to prevent any phase separation).

[0281] Once all the precursor materials were completely dissolved in the hydrogel precursor mixture, the active agent (when applicable to the target formulation) was added at the desired concentration (e.g., 1 - 350 mg / mL). The mixture was maintained at 37 °C with stirring until polymerization was ready.

[0282] Hydrogel disk samples were prepared by pipetting approximately 100 μL of the hydrogel precursor mixture into individual poly(dimethylsiloxane) (PDMS) cylinder-shaped molds placed within the wells of a 24-well untreated plate. The polymer composition was then photocrosslinked using a Dolan-Jenner high-intensity LED illumination device (MI-LED-US-B1) with a dual-arm gooseneck configuration (one arm on top and one arm on the bottom, allowing for dual light exposure from above and below, incident rays 90°). Each arm outputs an average light intensity of approximately 100 mW / cm2 (λmax = 450, 540 nm) with a light exposure time varying from 5 seconds to 4 minutes.

[0283] Hydrogel rod samples were prepared by immersing a 0.75 mm inner diameter borosilicate glass capillary into the hydrogel precursor mixture and then vibrating the capillary tube until it was filled to approximately 10 mm from the opening. The polymer composition was then photocrosslinked using a Dolan-Jenner high-intensity LED illumination device (MI-LED-US-B1) with a dual-arm gooseneck configuration (one arm on top and one arm on the bottom, allowing for dual light exposure from above and below, incident rays 90°). Each arm outputs an average light intensity of approximately 100 mW / cm2 (λmax = 450, 540 nm) with a light exposure time of approximately 4 minutes. The hydrogel rods were extruded from the capillary tubes using a 0.5 mm diameter quartz rod and then cut to a predetermined size using calipers.

[0284] Example 4: Study of Hydrogel Properties a) Degree of Crosslinking - Photopolymerization Time A study was completed to analyze the correlation of the degree of crosslinking within the hydrogel as a function of the photopolymerization time.

[0285] Hydrogels of HAMA only were prepared according to the general procedure of Example 3 with photo-crosslinking times of 15 seconds, 1 minute, 2 minutes, and 4 minutes. The resulting hydrogels were dried under vacuum, dissolved in deuterated DMSO, and then analyzed using proton NMR analysis (d-DMSO solvent). Other techniques such as Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy can also be used. For HAMA hydrogels, the change in the proton ratio between the methyl methacrylate group and the HA carbonylmethyl group was quantified as a function of the light exposure time and normalized to the ratio present in non-crosslinked HAMA to represent the degree of crosslinking (%). The results in Figure 4A show that the degree of crosslinking increases as the light exposure time increases.

[0286] Hydrogels of GelMA only were prepared according to the general procedure of Example 3 with photo-crosslinking times of 30 seconds, 1 minute, 2 minutes, and 4 minutes. The resulting hydrogels were dried under vacuum, dissolved in deuterated DMSO, and then analyzed using proton NMR analysis (d-DMSO solvent). Other techniques such as Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy can also be used. For GelMA hydrogels, the ratio of the ME methyl group to the GelMA lysine CH 2 group was analyzed. The results in Figure 4B show that as the light exposure time increases, the ratio of [ME methyl group to lysine CH 2 group] decreases.

[0287] b) Swelling ratio Studies were completed to analyze the swelling ratios of hydrogels with various GelMA, HAMA, and PEGDA concentrations.

[0288] G4-H M 1-P1, G7-H M 1, G4-H M 1, and H M 1-P1 hydrogels (described in Table 1) were prepared according to the general procedure of Example 3 with a photo-crosslinking time of 4 minutes. The resulting hydrogel cylinders had a diameter of 6 mm and a volume of 75 μL.

[0289] Two methods were adopted to evaluate swelling. In the first method, the weight of the hydrogel immediately after crosslinking was used as the "dry" hydrogel weight (Wd-1), and in the second method, the dry polymer weight (hydrogel dried in vacuum) was used as the dry hydrogel weight (Wd-2). In both cases, the "wet" hydrogel weight (Ws) refers to the hydrogel incubated in 1×PBS at 37 °C for 48 hours. The swelling ratio was calculated as follows. Swelling ratio = (Ws - Wd) / Wd

[0290] The results of the first measurement method are inconsistent as shown in Figure 5A. The results of the second measurement method are more consistent as shown in Figure 5B, indicating that the increase in GelMA concentration helps to reduce the swelling of the hydrogel.

[0291] G4-H M 1-P1, G7-H M 1, G4-H M 1, and H M The 1-P1 hydrogel was studied for the swelling / re-swelling effect. After drying and swelling the samples using the second method, a second re-drying and re-swelling were performed. The results presented in Figure 5C show that the swelling ratio decreases significantly when the hydrogel is exposed to multiple drying / swelling cycles.

[0292] G4-P1, G4-P0.1, G20, G10, G5, P20, and P5 hydrogels (described in Table 1) were prepared with a photocrosslinking time of 4 minutes according to the general procedure of Example 3. Swelling was evaluated using the "dry" polymer weight (hydrogel dried in vacuum) as the "dry" hydrogel weight (Wd) and the "wet" hydrogel weight (Ws) which refers to the hydrogel incubated in 1×PBS at 37 °C for 48 hours. The results presented in Figure 5D show that the inclusion of PEGDA significantly increases the swelling mass and the increase in GelMA concentration also increases the swelling mass of the hydrogel.

[0293] c) Swelling ratio with the active agent A study was completed to analyze the swelling ratios of hydrogels loaded with active agents and having various GelMA, HAMA, and PEGDA concentrations.

[0294] G4-H M 1-P1, G4-H M 1, G7-H M 1, H M The 1-P1, G4-P1, and G7-P1 hydrogels (described in Table 1) had a photocrosslinking time of 4 minutes according to the general procedure of Example 3. Samples of each hydrogel were also prepared with a corticosteroid active agent at 13.2 mg / mL.

[0295] Swelling was evaluated using the "dry" polymer weight (hydrogel dried in vacuo) as the "dry" hydrogel weight (Wd) and the "wet" hydrogel weight (Ws) of the hydrogel incubated in 1×PBS at 37°C for 48 hours. The swelling ratio was calculated as follows. Swelling ratio = (Ws - Wd) / Wd

[0296] The results presented in Figure 6A show that hydrogels loaded with active agents generally have a high swelling ratio, which is thought to be due to gel network crosslink disruption and low crosslink density associated with incorporating the active agent into the gel network.

[0297] G4-H M 1-P1, G4-H M 1, G7-H M 1, H M The 1-P1, G4-P1, and G7-P1 hydrogels (containing active agents) were also studied for the swelling / re-swelling effect. The samples were dried and swollen, then subjected to a second re-drying and re-swelling. The results shown in Figure 6B indicate that the swelling ratio of hydrogels containing MeHA decreased significantly when the hydrogels were exposed to two or more drying / swelling cycles, whereas hydrogels containing only GelMA + PEGDA had a minimal effect on re-swelling.

[0298] d) Enzymatic degradation A study was completed to analyze the enzymatic degradation stability of hydrogels with various GelMA, MeHA, and PEGDA concentrations.

[0299] G4-H G 3-P1, G4-H M 1-P0.67, G4-H G 3, G4-H M 1, G7-H G 3, G7-H M 1, H G 3-P1, and H M The 1-P0.67 hydrogel (described in Table 1) was prepared with a photo-crosslinking time of 4 minutes according to the general procedure of Example 3. The sample was then enzymatically digested with either hyaluronidase (Hy) and collagenase type I (C I ) or collagenase type II (C II ) at either 20 U / mL or 2 U / mL. The resulting degradation times are shown in Table 2.

Table 2

[0300] e) Drug release A study was completed to analyze the drug release rates of hydrogels with various GelMA, MeHA, and PEGDA concentrations.

[0301] G4-H M 1-P1 and G4-H G The 3-P1 hydrogel (described in Table 1) was prepared with a photo-crosslinking time of 4 minutes using a corticosteroid active agent at 13.2 mg / mL according to the general procedure of Example 3. The resulting hydrogel cylinders had a diameter of 6 mm and a volume of 75 μL.

[0302] In the release study, to simulate tears, the hydrogels were incubated statically (without physical stirring) at 37 °C in 1 mL of 1× PBS supplemented with 2% Triton X-100. At each time point (over 10 - 13 days), the incubation solution was completely removed and replaced with fresh 1× PBS + 2% Triton X-100. To quantify corticosteroid release, samples were diluted 1:2 with acetonitrile and analyzed using reverse-phase liquid chromatography. An Agilent Zorbax Eclipse (XDB-C18) 4.6×250 mm, 5 μm analytical column was used with an Agilent 1290 HPLC system equipped with a diode array detector. The column was equilibrated at 25 °C with 70% acetonitrile and 30% water. After injecting 20 μL of the sample, the solvent gradient increased from 70% to 90% ACN over 10 minutes. When the ACN gradient reached approximately 80%, the corticosteroid eluted in nearly 5 minutes. This peak was integrated and the concentration was determined by comparing it to the standard curve of the corticosteroid using the area under the curve. The results presented in Figure 7A show that hydrogels containing higher concentrations of MeHA provide a more accelerated release profile. These results correlate with corresponding findings indicating that higher concentrations of MeHA in the hydrogel result in an increase in hydrogel swelling.

[0303] Based on the results of the swelling rate study, as the concentration of MeHA in the hydrogel increases, the swelling of the hydrogel increases, and as a result, there is a high likelihood of a more accelerated burst release of the active agent. As the concentration of MeHA increases, phase separation with GelMA in the precursor solution may also occur, which can cause imperfections in the gel network (i.e., regions of higher and lower cross-linking density), resulting in a higher initial burst release.

[0304] G4-H M The release profile of 1-P1 was continued up to 35 days (Figure 7B) and 65 days (Figure 7C). G4-H MThe release profile of 1-P1 was also compared with G4-P1 and G7-P1 (Figure 7D), and again the presence of MeHA in the hydrogel was shown to increase the release rate of the active agent from the hydrogel.

[0305] f) Vacuum drying A study was completed to analyze the effect of vacuum drying on the drug release rate of hydrogels with various GelMA, MeHA, and PEGDA concentrations.

[0306] G4-H M 1-P1, G4-P1, and G7-P1 hydrogels (described in Table 1) were prepared with a 4-minute photocrosslinking time using a corticosteroid active agent at 13.2 mg / mL according to the general procedure of Example 3. Next, samples from each hydrogel were vacuum dried. Release studies were performed using wet and dry samples for each hydrogel and then completed according to the general study procedure of Example 3(e). G4-H M The results for the 1-P1 hydrogel (Figure 8A) show that the release profile of the hydrogel containing MeHA can be reduced by vacuum drying the hydrogel, such that including MeHA in the hydrogel formulation reduces the release profile for the dry sample while instead increasing swelling and the corresponding release profile in the non-dried sample. The results for the G4-P1 and G7-P1 hydrogels (Figure 8B) show that the release profile of the GelMA+PEGDA hydrogel without MeHA is generally not affected by vacuum drying of the hydrogel.

[0307] g) Rod vs. disk A study was completed to analyze the effect of hydrogel shape (i.e., rod vs. disk) on the drug release rate of hydrogels containing GelMA, MeHA, and PEGDA.

[0308] G4-H MThe 1-P1 hydrogel (described in Table 1) was prepared as both a disk and a rod using a corticosteroid active agent at 13.2 mg / mL according to the general procedure of Example 3 with a 4-minute photocrosslinking time.

[0309] G4-H M The 1-P1 hydrogel disk had a diameter (D) of 6 mm, a volume (V) of 75 μL, and a surface area (SA) of 107 mm 2 and the SA:V ratio was 1.4.

[0310] G4-H M The 1-P1 hydrogel rod had a diameter (D) of 2 mm, a volume (V) of 25 μL, and a surface area (SA) of 56 mm 2 and the SA:V ratio was 2.2.

[0311] Samples from the rod hydrogels were then vacuum dried or lyophilized (i.e., freeze dried). Release studies were performed using the resulting wet and dry samples and then completed according to the general study procedure of Example 3(e). The results of total drug release (Figure 9A) show that the cylinder disk provided a greater total release of the active agent (presumably as a result of the higher surface area), and Rod wet Rod lyo and Rod dry all had similar total release amounts. The results of drug release rate (Figure 9B) show that the wet hydrogels (cylinder disk and rod) released a higher percentage of the active agent than the vacuum dried or lyophilized rod hydrogels. Thus, the study results indicated that the swelling properties, surface area (i.e., shape), and hydration state of the hydrogel play a role in the drug release profile of the hydrogel composition.

[0312] h) Degree of crosslinking - degree of methacrylation Studies were completed to analyze the correlation between the release profile of the GelMA+PEGDA hydrogel and the degree of methacrylation of GelMA within the hydrogel.

[0313] G4(160P80)-P1(2K) and G4(160P40)-P1(35K) hydrogels (described in Table 1) were prepared using a corticosteroid active agent at 13.2 mg / mL with a 4-minute photocrosslinking time according to the general procedure of Example 3. Subsequently, according to the general research procedure of Example 3(e), each sample was exposed to 0.5 U / mL collagenase II conditions and non-enzymatic standard conditions to complete the release study. The results of total drug release (Figure 10) indicate that the lower 40% DoM in GelMA provides a faster release profile than the higher 80% DoM GelMA hydrogel.

[0314] Example 5: Cell Aggregation and Viability Studies A population of human umbilical vein endothelial cells (HUVECs) was encapsulated in two separate hydrogel formulations: (i) G7(40)P1 [7% GelMA(160P40), 1% PEGDA(35 kDa)], and (ii) P8(35 kDa) [8% PEGDA, 35 kDa]. After 16 days, the hydrogel samples were stained with calcein AM and ethidium homodimer-1 and imaged for cell aggregation and viability. The results are shown in Figure 11A. As shown in Figure 11A, hydrogels containing a combination of chemically modified gelatin (e.g., GelMA) and chemically modified PEG (e.g., PEGDA) reduced cell aggregation of 3D adherent cells (e.g., HUVEC cells) encapsulated within the hydrogel framework.

[0315] Both hydrogel formulations were also analyzed for the ratio of calcein AM (live) / [calcein AM (live) + ethidium homodimer-1 (dead)] on days 3, 8, and 16 to quantify the HUVEC cell viability in each hydrogel. The results are shown in Figure 11B. As shown in Figure 11B, hydrogels containing a combination of chemically modified gelatin (e.g., GelMA) and chemically modified PEG (e.g., PEGDA) increased the cell viability of 3D adherent cells (e.g., HUVEC cells) encapsulated within the hydrogel framework. The G7(40)P1 hydrogel maintained an average cell viability of 60 - 80% over 16 days, while P8(35 kDa) resulted in an average cell viability of 20 - 40%.

[0316] Example 6: Transwell Lower Cell Proliferation Study - HUVEC A study was completed to evaluate the ability of various hydrogel formulations to attach and deliver cells to a membrane under difficult gravity requirements.

[0317] A population of human umbilical vein endothelial cells (HUVECs) was added to two hydrogel precursor formulations as prepared in Example 3 in PBS: (i) G5(80) [5% GelMA, 80% DoM] and (ii) P5(35 kDa) [5% PEGDA, 35 kDa]. The cell concentration was approximately 10 million HUVECs (GFP+) per mL.

[0318] Next, PET Transwell inserts (pore size 0.4 μm) were incubated in serum containing endothelial growth medium for 15 minutes. The inserts were dried with an ice piper, and 13.2 μL of each hydrogel precursor solution (containing HUVECs) was added to the lower side of each Transwell and exposed to high-intensity white light for 1 minute (Dolan-Jenner high-intensity LED illuminator MI-LED-US-B1 with a dual-arm gooseneck configuration). Then, the Transwells were inverted onto the well plate so that the hydrogels on the lower side of each Transwell were immersed in endothelial growth medium supplemented with growth factors and serum. Free HUVECs (i.e., non-hydrogel-encapsulated cells) were also added as a control to the endothelial growth medium in the well plate. Cell proliferation of each formulation was analyzed several days later (e.g., day 4 and day 11) using GFP+ imaging and confocal imaging on day 11.

[0319] a) GFP+ imaging results The results of GFP+ imaging are shown in Figures 12A - 12I.

[0320] G5(80) -- Four days later, the sample showed that the HUVECs remained localized on the underside of the transwell (Figure 12A), had strong endothelial cell viability and distinct cell network formation (see arrows in Figure 12A), and it was found that only cell debris was localized on the bottom of the well plate. Eleven days later, the G5(80) sample continued to show HUVECs localized on the underside of the transwell (Figure 12B), continued to have strong endothelial cell viability and distinct cell network formation (see arrows in Figure 12B), and it was found that only cell debris was localized on the bottom of the well plate.

[0321] P5(35 kDa) -- Four days later, the sample showed partial hydrogel detachment from the underside of the transwell, some HUVECs remained localized on the underside of the transwell (Figure 12C), and other HUVECs had migrated to the bottom of the well plate (Figure 12D). The overall cell density was low at both locations. Eleven days later, the P5(35 kDa) sample continued to show low cell density and minimal cell network formation on the underside of the transwell (Figure 12E), there were few remaining live cells, and it was found that most of the cell debris was localized on the bottom of the well plate.

[0322] Free cell control sample (i.e., without hydrogel) -- Four days later, the control sample showed minimal HUVEC attachment to the underside of the transwell (Figure 12F), and most of the cells had migrated to the bottom of the well (Figure 12G). The overall cell density was low. Eleven days later, the control sample continued to show low cell density and minimal cell network formation on the underside of the transwell (Figure 12H), live cells remained localized on the bottom of the well plate (Figure 12I), with some cell elongation but minimal cell network formation.

[0323] b) Confocal imaging results On the 11th day, the hydrogel samples of G5(80) and P8(35 kDa) from Example 6(a) were stained for the following: actin (the cytoskeleton indicating cell spreading and network formation), PECAM / CD31 (an endothelial marker), and DAPI (the nucleus). The stained samples were then subjected to confocal imaging, and the results are shown in FIGS. 13A - 13E.

[0324] The G5(80) sample showed high HUVEC deposition on the lower side of the transwell (FIGS. 13A and 13B), had a good cell distribution, and included the formation of a high - density cell network without signs of cell aggregation.

[0325] The P8(35 kDa) sample showed limited HUVEC deposition on the lower side of the transwell (FIGS. 13C and 13D), had a low cell density in a non - uniform distribution, high cell debris content, restricted cell spreading and network formation, no CD31 expression (highly likely related to endothelial cell dysfunction), and localization of actin filaments on the cell surface.

[0326] When compared with the G5(80) sample, the free - cell (i.e., without hydrogel) control sample showed a lower cell density and insufficient network formation (FIG. 13E).

[0327] c) Additional formulation tests Additional hydrogel formulations were analyzed according to the procedure of Example 6(a). The results of transwell lower - side imaging and analysis are shown in Table 3.

Table 3

[0328] d) Research observations The results showed that HUVECs spread more effectively, created stronger cell networks, and aggregated less within the GelMA-based formulations compared to the PEGDA-based formulations. The results also showed that higher crosslinking densities (e.g., higher polymer concentrations, higher DOM) generally did not promote stronger cell spreading and network formation. The GelMA-based materials also provided stronger adhesion to the test surface (i.e., the bottom of the transwell), deposited cells more effectively, and formed high-density cell networks even in challenging environments and growth conditions (e.g., with respect to gravity).

[0329] Example 7: Sub-transwell Cell Proliferation Study - HRPEC A study was completed to evaluate the ability of various hydrogel formulations to attach and deliver cells to a membrane in a challenging gravity environment mimicking the subretinal space.

[0330] A population of human retinal pigment epithelial cells (HRPEC) was added to the following hydrogel precursor formulations as prepared in Example 3 in PBS: (i) G2(80)H2(500 kDa, 30) [2% GelMA, 80% DOM; 2% HAMA, 500 kDa, ~30% DOM], (ii) G5(80) [5% GelMA, 80% DoM], (iii) G4(80) [5% GelMA, 80% DoM], (iv) H2(500 kDa, 30) [2% HAMA, 500 kDa, ~30% DOM], (v) P5(35 kDa) [5% PEGDA, 35 kDa]. The cell concentration was approximately 20 million HRPEC per mL.

[0331] Next, a PET transwell insert (pore size 0.4 μm) was incubated for 15 minutes in serum (supplemented with 10% FBS) containing DMEM / F-12 medium. The insert was dried with an ice piper, 13.2 μL of each hydrogel precursor solution (containing HRPEC) was added to the lower side of each transwell, and exposed to light for 1 minute. Then, the transwell was inverted onto a well plate so that the hydrogel on the lower side of each transwell was immersed in DMEM / F-12 medium (supplemented with 10% FBS). Cell growth of each formulation was imaged and analyzed after several days (e.g., day 3, day 8, day 10, day 20), which included the use of calcein AM staining and imaging.

[0332] The results are shown in FIGS. 15A - 15G.

[0333] G2(80)H2(500 kDa, 30) - After 3 days, the sample remained localized within the hydrogel on the lower side of the transwell and showed HRPEC that was unable to form a cell monolayer across the surface of the transwell (FIG. 15A). Similar local results were observed after 10 days without HRPEC monolayer formation (FIG. 15B).

[0334] G5(80) - After 8 days, the sample showed well - dispersed round cells but also showed minimal cell spreading and no cell network formation (FIG. 15C).

[0335] H2(500 kDa, 30) - After 7 days, the sample showed well - dispersed round cells but also showed minimal cell spreading and no cell network formation (FIG. 15D).

[0336] G4(80) - After 8 days, the hydrogel was shown to have rapidly degraded / decomposed, but also shown to provide an HRPEC monolayer with good coverage and viability, including a zone of mature monolayer formation (FIG. 15E). After 20 days, it was shown that a mature HRPEC monolayer was formed across the entire surface of the transwell membrane (FIG. 15F).

[0337] P5 (35 kDa) -- After 8 days, the hydrogel was shown to rapidly degrade / decompose and also to provide an HRPEC monolayer with incomplete coverage and inconsistent monolayer formation (Figure 15G).

[0338] a) Research observations The research results showed that HRPECs had minimal spread and networking within hydrogel formulations having high levels of methacrylation, high molecular weights, and / or high polymer concentrations of hydrogels (e.g., G2(80)H2(500 kDa, 30)). HRPECs improved the spread and networking within hydrogel formulations having lower levels of methacrylation and lower polymer concentrations of hydrogels, thus enabling effective HRPEC deposition and monolayer formation.

[0339] Also, PEG-based hydrogels such as PEGDA were shown to generally have poor biocompatibility and poor biodegradability (often removed from under the transwell in just 24 - 48 hours). The remaining HRPECs on the transwell surface are thought to be those captured on the surface of the PEGDA hydrogel during the photopolymerization process. Cells were unable to attach or migrate through the PEGDA polymer network and remained trapped, able to attach only to adjacent cells.

[0340] Example 8: Transwell upper cell growth study - HRPEC A study was completed to evaluate the ability of various hydrogel formulations to dissociate cells and deliver them to the membrane.

[0341] A population of human retinal pigment epithelial cells (HRPECs) was added to two hydrogel precursor formulations: (i) G5(10) [5% GelMA, 10% DoM] prepared in Example 3 in PBS, and (ii) G5(80) [5% GelMA, 80% DoM]. The cell concentration was approximately 20 million HRPECs per mL.

[0342] Next, a PET transwell insert (pore size 0.4 μm) was incubated in serum (supplemented with 10% FBS) containing DMEM / F-12 medium for 15 minutes. The insert was dried with an ice pippette, and approximately 10 μL of each hydrogel precursor solution (containing HRPEC) was added to the upper side of each transwell and exposed to light for 1 minute. The transwell was then immersed in DMEM / F-12 medium (supplemented with 10% FBS). Free HRPEC (i.e., without hydrogel) was also added to the medium in the well plate as a control.

[0343] After 24 hours, the hydrogel samples were stained with calcein AM and imaged for cell aggregation and viability (Figure 16A). The samples were analyzed similarly after 3 days (Figure 16B) and after 6 days (Figure 16C).

[0344] After 24 hours (Figure 16A) - The cell-only control samples showed uninhibited cell growth and monolayer formation. The G5(10) samples showed that the hydrogel began to degrade, deposited HRPEC, and began to form a monolayer network. The G5(80) samples showed no hydrogel degradation, no monolayer formation, and the cells generally remained rounded and suspended within the hydrogel.

[0345] On day 3 (Figure 16B) - The cell-only control samples continued to show uninhibited cell growth and monolayer formation, indicating cell confluence. The G5(10) samples showed that the hydrogel was almost completely degraded, and the deposited HRPEC formed a transparent monolayer network similar to the control samples after 24 hours, with slightly rounded, encapsulated cells remaining on the monolayer. The G5(80) samples showed little hydrogel degradation, little monolayer formation, and the cells generally remained rounded and suspended within the hydrogel.

[0346] Day 6 (Figure 16C) - The cell-only control sample continued to show uninhibited cell growth and monolayer formation with a dense cell density and networking. The G5(10) sample showed a degraded hydrogel, and the deposited HRPEC monolayer network was similar in density and morphology to the control sample, with few rounded cells remaining on the monolayer. The G5(80) sample showed a hydrogel that had started to degrade in a specific region (upper right of the Figure 16C image for G5(80)) and monolayer formation, while other regions remained undegraded without monolayer formation (lower left of the Figure 16C image for G5(80)), and the HRPEC were rounded and remained suspended within the hydrogel.

[0347] Example 9: In Vitro Retinal Cell Proliferation Study A study was completed to evaluate the G5(10) [5% GelMA, 10% DoM] hydrogel formulation for promoting the in vitro delivery and cell proliferation of stem cell-derived retinal cells, including retinal pigment epithelial (RPE) cells and rod photoreceptor cells.

[0348] a) Monolayer Formation Study The G5(10) hydrogel precursor formulation [5% GelMA, 10% DoM] was prepared according to the general procedure of Example 3 with a reduced concentration of the photoinitiator mixture. Then, a population of human retinal pigment epithelial cells (HRPEC) derived from induced pluripotent stem cells (iPSC) was added to the G5(10) hydrogel precursor formulation (about 20 million cells / mL). The iPSC-derived RPE was also added to physiological saline as a control. Approximately 200 μL of each formulation was injected through a subretinal cannula (including a standard 1 mL Luer lock plastic syringe) of MedOne Subretinal PolyTip® 38G (outer diameter) (41G inner diameter) (in PBS) at a controlled rate of 200 - 300 μL / min (flow rate controlled by a syringe pump) onto a 6-well Transwell plate with a 0.4 μm polyester (polyethylene terephthalate) membrane. The polyester membrane was included to mimic the natural Bruch's membrane and is permeable to nutrients and proteins but not to cells. The injected formulation was then photocrosslinked in 10 μL drops using white light.

[0349] One week later, live cell imaging was completed using Calcein AM. The results are shown in Figure 17A. The study results showed that after crosslinking, the RPE cells could slowly degrade the G5(10) hydrogel substrate and then migrate to the Transwell membrane. The RPE cells formed a transparent cell monolayer after one week, mimicking the target anatomical pattern of functional RPE cells (see Figure 17A). The RPE cells delivered in the physiological saline (control) formulation did not form an observable cell monolayer after one week, even though they were delivered at the same seeding density as the cells in the hydrogel formulation.

[0350] The fluid shear rate associated with extrusion through a 38G (inner 41G) small diameter needle was measured during the sampled delivery. The results (Figure 17B) showed that the GelMA prepolymer may be shear-thinning in the precursor polymer solution, thereby enabling plug flow dynamics through the small diameter needle that protects the passenger cells.

[0351] b) iPSC Dedifferentiation Study The precursor formulation from Example 9a was prepared at approximately 1 million cells / mL and then injected (in PBS) onto a 6-well Transwell plate with a 0.4 μm polyester (polyethylene terephthalate) membrane using the same delivery system and conditions as in Example 9A. The injected formulation was then photocrosslinked in 10 μL drops using white light. After one week, live cell imaging was completed using Calcein AM. The results are shown in Figure 17C. The study results showed that the RPE cells formed a monolayer and the cells retained the cuboidal morphology and pigmentation pattern that are characteristic of a standard and healthy RPE morphology. RPE cells delivered in a saline (control) formulation were elongated, highly proliferative, and unable to retain the cuboidal morphology and RPE pigmentation pattern indicative of dedifferentiation.

[0352] Without being bound by theory, the study observations indicate that the hydrogel matrix provides a localized and condensed environment for the RPE cells and thus enables monolayer formation with a characteristic RPE morphology by reducing RPE dedifferentiation due to environmental factors.

[0353] c) Rod Photoreceptor Delivery Study The G5(10) [5% GelMA, 10% DoM] hydrogel precursor formulation was prepared according to the general procedure of Example 3 with a reduced concentration of the photoinitiator mixture. A population of rod photoreceptor cells derived from human retinal organoids (Lako Lab, University of Newcastle) was then added to the G5(10) hydrogel precursor formulation. The formulation was then injected (in PBS) onto the RPE monolayer in a 6-well Transwell plate with a 0.4 μm polyester (polyethylene terephthalate) membrane using the same delivery system and conditions as in Example 9A. The injected formulation was then photocrosslinked using white light.

[0354] Two weeks later, live cell imaging was completed using anti-CD73 and anti-rhodopsin imaging. The results are shown in Figure 17D. The findings indicate that rod photoreceptor cells (upper left in Figure 17D) are localized on the RPE monolayer (lower left in Figure 17D), are viable for two weeks, and both rod photoreceptor cells and RPE cells express CD73. Only rod photoreceptor cells were shown to express rhodopsin (Figure 17D).

[0355] Example 10: In Vivo Retinal Cell Proliferation Study A study was completed to evaluate the cellular localization and viability of retinal pigment epithelial cells (RPE) following a single subretinal (SR) injection in pigs with one of three G5 [5% GelMA] hydrogel formulations polymerized in situ via intraocular treatment light.

[0356] Four test groups included 200 μl injections into 6 eyes (for each group, 24 eyes and a total of 12 animals):

[0357] Group 1 (control): Saline + 200k RPE cells, Group 2: G5(10) [5% GelMA, 10% DoM] + 200k RPE cells, Group 3: G5(80) [5% GelMA, 80% DoM] + 200k RPE cells, Group 4: [5% GelMA, 80% DoM]G5(80) + 1m RPE cells.

[0358] Animals in Groups 2 - 4 received a single SR injection of RPE using the designated hydrogel formulation. Group 1 received a single injection of RPE in saline as a control. Oral administration of prednisone was performed preoperatively and throughout the study. A 38G / 41G MedOne subretinal needle with a 6 - 7 inch extension tube was used for the injection. The hydrogel precursor and cell mixture were freshly prepared prior to each group. A Constellation 23G light pipe was used for radiation emitting a light power of approximately 27 mW at maximum output (measured at 517 nm).

[0359] All animals underwent eye examinations, color fundus examinations, and optical coherence tomography (OCT) after injection and on days 1, 3, 7, 14, 21, and 28 after injection. All registered eyes were stained for STEM121 (transplanted human cells), Iba-1 (microglia / leukocytes), RPE65 (RPE cells), and DAPI (all nuclei).

[0360] For immunofluorescence staining, two eyes from each group were also selected. Blocks were sectioned sagittally (14 μm) at five levels across the eye: nasal, peripheral, mid-nasal and optic nerve head (ONH), ONH, mid-ONH and temporal, and temporal peripheral. At least 10 slides / level were collected. Slides were scanned during acquisition of cell deposits. For IHC, a series of 8 - 10 slides / eye were selected.

[0361] Observations related to cell migration to the retinal surface are shown in Figure 18A. The saline control formulation presented more cell migration (over 80%) and corresponding supraretinal adverse events due to inadequate cell localization (i.e., cells outside the subretinal region). This finding was confirmed by histological examination of eyes at day 28, in which vitreous cells were found, and the subretinal hydrogel formulation presented minimal cell migration of approximately 20% or less.

[0362] Observations related to hydrogel degradation after 28 days are shown in Figure 18B. Group 1 (control) did not contain hydrogel. For Group 2, only approximately 33% of the hydrogel remained after 28 days (i.e., approximately 67% was lost). Groups 3 and 4 had approximately 100% of the hydrogel remaining after 28 days (i.e., approximately 0% loss). After 28 days, hydrogels with high 80% methacrylation (Groups 3 and 4) showed detachment of the retina from the underlying layer, as shown in Figures 18C and 18D. The staining results indicated successful transplantation of human stem cells onto the native porcine RPE layer, which was not disrupted (see Figure 18E, arrows represent human cells).

[0363] Example 11: Radical Propagator Study A study was completed to evaluate N-vinylcaprolactam (NVC), N-vinylpyrrolidone (NVP), and ethylene glycol diacrylate (EGDA) in hydrogel formation.

[0364] Each of the three propagators (NVC, NVP, EGDA) was combined with G5(80) [5% GelMA, 80% DoM] at a propagator concentration of 10 μL / mL and then gelled. The average Young's modulus (kPa) of the resulting hydrogels was as follows: EGDA [7.2494 kPa], NVP [6.4995 kPa], NVC [3.4744 kPa]. NVP and EGDA were also tested for the minimum concentrations of G5(10) and G5(80) that would still allow hydrogel formation (crosslinking for 1 minute at approximately 0.43 W). The minimum propagator concentrations for hydrogel formation were as follows. G5(10) [NVP - 5 μL / mL, EGDA - 1.5 μL / mL], G5(80) [NVP - 0.3 μL / mL, EGDA - 0.1 μL / mL]

[0365] Next, G5(10) - 1.5 μL / mL of EGDA and G5(10) - 5 μL / mL of NVP were studied for encapsulation of RPE cells by needle extrusion (with PBS as a control). After initial extrusion, all three samples showed cells that retained pigmentation and morphology. After 24 hours, the PBS cells lost pigment and showed early signs of epithelial-mesenchymal transition (EMT). Both hydrogel formulations showed cells that retained pigmentation and morphology. The same results were observed on day 6.

[0366] Cytotoxicity studies were also completed using RPE cells for the following: NVC, NVP, EGDA, and phenylacetyl bromide (BAP), with 10% DMSO as a control. The results of cell viability after 18 hours of cell incubation in each of the four propagators are shown in Figure 19. The results showed that at all relative concentrations, NVP and BAP had higher average cell viabilities than NVC, and EGDA had lower cell viability results.

[0367] Example 12: Research on G5 formulation using N - vinylpyrrolidone (NVP) G5 formulations [5% GelMA, various DoM and MW] with different average molecular weights were studied for their gelation and cell encapsulation properties. Specifically, G5(10)160 kDa, G5(60)90 kDa, and G5(45)160 kDa were mixed with a standard photo - initiator concentration (having 1% NVP instead of NVC) and a formulation of 5M ARPE - 19 cells / mL (a total of approximately 200k cells per gel). Samples (containing extruded saline with cells) were injected into sample containers using a MedOne 38 / 41G sub - retinal needle (injection rate of 300 μL / min), and a light pipe (set to 26 - 27 mW) was used to mimic the settings of stellate vitrectomy (30 - second exposure). Saline (non - extruded) was used as a normalized control.

[0368] The results of cell viability using Calcein AM (normalized to non - extruded saline) are shown in Figure 20. All formulations showed at least 150% normalized cell viability, and G5(10)160 kDa and G5(60)90 kDa showed more than 200% normalized cell viability. After 48 hours, gel degradation was evident for all three hydrogel samples, and cells adhered to the transwell surface with early monolayer formation. There were few rounded non - adherent cells.

[0369] Additional studies were completed to analyze the gelation properties of various G4 formulations [4% GelMA, various DoM and MW] and G5 formulations [5% GelMA, various DoM and MW] using NVP as a radical propagator. Table 4 shows the research conditions and results of various formulations.

Table 4

[0370] Additional polymer blends using GelMA concentrations of 0.5% - 3.0% were also tested for their gelation and cell encapsulation properties. Table 5 shows the research conditions and results of various formulations.

Table 5

[0371] Example 13: Combined Preparation Research The G1(160 / 10)G2.5(90 / 60) blend was studied with various photoinitiator formulations (including 1.5% v / v triethanolamine (TEOA)) and polymerization conditions. Table 6 shows the research conditions and results of various formulations.

Table 6

[0372] Rxn3 and Rxn4 were further studied. 10M cells / mL were added to each of Rxn3, Rxn4, and PBS (as a control). Portions of the Rxn 3 sample, the Rxn 4 sample, and the PBS control were extruded onto the Transwell surface and onto unextruded PBS. After two weeks, both Rxn3 and Rxn4 showed healthy RPE monolayer growth, similar to the PBS control, and the Rxn3 sample showed a higher cell count than either PBS control sample. The cell count results are shown in Figure 21.

Claims

1. A polymer composition, (i) At least one chemically modified gelatin containing methacrylated gelatin (GelMA) in an amount of approximately 0.5% to approximately 4% w / v, (ii) at least one crosslinking agent, and (iii) at least one cell, A polymer composition containing the following:

2. The polymer composition according to claim 1, wherein the polymer composition comprises eosin Y or a salt thereof, N-vinylcaprolactam (NVC), N-vinylpyrrolidone (NVP), triethanolamine, or methylenebisacrylamide.

3. The polymer composition according to claim 2, wherein the polymer composition comprises (i) eosin Y, N-vinylcaprolactam (NVC), triethanolamine, or any combination thereof; (ii) eosin Y disodium salt (EYDS), N-vinylcaprolactam (NVC), triethanolamine, or any combination thereof; (iii) eosin Y disodium salt (EYDS), N-vinylpyrrolidone (NVP), triethanolamine, or any combination thereof; or (iv) eosin Y, methylenebisacrylamide, triethanolamine, or any combination thereof.

4. The polymer composition according to claim 1, wherein the at least one chemically modified gelatin is GelMA.

5. The polymer composition according to claim 4, wherein the GelMA has a degree of methacrylate of about 5 to 60%.

6. The polymer composition according to claim 4, wherein the polymer composition contains about 1% w / v or about 2% w / v of GelMA.

7. The polymer composition according to claim 1, wherein the polymer composition comprises a combination of a first GelMA mixture and a second GelMA mixture.

8. The polymer composition according to claim 7, wherein the polymer composition comprises about 0.5% to about 3% w / v of the first GelMA mixture and about 0.5% to about 3% w / v of the second GelMA mixture, optionally about 0.5% to about 1.5% w / v of the first GelMA mixture and about 1.5% to about 3% w / v of the second GelMA mixture, optionally about 1% w / v of the first GelMA mixture and about 2.5% w / v of the second GelMA mixture.

9. The polymer composition according to claim 7 or claim 8, wherein the first GelMA mixture comprises GelMA having a high average molecular weight and a low degree of methacrylate (DOM), the second GelMA mixture comprises GelMA having a low average molecular weight and a high DOM, optionally the first GelMA mixture comprises GelMA having an average molecular weight of 140 to 180 kDa and 5% to 40% DOM, the second GelMA mixture comprises GelMA having an average molecular weight of 75 to 115 kDa and 50% to 80% DOM, optionally the first GelMA mixture comprises GelMA having an average molecular weight of 140 to 180 kDa and 5% to 20% DOM, and the second GelMA mixture comprises GelMA having an average molecular weight of 80 to 100 kDa and 50% to 70% DOM.

10. The polymer composition according to claim 7 or claim 8, wherein the first GelMA mixture comprises GelMA having an average molecular weight of about 160 kDa and a degree of methacrylate (DOM) of about 10%, and the second GelMA mixture comprises GelMA having an average molecular weight of about 90 kDa and a degree of DOM of about 60%.

11. The polymer composition according to claim 7, wherein the polymer composition comprises a first GelMA mixture in an amount of about 1% w / v containing GelMA having an average molecular weight of about 160 kDa and a degree of methacrylate (DOM) of about 10%, and a second GelMA mixture in an amount of about 2.5% w / v containing GelMA having an average molecular weight of about 90 kDa and a degree of DOM of about 60%.

12. The polymer composition according to claim 1, wherein the at least one cell comprises an endothelial cell, optionally a human umbilical vein endothelial cell (HUVEC).

13. The polymer composition according to claim 1, wherein the at least one cell comprises epithelial cells, optionally human retinal pigment epithelial cells (HRPEC), optionally embryonic stem cells, or induced pluripotent stem cells (iPSC) derived from HRPEC.

14. The polymer composition according to claim 1, wherein the at least one cell comprises an ophthalmic cell, optionally an ophthalmic cell derived from a pluripotent stem cell or an embryonic stem cell.

15. A precursor polymer composition comprising the polymer composition described in claim 1.

16. A gel polymer composition wherein the gel polymer composition is formed by photocrosslinking a precursor polymer composition according to claim 15, and optionally the gel polymer composition is a hydrogel.

17. The precursor polymer composition is for treating and / or repairing defects, damage, and / or diseases in a target soft tissue, The precursor polymer composition is administered, optionally, to the surface or beneath the target soft tissue, at the site of a defect, injury, and / or disease in the soft tissue. The precursor polymer composition is crosslinked by exposure of at least one crosslinking agent in the precursor polymer composition to crosslinking conditions, and the crosslinking of the precursor polymer composition generates a gel polymer composition. The precursor polymer composition according to claim 15.

18. The gel polymer composition is for treating defects, injuries, and / or diseases in a target soft tissue, The gel polymer composition is administered to the surface of, beneath, or near the target soft tissue, optionally at the site of a defect, injury, and / or disease in the soft tissue. The gel polymer composition according to claim 16.

19. A method for producing a gel polymer composition, To provide the precursor polymer composition described in claim 15, and The precursor polymer composition is crosslinked by exposing at least one crosslinking agent in the precursor polymer composition to crosslinking conditions, and the gel polymer composition is produced by the crosslinking of the precursor polymer composition. Methods that include...

20. A gel polymer composition produced by the method described in Claim 19.

21. (i) at least one chemically modified gelatin comprising about 0.5% to about 4.0% w / v of methacrylated gelatin (GelMA), (ii) at least one crosslinking agent, and (iii) at least one cell A method for producing a polymer composition containing, (a) To provide at least one chemically modified gelatin containing GelMA in solution, (b) Adding at least one of the crosslinking agents to the solution, (c) Adding at least one of the cells to the solution, (d) optionally crosslinking the solution to produce a gel polymer composition, Methods that include...

22. The polymer composition according to claim 5, wherein the GelMA has a degree of methacrylate comprising about 5%, about 10%, about 15%, about 20%, about 30%, about 40%, about 50%, or about 60%.

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