Photocurable hydrogel composition

Abstract

The present disclosure provides photocurable hydrogel compositions and methods of use thereof for the treatment of tympanic membrane perforation.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 390,837, filed on July 20, 2022, the contents of which are hereby incorporated by reference in their entirety for all purposes. Government support

[0002] This invention was made with government support under Grant No. 1R44DC01855-01 awarded by the National Institute on Deafness and Other Communication Disorders (NIDCD). The government has certain rights in this invention. Background Art

[0003] Tympanostomy tube (TT) insertion is a very common procedure in children and adults. During TT insertion, a thin tube is placed into the tympanic membrane (TM) to allow fluid to leave the middle ear. It also allows air to enter the middle ear through the TM. It is most commonly used to improve hearing due to chronic fluid behind the tympanic membrane, and to reduce the frequency of infections and direct topical antibiotics for recurrent middle ear infections. In most cases, tympanostomy tubes will extrude spontaneously after 6 to 18 months of placement, and the tympanic membrane (TM) will repair completely. However, approximately 15% of patients will develop a TM perforation following TT extrusion.

[0004] To treat a TM piercing, the fibrous tissue surrounding the piercing is abraded to induce an inflammatory response. If the piercing is small, such as approximately 20% of the TM cross-sectional area or less, the piercing may be filled with paper or fat taken from the earlobe. Larger piercings require more robust filling materials, such as autografts of cartilage or fascia, surgical gelatin, or synthetic polymer patches. All of these therapies require some type of surgery, and the success rate of the treatment is highly dependent on the surgeon's skill.

[0005] To date, no photopolymerizable hydrogels have been used as otologic scaffolds, where a semi-gel polymer is applied to the desired site before solidifying. The use of scaffolds that are initially liquid and only form a scaffold when exposed to specific conditions is a radical departure from current conventional therapies for repairing TM perforations. Summary of the invention

[0006] The present disclosure provides a pre-lyophilized composition comprising: (i) a substrate comprising about 40 wt % to about 80 wt % of methacrylated chitosan, about 20 wt % to about 40 wt % of hyaluronic acid, and about 0.01 wt % to about 0.1 wt % of a photoinitiator; and (ii) diluent.

[0007] The present disclosure provides a pre-lyophilized composition comprising: (i) a substrate comprising about 40 wt % to about 80 wt % of methacrylated chitosan, about 20 wt % to about 40 wt % of hyaluronic acid, and about 0.01 wt % to about 0.5 wt % of a photoinitiator; and (ii) diluent.

[0008] In embodiments, the photoinitiator is riboflavin.

[0009] In embodiments, the composition further comprises from about 0 wt% to about 12 wt% arginine. In embodiments, the composition further comprises from about 0 wt% to about 20 wt% arginine.

[0010] In embodiments, the methacrylated chitosan is from about 30% to about 40% methacrylated. In embodiments, the methacrylated chitosan is from about 25% to about 40% methacrylated.

[0011] The present disclosure provides a lyophilized composition prepared by lyophilizing any composition of the present invention, wherein the lyophilized composition comprises about 0% to about 2% water, and wherein the lyophilized composition can be reconstituted in less than 1 minute.

[0012] The present disclosure provides a semi-gel composition prepared by mixing a lyophilized composition of the present disclosure with a diluent comprising water until homogeneous; wherein the mixing time is less than 60 seconds; wherein the viscosity of the semi-gel composition is from about 0.01 kPa*s to about 1.0 kPa*s, as measured by rheometry; wherein the composition comprises from about 80 wt % to about 99 wt % water.

[0013] The present disclosure provides a cured composition produced by irradiating the semi-gel composition of the present invention with light, wherein the cured composition has an elastic modulus of about 0.05 kPa to about 20 kPa as measured by rheometry.

[0014] In an embodiment, the light radiation comprises a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 About 1000mW / cm 2 In an embodiment, the light radiation comprises irradiating the composition with a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 To about 100000mW / cm 2 The light radiation intensity of the radiation composition.

[0015] The present invention provides a medicine kit, which comprises a lyophilized composition, a gel dispensing tray and a light guide.

[0016] The present disclosure provides a method for treating tympanic membrane perforation, comprising: (a) applying the semi-gel composition of the present disclosure to the tympanic membrane of a patient in need thereof, (b) Insert the light guide into the ear canal, and (c) Curing the composition by irradiating it with light.

[0017] In an embodiment, the light radiation comprises a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 About 1000mW / cm 2 In an embodiment, the light radiation comprises irradiating the composition with a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 To about 100000mW / cm 2 The light radiation intensity of the radiation composition.

[0018] In an embodiment, the present disclosure provides a method of treating a tympanic membrane perforation in a patient in need thereof, comprising: (a) mixing a solvent (e.g., saline) with the lyophilized composition of the present disclosure to provide a semi-gel composition; (b) applying the semi-gel composition to the site of the tympanic membrane perforation of the patient; and (c) curing the semi-gel composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1A -D shows the components of the perforation treatment kit described in Example 4. A gel pack with dry reagents ( Figure 1A ); a gel application kit (with a dispensing tray (1), a light guide (2) and a V-shaped bag (3) Figure 1B ); V-shaped bag with gel pack ( Figure 1C ); and an outer tray with a lid, a dispensing tray with a light guide, a gel bag with a lyophilized composition, and a V-shaped bag ( Figure 1D ). DETAILED DESCRIPTION definition

[0020] Any term or expression not expressly defined herein shall have a generally accepted definition understood by those skilled in the art. To the extent that the following description is directed to a particular embodiment or particular use of the invention, the description is intended to be illustrative only and does not limit the claimed invention. The following description is intended to cover all substitutions, modifications, and equivalents included within the spirit and scope of the invention as defined by the appended claims.

[0021] The term "about" when immediately preceding a numerical value means a range (e.g., the value plus or minus 10%). For example, "about 50" may mean 45 to 55, "about 25,000" may mean 22,500 to 27,500, etc., unless the context of the present disclosure indicates otherwise or is inconsistent with such an interpretation.

[0022] The term "semi-gel composition" refers to a composition (comprising at least one polymer) that is capable of being activated to assemble into a more viscous form but has not yet assembled. In embodiments, the semi-gel composition has sufficient viscosity so that it can be positioned to cover a perforation without falling into the middle ear cavity. In embodiments, the semi-gel composition can be prepared from a dehydrated composition (e.g., a lyophilized composition).

[0023] As used herein, the term "chitosan" means chitosan and its derivatives and analogs (eg, methacrylated chitosan). As used herein, the term "hyaluronic acid" means hyaluronic acid, its derivatives and analogs.

[0024] With respect to a patient, the term "treat" refers to improving at least one symptom of a condition in the patient. Treatment may improve or at least partially alleviate the condition. Composition

[0025] The present disclosure provides semi-gel compositions that can be used to treat ruptured tympanic membranes and the like in patients in need thereof after solidification, and compositions that can be used as intermediates for making semi-gel compositions. Thus, the present disclosure provides pre-lyophilized compositions; lyophilized compositions; semi-gel compositions and solidified compositions.

[0026] The pre-lyophilized composition of the present disclosure is a flowable mixture that provides a shelf-stable lyophilized composition after lyophilization (or similar techniques, such as spray drying). The lyophilized composition of the present disclosure can be reconstituted in a suitable solvent to provide a semi-gel composition having suitable viscosity properties to be applied to, for example, a ruptured tympanic membrane of a patient in need. After application to a patient in need, the semi-gel composition can be solidified using the methods disclosed herein to provide a solidified composition (or scaffold).

[0027] In embodiments, the compositions of the present disclosure include ingredients that are biocompatible and suitable for use in the ear (e.g., the external auditory canal, the middle ear, or both). In embodiments, the ingredients are selected so that the resulting polymer matrix is ​​biodegradable and can be processed and cleared by the body (e.g., by the kidneys).

[0028] Compositions according to the present disclosure are initially semi-gel compositions for application to a desired location in a patient's ear, and are configured to solidify into a polymer matrix when activated in the patient's ear. Therefore, the semi-gel composition includes one or more polymers and diluents that can selectively assemble into a polymer matrix at a desired time under specific conditions (e.g., conditions found in the ear, such as the middle ear). In an embodiment, the semi-gel composition includes an initiator (e.g., a photoinitiator), and the semi-gel composition is solidified by exposing the composition to certain conditions (e.g., light) to activate the initiator and produce a polymer matrix. In an embodiment, changes in optical density, pH, temperature, or ion concentration trigger spontaneous assembly of the polymer matrix. In an embodiment, the polymer is functionalized to give them cross-linking ability (e.g., methacrylate) after activation by an initiator (e.g., riboflavin).

[0029] In embodiments, the semi-gel composition is designed to solidify into a polymer matrix or scaffold whose porosity and average pore size are compatible with loading the therapeutic agent into the resulting polymer matrix or scaffold. In embodiments, the porosity and average pore size are selected to provide a desired time-dependent release profile of the loaded therapeutic agent.

[0030] In embodiments, the polymer matrix is ​​a scaffold having sufficient stiffness and / or adhesiveness (e.g., as a result of functional groups providing cell adhesion domains) such that epithelial cells in the TM can migrate into and / or onto the scaffold and adhere and / or grow. In embodiments, the polymer matrix or scaffold ultimately degrades in situ, i.e., biodegrades, e.g., after epithelial cells have grown sufficiently to repair the perforation. Pre-freeze dried composition

[0031] The present disclosure provides a pre-lyophilized composition comprising: (i) a substrate comprising methacrylated chitosan, hyaluronic acid and a photoinitiator; and (ii) a diluent. In an embodiment, the diluent comprises water.

[0032] In embodiments, the pre-lyophilized composition provides a shelf-stable lyophilized composition following lyophilization (or similar techniques, such as spray drying).

[0033] In an embodiment, the present disclosure provides a pre-lyophilized composition comprising: (i) a substrate comprising about 40 wt % to about 80 wt % of methacrylate, about 20 wt % to about 40 wt % of hyaluronic acid, about 0.01 wt % to about 0.1 wt % of a photoinitiator, and 0 wt % to about 20 wt % of arginine; and (ii) diluent.

[0034] In an embodiment, the present disclosure provides a pre-lyophilized composition comprising: (i) a substrate comprising about 40 wt % to about 80 wt % of methacrylated chitosan, about 20 wt % to about 40 wt % of hyaluronic acid, about 0.01 wt % to about 0.5 wt % of a photoinitiator, and 0 wt % to about 20 wt % of arginine; and (ii) diluent.

[0035] In embodiments, the substrate comprises about 30 wt % to about 90 wt % of methacrylated chitosan, such as about 30 wt %, about 32 wt %, about 34 wt %, about 36 wt %, about 38 wt %, about 40 wt %, about 42 wt %, about 44 wt %, about 46 wt %, about 48 wt %, about 50 wt %, about 52 wt %, about 54 wt %, about 56 wt %, about 58 wt %, about 60 wt %, about 62 wt %, about 64 wt %, about 66 wt %, about 68 wt %, about 70 wt %, about 72 wt %, about 74 wt %, about 76 wt %, about 78 wt %, about 80 wt %, about 82 wt %, about 84 wt %, about 86 wt %, about 88 wt % or about 90 wt %, including all ranges and values ​​therebetween. In embodiments, the composition comprises about 40 wt % to about 80 wt % of methacrylated chitosan. In embodiments, the composition comprises about 60 wt% methacrylated chitosan.

[0036] In some embodiments, the methacrylated chitosan is about 10% to about 60% methacrylated, for example, 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 28%, 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 78%, 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%, about 99%, about 100%, about 10 %, 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%, including all ranges and values ​​therebetween. In embodiments, the methacrylated chitosan is about 21% to about 32% methacrylated. In embodiments, the methacrylated chitosan is about 21% to about 34% methacrylated. In embodiments, the methacrylated chitosan is about 21% to about 36% methacrylated. In embodiments, the methacrylated chitosan is about 25% to about 40% methacrylated. In embodiments, the methacrylated chitosan is about 30% methacrylated.

[0037] In embodiments, the molecular weight of chitosan may range from about 100 to about 1,000,000 g / mol. In embodiments, the degree of deacetylation of chitosan ranges from about 0 to about 100%.

[0038] In embodiments, substrate comprises the hyaluronic acid of about 10wt% to about 50wt%, for example, about 10wt%, about 12wt%, about 14wt%, about 16wt%, about 18wt%, about 20wt%, about 22wt%, about 24wt%, about 26wt%, about 28wt%, about 30wt%, about 32wt%, about 34wt%, about 36wt%, about 38wt%, about 40wt%, about 42wt%, about 44wt%, about 46wt%, about 48wt% or about 50wt%, including all ranges and values ​​therebetween. In embodiments, composition comprises the hyaluronic acid of about 20wt% to about 40wt%. In embodiments, composition comprises the hyaluronic acid of about 30wt%.

[0039] In embodiments, the molecular weight of the hyaluronic acid may range from about 100 g / mol to about 1,000,000 g / mol.

[0040] "Photoinitiators" generally include agents that form free radicals under light irradiation of appropriate wavelengths. In embodiments, the photoinitiator is an aromatic carbonyl compound (e.g., benzoin derivatives, benzoketals, acetophenone derivatives, hydroxyalkyl phenones) or an aromatic ketone (e.g., benzophenone and thioxanthone). In embodiments, the photoinitiator is benzophenone, dimethoxyphenylacetophenone, 2,2-dimethoxy-2-phenylacetophenone and 2,2-diethoxyacetophenone, 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetophenone, 12959, camphorquinone, rose Bengal, methylene blue, erythrosine, phloxine dye, thionine, riboflavin and methyl green.Other photoinitiators include 1-(4-fluorophenyl)-2-methyl-2-morpholino-1-propanone, 1,7-bis(9-acridinyl)heptane, 1-chloro-4-propoxythioxanthone, 1-hydroxycyclohexyl phenyl ketone, 2,2-diethoxyacetophenone, 2,3,4,4′-tetrahydroxybenzophenone, 2,3,4-trihydroxybenzophenone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 2 / 4-diethylthioxanthone, 2 / 4-isopropylthioxanthone, 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholino)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminobenzoic acid, and 1-[4-(4-morpholino)phenyl]-1-butanone. Ethyl ester, 2-ethylhexyl 4-dimethylaminobenzoate, 2-hydroxy-2-methylphenyl-propan-1-one, 2-hydroxy-4′-hydroxyethoxy-2-methylphenylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropiotone-1,4-(4-methylphenylthiophenyl)-phenyl ketone, 4,4′-difluorobenzophenone, 4,4′-dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzil dimethyl ketal, benzophenone, benzophenone hydrazone, bis(p-tolyl)iodonium hexafluorophosphate, dimethyl sebacate , diphenyliodonium hexafluorophosphate, ethyl (2,4,6-trimethylbenzoyl)phenylphosphinate, ethyl 4-(dimethylamino)benzoate, methyl o-benzoylbenzoate, methyl phenylglyoxylate, N,N,N′,N′-tetraethyl-4,4-diaminobenzophenone, phenyltribromomethyl sulfone, acylphosphine oxide (APO) and bisacylphosphine oxide (BAPO), 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, 2,2-dimethoxy-1,2-diphenylethan-1-one, hydroxycyclohexyl-phenyl-ketone, methyl benzoylformate, 2-[2-oxo-phenyl-acetoxy]-acetate, 2-[4-(4-morpholinyl)phenyl]-1-butanone, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide, phosphine oxide, bis(η5-2,4-cyclopentadien-1-yl), bis[2,6-difluoro-3-(1H-pyrrol-1-yl)phenyl]titanium, iodonium, (4-methylphenyl)[4-(2-methylpropyl)phenyl]-hexafluorophosphate (1-), bis(2,6-dimethoxybenzoyl)-2,4,4-trimethylpentylphosphine oxide or its derivatives.

[0041] In embodiments, the photoinitiator is riboflavin. In embodiments, the riboflavin is riboflavin 5' monophosphate sodium salt.

[0042] In an embodiment, the substrate comprises about 0.01 wt% to about 0.10 wt% of a photoinitiator. In an embodiment, the substrate comprises about 0.01 wt% to about 0.5 wt% of a photoinitiator, for example, about 0.01 wt%, about 0.02 wt%, about 0.03 wt%, about 0.04 wt%, about 0.05 wt%, about 0.06 wt%, about 0.07 wt%, about 0.08 wt%, about 0.09 wt%, about 0.10 wt%, about 0.11 wt%, 0.12 wt%, about 0.13 wt%, about 0.14 wt%, about 0.15 wt%, about 0.16 wt%, about 0.17 wt%, about 0.18 wt%, about 0.19 wt%, about 0.20 wt%, about 0.21 wt%, 0.22 wt%, about 0.23 wt%, about 0.24 wt%. %, about 0.25 wt%, about 0.26 wt%, about 0.27 wt%, about 0.28 wt%, about 0.29 wt%, about 0.30 wt%, about 0.31 wt%, about 0.32 wt%, about 0.33 wt%, about 0.34 wt%, about 0.35 wt%, about 0.36 wt%, about 0.37 wt%, about 0.38 wt%, about 0.39 wt%, about 0.40 wt%, about 0.41 wt%, 0.42 wt%, about 0.43 wt%, about 0.44 wt%, about 0.45 wt%, about 0.46 wt%, about 0.47 wt%, about 0.48 wt%, about 0.49 wt%, or about 0.5 wt%, including all ranges and values ​​therebetween. In an embodiment, the composition comprises from about 0.08 wt% to about 0.016 wt% of the photoinitiator. In an embodiment, the composition comprises from about 0.08 wt% to about 0.40 wt% of a photoinitiator. In an embodiment, the composition comprises about 0.08 wt% of a photoinitiator. In an embodiment, the composition comprises about 0.50 wt% of a photoinitiator. In an embodiment, the composition comprises about 0.12 wt% of a photoinitiator. In an embodiment, the composition comprises about 0.40 wt% of a photoinitiator.

[0043] Applicants have discovered that the addition of arginine to the compositions of the present disclosure improves the formation of the gel and the distribution of other agents within the gel. In embodiments, arginine acts as a co-initiator for cross-linking.

[0044] In embodiments, the substrate comprises arginine. In embodiments, arginine is L-arginine. In embodiments, the substrate comprises about 0wt% to about 20wt% arginine, such as about 0wt%, about 1wt%, about 2wt%, about 3wt%, about 4wt%, about 5wt%, about 6wt%, about 7wt%, about 8wt%, about 9wt%, about 10wt%, about 11wt%, about 12wt%, about 13wt%, about 14wt%, about 15wt%, about 16wt%, about 17wt%, about 18wt%, about 19wt% or about 20wt%, including all ranges and values ​​therebetween. In embodiments, the substrate comprises about 6wt% to about 12wt% arginine. In embodiments, the substrate comprises about 6wt% arginine. In embodiments, the substrate comprises about 9wt% arginine.

[0045] In embodiments, the substrate comprises an amine catalyst, for example, ethyl 4-dimethylaminobenzoate (EDMAB).

[0046] In an embodiment, the substrate comprises a free radical inhibitor, such as 2,6-di-tert-butyl-4-methylphenol (BHT).

[0047] In embodiments, the diluent comprises about 0 wt % to about 99 wt % of the composition. In embodiments, the diluent comprises about 1 wt % to about 99 wt % of the composition, for example, about 1 wt %, about 2 wt %, about 3 wt %, about 4 wt %, about 5 wt %, about 6 wt %, about 7 wt %, about 8 wt %, about 9 wt %, about 10 wt %, about 11 wt %, about 12 wt %, about 13 wt %, about 14 wt %, about 15 wt %, about 16 wt %, about 17 wt %, about 18 wt %, about 19 wt %, about 20 wt %, about 21 wt %, about 22 wt %, about 23 wt % %, about 24wt%, about 25wt%, about 26wt%, about 27wt%, about 28wt%, about 29wt%, about 30wt%, about 31wt%, about 32wt%, about 33wt%, about 34wt%, about 35wt%, about 36wt%, about 37wt%, about 38wt%, about 39wt%, about 40wt%, about 41wt%, about 42wt%, about 43wt%, about 44wt%, about 45wt%, about 46wt%, about 47wt%, about 48wt%, about 49wt% %, about 50wt%, about 51wt%, about 52wt%, about 53wt%, about 54wt%, about 55wt%, about 56wt%, about 57wt%, about 58wt%, about 59wt%, about 60wt%, about 61wt%, about 62wt%, about 63wt%, about 64wt%, about 65wt%, about 66wt%, about 67wt%, about 68wt%, about 69wt%, about 70wt%, about 71wt%, about 72wt%, about 73wt%, about 74wt%, about 75wt %, about 76wt%, about 77wt%, about 78wt%, about 79wt%, about 80wt%, about 81wt%, about 82wt%, about 83wt%, about 84wt%, about 85wt%, about 86wt%, about 87wt%, about 88wt%, about 89wt%, about 90wt%, about 91wt%, about 92wt%, about 93wt%, about 94wt%, about 95wt%, about 96wt%, about 97wt%, about 98wt% or about 99wt%, including all ranges and values ​​therebetween. In embodiments, the diluent accounts for about 0wt% to about 95wt% of the composition. In embodiments, the diluent accounts for about 1wt% to about 95wt% of the composition. In embodiments, the diluent accounts for about 97wt% of the composition. In embodiments, the diluent accounts for about 1wt% of the composition. In embodiments, the diluent accounts for about 2wt% of the composition. In embodiments, the diluent includes water. In embodiments, the diluent includes water and sodium chloride. In embodiments, the diluent comprises water and phosphate buffer. In embodiments, the diluent comprises water, sodium chloride, and phosphate buffer. In embodiments, the diluent is 0.9% sodium chloride in water (ie, saline). Freeze-dried composition

[0048] The pre-lyophilized compositions of the present disclosure can be lyophilized to produce a lyophilized composition. Without being limited by theory, lyophilization can increase the shelf life of the compositions of the present disclosure. In embodiments, lyophilization of the composition provides a shelf life of greater than 1 year. In embodiments, lyophilization of the composition provides a shelf life of about 1 year.

[0049] In embodiments, the lyophilized composition is porous to allow for faster reconstitution times. In embodiments, porosity is introduced by freeze drying, aeration, or the introduction of ice particles during freezing. In embodiments, freeze drying parameters are critical to prevent the material from collapsing during the freeze drying process. Inadequate drying can result in gel collapse, forming a dense, hard mass that becomes difficult to reconstitute and has a poor gel consistency.

[0050] In an embodiment, the freeze-dried composition comprises water, methacrylated chitosan, hyaluronic acid and a photoinitiator. In an embodiment, the freeze-dried composition further comprises arginine. In an embodiment, arginine is L-arginine. In an embodiment, the photoinitiator is riboflavin. In an embodiment, riboflavin is riboflavin 5' monophosphate sodium salt. In an embodiment, the present disclosure provides a freeze-dried composition comprising about 0wt% to about 10wt% of water, 50wt% to about 65wt% of methacrylated chitosan, 25wt% to about 35wt% of hyaluronic acid, 0.10wt% to about 0.15wt% of riboflavin and 0wt% to about 10wt% of arginine. In an embodiment, the present disclosure provides a freeze-dried composition comprising about 0wt% to about 10wt% of water, 40wt% to about 80wt% of methacrylated chitosan, 20wt% to about 40wt% of hyaluronic acid, 0.01wt% to about 0.5wt% of riboflavin and 0wt% to about 20wt% of arginine.

[0051] In embodiments, the freeze-dried composition comprises about 0wt% to about 10wt% water, such as about 0wt%, about 0.5wt%, about 1.0wt%, about 1.5wt%, about 2.0wt%, about 2.5wt%, about 3.0wt%, about 3.5wt%, about 4.0wt%, about 4.5wt%, about 5.0wt%, about 5.5wt%, about 6.0wt%, about 6.5wt%, about 7.0wt%, about 7.5wt%, about 8.0wt%, about 8.5wt%, about 9.0wt%, about 9.5wt% or about 10.0wt%, including all ranges and values ​​therebetween. In embodiments, the freeze-dried composition comprises about 0wt% to about 2wt% water. In embodiments, the freeze-dried composition comprises about 0wt% water. In embodiments, the freeze-dried composition comprises about 1wt% water. In embodiments, the freeze-dried composition comprises about 2wt% water.

[0052] In embodiments, the lyophilized composition comprises about 50 wt % to about 65 wt % of methacrylated chitosan. In embodiments, the lyophilized composition comprises about 30 wt % to about 90 wt % of methacrylated chitosan, for example, about 30 wt %, about 31 wt %, about 32 wt %, about 33 wt %, about 34 wt %, about 35 wt %, about 36 wt %, about 37 wt %, about 38 wt %, about 39 wt %, about 40 wt %, about 41 wt %, about 42 wt %, about 43 wt %, about 44 wt %, about 45 wt %, about 46 wt %, about 47 wt %, about 48 wt %, about 49 wt %, about 50 wt %, about 51 wt %, about 52 wt %, about 53 wt %, about 54 wt %, about 55 wt %, about 56 wt %, about 57 wt %, about 58 wt %, about 59 wt %, about 60 wt %, about 61 wt %, about 62 wt %, about 63 wt %, about 64 wt %, about 65 wt %, about 66 wt %, about 67 wt %, about 68 wt %, about 69 wt %, about 70 wt %, about 71 wt %, about 72 wt %, about 73 wt %, about 74 wt %, about 75 wt %, about 76 wt %, about 77 wt %, about 78 wt %, about 79 wt %, about 80 wt %, about 81 wt %, about 82 wt %, about 83 wt %, about 84 %, about 8wt%, about 59wt%, about 60wt%, about 61wt%, about 62wt%, about 63wt%, about 64wt%, about 65wt%, about 66wt%, about 67wt%, about 68wt%, about 69wt%, about 70wt%, about 71wt%, about 72wt%, about 73wt%, about 74wt%, about 75wt%, about 76wt%, about 77wt%, about 78wt%, about 79wt%, about 80wt%, about 81wt%, about 82wt%, about 83wt%, about 84wt%, about 85wt%, about 86wt%, about 87wt%, about 88wt%, about 89wt% or about 90wt%, including all ranges and values ​​therebetween. In embodiments, the lyophilized composition comprises about 59wt% methacrylated chitosan. In embodiments, the lyophilized composition comprises about 60wt% methacrylated chitosan. In embodiments, the lyophilized composition comprises about 61 wt% methacrylated chitosan. In embodiments, the lyophilized composition comprises about 40 wt% to about 80 wt% methacrylated chitosan. In embodiments, the lyophilized composition comprises about 50 wt% to about 65 wt% methacrylated chitosan.

[0053] In embodiments, the freeze-dried composition comprises about 25wt% to about 35wt% hyaluronic acid. In embodiments, the freeze-dried composition comprises about 10wt% to about 50wt% hyaluronic acid, such as about 10wt%, about 11wt%, about 12wt%, about 13wt%, about 14wt%, about 15wt%, about 16wt%, about 17wt%, about 18wt%, about 19wt%, about 20wt%, about 21wt%, about 22wt%, about 23wt%, about 24wt%, about 25wt%, about 26wt%, about 27wt%, about 28wt%. , about 29wt%, about 30wt%, about 31wt%, about 32wt%, about 33wt%, about 34wt%, about 35wt%, about 36wt%, about 37wt%, about 38wt%, about 39wt%, about 40wt%, about 41wt%, about 42wt%, about 43wt%, about 44wt%, about 45wt%, about 46wt%, about 47wt%, about 48wt%, about 49wt% or about 50wt%, including all ranges and values ​​therebetween. In embodiments, the lyophilized composition comprises about 29wt% hyaluronic acid. In embodiments, the lyophilized composition comprises about 30wt% hyaluronic acid. In embodiments, the lyophilized composition comprises about 31wt% hyaluronic acid. In embodiments, the lyophilized composition comprises about 20wt% to about 40wt% hyaluronic acid. In embodiments, the lyophilized composition comprises about 25wt% to about 35wt% hyaluronic acid.

[0054] In an embodiment, the photoinitiator is an aromatic carbonyl compound (e.g., a benzoin derivative, a benzoketal, an acetophenone derivative, a hydroxyalkyl phenone) or an aromatic ketone (e.g., benzophenone and thioxanthone). In an embodiment, the photoinitiator is benzophenone, dimethoxyphenylacetophenone, 2,2-dimethoxy-2-phenylacetophenone and 2,2-diethoxyacetophenone, 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetophenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, phloxine dye, thionine, riboflavin and methyl green.Other photoinitiators include 1-(4-fluorophenyl)-2-methyl-2-morpholino-1-propanone, 1,7-bis(9-acridinyl)heptane, 1-chloro-4-propoxythioxanthone, 1-hydroxycyclohexyl phenyl ketone, 2,2-diethoxyacetophenone, 2,3,4,4′-tetrahydroxybenzophenone, 2,3,4-trihydroxybenzophenone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 2 / 4-diethylthioxanthone, 2 / 4-isopropylthioxanthone, 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholino)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminobenzoic acid, and 1-[4-(4-morpholino)phenyl]-1-butanone. Ethyl ester, 2-ethylhexyl 4-dimethylaminobenzoate, 2-hydroxy-2-methylphenyl-propan-1-one, 2-hydroxy-4′-hydroxyethoxy-2-methylphenylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropiotone-1,4-(4-methylphenylthiophenyl)-phenyl ketone, 4,4′-difluorobenzophenone, 4,4′-dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzil dimethyl ketal, benzophenone, benzophenone hydrazone, bis(p-tolyl)iodonium hexafluorophosphate, dimethyl sebacate , diphenyliodonium hexafluorophosphate, ethyl (2,4,6-trimethylbenzoyl)phenylphosphinate, ethyl 4-(dimethylamino)benzoate, methyl o-benzoylbenzoate, methyl phenylglyoxylate, N,N,N′,N′-tetraethyl-4,4-diaminobenzophenone, phenyltribromomethyl sulfone, acylphosphine oxide (APO) and bisacylphosphine oxide (BAPO), 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, 2,2-dimethoxy-1,2-diphenylethan-1-one, hydroxycyclohexyl-phenyl-ketone, methyl benzoylformate, 2-[2-oxo-phenyl-acetoxy]-acetate, 2-[4-(4-morpholinyl)phenyl]-1-butanone, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide, phosphine oxide, bis(η5-2,4-cyclopentadien-1-yl), bis[2,6-difluoro-3-(1H-pyrrol-1-yl)phenyl]titanium, iodonium, (4-methylphenyl)[4-(2-methylpropyl)phenyl]-hexafluorophosphate (1-), bis(2,6-dimethoxybenzoyl)-2,4,4-trimethylpentylphosphine oxide or its derivatives.

[0055] In embodiments, the photoinitiator is riboflavin. In embodiments, the riboflavin is riboflavin 5' monophosphate sodium salt. In embodiments, the lyophilized composition comprises about 0.10 wt% to about 0.15 wt% riboflavin. In embodiments, the lyophilized composition comprises about 0.10 wt% to about 0.7 wt% riboflavin, for example, about 0.10 wt%, about 0.11 wt%, about 0.12 wt%, about 0.13 wt%, about 0.14 wt%, about 0.15 wt%, about 0.16 wt%, about 0.17 wt%, about 0.18 wt%, about 0.19 wt%, about 0.20 wt%, about 0.21 wt%, about 0.2 2wt%, about 0.23wt%, about 0.24wt%, about 0.25wt%, about 0.26wt%, about 0.27wt%, about 0.28wt%, about 0.29wt%, about 0.30wt%, about 0.31wt%, 0.32wt%, about 0.33wt%, about 0.34wt%, about 0.35wt%, about 0.36wt%, about 0.37wt%, about 0.38wt%, About 0.39wt%, about 0.40wt%, about 0.41wt%, 0.42wt%, about 0.43wt%, about 0.44wt%, about 0.45wt%, about 0.46wt%, about 0.47wt%, about 0.48wt%, about 0.49wt%, about 0.50wt%, about 0.51wt%, 0.52wt%, about 0.53wt%, about 0.54wt%, about 0.55wt t%, about 0.56wt%, about 0.57wt%, about 0.58wt%, about 0.59wt%, about 0.60wt%, about 0.61wt%, 0.62wt%, about 0.63wt%, about 0.64wt%, about 0.65wt%, about 0.66wt%, about 0.67wt%, about 0.68wt%, about 0.69wt% or about 0.7wt%, including all ranges and values ​​therebetween. In embodiments, riboflavin is riboflavin 5' monophosphate sodium salt. In embodiments, the lyophilisate comprises about 0.11wt% riboflavin. In embodiments, the lyophilized composition comprises about 0.12wt% riboflavin. In embodiments, the lyophilized composition comprises about 0.13wt% riboflavin. In embodiments, the lyophilized composition comprises about 0.01wt% to about 0.5wt% riboflavin. In embodiments, the lyophilized composition comprises from about 0.01 wt% to about 0.15 wt% riboflavin.

[0056] In embodiments, the lyophilized composition comprises about 0wt% to about 10wt% arginine. In embodiments, the lyophilized composition comprises about 0wt% to about 30wt% arginine, such as about 0wt%, about 1wt%, about 2wt%, about 3wt%, about 4wt%, about 5wt%, about 6wt%, about 7wt%, about 8wt%, about 9wt%, about 10wt%, about 11wt%, about 12wt%, about 13wt%, about 14wt%, about 15wt%, about 16wt%, about 17wt%, about 18wt%, about 19wt%, about 20wt%, about 21wt%, about 22wt%, about 23wt%, about 24wt%, about 25wt%, about 26wt%, about 27wt%, about 28wt%, about 29wt% or about 30wt%, including all ranges and values ​​therebetween. In embodiments, the lyophilized composition comprises about 8wt% arginine. In an embodiment, the arginine is L-arginine. In an embodiment, the lyophilized composition comprises about 9 wt% arginine. In an embodiment, the lyophilized composition comprises about 10 wt% arginine. In an embodiment, the lyophilized composition comprises about 0 wt% to about 10 wt% arginine. In an embodiment, the lyophilized composition comprises about 0 wt% to about 20 wt% arginine. In an embodiment, the arginine is L-arginine.

[0057] In an embodiment, the composition is a lyophilized composition prepared by lyophilizing any pre-lyophilized composition of the present disclosure, comprising about 0 wt % to about 2 wt % water, wherein the lyophilized composition can be reconstituted in less than 60 seconds to provide a semi-gel composition of the present disclosure.

[0058] In an embodiment, the lyophilized composition comprises about 1 wt% water, about 60 wt% methacrylated chitosan, about 30 wt% hyaluronic acid, about 9 wt% L-arginine, and 0.12 wt% riboflavin 5' monophosphate sodium salt.

[0059] In an embodiment, the lyophilized composition is sterilized, wherein the sterilization is achieved by ionizing radiation. In an embodiment, the ionizing radiation is electron beam radiation. In an embodiment, the ionizing radiation is X-ray radiation. In an embodiment, the ionizing radiation is gamma radiation.

[0060] In embodiments, sterilization partially solidifies the composition. In embodiments, sterilization is required to fully solidify the composition. In embodiments, electron beam radiation partially solidifies the gel. In embodiments, x-ray radiation partially solidifies the gel. In embodiments, the lyophilized composition is sterilized, wherein sterilization is achieved by ionizing radiation. In embodiments, the ionizing radiation is gamma radiation. In embodiments, gamma radiation partially solidifies the composition.

[0061] In embodiments, the dose of ionizing radiation is about 10 kGy to about 70 kGy, about 10 kGy, about 12 kGy, about 14 kGy, about 16 kGy, about 18 kGy, about 20 kGy, about 22 kGy, about 24 kGy, about 25 kGy, about 26 kGy, about 28 kGy, about 30 kGy, about 32 kGy, about 34 kGy, about 36 kGy In some embodiments, the dosage of ionizing radiation is about 25 kGy, about 38 kGy, about 40 kGy, about 42 kGy, about 44 kGy, about 46 kGy, about 48 kGy, about 50 kGy, about 52 kGy, about 54 kGy, about 56 kGy, about 58 kGy, about 60 kGy, about 62 kGy, about 64 kGy, about 66 kGy, about 68 kGy or about 70 kGy, including all ranges and values ​​therebetween. In embodiments, the dosage of ionizing radiation is about 25 kGy.

[0062] In embodiments, after sterilization, the log reduction in colony forming units is between about 2 and about 4, e.g., about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, or about 5.0, including all ranges and values ​​therebetween. In embodiments, after sterilization, the log reduction in colony forming units is between about 3.4 and about 3.6. In embodiments, after sterilization, the log reduction in colony forming units is about 3.4. Semi-gel composition

[0063] In embodiments, the lyophilized composition of the present disclosure is reconstituted to provide a semi-gel composition. In embodiments, the semi-gel composition is prepared by mixing the lyophilized composition of the present disclosure with a diluent until homogeneous.

[0064] As used herein, the term "reconstitution time" means the mixing time required to form a homogeneous mixture during reconstitution. In an embodiment, the mixing time is from about 5 seconds to about 120 seconds, for example, about 1 second, about 2 seconds, about 3 seconds, about 4 seconds, about 5 seconds, about 6 seconds, about 7 seconds, about 8 seconds, about 9 seconds, about 10 seconds, about 12 seconds, about 14 seconds, about 16 seconds, about 18 seconds, about 20 seconds, about 22 seconds, about 24 seconds, about 26 seconds, about 28 seconds, about 30 seconds, about 32 seconds, about 34 seconds, about 36 seconds, about 38 seconds, about 40 seconds, about 42 seconds, about 44 seconds, about 46 seconds, about 48 seconds, about 50 seconds, about 52 seconds, about 54 seconds, about 56 seconds, about 58 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 110 seconds, or about 120 seconds, including all values ​​or ranges therebetween. In an embodiment, recovery takes about 30 seconds. In an embodiment, recovery takes less than about 60 seconds. In an embodiment, recovery takes less than about 90 seconds.

[0065] In an embodiment, a semi-gel composition comprises a diluent, methacrylated chitosan, hyaluronic acid, and a photoinitiator. In an embodiment, the semi-gel composition further comprises arginine. In an embodiment, the arginine is L-arginine. In an embodiment, the photoinitiator is riboflavin. In an embodiment, riboflavin is riboflavin 5' monophosphate sodium salt. In an embodiment, the present disclosure provides a semi-gel composition comprising about 80 wt% to about 99 wt% of a diluent, about 1 wt% to about 3 wt% of methacrylated chitosan, about 0.5 wt% to about 2 wt% of hyaluronic acid, about 0.0010 wt% to about 0.01 wt% of a photoinitiator, and about 0 wt% to about 0.5 wt% of arginine.

[0066] In an embodiment, the final concentration of the diluent in the semi-gel composition is about 80 wt% to about 99 wt%, such as about 80 wt%, about 81 wt%, about 82 wt%, about 83 wt%, about 84 wt%, about 85 wt%, about 86 wt%, about 87 wt%, about 88 wt%, about 89 wt%, about 90 wt%, about 91 wt%, about 92 wt%, about 93 wt%, about 94 wt%, about 95 wt%, about 96 wt%, about 97 wt%, about 98 wt% or about 99 wt%, including all ranges and values ​​therebetween. In an embodiment, the final concentration of the diluent in the semi-gel composition is about 96 wt%. In an embodiment, the final concentration of the diluent in the semi-gel composition is about 97 wt%. In an embodiment, the final concentration of the diluent in the semi-gel composition is about 98 wt%. In an embodiment, the diluent comprises water. In an embodiment, the diluent comprises water and sodium chloride. In an embodiment, the diluent comprises water and phosphate buffer. In embodiments, the diluent comprises water, sodium chloride, and phosphate buffer. In embodiments, the diluent is 0.9% sodium chloride in water (ie, saline).

[0067] In an embodiment, the semi-gel composition is prepared by mixing any of the compositions of the present disclosure with a diluent until homogeneous; wherein the final concentration of the diluent is about 97 wt %; wherein the mixing time is less than about 60 seconds; wherein the viscosity of the semi-gel composition is from about 0.01 kPa*s to about 1.0 kPa*s, as measured by rheometry.

[0068] In an embodiment, the semi-gel composition comprises about 80 wt% to about 99 wt% water, such as about 80 wt%, about 81 wt%, about 82 wt%, about 83 wt%, about 84 wt%, about 85 wt%, about 86 wt%, about 87 wt%, about 88 wt%, about 89 wt%, about 90 wt%, about 91 wt%, about 92 wt%, about 93 wt%, about 94 wt%, about 95 wt%, about 96 wt%, about 97 wt%, about 98 wt% or about 99 wt%, including all ranges and values ​​therebetween. In an embodiment, the semi-gel composition comprises about 90 wt% to about 97 wt% water. In an embodiment, the semi-gel composition comprises about 97 wt% water.

[0069] In embodiments, the semi-gel composition comprises about 1 wt% to about 3 wt% methacrylated chitosan, such as about 1 wt%, about 1.1 wt%, about 1.2 wt%, about 1.3 wt%, about 1.4 wt%, about 1.5 wt%, about 1.6 wt%, about 1.7 wt%, about 1.8 wt%, about 1.9 wt%, about 2 wt%, about 2.1 wt%, about 2.2 wt%, about 2.3 wt%, about 2.4 wt%, about 2.5 wt%, about 2.6 wt%, about 2.7 wt%, about 2.8 wt%, about 2.9 wt%, or about 3 wt%, including all ranges and values ​​therebetween. In embodiments, the semi-gel composition comprises about 2 wt% methacrylated chitosan. In embodiments, the methacrylated chitosan is about 25% to about 40% methacrylated.

[0070] In an embodiment, the semi-gel composition comprises about 0.5wt% to about 2wt% hyaluronic acid, such as about 0.5wt%, about 0.6wt%, about 0.7wt%, about 0.8wt%, about 0.9wt%, about 1wt%, about 1.1wt%, about 1.2wt%, about 1.3wt%, about 1.4wt%, about 1.5wt%, about 1.6wt%, about 1.7wt%, about 1.8wt%, about 1.9wt% or about 2wt%, including all ranges and values ​​therebetween. In an embodiment, the semi-gel composition comprises about 1wt% hyaluronic acid.

[0071] In an embodiment, the photoinitiator is an aromatic carbonyl compound (e.g., a benzoin derivative, a benzoketal, an acetophenone derivative, a hydroxyalkyl phenone) or an aromatic ketone (e.g., benzophenone and thioxanthone). In an embodiment, the photoinitiator is benzophenone, dimethoxyphenylacetophenone, 2,2-dimethoxy-2-phenylacetophenone and 2,2-diethoxyacetophenone, 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetophenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, phloxine dye, thionine, riboflavin and methyl green.Other photoinitiators include 1-(4-fluorophenyl)-2-methyl-2-morpholino-1-propanone, 1,7-bis(9-acridinyl)heptane, 1-chloro-4-propoxythioxanthone, 1-hydroxycyclohexyl phenyl ketone, 2,2-diethoxyacetophenone, 2,3,4,4′-tetrahydroxybenzophenone, 2,3,4-trihydroxybenzophenone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 2 / 4-diethylthioxanthone, 2 / 4-isopropylthioxanthone, 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholino)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminobenzoic acid, and 1-[4-(4-morpholino)phenyl]-1-butanone. Ethyl ester, 2-ethylhexyl 4-dimethylaminobenzoate, 2-hydroxy-2-methylphenyl-propan-1-one, 2-hydroxy-4′-hydroxyethoxy-2-methylphenylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropiotone-1,4-(4-methylphenylthiophenyl)-phenyl ketone, 4,4′-difluorobenzophenone, 4,4′-dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzil dimethyl ketal, benzophenone, benzophenone hydrazone, bis(p-tolyl)iodonium hexafluorophosphate, dimethyl sebacate , diphenyliodonium hexafluorophosphate, ethyl (2,4,6-trimethylbenzoyl)phenylphosphinate, ethyl 4-(dimethylamino)benzoate, methyl o-benzoylbenzoate, methyl phenylglyoxylate, N,N,N′,N′-tetraethyl-4,4-diaminobenzophenone, phenyltribromomethyl sulfone, acylphosphine oxide (APO) and bisacylphosphine oxide (BAPO), 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, 2,2-dimethoxy-1,2-diphenylethan-1-one, hydroxycyclohexyl-phenyl-ketone, methyl benzoylformate, 2-[2-oxo-phenyl-acetoxy]-acetate, 2-[4-(4-morpholinyl)phenyl]-1-butanone, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide, phosphine oxide, bis(η5-2,4-cyclopentadien-1-yl), bis[2,6-difluoro-3-(1H-pyrrol-1-yl)phenyl]titanium, iodonium, (4-methylphenyl)[4-(2-methylpropyl)phenyl]-hexafluorophosphate (1-), bis(2,6-dimethoxybenzoyl)-2,4,4-trimethylpentylphosphine oxide or its derivatives.

[0072] In embodiments, the photoinitiator is riboflavin. In embodiments, the riboflavin is riboflavin 5' monophosphate sodium salt. In embodiments, the semi-gel composition comprises about 0.001 wt% to about 0.01 wt% riboflavin, such as about 0.0010 wt%, about 0.0015 wt%, about 0.0020 wt%, about 0.0025 wt%, about 0.0030 wt%, about 0.0035 wt%, about 0.0040 wt%, about 0.0045 wt%, about 0.0050 wt%, about 0.0055 wt%, about 0.0060 wt%, about 0.0065 wt%, about 0.0070 wt%, about 0.0075 wt%, about 0.0080 wt%, about 0.0085 wt%, about 0.0090 wt%, about 0.0095 wt%, or about 0.0100 wt%, including all ranges and values ​​therebetween. In embodiments, the semi-gel composition comprises about 0.004 wt% riboflavin. In an embodiment, the riboflavin is riboflavin 5' monophosphate sodium salt.

[0073] In an embodiment, the semi-gel composition comprises about 0 wt% to about 0.5 wt% arginine, such as about 0 wt%, about 0.10 wt%, about 0.15 wt%, about 0.20 wt%, about 0.25 wt%, about 0.30 wt%, about 0.35 wt%, about 0.40 wt%, about 0.45 wt%, or about 0.50 wt%, including all ranges and values ​​therebetween. In an embodiment, the semi-gel composition comprises about 0.3 wt% arginine. In an embodiment, the arginine is L-arginine.

[0074] In an embodiment, the semi-gel composition comprises about 97 wt% water, about 2 wt% methacrylated chitosan, about 1 wt% hyaluronic acid, about 0.3 wt% L-arginine, and about 0.004 wt% riboflavin 5' monophosphate sodium salt.

[0075] In an embodiment, the viscosity of the semi-gel composition is from about 0.01 kPa*s to about 1 kPa*s, as measured by rheometry, e.g., about 0.01 kPa*s, about 0.02 kPa*s, about 0.03 kPa*s, about 0.04 kPa*s, about 0.05 kPa*s, about 0.06 kPa*s, about 0.07 kPa*s, about 0.08 kPa*s, about 0.09 kPa*s, about 0.1 kPa*s, about 0.2 kPa*s, about 0.3 kPa*s, about 0.4 kPa*s, about 0.5 kPa*s, about 0.6 kPa*s, about 0.7 kPa*s, about 0.8 kPa*s, about 0.9 kPa*s, or about 1.0 kPa*s, including all ranges and values ​​therebetween. In an embodiment, the viscosity of the semi-gel composition is from about 0.01 kPa*s to about 1.0 kPa*s, as measured by rheometry. In an embodiment, the viscosity of the semi-gel composition is from about 0.01 kPa*s to about 0.3 kPa*s, as measured by rheometry. In an embodiment, the viscosity of the semi-gel composition is from about 0.5 kPa*s to about 1.0 kPa*s, as measured by rheometry.

[0076] In an embodiment, the semi-gel composition is sterilized, wherein the sterilization is achieved by ionizing radiation. In an embodiment, the ionizing radiation is electron beam radiation. In an embodiment, the ionizing radiation is X-ray radiation. In an embodiment, the ionizing radiation is gamma radiation.

[0077] In embodiments, sterilization partially cures the composition. In embodiments, sterilization is required to fully cure the composition. In embodiments, electron beam radiation partially cures the gel. In embodiments, x-ray radiation partially cures the gel. In embodiments, gamma radiation partially cures the composition.

[0078] In embodiments, the dose of ionizing radiation is about 10 kGy to about 70 kGy, about 10 kGy, about 12 kGy, about 14 kGy, about 16 kGy, about 18 kGy, about 20 kGy, about 22 kGy, about 24 kGy, about 25 kGy, about 26 kGy, about 28 kGy, about 30 kGy, about 32 kGy, about 34 kGy, about 36 kGy In some embodiments, the dosage of ionizing radiation is about 25 kGy, about 38 kGy, about 40 kGy, about 42 kGy, about 44 kGy, about 46 kGy, about 48 kGy, about 50 kGy, about 52 kGy, about 54 kGy, about 56 kGy, about 58 kGy, about 60 kGy, about 62 kGy, about 64 kGy, about 66 kGy, about 68 kGy or about 70 kGy, including all ranges and values ​​therebetween. In embodiments, the dosage of ionizing radiation is about 25 kGy.

[0079] In embodiments, after sterilization, the log reduction in colony forming units is between about 2 and about 4, e.g., about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, or about 5.0, including all ranges and values ​​therebetween. In embodiments, after sterilization, the log reduction in colony forming units is between about 3.4 and about 3.6. In embodiments, after sterilization, the log reduction in colony forming units is about 3.4.

[0080] In embodiments, the semi-gel adheres to epithelial and mucosal tissues with a strength ranging from about 1 kPa to about 20 kPa, as measured using a lap shear test. In embodiments, the semi-gel adheres to epithelial and mucosal tissues with a strength ranging from about 0.1 kPa to about 20 kPa, as measured using a lap shear test, e.g., about 0.1 kPa, about 0.2 kPa, about 0.3 kPa, about 0.4 kPa, about 0.5 kPa, about 0.6 kPa, about 0.7 kPa, about 0.8 kPa, about 0.9 kPa, about 1.0 kPa, about 1.5 kPa, about 2.0 kPa, about 2.5 kPa, about 3.0 kPa, about 3.5 kPa, about 4.0 kPa, about 4.5 kPa, about 5.0 kPa, about 5.5 kPa, about 6.0 kPa, about 6.5 kPa, about 7.0 kPa, about 7.5 kPa, about 8.0 kPa, about 8.5 kPa, about 9.0 kPa, about 9.5 kPa, about 10.0 kPa, about 10.5 kPa, about 11.0 kPa, about 11.5 kPa, about 12.0 kPa, about 12.5 kPa, about 13.0 kPa, about 13.5 kPa, about 14.0 kPa, about 14.5 kPa, about 15.0 kPa, about 15.5 kPa, about 16.0 kPa, about 16.5 kPa, about 17.0 kPa, about 17.5 kPa, about 18.0 kPa, about 18.5 kPa, about 19.0 kPa, about 19.5 kPa or about 20.0 kPa, including all ranges and values ​​therebetween.

[0081] In embodiments, the semi-gel is a shear-thinning thixotropic material. Cured composition

[0082] Compositions according to the present disclosure include cured compositions, which are compositions that have been cured to form a polymer matrix. In embodiments, the cured composition has sufficient rigidity and / or adhesion (e.g., as a result of functional groups that provide cell adhesion domains) so that epithelial cells in the TM can migrate into and / or onto the scaffold and adhere and / or grow. In embodiments, the compositions of the present disclosure are cured compositions produced by light radiation. In embodiments, the cured composition is prepared by light radiation of a semi-gel composition.

[0083] In embodiments, the viscosity or stiffness of the compositions of the present disclosure is described by the elastic modulus measured by rheometry. In embodiments, the elastic modulus can be measured as described in the literature. For example, the elastic modulus can be measured using a parallel plate rheometer test that applies an oscillating shear force to the composition.

[0084] In embodiments, the elastic modulus of the cured composition is from about 0.05 kPa to about 100 kPa, as measured by rheometry, e.g., about 0.05 kPa, about 0.10 kPa, about 0.15 kPa, about 0.20 kPa, about 0.25 kPa, about 0.30 kPa, about 0.35 kPa, about 0.40 kPa, about 0.45 kPa, about 0.50 kPa, about 0.55 kPa, about 0.60 kPa, about 0.65 kPa, about 0.70 kPa, about 0.75 kPa, about 0.80 kPa, about 0.85 kPa, about 0.90 kPa, about 100 kPa, about 110 kPa, about 120 kPa, about 130 kPa, about 140 kPa, about 150 kPa, about 160 kPa, about 170 kPa, about 180 kPa, about 190 kPa, about 200 kPa, about 210 kPa, about 220 kPa, about 230 kPa, about 240 kPa, about 250 kPa kPa, about 0.80 kPa, about 0.85 kPa, about 0.90 kPa, about 0.95 kPa, about 1.0 kPa, about 1.5 kPa, about 2.0 kPa, about 2.5 kPa, about 3.0 kPa, about 3.5 kPa, about 4.0 kPa, about 4.5 kPa, about 5.0 kPa, about 5.50 kPa, about 6.0 kPa, about 6.5 kPa, about 7.0 kPa, about 7.5 kPa, about 8.0 kPa, about 8.5 kPa, about 9.0 kPa, about 9.5kPa, about 10kPa, about 11kPa, about 12kPa, about 13kPa, about 14kPa, about 15kPa, about 16kPa, about 17kPa, about 18kPa, about 19kPa, about 20kPa, about 21kPa, about 22kPa, about 23kPa, about 24kPa, about 25kPa, about 26kPa, about 27kPa, about 28kPa, about 29kPa, about 30kPa, about 31kPa, about 32k Pa, about 33kPa, about 34kPa, about 35kPa, about 36kPa, about 37kPa, about 38kPa, about 39kPa, about 40kPa, about 41kPa, about 42kPa, about 43kPa, about 44kPa, about 45kPa, about 46kPa, about 47kPa, about 48kPa, about 49kPa, about 50kPa, about 51kPa, about 52kPa, about 53kPa, about 54kPa, about 55kPa, about 56kPa, about 57kPa, about 58kPa, about 59kPa, about 60kPa, about 61kPa, about 62kPa, about 63kPa, about 64kPa, about 65kPa, about 66kPa, about 67kPa, about 68kPa, about 69kPa, about 70kPa, about 71kPa, about 72kPa, about 73kPa, about 74kPa, about 75kPa, about 76kPa, about 77kPa, about 78kPa, about 79kPa kPa, about 80 kPa, about 81 kPa, about 82 kPa, about 83 kPa, about 84 kPa, about 85 kPa, about 86 kPa, about 87 kPa, about 88 kPa, about 89 kPa, about 90 kPa, about 91 kPa, about 92 kPa, about 93 kPa, about 94 kPa, about 95 kPa, about 96 kPa, about 97 kPa, about 98 kPa, about 99 kPa, or about 100 kPa, including all ranges and values ​​therebetween. In embodiments, the elastic modulus of the cured composition is from about 0.4 kPa to about 3 kPa, as measured by rheometry. .

[0085] In an embodiment, the curing composition comprises a diluent, methacrylated chitosan, hyaluronic acid, and a photoinitiator. In an embodiment, the curing composition further comprises arginine. In an embodiment, arginine is L-arginine. In an embodiment, the photoinitiator is riboflavin. In an embodiment, riboflavin is riboflavin 5' monophosphate sodium salt. In an embodiment, the diluent comprises water. In an embodiment, the curing composition comprises about 80wt% to about 99wt% of a diluent, 1wt% to about 3wt% of methacrylated chitosan, 0.5wt% to about 2wt% of hyaluronic acid, 0.001wt% to about 0.01wt% of riboflavin, and 0wt% to about 0.5wt% of arginine.

[0086] In an embodiment, the solidified composition comprises about 80 wt% to about 99 wt% of a diluent, such as about 80 wt%, about 81 wt%, about 82 wt%, about 83 wt%, about 84 wt%, about 85 wt%, about 86 wt%, about 87 wt%, about 88 wt%, about 89 wt%, about 90 wt%, about 91 wt%, about 92 wt%, about 93 wt%, about 94 wt%, about 95 wt%, about 96 wt%, about 97 wt%, about 98 wt% or about 99 wt%, including all ranges and values ​​therebetween. In an embodiment, the semi-gel composition comprises about 90 wt% to about 97 wt% of water. In an embodiment, the semi-gel composition comprises about 97 wt% of water. In an embodiment, the diluent comprises water. In an embodiment, the diluent comprises water and sodium chloride. In an embodiment, the diluent comprises water and phosphate buffer. In an embodiment, the diluent comprises water, sodium chloride and phosphate buffer. In an embodiment, the diluent is a 0.9% sodium chloride aqueous solution (i.e., saline).

[0087] In embodiments, the curing composition comprises about 1 wt% to about 3 wt% methacrylated chitosan, such as about 1 wt%, about 1.1 wt%, about 1.2 wt%, about 1.3 wt%, about 1.4 wt%, about 1.5 wt%, about 1.6 wt%, about 1.7 wt%, about 1.8 wt%, about 1.9 wt%, about 2 wt%, about 2.1 wt%, about 2.2 wt%, about 2.3 wt%, about 2.4 wt%, about 2.5 wt%, about 2.6 wt%, about 2.7 wt%, about 2.8 wt%, about 2.9 wt%, or about 3 wt%, including all ranges and values ​​therebetween. In embodiments, the semi-gel composition comprises about 2 wt% methacrylated chitosan. In embodiments, the methacrylated chitosan is about 25% to about 40% methacrylated.

[0088] In embodiments, the solidified composition comprises about 0.5wt% to about 2wt% hyaluronic acid, such as about 0.5wt%, about 0.6wt%, about 0.7wt%, about 0.8wt%, about 0.9wt%, about 1wt%, about 1.1wt%, about 1.2wt%, about 1.3wt%, about 1.4wt%, about 1.5wt%, about 1.6wt%, about 1.7wt%, about 1.8wt%, about 1.9wt% or about 2wt%, including all ranges and values ​​therebetween. In embodiments, the semi-gel composition comprises about 1wt% hyaluronic acid.

[0089] In an embodiment, the photoinitiator is an aromatic carbonyl compound (e.g., a benzoin derivative, a benzoketal, an acetophenone derivative, a hydroxyalkyl phenone) or an aromatic ketone (e.g., benzophenone and thioxanthone). In an embodiment, the photoinitiator is benzophenone, dimethoxyphenylacetophenone, 2,2-dimethoxy-2-phenylacetophenone and 2,2-diethoxyacetophenone, 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetophenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, phloxine dye, thionine, riboflavin and methyl green.Other photoinitiators include 1-(4-fluorophenyl)-2-methyl-2-morpholino-1-propanone, 1,7-bis(9-acridinyl)heptane, 1-chloro-4-propoxythioxanthone, 1-hydroxycyclohexyl phenyl ketone, 2,2-diethoxyacetophenone, 2,3,4,4′-tetrahydroxybenzophenone, 2,3,4-trihydroxybenzophenone, 2,4,6-trimethylbenzoyldiphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 2 / 4-diethylthioxanthone, 2 / 4-isopropylthioxanthone, 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholino)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminobenzoic acid, and 1-[4-(4-morpholino)phenyl]-1-butanone. Ethyl ester, 2-ethylhexyl 4-dimethylaminobenzoate, 2-hydroxy-2-methylphenyl-propan-1-one, 2-hydroxy-4′-hydroxyethoxy-2-methylphenylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropiotone-1,4-(4-methylphenylthiophenyl)-phenyl ketone, 4,4′-difluorobenzophenone, 4,4′-dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzil dimethyl ketal, benzophenone, benzophenone hydrazone, bis(p-tolyl)iodonium hexafluorophosphate, dimethyl sebacate , diphenyliodonium hexafluorophosphate, ethyl (2,4,6-trimethylbenzoyl)phenylphosphinate, ethyl 4-(dimethylamino)benzoate, methyl o-benzoylbenzoate, methyl phenylglyoxylate, N,N,N′,N′-tetraethyl-4,4-diaminobenzophenone, phenyltribromomethyl sulfone, acylphosphine oxide (APO) and bisacylphosphine oxide (BAPO), 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one, 2,2-dimethoxy-1,2-diphenylethan-1-one, hydroxycyclohexyl-phenyl-ketone, methyl benzoylformate, 2-[2-oxo-phenyl-acetoxy]-acetate, 2-[4-(4-morpholinyl)phenyl]-1-butanone, diphenyl(2,4,6-trimethylbenzoyl)phosphine oxide, phosphine oxide, bis(η5-2,4-cyclopentadien-1-yl), bis[2,6-difluoro-3-(1H-pyrrol-1-yl)phenyl]titanium, iodonium, (4-methylphenyl)[4-(2-methylpropyl)phenyl]-hexafluorophosphate (1-), bis(2,6-dimethoxybenzoyl)-2,4,4-trimethylpentylphosphine oxide or its derivatives.

[0090] In embodiments, the photoinitiator is riboflavin. In embodiments, the riboflavin is riboflavin 5' monophosphate sodium salt. In embodiments, the cured composition comprises about 0.001 wt% to about 0.01 wt% riboflavin, such as about 0.0010 wt%, about 0.0015 wt%, about 0.0020 wt%, about 0.0025 wt%, about 0.0030 wt%, about 0.0035 wt%, about 0.0040 wt%, about 0.0045 wt%, about 0.0050 wt%, about 0.0055 wt%, about 0.0060 wt%, about 0.0065 wt%, about 0.0070 wt%, about 0.0075 wt%, about 0.0080 wt%, about 0.0085 wt%, about 0.0090 wt%, about 0.0095 wt%, or about 0.0100 wt%, including all ranges and values ​​therebetween. In embodiments, the cured composition comprises about 0.004 wt% riboflavin. In an embodiment, the riboflavin is riboflavin 5' monophosphate sodium salt.

[0091] In an embodiment, the solidified composition comprises about 0 wt% to about 0.5 wt% arginine, such as about 0 wt%, about 0.10 wt%, about 0.15 wt%, about 0.20 wt%, about 0.25 wt%, about 0.30 wt%, about 0.35 wt%, about 0.40 wt%, about 0.45 wt%, or about 0.50 wt%, including all ranges and values ​​therebetween. In an embodiment, the semi-gel composition comprises about 0.3 wt% arginine. In an embodiment, the arginine is L-arginine.

[0092] In an embodiment, the solidified composition comprises about 96.8 wt% water, about 1.936 wt% methacrylated chitosan, about 0.968 wt% hyaluronic acid, about 0.29 wt% L-arginine, and 0.004 wt% riboflavin 5' monophosphate sodium salt. The medicine box of the present disclosure

[0093] In an embodiment, the present disclosure provides a kit comprising: a composition of the present disclosure (eg, a lyophilized composition or a semi-gel composition), a gel dispensing tray, and a light guide. In an embodiment, the kit is enclosed in a sealed tray.

[0094] In an embodiment, the kit is sterilized after sealing the tray. In an embodiment, sterilization is achieved by ionizing radiation. In an embodiment, the ionizing radiation is electron beam radiation. In an embodiment, the ionizing radiation is X-ray radiation. In an embodiment, the ionizing radiation is gamma radiation.

[0095] In embodiments, the dose of ionizing radiation is about 10 kGy to about 70 kGy, about 10 kGy, about 12 kGy, about 14 kGy, about 16 kGy, about 18 kGy, about 20 kGy, about 22 kGy, about 24 kGy, about 25 kGy, about 26 kGy, about 28 kGy, about 30 kGy, about 32 kGy, about 34 kGy, about 36 kGy In some embodiments, the dosage of ionizing radiation is about 25 kGy, about 38 kGy, about 40 kGy, about 42 kGy, about 44 kGy, about 46 kGy, about 48 kGy, about 50 kGy, about 52 kGy, about 54 kGy, about 56 kGy, about 58 kGy, about 60 kGy, about 62 kGy, about 64 kGy, about 66 kGy, about 68 kGy or about 70 kGy, including all ranges and values ​​therebetween. In embodiments, the dosage of ionizing radiation is about 25 kGy.

[0096] In embodiments, after sterilization, the log reduction in colony forming units is between about 2 and about 4, e.g., about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, or about 5.0, including all ranges and values ​​therebetween. In embodiments, after sterilization, the log reduction in colony forming units is between about 3.4 and about 3.6. In embodiments, after sterilization, the log reduction in colony forming units is about 3.4.

[0097] In an embodiment, a sterilized composition (e.g., a sterilized lyophilized composition or a sterilized semi-gel composition) is contained within a bag. In an embodiment, a lyophilized composition or a semi-gel composition is contained within a bag. In an embodiment, the bag comprises a moisture barrier. In an embodiment, the bag comprises a transparent portion. In an embodiment, the bag comprises a moisture barrier and a transparent portion. In an embodiment, the bag comprises a Luer lock connector. In an embodiment, the area of ​​the bag is about 9 cm 2 About 30cm 2 , for example about 9cm 2 , about 10cm 2 、Approx. 11cm 2 、Approx. 12cm 2 、Approx. 13cm 2 、Approx. 14cm 2 、Approx. 15cm 2 、Approx. 16cm 2 、Approx. 17cm 2 、Approx. 18cm 2 、Approx. 19cm 2 、About 20cm2 、Approx. 21cm 2 、Approx. 22cm 2 、Approx. 23cm 2 、Approx. 24cm 2 、Approx. 25cm 2 、Approx. 26cm 2 、Approx. 27cm 2 、Approx. 28cm 2 、Approx. 29cm 2 or about 30cm 2 , including all ranges and values ​​in between.

[0098] In an embodiment, the bag contains about 0.2 mL to about 10 mL of the composition, e.g., about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 0.6 mL, about 0.7 mL, about 0.8 mL, about 0.9 mL, about 1.0 mL, about 1.1 mL, about 1.2 mL, about 1.3 mL, about 1.4 mL, about 1.5 mL, about 1.6 mL, about 1.7 mL, about 1.8 mL, about 1.9 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL, including all ranges and values ​​therebetween.

[0099] In an embodiment, the composition is contained within a mixing container (eg, a syringe, a pouch, a vial, or a tube). In an embodiment, the composition is contained within a mixing syringe.

[0100] In embodiments, the shelf life of the kit is greater than about 1 year. In embodiments, the shelf life of the kit is about 1 year.

[0101] In an embodiment, the present disclosure provides a kit comprising: a lyophilized composition, a gel dispensing tray, and a light guide. In an embodiment, the lyophilized composition is contained within a bag. In an embodiment, the kit is enclosed in a sealed tray. Methods of the present disclosure

[0102] The present disclosure provides compositions and methods for treating tympanic membrane perforation. In embodiments, the perforation is a chronic perforation of the tympanic membrane. In embodiments, the otologic material comprises a semi-gel composition that cures in situ to produce a polymer matrix (e.g., a hydrogel scaffold). In embodiments, the method comprises applying the semi-gel to a desired portion of a patient's ear and curing the material in situ. In embodiments, the method comprises applying the semi-gel composition to an otologic filling material, and curing the semi-gel composition after application to the filling material.

[0103] In an embodiment, the present disclosure provides a method of treating a tympanic membrane perforation, comprising: (i) applying the semi-gel composition of the present disclosure to the tympanic membrane, (ii) inserting the light guide into the ear canal, and (iii) curing the composition by irradiation with light.

[0104] In an embodiment, the light radiation comprises a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 To about 100000mW / cm 2 The light radiation intensity of the radiation composition.

[0105] In embodiments, about 50 μL to about 500 μL of the composition is applied to the tympanic membrane. In an embodiment, about 10 μL to about 500 μL of the composition is applied to the tympanic membrane, for example, about 10 μL, about 20 μL, about 30 μL, about 40 μL, about 50 μL, about 60 μL, about 70 μL, about 80 μL, about 90 μL, about 100 μL, about 120 μL, about 140 μL, about 160 μL, about 180 μL, about 200 μL, about 220 μL, about 240 μL, about 260 μL, about 280 μL, about 300 μL, about 320 μL, about 340 μL, about 360 μL, about 380 μL, about 400 μL, about 420 μL, about 440 μL, about 460 μL, about 480 μL, or about 500 μL, including all ranges and values ​​therebetween. In embodiments, about 200 μL of the composition is applied to the tympanic membrane.

[0106] In an embodiment, the light radiation comprises a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 About 1000mW / cm 2 In an embodiment, the light radiation comprises irradiating the semi-gel composition with a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 About 10000mW / cm 2 In an embodiment, the light radiation comprises irradiating the semi-gel composition with a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 To about 100000mW / cm 2In an embodiment, the light irradiation comprises irradiating the composition (e.g., a semi-gel composition) at a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a light radiation power of about 2 mW to about 1 W. In an embodiment, the light irradiation comprises irradiating the composition (e.g., a semi-gel composition) at a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a light radiation power of about 0.2 mW to about 1 W.

[0107] In an embodiment, the wavelength is from about 200 nm to about 800 nm, for example, about 200 nm, about 205 nm, about 210 nm, about 215 nm, about 220 nm, about 225 nm, about 230 nm, about 235 nm, about 240 nm, about 245 nm, about 250 nm, about 255 nm, about 260 nm, about 265 nm, about 270 nm, about 275 nm, about 280 nm, about 285 nm, about 290 nm, about 295 nm, about 300 nm, about 305 nm, about 310 nm, about 315 nm, about 320 nm, about 325 nm, about 330 nm, about 335 nm , about 340nm, about 345nm, about 350nm, about 355nm, about 360nm, about 365nm, about 370nm, about 375nm, about 380nm, about 385nm, about 390nm, about 395nm, about 400nm, about 405nm, about 410nm, about 415nm, about 420nm, about 425nm, about 430nm, about 435nm, about 440nm, about 445nm, about 450nm, about 455nm, about 460nm, about 465nm, about 470nm, about 475nm, about 480nm, about 485nm, about 490nm, about 495nm 5nm, about 500nm, about 505nm, about 510nm, about 515nm, about 520nm, about 525nm, about 530nm, about 535nm, about 540nm, about 545nm, about 550nm, about 555nm, about 560nm, about 565nm, about 570nm, about 575nm, about 580nm, about 585nm, about 590nm, about 595nm, about 600nm, about 605nm, about 610nm, about 615nm, about 620nm, about 625nm, about 630nm, about 635nm, about 640nm, about 645nm, about 650nm, In some embodiments, the radiation wavelength is about 655nm, about 660nm, about 665nm, about 670nm, about 675nm, about 680nm, about 685nm, about 690nm, about 695nm, about 700nm, about 705nm, about 710nm, about 715nm, about 720nm, about 725nm, about 730nm, about 735nm, about 740nm, about 745nm, about 750nm, about 755nm, about 760nm, about 765nm, about 770nm, about 775nm, about 780nm, about 785nm, about 790nm, about 795nm or about 800nm, including all ranges and values ​​therebetween. In embodiments, the radiation wavelength is about 420nm to about 530nm. In embodiments, the radiation wavelength is about 450nm.

[0108] In an embodiment, the optical radiation power is about 2 mW to about 1 W. In an embodiment, the optical radiation power is about 0.2 mW to about 1 W, for example, about 0.2 mW, about 0.4 mW, about 0.6 mW, about 0.8 mW, about 1.0 mW, about 1.2 mW, about 1.4 mW, about 1.6 mW, about 1.8 mW, about 2 mW, about 4 mW, about 5 mW, about 6 mW, about 7 mW, about 8 mW, about 9 mW, about 10 mW, about 11 mW, about 12 mW, about 14 mW, about 16 mW, about 18 mW, about 20 mW, about 22 mW, about 24 mW, about 26 mW, about 28 mW, about 30 mW, about 32 mW, about 34 mW, about 36 mW. In some embodiments, the light radiation power is about 15mW to about 50mW. In some embodiments, the light radiation power is about 25mW.

[0109] In an embodiment, the light radiation intensity is about 100 mW / cm 2 About 1000mW / cm 2 In an embodiment, the light radiation intensity is about 100 mW / cm 2 About 10000mW / cm 2 。 In an embodiment, the light radiation intensity is about 100 mW / cm 2 To about 100000mW / cm 2 , for example, about 100 mW / cm 2 , about 110mW / cm 2 , about 120mW / cm 2 , about 130mW / cm 2 , about 140mW / cm 2 , about 150mW / cm 2 , about 160mW / cm 2 , about 170mW / cm 2 , about 180mW / cm 2 , about 190mW / cm 2 , about 200mW / cm 2 , about 220mW / cm 2 , about 240mW / cm2 , about 260mW / cm 2 , about 280mW / cm 2 , about 300mW / cm 2 , about 320mW / cm 2 , about 340mW / cm 2 , about 360mW / cm 2 , about 380mW / cm 2 , about 400mW / cm 2 , about 420mW / cm 2 , about 440mW / cm 2 , about 460mW / cm 2 , about 480mW / cm 2 , about 500mW / cm 2 , about 550mW / cm 2 , about 600mW / cm 2 , about 650mW / cm 2 , about 700mW / cm 2 , about 750mW / cm 2 , about 800mW / cm 2 , about 850mW / cm 2 , about 900mW / cm 2 , about 950mW / cm 2 , about 1000mW / cm 2 , about 1100mW / cm 2 , about 1200mW / cm 2 , about 1300mW / cm 2 , about 1400mW / cm 2 , about 1500mW / cm 2 , about 1600mW / cm 2 , about 1700mW / cm 2 , about 1800mW / cm 2 , about 1900mW / cm 2 , about 2000mW / cm 2 , about 2200mW / cm 2 , about 2400mW / cm 2 , about 2600mW / cm 2 , about 2800mW / cm 2 , about 3000mW / cm 2 , about 3200mW / cm 2 , about 3400mW / cm 2 , about 3600mW / cm 2 , about 3800mW / cm 2, about 4000mW / cm 2 , about 5000mW / cm 2 , about 6000mW / cm 2 , about 7000mW / cm 2 , about 8000mW / cm 2 , about 9000mW / cm 2 , about 10000mW / cm 2 , about 11000mW / cm 2 , about 12000mW / cm 2 , about 13000mW / cm 2 , about 14000mW / cm 2 , about 15000mW / cm 2 , about 16000mW / cm 2 , about 17000mW / cm 2 , about 18000mW / cm 2 , about 19000mW / cm 2 , about 20000mW / cm 2 , about 30000mW / cm 2 , about 40000mW / cm 2 , about 50000mW / cm 2 , about 60000mW / cm 2 , about 70000mW / cm 2 , about 80000mW / cm 2 , about 90000mW / cm 2 or about 100000mW / cm 2 , including all ranges and values ​​therebetween. In an embodiment, the power of the radiation is about 178 mW / cm 2 .

[0110] In an embodiment, the curing time is from about 1 second to about 300 seconds (sec), for example, from about 1 second, about 2 seconds, about 3 seconds, about 4 seconds, about 5 seconds, about 6 seconds, about 7 seconds, about 8 seconds, about 9 seconds, about 10 seconds, about 12 seconds, about 14 seconds, about 16 seconds, about 18 seconds, about 20 seconds, about 22 seconds, about 24 seconds, about 26 seconds, about 28 seconds, about 30 seconds, about 32 seconds, about 34 seconds, about 36 seconds, about 38 seconds, about 40 seconds, about 42 seconds, about 44 seconds, about 46 seconds, about 48 seconds, about 50 seconds, about 52 seconds, about 54 seconds, about 56 seconds In some embodiments, the curing time is about 5 seconds to about 120 seconds. In some embodiments, the curing time is about 12 seconds to about 300 seconds. In some embodiments, the curing time is about 90 seconds to about 150 seconds. In some embodiments, the curing time is about 150 seconds to about 160 seconds. In some embodiments, the curing time is about 190 seconds. In some embodiments, the curing time is about 200 seconds. In some embodiments, the curing time is about 210 seconds, about 220 seconds, about 230 seconds, about 240 seconds, about 250 seconds, about 260 seconds, about 270 seconds, about 280 seconds, about 290 seconds or about 300 seconds, including all ranges and values ​​therebetween. In embodiments, the curing time is about 5 seconds to about 120 seconds. In embodiments, the curing time is about 12 seconds to about 300 seconds. In embodiments, the curing time is 90 seconds.

[0111] In embodiments, light radiation (i.e., curing time) is continuous. In embodiments, light radiation (i.e., curing time) is applied with one or more bursts, for example, 1 burst, 2 bursts, 3 bursts, 4 bursts or 5 bursts. For example, in embodiments, curing time is about 90 seconds, and light radiation is applied in three burst forms, about 30 seconds each time. In embodiments, light radiation is applied in a burst mode. In embodiments, light radiation is applied in two burst modes. In embodiments, light radiation is applied in three burst modes. In embodiments, the time between bursts is about 5 seconds to about 30 seconds, for example, about 1 second, about 2 seconds, about 3 seconds, about 4 seconds, about 5 seconds, about 6 seconds, about 7 seconds, about 8 seconds, about 9 seconds, about 10 seconds, about 12 seconds, about 14 seconds, about 16 seconds, about 18 seconds, about 20 seconds, about 22 seconds, about 24 seconds, about 26 seconds, about 28 seconds or about 30 seconds, including all ranges and values ​​therebetween. In embodiments, the time between bursts is about 10 seconds.

[0112] In embodiments, the present disclosure provides a method for treating a perforated tympanic membrane of a patient in need thereof, comprising: (a) adding and mixing a solvent (e.g., saline) with any of the lyophilized compositions disclosed herein to form a semi-gel composition, (b) applying the semi-gel composition to the perforated tympanic membrane of the patient, and (c) curing the applied semi-gel composition with light radiation. In embodiments, the lyophilized composition is mixed with about 0.1 mL to about 2 mL of a solvent (e.g., saline), such as about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 0.6 mL, about 0.7 mL, about 0.8 mL, about 0.9 mL, about 1.0 mL, about 1.1 mL, about 1.2 mL, about 1.3 mL, about 1.4 mL, about 1.5 mL, about 1.6 mL, about 1.7 mL, about 1.8 mL, about 1.9 mL, or about 2.0 mL, including all ranges and values ​​therebetween. In embodiments, the lyophilized composition is mixed with about 1 mL of saline. In an embodiment, the semi-gel composition is applied directly to the perforation site of the patient's tympanic membrane, where it is cured by any of the photoirradiation methods described in the present disclosure. In an embodiment, the cured semi-gel composition provides a scaffold for endogenous cell migration and is replaced by newly formed tympanic membrane tissue.

[0113] In embodiments, the patient in need is a child. In embodiments, the patient in need is an adolescent 18 years of age or younger. In embodiments, the patient in need is an adult. In embodiments, the patient is 5 years of age or older or about at least 5 years of age. In embodiments, the patient is 5-11 years of age, such as about 5 years of age, about 6 years of age, about 7 years of age, about 8 years of age, about 9 years of age, about 10 years of age, or about 11 years of age, including all values ​​or ranges therebetween.

[0114] In some embodiments, the methods of the present disclosure provide treatment for perforations in patients in need thereof, wherein at least about 1% (e.g., at least 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 78%, 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 9 %, 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 78%, about 79%, about 80%, including all values ​​or ranges therebetween) of the tympanic membrane are affected by a perforation. In embodiments, the methods of the present disclosure provide treatment for perforations in patients in need thereof, wherein at least about 5% of the patient's tympanic membrane is affected by the perforation. In embodiments, the methods of the present disclosure provide treatment for perforations in patients in need thereof, wherein at least about 10% of the patient's tympanic membrane is affected by the perforation. In embodiments, at least about 15% of the patient's tympanic membrane is affected by the perforation. In embodiments, at least about 20% of the patient's tympanic membrane is affected by the perforation. In embodiments, at least about 25% of the patient's tympanic membrane is affected by the perforation. In embodiments, at least about 30% of the patient's tympanic membrane is affected by the perforation. In embodiments, at least about 50% of the patient's tympanic membrane is affected by the perforation. In embodiments, at least about 80% of the patient's tympanic membrane is affected by the perforation.

[0115] In embodiments, the methods of the present disclosure provide a tympanic membrane perforation closure rate of at least about 40%. In embodiments, the methods of the present disclosure provide a tympanic membrane perforation closure rate of about ≥50%. In embodiments, the methods of the present disclosure provide a tympanic membrane perforation closure rate of about ≥70%. In embodiments, the methods of the present disclosure provide a tympanic membrane perforation closure rate of about ≥75%. In embodiments, the methods of the present disclosure provide a tympanic membrane perforation closure rate of>75%. In embodiments, the perforation closure rate is determined by visual assessment. In embodiments, visual assessment is performed using a microscope or endoscope.

[0116] In an embodiment, the method of the present disclosure improves the pure tone mean score and / or air-bone difference by about ≥40% compared to a baseline. In an embodiment, the method of the present disclosure provides an improvement of about ≥50% in the pure tone mean score compared to a baseline. In an embodiment, the method of the present disclosure provides an improvement of about ≥70% in the pure tone mean score compared to a baseline. In an embodiment, the method of the present disclosure provides an improvement of about ≥75% in the pure tone mean score compared to a baseline.

[0117] In embodiments, the method provides a tympanic membrane perforation closure rate or pure tone mean score of> 75% compared to baseline by a single application. In embodiments, the method increases the tympanic membrane perforation closure rate within about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 3 months, about 4 months, about 5 months, or about 6 months after administration. In embodiments, the treatment increases the tympanic membrane perforation closure rate within about 3 months after administration. Example Example 1: Sterilization of lyophilized composition program

[0118] The kit was sterilized by radiation at a dose of 25 kGy. The irradiated samples were incubated in culture medium at 37 ° C for 24 h and 1 week. The samples were then placed in the extraction medium in the biosafety cabinet using sterilized instruments. After extraction, an aliquot of the extraction medium was placed on selective agar medium and incubated overnight at 37 ° C to identify various microorganisms (bacteria and fungi) that may be present. result

[0119] After 24 h and 1 week of incubation, no microorganisms appeared on any of the selective media tested (Figures 1 and 2). In contrast, the non-irradiated samples showed hundreds of colonies, indicating the presence of bacteria. This suggests that the radiation level was sufficient to kill any microorganisms that might be present before sterilization. in conclusion

[0120] Irradiation appeared to be sufficient for sterilization. The log reduction between non-sterile and sterilized samples was between 3.4 and 3.6 compared to non-irradiated controls. Example 2: Curing and characterization of semi-gel program

[0121] In an office setting, the lyophilized composition was rehydrated with 1 mL of saline and mixed for 20-30 seconds to form a semi-gel composition. After preparing the treatment site, the semi-gel composition was applied to cover the tympanic membrane perforation with a 1 mm overlap at the edges. The semi-gel was irradiated for 1.5 min using a light guide mounted on a curing light. result

[0122] Flow curve tests were performed on the semi-gels at shear rates ranging from 0.01 s-1 to 10 s-1 for samples reconstituted in n=4 bags. At a shear rate of 1 s-1, viscosity values ​​ranging from 0.01 kPa*s to 0.3 kPa*s were obtained. Example 3: In vivo animal model establishment and testing program

[0123] The eardrums of adult male chinchillas were perforated with a laser to create perforations of similar size. Perforations that remained open for 8 weeks were considered chronic perforations. The semi-gel composition was applied to the eardrum perforation and cured with a curing light and a light guide to form a cured composition. result

[0124] The perforations healed within three weeks. After fourteen weeks of treatment with the solidified composition, the tympanic membrane and cochlea were evaluated by histopathological analysis. The tympanic membrane perforations treated with the solidified composition healed, and the regenerated tissue had the same architecture as the native tympanic membrane tissue. The hair cells in the cochlea were intact, indicating that the solidified composition was not ototoxic. Example 4: Evaluation of the ototoxic potential of the cured compositions program

[0125] The eardrum of an adult guinea pig was perforated with a scalpel and then immediately treated with the semi-gel composition. The semi-gel composition was then cured using a light guide and a curing lamp. result

[0126] The perforations were monitored for 4 weeks, during which time the perforations healed. Ototoxicity was assessed by auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAE). The results of the ABR and DPOAE tests showed no loss of hearing and hair cell integrity, indicating that the cured composition did not have any ototoxic potential. Example 5: Otological Gel Kit

[0127] Figure 1A -C shows an exemplary otological gel kit of the present disclosure. The components contained in the kit are shown in Table 1. Table 1. Otologic Gel Kit Components Parts, components, technology, software, accessories Otology Gel Patch Gel Application Kit (Sterile) 1 gel sachet containing gel pack 1 distribution tray 1 light guide Gel Curing Light Package 1 curing light 1 foot pedal 1 power cord 2 safety goggles Sterile saline Cleaning Application Equipment

[0128] The otological gel kit (i.e., otological gel patch kit) provides a composition of the present disclosure, which comprises hyaluronic acid (HA), methacrylated chitosan (MeCS) and riboflavin monophosphate (RM). The composition is supplied in a bag. Before the contents in the gel bag are applied to the patient, the contents are mixed with saline (1 mL saline). After adding the saline, the bag is re-covered and then kneaded to mix the dry reagent with the saline. After mixing, the end of the bag is then torn off and the contents are emptied into the supplied tray. The uncured composition is a semi-gel that can be applied directly to the perforation through the external auditory canal using a curette or other preferred instrument. After exposure to the blue light of the curing lamp, the semi-gel of the composition forms a hard three-dimensional gel patch to fill the perforation. Animal studies have shown that the gel material stays in situ for less than 4 weeks. Example 6: Clinical study on the safety and effectiveness of otologic gel

[0129] The following example provides a clinical study to evaluate the safety and efficacy of the disclosed composition in repairing chronic tympanic membrane perforations. The study will: i) evaluate the effectiveness of the composition as a gel patch in promoting the natural healing process, closing chronic, >25% tympanic membrane perforations with a single application; and ii) evaluate the safety of the composition and application procedures.

[0130] The study will also i) evaluate the success rate of the gel application procedure (e.g., in the presence of variations in anatomical conditions, perforation location, TMP size, and / or patient age); ii) measure the time for a single gel application; iii) measure the time from gel patch application to tympanic membrane healing; iv) evaluate hearing changes in all patient groups; v) evaluate the safety of the gel patch for its intended use in the target population; and vi) assess pain and discomfort using a visual analog scale (VAS).

[0131] The primary treatment endpoint will be to measure the rate of complete closure of tympanic membrane perforations, which will be reported by clinical visual assessment to be greater than 75% at the 5th visit (3 months) after application of the gel composition. Safety endpoints of the incidence of serious adverse events related to the device and procedure during and after application will also be measured. Additional secondary endpoints include: visual assessment of complete closure of tympanic membrane perforations at the 5th visit after successful application of the gel composition and sedation; surgical success / failure rate; median time (in minutes) to successfully complete an application procedure; comparison of audiogram data at baseline and visit 5 for pure tone mean scores and air-bone difference (ABG); comparison of word recognition rate data at baseline and visit 5; comparison of tympanometric measurement data of tympanometric activity at baseline and visit 5; and median time (in weeks) to closure of tympanic membrane perforations after treatment.

[0132] Inclusion Criteria: Patients will be eligible for this study if they meet all of the following criteria at the baseline visit: 1. Be willing and able to provide informed consent, Legally Authorized Representative (LAR) agreement, and assent when age appropriate. 2. Females and males must be at least 5 years old. 3. Criteria for affected ears: a. Perforation involves more than 25% of the tympanic membrane; b. After 4 weeks of observation and waiting, the perforation has not closed on its own; c. The perforation did not heal actively; d. The perforation can be visualized by endoscopy or microscopy; and e.Earwax will not clog the perforation.

[0133] Exclusion Criteria: Patients will be eligible for this study if they meet any of the following criteria: 1. The perforation is marginal (perforation of the tympanic membrane area between the perforation and the bone canal). 2. Active otitis media, with or without effusion. 3. Middle ear leakage lasts for more than 3 months. 4. History of cleft palate. 5. Receiving radiation therapy or taking corticosteroids, immunosuppressants or chemotherapy. 6. Currently taking systemic antibiotics, antibiotic ear drops, and / or steroid ear drops. 7.Currently have bacterial or viral infection. 8. Fever (temperature > 100°F) during index procedure. 9. The mass was diagnosed as tympanic cholesteatoma. 10. Known history of malignant ear canal tumor within 3 years of screening. 11. Abrasions / lacerations of the external auditory canal. 12. Have a serious medical condition that may prevent full participation in the procedures required for the study. 13. The researcher believes that the patient will not be able to cooperate with the procedure. 14. The parent / LAR believes that the patient will not be able to cooperate with the procedure. 15. Allergy to shellfish. 16. Known to be pregnant or may be pregnant. 17. Adults who lack the capacity to consent.

[0134] Study Design: Patients will be screened at Visit 1. Eligible patients will be treated with otologic gel at Visit 2 (Day 0) and will provide postoperative pain scores. Perforations will be monitored one week (Visit 3) and three weeks (Visit 4) postoperatively to determine perforation closure rate. Perforation closure and hearing ability will be assessed 12 weeks after surgery (Visit 5). Audiometry and tympanometry will be performed at Visit 5 (end of study visit). Adverse events will be observed at each visit.

[0135] Otologic Gel Application Procedure: The patient will lie supine on the exam room chair and the ear will be placed under the microscope or endoscope. The ENT clinician will apply the uncured semi-gel composition material using clean microsurgical tools. The semi-gel composition is applied to the piercing so that the uncured semi-gel composition and the TM tissue surrounding the piercing overlap by approximately 1 mm. Because the procedure is performed under the microscope or endoscope, the ENT is able to precisely cover the piercing with gel. The procedure is a clean procedure and does not require aseptic application. The edges of the piercing are not manipulated or abraded prior to applying the gel composition. Once the uncured gel composition material is applied and covers the piercing site, the ENT will cure the material using the provided reusable curing light and disposable light guide. The patient, clinician, and others in the room will wear the provided safety glasses. The curing light will beep intermittently during the 90-second curing period. The entire procedure to treat the ear takes approximately 10 minutes. Incorporated by Reference

[0136] All references, articles, publications, patents, patent publications and patent and patent applications cited herein are incorporated by reference in their entirety for all purposes. However, the mention of any reference, article, publication, patent, patent publication and patent application cited herein does not and should not be taken as an admission or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world.

Claims

1. A pre-freeze-dried composition comprising: (i) a substrate comprising about 40 wt % to about 80 wt % of methacrylated chitosan, about 20 wt % to about 40 wt % of hyaluronic acid, about 0.01 wt % to about 0.5 wt % of a photoinitiator, and 0 wt % to about 20 wt % of arginine; and (ii) diluent.

2. The pre-lyophilized composition of claim 1, wherein the substrate comprises about 60 wt% of methacrylated chitosan.

3. The pre-lyophilized composition according to any one of claims 1 to 2, wherein the base material comprises about 30 wt% of hyaluronic acid.

4. The pre-lyophilized composition according to any one of claims 1 to 3, wherein the substrate comprises about 0.12 wt% of a photoinitiator, wherein the photoinitiator is riboflavin.

5. The pre-lyophilized composition of any one of claims 1 to 4, wherein the base comprises about 9 wt% arginine.

6. The pre-lyophilized composition of any one of claims 1 to 5, wherein the methacrylated chitosan is about 25% to about 40% methacrylated.

7. The pre-lyophilized composition of any one of claims 1 to 6, wherein the diluent comprises water, and wherein the diluent comprises about 0 wt% to about 95 wt% of the composition.

8. A freeze-dried composition comprising: about 0 wt % to about 10 wt % water, about 40 wt % to about 80 wt % of methacrylated chitosan, about 20 wt % to about 40 wt % of hyaluronic acid, about 0.01 wt % to about 0.5 wt % of a photoinitiator, and From about 0 wt % to about 20 wt % arginine.

9. The lyophilized composition of claim 8, wherein the composition comprises about 1 wt% water.

10. The freeze-dried composition of claim 8 or 9, wherein the composition comprises about 60 wt% methacrylated chitosan.

11. The lyophilized composition of claim 8 or 10, wherein the methacrylated chitosan is about 25% to about 40% methacrylated.

12. The freeze-dried composition of any one of claims 8 to 11, wherein the composition comprises about 30 wt% hyaluronic acid.

13. The lyophilized composition of any one of claims 8 to 12, wherein the composition comprises about 0.12 wt% of a photoinitiator, wherein the photoinitiator is riboflavin.

14. The lyophilized composition of any one of claims 8 to 13, wherein the composition comprises about 9 wt% arginine.

15. The lyophilized composition of any one of claims 8 to 14, wherein the composition can be reconstituted in less than 1 minute.

16. A semi-gel composition comprising: about 80 wt % to about 99 wt % of a diluent, about 1 wt % to about 3 wt % of methacrylated chitosan, about 0.5 wt % to about 2 wt % of hyaluronic acid, about 0.0010 wt % to about 0.01 wt % of a photoinitiator, and about 0 wt % to about 0.5 wt % arginine.

17. The semi-gel composition of claim 16, wherein the composition comprises about 2 wt% methacrylated chitosan.

18. The semi-gel composition of claim 16 or 17, wherein the methacrylated chitosan is about 25% to about 40% methacrylated.

19. The semi-gel composition of any one of claims 16 to 18, wherein the composition comprises about 1 wt% hyaluronic acid.

20. The semi-gel composition of any one of claims 16 to 19, wherein the composition comprises about 0.004 wt% of a photoinitiator, wherein the photoinitiator is riboflavin.

21. The semi-gel composition of any one of claims 16 to 20, wherein the composition comprises about 0.3 wt% arginine.

22. The semi-gel composition of any one of claims 16 to 21, wherein the composition comprises about 97 wt% of diluent.

23. The semi-gel composition of claim 16 or 22, wherein the diluent is saline.

24. The semi-gel composition of any one of claims 16 to 23, wherein the composition has a viscosity of about 0.01 kPa*s to about 0.3 kPa*s as measured by rheometry.

25. The semi-gel composition of any one of claims 16 to 24, wherein the composition is prepared by mixing a lyophilized composition with a diluent until homogeneous, and wherein the mixing time of the semi-gel composition and the lyophilized composition is less than about 60 seconds.

26. A curing composition comprising: about 80 wt % to about 99 wt % of a diluent; about 1 wt % to about 3 wt % of methacrylated chitosan, about 0.5 wt % to about 2 wt % of hyaluronic acid, From about 0.0010 wt % to about 0.01 wt % of a photoinitiator; and From about 0 wt % to about 0.5 wt % arginine.

27. The curing composition of claim 26, wherein the composition comprises about 2 wt% methacrylated chitosan.

28. The curing composition of claim 26 or 27, wherein the methacrylated chitosan is about 25% to about 40% methacrylated.

29. The solidified composition of any one of claims 26 to 28, wherein the composition comprises about 1 wt% hyaluronic acid.

30. The cured composition of any one of claims 26 to 29, wherein the composition comprises about 0.004 wt% of a photoinitiator, wherein the photoinitiator is riboflavin.

31. The curing composition of any one of claims 26 to 30, wherein the composition comprises about 0.3 wt% arginine.

32. The curing composition of any one of claims 26 to 31, wherein the composition comprises about 97 wt% of a diluent.

33. The curing composition of claim 26 or 32, wherein the diluent is brine.

34. The cured composition of any one of claims 26 to 33, wherein the composition has an elastic modulus of from about 0.4 kPa to about 5 kPa as measured by rheometry.

35. The cured composition of any one of claims 26 to 34, wherein the composition is cured by heating at a wavelength of about 200 nm to about 800 nm and at about 100 mW / cm 2 To about 100000mW / cm 2 The semi-gel composition is prepared by subjecting the semi-gel composition to light irradiation for a curing time of about 1 second to about 300 seconds with a light radiation intensity of about 1 second.

36. The cured composition of claim 35, wherein the light radiation comprises a curing time of about 90 seconds.

37. A composition as described in any one of claims 8 to 25, wherein the composition is sterilized with ionizing radiation.

38. The sterilizing composition of claim 37, wherein the ionizing radiation is electron beam radiation.

39. A sterile composition as claimed in claim 37 or 38, wherein after sterilization, the log reduction in colony forming units is between 3.4 and 3.

6.

40. A medicine kit comprising: The composition, gel dispensing tray and light guide of any one of claims 8 to 25.

41. The kit of claim 40, wherein the sterilizing composition is contained within a bag.

42. A kit as claimed in claim 40 or 41, wherein the kit is enclosed in a sealed tray.

43. A kit as claimed in any one of claims 40 to 42, wherein the bag comprises a moisture barrier and a transparent portion.

44. A method for treating a perforated tympanic membrane, comprising: (a) applying the semi-gel composition according to any one of claims 16 to 25 to the tympanic membrane, (b) Insert the light guide into the ear canal, and (c) curing the composition by light radiation; The light radiation comprises a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a curing time of about 100 mW / cm 2 To about 100000mW / cm 2 The composition is irradiated with light radiation of an intensity of.

45. The method of claim 44, wherein the curing time lasts for about 90 seconds.

46. ​​A method of treating a tympanic membrane perforation in a patient in need thereof, comprising: (a) adding saline to and mixing with the lyophilized composition of any one of claims 8 to 15 to form a semi-gel composition; (b) applying the semi-gel composition to the site of the tympanic membrane perforation of a patient in need thereof; and (c) curing the applied semi-gel composition by irradiating it with light.

47. The method of any one of claims 44-46, wherein the patient is 5 years of age or older.

48. The method of claim 47, wherein the patient is 5-11 years old.

49. The method of any one of claims 44-48, wherein the patient has at least about 25% of the tympanic membrane affected by a perforation.

50. The method of any one of claims 44-49, wherein the patient has had a tympanic membrane perforation for at least 4 weeks.

51. The method of any one of claims 44-50, wherein the lyophilized composition is mixed with about 0.1 mL to about 2 mL of saline.

52. The method of claim 51, wherein the lyophilized composition is mixed with about 1 mL of saline.

53. The method of any one of claims 44-52, wherein the curing time lasts for about 90 seconds.

54. The method of any one of claims 44-53, wherein the solidified semi-gel is biodegradable.

55. The method of any one of claims 44-54, wherein the cured semi-gel remains in the applied tympanic membrane for less than about 4 weeks.

56. The method of any one of claims 44-55, wherein the treatment provides a tympanic membrane perforation closure rate of about ≥ 75%.

57. The method of any one of claims 44-56, wherein the treatment provides a tympanic membrane perforation closure rate of >75% with a single application.

58. The method of any one of claims 44-57, wherein the treatment increases the rate of closure of tympanic membrane perforations within about 3 months after administration.

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