Photocurable hydrogel composition

Abstract

The present disclosure provides a photocurable hydrogel composition and a method for use in treating tympanic membrane perforations.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims priority to U.S. Provisional Application No. 63 / 390,837, filed on July 20, 2022, the content of which is hereby incorporated by reference in its entirety for all purposes.

[0002] Government Support This invention was made with government support under grant number 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] The insertion of tympanostomy tubes (TTs) is a very common procedure in both children and adults. During TT insertion, a small tube is placed into the tympanic membrane (TM), allowing fluid to drain from the middle ear and air to enter the middle ear through the TM. It is most commonly performed for the purpose of improving hearing due to chronic effusion behind the tympanic membrane, as well as in recurrent otitis media to reduce the frequency of infections and for the direct topical application of antibiotics. In most cases, the tympanostomy tubes fall out naturally 6 - 18 months after placement, and the tympanic membrane (TM) is completely repaired. However, in about 15% of patients, TM perforation occurs after the TT has fallen out.

[0004] To treat TM perforation, the fibrous tissue around the perforation is abraded to induce an inflammatory response. When the perforation is small, such as less than about 20% of the cross - sectional area of the TM, the perforation can also be filled with paper or fat taken from the earlobe. When the perforation is large, more powerful packing materials such as autologous grafts of cartilage or fascia, surgical gelatin, or synthetic polymer patches are required. All of these treatment methods require some kind of surgery, and the success rate of treatment depends greatly on the surgeon's skill.

[0005] Previously, photo-polymerizable hydrogels have not been used as scaffolds for otology that apply semi-gel polymers to desired sites before curing. Using a scaffold that is initially liquid and forms a scaffold only when exposed to specific conditions is completely different from current and conventional treatment methods for repairing TM perforations.

Summary of the Invention

[0006] The present disclosure provides a pre-freeze-drying composition, which (i) a base 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) a diluent.

[0007] The present disclosure provides a pre-freeze-drying composition, which (i) a base 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) a diluent.

[0008] In an embodiment, the photoinitiator is riboflavin.

[0009] In an embodiment, the composition further comprises about 0 wt% to about 12 wt% of arginine. In an embodiment, the composition further comprises about 0 wt% to about 20 wt% of arginine.

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

[0011] The present disclosure provides a freeze-dried composition prepared by freeze-drying any of the compositions of the present invention, the freeze-dried composition containing about 0% to about 2% water, and the freeze-dried composition being reconstitutable 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 containing water until homogeneous, with a mixing time of less than 60 seconds, the viscosity of the semi-gel composition being from about 0.01 kPa·s to about 1.0 kPa·s as measured by rheometry, and the composition containing from about 80% to about 99% by weight of water.

[0013] The present disclosure provides a curable composition produced by light irradiation of the semi-gel composition of the present invention, the elastic modulus of the curable composition being from about 0.05 kPa to about 20 kPa as measured by rheometry.

[0014] In an embodiment, the light irradiation includes irradiating the 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 irradiation intensity of about 100 mW / cm 2 ~ about 1000 mW / cm 2 In an embodiment, the light irradiation includes irradiating the 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 irradiation intensity of about 100 mW / cm 2 ~ about 100000 mW / cm 2 In an embodiment, the light irradiation includes irradiating the 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 irradiation intensity of about 100 mW / cm

[0015] The present disclosure provides a kit including a lyophilized composition, a gel dispensing tray, and a light guide.

[0016] The present disclosure provides a method for treating a tympanic membrane perforation, the method comprising (a) applying the semi-gel composition of the present disclosure to the tympanic membrane of a patient in need of treatment; (b) inserting a light guide into the external auditory canal; (c) curing the composition by light irradiation.

[0017] In an embodiment, the light irradiation includes irradiating the 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 irradiation intensity of about 100 mW / cm 2 ~ about 1000 mW / cm 2irradiating the composition with a light irradiation intensity. In an embodiment, the light irradiation has a wavelength of about 200 nm to about 800 nm, a curing time of about 1 second to about 300 seconds, and a light irradiation intensity of about 100 mW / cm 2 ~ about 100,000 mW / cm 2 irradiating the composition.

[0018] In an embodiment, the present disclosure provides a method for treating a tympanic membrane perforation in a patient in need of treatment, the method comprising (a) mixing a solvent (such as physiological saline) and the lyophilized composition of the present disclosure to obtain a semi-gel composition; (b) administering the semi-gel composition to the tympanic membrane perforation site of the patient; (c) curing the semi-gel composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0019]

Fig. 1A

Fig. 1B

Fig. 1C

Fig. 1D

[0020] Definitions Terms or expressions not explicitly defined in this specification should be understood to have definitions generally accepted by those skilled in the art. The following description relates to specific embodiments or specific applications of the present invention, but it is for illustrative purposes only and does not limit the claimed invention. The following description is intended to cover all alternatives, modifications, and equivalents included within the spirit and scope of the present invention as defined in the appended claims.

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

[0022] The term "semi - gel composition" refers to a composition (including at least one polymer) that can be activated to assemble into a more viscous form but has not yet been assembled. In embodiments, the semi - gel composition is viscous enough to be placed to cover a perforation without falling into the middle ear cavity. In embodiments, the semi - gel composition can be prepared from a dehydrated composition such as a lyophilized composition.

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

[0024] The term "treatment" with respect to a patient refers to improving at least one symptom of the patient's disorder. Treatment can be improving the disorder or at least partially improving it.

[0025] Composition The present disclosure provides, upon hardening, a semi-gel composition that is useful, inter alia, for treating tympanic membrane rupture in patients in need of treatment, as well as a composition useful as an intermediate in the manufacture of the semi-gel composition. Accordingly, the present disclosure provides a pre-lyophilization composition, a lyophilized composition, a semi-gel composition, and a sclerosing composition.

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

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

[0028] The composition according to the present disclosure is a semi - gel composition for initially administering to a desired site of a patient's ear and is configured to harden into a polymer matrix in the patient's ear when activated. Thus, the semi - gel composition comprises one or more polymers that can be selectively assembled into a polymer matrix at a desired time under specific conditions (such as those found in the ear, e.g., the middle ear), and a diluent. In embodiments, the semi - gel composition comprises an initiator (e.g., a photo - initiator), and the semi - gel composition is hardened by exposing the composition to a condition (e.g., light), thereby activating the initiator and obtaining a polymer matrix. In embodiments, a change in light density, pH, temperature, or ionic concentration initiates the spontaneous assembly of the polymer matrix. In embodiments, the polymer is functionalized to confer cross - linking ability (e.g., methacrylation) upon activation of an initiator (e.g., riboflavin).

[0029] In embodiments, the semi - gel composition is designed to harden into a polymer matrix or scaffold having porosity and an average pore size suitable for loading a 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 for the loaded therapeutic agent.

[0030] In embodiments, the polymer matrix is a scaffold having sufficient rigidity and / or adhesiveness (e.g., as a result of functional groups providing cell - adhesion domains) such that epithelial cells within the TM can migrate onto and / or adhere and / or grow on the scaffold. In embodiments, the polymer matrix or scaffold is ultimately degraded in situ, i.e., biodegradable, for example, after the epithelial cells have grown sufficiently to repair the perforation.

[0031] Composition before lyophilization The present disclosure provides a composition before lyophilization comprising (i) a base comprising methacrylated chitosan, hyaluronic acid, and a photo - initiator, and (ii) a diluent. In embodiments, the diluent comprises water.

[0032] In an embodiment, the pre - lyophilization composition provides a lyophilized composition with storage stability by lyophilization (or similar techniques such as spray drying).

[0033] In an embodiment, the present disclosure provides a pre - lyophilization composition, which (i) a base containing about 40 wt% to about 80 wt% methacrylic acid, about 20 wt% to about 40 wt% hyaluronic acid, about 0.01 wt% to about 0.1 wt% photoinitiator, and 0 wt% to about 20 wt% arginine, and (ii) a diluent.

[0034] In an embodiment, the present disclosure provides a pre - lyophilization composition, which (i) a base containing about 40 wt% to about 80 wt% methacrylated chitosan, about 20 wt% to about 40 wt% hyaluronic acid, about 0.01 wt% to about 0.5 wt% photoinitiator, and 0 wt% to about 20 wt% arginine, and (ii) a diluent.

[0035] In an embodiment, the base contains about 30 wt% to about 90 wt% methacrylated chitosan, which is, for example, 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 an embodiment, the composition contains about 40 wt% to about 80 wt% methacrylated chitosan. In an embodiment, the composition contains about 60 wt% methacrylated chitosan.

[0036] In embodiments, the methacrylated chitosan is methacrylated at about 10% to about 60%, which can be, 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 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, or about 60% (including all ranges and values therebetween). In embodiments, the methacrylated chitosan is methacrylated at about 21% to about 32%. In embodiments, the methacrylated chitosan is methacrylated at about 21% to about 34%. In embodiments, the methacrylated chitosan is methacrylated at about 21% to about 36%. In embodiments, the methacrylated chitosan is methacrylated at about 25% to about 40%. In embodiments, the methacrylated chitosan is methacrylated at about 30%.

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

[0038] In embodiments, the base comprises about 10 wt% to about 50 wt% hyaluronic acid, which can be, for example, about 10 wt%, about 12 wt%, about 14 wt%, about 16 wt%, about 18 wt%, about 20 wt%, about 22 wt%, about 24 wt%, about 26 wt%, about 28 wt%, 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%, or about 50 wt% (including all ranges and values therebetween). In embodiments, the composition comprises about 20 wt% to about 40 wt% hyaluronic acid. In embodiments, the composition comprises about 30 wt% hyaluronic acid.

[0039] In an embodiment, the molecular weight of hyaluronic acid can be in the range of about 100 g / mol to about 1,000,000 g / mol.

[0040] "Photoinitiator" usually includes substances that form free radicals when irradiated with light of an appropriate wavelength. In an embodiment, the photoinitiator is an aromatic carbonyl compound (e.g., benzoin derivative, benzil ketal, acetophenone derivative, hydroxyalkylphenone) or an aromatic ketone (e.g., benzophenone, 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-propan-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetophenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, floxim, thionin, riboflavin, and methyl green. Further other photoinitiators are 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-trimethylbenzoyl diphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 2 / 4-diethylthioxanthone, 2 / 4-isopropylthioxanthone, 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholinyl)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminoethyl benzoate, 2-ethylhexyl-4-dimethylaminobenzoate, 2-hydroxy-2-methyl-phenyl-propan-1-one, 2-hydroxy-4'-hydroxyethoxy-2-methylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropanone-1,4-(4-methylphenylthiophenyl)-phenylmethanone, 4,4'-difluorobenzophenone, 4,4'-Dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzyldimethylketal, 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, phenyltribromomethylsulfone, acylphosphine oxide (APO) and bisacylphosphine oxide (BAPO), 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propan-1-one, 2,2-dimethoxy-1,2-diphenylethane-1-one, hydroxy-cyclohexyl-phenyl-ketone, methyl benzoylformate, oxy-phenyl-acetic acid 2-[2-oxo-2-phenyl-acetoxy-ethoxy]ethyl ester, oxy-phenyl-acetic acid 2-[2-hydroxy-ethoxy]-ethyl ester, α-dimethoxy-α-phenylacetophenone, 2-benzyl-2-(dimethylamino)-1-[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 derivatives thereof.,

[0041] In an embodiment, the photoinitiator is riboflavin. In an embodiment, riboflavin is sodium riboflavin 5'-phosphate salt.

[0042] In embodiments, the base comprises from about 0.01 wt% to about 0.10 wt% photoinitiator. In embodiments, the base comprises from about 0.01 wt% to about 0.5 wt% photoinitiator, which can be, 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 embodiments, the composition comprises from about 0.08 wt% to about 0.016 wt% photoinitiator. In embodiments, the composition comprises from about 0.08 wt% to about 0.40 wt% photoinitiator. In embodiments, the composition comprises about 0.08 wt% photoinitiator. In embodiments, the composition comprises about 0.50 wt% photoinitiator. In embodiments, the composition comprises about 0.12 wt% photoinitiator. In embodiments, the composition comprises about 0.40 wt% photoinitiator.

[0043] Applicants have discovered that adding arginine to the compositions of the present disclosure improves gel formation and the distribution of other reagents within the gel. In embodiments, arginine functions as a co-initiator for crosslinking.

[0044] In an embodiment, the base contains arginine. In an embodiment, the arginine is L-arginine. In an embodiment, the base contains from about 0 wt% to about 20 wt% arginine, which can be, for example, about 0 wt%, 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%, or about 20 wt% (including all ranges and values therebetween). In an embodiment, the base contains from about 6 wt% to about 12 wt% arginine. In an embodiment, the base contains about 6 wt% arginine. In an embodiment, the base contains about 9 wt% arginine.

[0045] In an embodiment, the base contains an amine catalyst such as, for example, ethyl 4-dimethylaminobenzoate (EDMAB).

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

[0047] In an embodiment, the diluent is from about 0 wt% to about 99 wt% of the composition. In an embodiment, the diluent is from about 1 wt% to about 99 wt% of the composition, such as 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 24 wt%, about 25 wt%, about 26 wt%, about 27 wt%, about 28 wt%, about 29 wt%, 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 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 diluent is from about 0 wt% to about 95 wt% of the composition. In an embodiment, the diluent is from about 1 wt% to about 95 wt% of the composition. In an embodiment, the diluent is about 97 wt% of the composition. In an embodiment, the diluent is about 1 wt% of the composition. In an embodiment, the diluent is about 2 wt% of the composition. In an embodiment, the diluent contains water. In an embodiment, the diluent contains water and sodium chloride. In an embodiment, the diluent contains water and a phosphate buffer.In an embodiment, the diluent includes water, sodium chloride, and a phosphate buffer. In an embodiment, the diluent is a 0.9% aqueous sodium chloride solution (i.e., physiological saline).

[0048] Lyophilized composition The pre-lyophilization composition of the present disclosure can be lyophilized to produce a lyophilized composition. Without being limited by theory, lyophilization extends the storage period of the composition of the present disclosure. In an embodiment, the lyophilization of the composition provides a storage period exceeding one year. In an embodiment, the lyophilization of the composition provides a storage period of about one year.

[0049] In an embodiment, the lyophilized composition is porous, so the reconstitution time is shortened. In an embodiment, the porosity is introduced by lyophilization, gas treatment, or introduction of ice particles during freezing. In an embodiment, the lyophilization parameters are important to prevent the material from collapsing during the lyophilization process. Insufficient drying causes the gel to collapse and a dense and hard substance to be produced, making reconstitution very difficult and the viscosity of the gel very poor.

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

[0051] In embodiments, the lyophilized composition contains from about 0 wt% to about 10 wt% water, which can be, for example, about 0 wt%, about 0.5 wt%, about 1.0 wt%, about 1.5 wt%, about 2.0 wt%, about 2.5 wt%, about 3.0 wt%, about 3.5 wt%, about 4.0 wt%, about 4.5 wt%, about 5.0 wt%, about 5.5 wt%, about 6.0 wt%, about 6.5 wt%, about 7.0 wt%, about 7.5 wt%, about 8.0 wt%, about 8.5 wt%, about 9.0 wt%, about 9.5 wt%, or about 10.0 wt% (including all ranges and values therebetween). In embodiments, the lyophilized composition contains from about 0 wt% to about 2 wt% water. In embodiments, the lyophilized composition contains about 0 wt% water. In embodiments, the lyophilized composition contains about 1 wt% water. In embodiments, the lyophilized composition contains about 2 wt% water.

[0052] In an embodiment, the lyophilized composition comprises from about 50 wt% to about 65 wt% of methacrylated chitosan. In an embodiment, the lyophilized composition comprises from about 30 wt% to about 90 wt% of methacrylated chitosan, which can be, 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 wt%, about 85 wt%, about 86 wt%, about 87 wt%, about 88 wt%, about 89 wt%, or about 90 wt% (including all ranges and values therebetween). In an embodiment, the lyophilized composition comprises about 59 wt% of methacrylated chitosan. In an embodiment, the lyophilized composition comprises about 60 wt% of methacrylated chitosan. In an embodiment, the lyophilized composition comprises about 61 wt% of methacrylated chitosan. In an embodiment, the lyophilized composition comprises from about 40 wt% to about 80 wt% of methacrylated chitosan. In an embodiment, the lyophilized composition comprises from about 50 wt% to about 65 wt% of methacrylated chitosan.

[0053] In an embodiment, the lyophilized composition comprises from about 25 wt% to about 35 wt% hyaluronic acid. In an embodiment, the lyophilized composition comprises from about 10 wt% to about 50 wt% hyaluronic acid, which can be, for example, 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 24 wt%, about 25 wt%, about 26 wt%, about 27 wt%, about 28 wt%, about 29 wt%, 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%, or about 50 wt% (including all ranges and values therebetween). In an embodiment, the lyophilized composition comprises about 29 wt% hyaluronic acid. In an embodiment, the lyophilized composition comprises about 30 wt% hyaluronic acid. In an embodiment, the lyophilized composition comprises about 31 wt% hyaluronic acid. In an embodiment, the lyophilized composition comprises from about 20 wt% to about 40 wt% hyaluronic acid. In an embodiment, the lyophilized composition comprises from about 25 wt% to about 35 wt% hyaluronic acid.

[0054] In an embodiment, the photoinitiator is an aromatic carbonyl compound (e.g., benzoin derivatives, benzil ketals, acetophenone derivatives, hydroxyalkylphenones) or an aromatic ketone (e.g., benzophenone, 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-propan-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetone phenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, floxim, thionin, riboflavin, and methyl green. Further other photoinitiators are 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-trimethylbenzoyl diphenylphosphine oxide, 2,4,6-trimethylbenzophenone, 2 / 4-diethylthioxanthone, 2 / 4-isopropylthioxanthone, 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholinyl)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminoethyl benzoate, 2-ethylhexyl-4-dimethylaminobenzoate, 2-hydroxy-2-methyl-phenyl-propan-1-one, 2-hydroxy-4'-hydroxyethoxy-2-methylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropanone-1,4-(4-methylphenylthiophenyl)-phenylmethanone, 4,4'-difluorobenzophenone, 4,4'-Dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzyl 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, phenyl tribromomethyl sulfone, acylphosphine oxide (APO) and bisacylphosphine oxide (BAPO), 1-[4-(2-hydroxyethoxy)phenyl]-2-hydroxy-2-methyl-1-propan-1-one, 2,2-dimethoxy-1,2-diphenylethane-1-one, hydroxy-cyclohexyl-phenyl-ketone, methyl benzoylformate, oxy-phenyl-acetic acid 2-[2-oxo-2-phenyl-acetoxy-ethoxy]ethyl ester, oxy-phenyl-acetic acid 2-[2-hydroxy-ethoxy]-ethyl ester, α-dimethoxy-α-phenylacetophenone, 2-benzyl-2-(dimethylamino)-1-[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 a derivative thereof.,

[0055] In an embodiment, the photoinitiator is riboflavin. In an embodiment, the riboflavin is sodium riboflavin 5'-phosphate. In an embodiment, the lyophilized composition contains from about 0.10 wt% to about 0.15 wt% riboflavin. In an embodiment, the lyophilized composition contains from about 0.10 wt% to about 0.7 wt% riboflavin, which can be, 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%, 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%, 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%, about 0.50 wt%, about 0.51 wt%, 0.52 wt%, about 0.53 wt%, about 0.54 wt%, about 0.55 wt%, about 0.56 wt%, about 0.57 wt%, about 0.58 wt%, about 0.59 wt%, about 0.60 wt%, about 0.61 wt%, 0.62 wt%, about 0.63 wt%, about 0.64 wt%, about 0.65 wt%, about 0.66 wt%, about 0.67 wt%, about 0.68 wt%, about 0.69 wt%, or about 0.7 wt% (including all ranges and values therebetween). In an embodiment, the riboflavin is sodium riboflavin 5'-phosphate. In an embodiment, the lyophilized product contains about 0.11 wt% riboflavin. In an embodiment, the lyophilized composition contains about 0.12 wt% riboflavin. In an embodiment, the lyophilized composition contains about 0.13 wt% riboflavin. In an embodiment, the lyophilized composition contains from about 0.01 wt% to about 0.5 wt% riboflavin. In an embodiment, the lyophilized composition contains from about 0.01 wt% to about 0.15 wt% riboflavin.

[0056] In embodiments, the lyophilized composition contains from about 0 wt% to about 10 wt% arginine. In embodiments, the lyophilized composition contains from about 0 wt% to about 30 wt% arginine, which can be, for example, about 0 wt%, 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 24 wt%, about 25 wt%, about 26 wt%, about 27 wt%, about 28 wt%, about 29 wt%, or about 30 wt% (including all ranges and values therebetween). In embodiments, the lyophilized composition contains about 8 wt% arginine. In embodiments, the arginine is L-arginine. In embodiments, the lyophilized composition contains about 9 wt% arginine. In embodiments, the lyophilized composition contains about 10 wt% arginine. In embodiments, the lyophilized composition contains from about 0 wt% to about 10 wt% arginine. In embodiments, the lyophilized composition contains from about 0 wt% to about 20 wt% arginine. In embodiments, the arginine is L-arginine.

[0057] In embodiments, the composition is a lyophilized composition prepared by lyophilizing any one of the pre-lyophilization compositions of the present disclosure, contains from about 0 wt% to about 2 wt% water, and the lyophilized composition can be reconstituted in less than 60 seconds to provide the semi-gel composition of the present disclosure.

[0058] In embodiments, the lyophilized composition contains 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'-sodium phosphate salt.

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

[0060] In an embodiment, sterilization partially cures the composition. In an embodiment, sterilization is necessary to fully cure the composition. In an embodiment, ionizing radiation partially cures the gel. In an embodiment, X-ray radiation partially cures the gel. In an embodiment, the lyophilized composition is sterilized, and the sterilization is achieved by ionizing radiation. In an embodiment, the ionizing radiation is gamma rays. In an embodiment, gamma irradiation partially cures the composition.

[0061] In an embodiment, the dose of ionizing radiation is from 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, 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 an embodiment, the dose of ionizing radiation is about 25 kGy.

[0062] In an embodiment, after sterilization, the log reduction of colony forming units is from about 2 to about 4, such as 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 an embodiment, after sterilization, the log reduction of colony forming units is from about 3.4 to about 3.6. In an embodiment, after sterilization, the log reduction of colony forming units is about 3.4.

[0063] Semi-gel composition In an embodiment, the lyophilized composition of the present disclosure is reconstituted to provide a semi-gel composition. In an embodiment, 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, such as 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, or about 120 seconds (including all values and ranges therebetween). In an embodiment, reconstitution takes about 30 seconds. In an embodiment, reconstitution takes less than about 60 seconds. In an embodiment, reconstitution takes less than about 90 seconds.

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

[0066] In an embodiment, the final concentration of the diluent in the semi-gel composition is from about 80 wt% to about 99 wt%, for example, 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 contains water. In an embodiment, the diluent contains water and sodium chloride. In an embodiment, the diluent contains water and a phosphate buffer. In an embodiment, the diluent contains water, sodium chloride and a phosphate buffer. In an embodiment, the diluent is a 0.9% aqueous sodium chloride solution (i.e., physiological saline).

[0067] In an embodiment, the semi-gel composition is prepared by mixing any one of the compositions of the present disclosure with a diluent until homogeneous, the final concentration of the diluent is about 97 wt%, the mixing time is less than about 60 seconds, and 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 contains from about 80 wt% to about 99 wt% water, which is, for example, 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 contains from about 90 wt% to about 97 wt% water. In an embodiment, the semi-gel composition contains about 97 wt% water.

[0069] In an embodiment, the semi-gel composition comprises from about 1 wt% to about 3 wt% of methacrylated chitosan, which can be, for example, 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 an embodiment, the semi-gel composition comprises about 2 wt% of methacrylated chitosan. In an embodiment, the methacrylated chitosan is about 25% to about 40% methacrylated.

[0070] In an embodiment, the semi-gel composition comprises from about 0.5 wt% to about 2 wt% of hyaluronic acid, which can be, for example, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, 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%, or about 2 wt% (including all ranges and values therebetween). In an embodiment, the semi-gel composition comprises about 1 wt% of hyaluronic acid.

[0071] In the embodiment, the photoinitiator is an aromatic carbonyl compound (for example, benzoin derivatives, benzil ketals, acetophenone derivatives, hydroxyalkylphenones) or an aromatic ketone (for example, benzophenone, thioxanthone). In the 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-propan-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetonephenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, floxim, thionin, riboflavin, and methyl green. Further other photoinitiators are 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-morpholinyl)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminoethyl benzoate, 2-ethylhexyl-4-dimethylaminobenzoate, 2-hydroxy-2-methyl-phenyl-propan-1-one, 2-hydroxy-4'-hydroxyethoxy-2-methylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropanone-1,4-(4-methylphenylthiophenyl)-phenylmethanone, 4,4'-difluorobenzophenone, 4,4'-Dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzyl 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-propan-1-one, 2,2-dimethoxy-1,2-diphenylethane-1-one, hydroxy-cyclohexyl-phenyl-ketone, methyl benzoylformate, oxy-phenyl-acetic acid 2-[2-oxo-2-phenyl-acetoxy-ethoxy]ethyl ester, oxy-phenyl-acetic acid 2-[2-hydroxy-ethoxy]-ethyl ester, α-dimethoxy-α-phenylacetophenone, 2-benzyl-2-(dimethylamino)-1-[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 a derivative thereof.,

[0072] In an embodiment, the photoinitiator is riboflavin. In an embodiment, riboflavin is sodium riboflavin 5'-phosphate. In an embodiment, the semi-gel composition contains from about 0.001 wt% to about 0.01 wt% riboflavin, which is, for example, 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 an embodiment, the semi-gel composition contains about 0.004 wt% riboflavin. In an embodiment, riboflavin is sodium riboflavin 5'-phosphate.

[0073] In an embodiment, the semi-gel composition contains from about 0 wt% to about 0.5 wt% arginine, which is, for example, 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 contains about 0.3 wt% arginine. In an embodiment, arginine is L-arginine.

[0074] In an embodiment, the semi-gel composition contains 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% sodium riboflavin 5'-phosphate.

[0075] In an embodiment, when the viscosity of the semi-gel composition is measured by rheometry, it is from about 0.01 kPa·s to about 1 kPa·s, for example, 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, when the viscosity of the semi-gel composition is measured by rheometry, it is from about 0.01 kPa·s to about 1.0 kPa·s. In an embodiment, when the viscosity of the semi-gel composition is measured by rheometry, it is from about 0.01 kPa·s to about 0.3 kPa·s. In an embodiment, when the viscosity of the semi-gel composition is measured by rheometry, it is from about 0.5 kPa·s to about 1.0 kPa·s.

[0076] In an embodiment, the semi-gel composition is sterilized, and 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 ray.

[0077] In an embodiment, the sterilization partially cures the composition. In an embodiment, sterilization is necessary to fully cure the composition. In an embodiment, the ionizing radiation partially cures the gel. In an embodiment, the X-ray radiation partially cures the gel. In an embodiment, the gamma ray irradiation partially cures the composition.

[0078] In an embodiment, 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, 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 an embodiment, the dose of ionizing radiation is about 25 kGy.

[0079] In an embodiment, after sterilization, the log reduction of colony forming units is about 2 to about 4, for example, 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 an embodiment, after sterilization, the log reduction of colony forming units is about 3.4 to about 3.6. In an embodiment, after sterilization, the log reduction of colony forming units is about 3.4.

[0080] In embodiments, the semi-gel adheres to epithelial and mucosal tissues with a strength in the range of about 1 kPa to about 20 kPa when measured by a lap shear test. In embodiments, the semi-gel adheres to epithelial and mucosal tissues with a strength in the range of about 0.1 kPa to about 20 kPa when measured by a lap shear test, which is, for example, 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.

[0082] Sclerosing composition The compositions according to the present disclosure include a sclerosing composition that is a composition that hardens into a polymer matrix. In embodiments, the sclerosing composition has sufficient rigidity and / or adhesiveness (e.g., as a result of functional groups that provide cell adhesion domains) such that epithelial cells within the TM can move onto and / or on the scaffold and adhere and / or grow. In embodiments, the compositions of the present disclosure are sclerosing compositions produced by light irradiation. In embodiments, the sclerosing composition is prepared from light irradiation of a semi-gel composition.

[0083] In an embodiment, when the viscosity or rigidity of the composition of the present disclosure is measured by rheometry, it is represented by the elastic modulus. In an embodiment, 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 oscillatory shear force to the composition.

[0084] In an embodiment, when the elastic modulus of the curable composition is measured by rheometry, it is about 0.05 kPa to about 100 kPa, for example, 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 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 9.5 kPa, about 10 kPa, about 11 kPa, about 12 kPa, about 13 kPa, about 14 kPa, about 15 kPa, about 16 kPa, about 17 kPa, about 18 kPa, about 19 kPa, about 20 kPa, about 21 kPa, about 22 kPa, about 23 kPa, about 24 kPa, about 25 kPa, about 26 kPa, about 27 kPa, about 28 kPa, about 29 kPa, about 30 kPa, about 31 kPa, about 32 kPa, about 33 kPa, about 34 kPa, about 35 kPa, about 36 kPa, about 37 kPa, about 38 kPa, about 39 kPa, about 40 kPa, about 41 kPa, about 42 kPa, about 43 kPa, about 44 kPa, about 45 kPa, about 46 kPa, about 47 kPa, about 48 kPa, about 49 kPa, about 50 kPa, about 51 kPa, about 52 kPa, about 53 kPa, about 54 kPa, about 55 kPa, about 56 kPa, about 57 kPa, about 58 kPa, about 59 kPa, about 60 kPa, about 61 kPa, about 62 kPa, about 63 kPa, about 64 kPa, about 65 kPa, about 66 kPa, about 67 kPa, about 68 kPa, about 69 kPa, about 70 kPa, about 71 kPa, about 72 kPa, about 73 kPa, about 74 kPa, about 75 kPa, about 76 kPa, about 77 kPa, about 78 kPa, about 79 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 an embodiment, the elastic modulus of the curable composition is about 0.4 kPa to about 3 kPa when measured by rheometry.

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

[0086] In an embodiment, the curable composition includes about 80 wt% to about 99 wt% diluent, which is, for example, 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 includes about 90 wt% to about 97 wt% water. In an embodiment, the semi-gel composition includes about 97 wt% water. In an embodiment, the diluent includes water. In an embodiment, the diluent includes water and sodium chloride. In an embodiment, the diluent includes water and a phosphate buffer. In an embodiment, the diluent includes water, sodium chloride, and a phosphate buffer. In an embodiment, the diluent is a 0.9% aqueous sodium chloride solution (i.e., physiological saline).

[0087] In an embodiment, the curable composition comprises from about 1 wt% to about 3 wt% of methacrylated chitosan, which is, for example, 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 an embodiment, the semi-gel composition comprises about 2 wt% of methacrylated chitosan. In an embodiment, the methacrylated chitosan is about 25% to about 40% methacrylated.

[0088] In an embodiment, the curable composition comprises from about 0.5 wt% to about 2 wt% of hyaluronic acid, which is, for example, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, 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%, or about 2 wt% (including all ranges and values therebetween). In an embodiment, the semi-gel composition comprises about 1 wt% of hyaluronic acid.

[0089] In the embodiment, the photoinitiator is an aromatic carbonyl compound (e.g., benzoin derivative, benzil ketal, acetophenone derivative, hydroxyalkylphenone) or an aromatic ketone (e.g., benzophenone, thioxanthone). In the 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-propan-1-one, ethyl eosin, eosin Y, fluorescein, 2,2-dimethoxy-2-phenylacetophenone, 2-methyl-2-phenylacetonephenone, 12959, camphorquinone, rose bengal, methylene blue, erythrosine, floxim, thionin, riboflavin, and methyl green. Further other photoinitiators are 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-morpholinyl)phenyl]-1-butanone, 2-chlorothioxanthone, 2-dimethyl-aminoethyl benzoate, 2-ethylhexyl-4-dimethylaminobenzoate, 2-hydroxy-2-methyl-phenyl-propan-1-one, 2-hydroxy-4'-hydroxyethoxy-2-methylpropiophenone, 2-isopropylthioxanthone, 2-methylbenzophenone, 2-methyl-1-[4-(methylthio)phenyl]-2-morpholinopropanone-1,4-(4-methylphenylthiophenyl)-phenylmethanone, 4,4'-difluorobenzophenone, 4,4'-Dimethoxybenzophenone, 4-chlorobenzophenone, 4-methylacetophenone, 4-methylbenzophenone, 4-phenylbenzophenone, benzyl 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-propan-1-one, 2,2-dimethoxy-1,2-diphenylethane-1-one, hydroxy-cyclo-hexyl-phenyl-ketone, methyl benzoylformate, oxy-phenyl-acetic acid 2-[2-oxo-2-phenyl-acetoxy-ethoxy]ethyl ester, oxy-phenyl-acetic acid 2-[2-hydroxy-ethoxy]-ethyl ester, α-dimethoxy-α-phenylacetophenone, 2-benzyl-2-(dimethylamino)-1-[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 a derivative thereof.,

[0090] In an embodiment, the photoinitiator is riboflavin. In an embodiment, riboflavin is sodium riboflavin 5'-phosphate. In an embodiment, the curable composition contains from about 0.001 wt% to about 0.01 wt% riboflavin, which can be, for example, 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 an embodiment, the curable composition contains about 0.004 wt% riboflavin. In an embodiment, riboflavin is sodium riboflavin 5'-phosphate.

[0091] In an embodiment, the curable composition contains from about 0 wt% to about 0.5 wt% arginine, which can be, for example, 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 contains about 0.3 wt% arginine. In an embodiment, arginine is L-arginine.

[0092] In an embodiment, the curable composition contains 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% sodium riboflavin 5'-phosphate.

[0093] The kit of the present disclosure In an embodiment, the present disclosure provides a kit including the composition of the present disclosure (e.g., 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, after the tray is sealed, the kit is sterilized. 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 ray.

[0095] In an embodiment, the dose of ionizing radiation is from 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, 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 an embodiment, the dose of ionizing radiation is about 25 kGy.

[0096] In an embodiment, after sterilization, the log reduction of colony forming units is from about 2 to about 4, such as 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 an embodiment, after sterilization, the log reduction of colony forming units is from about 3.4 to about 3.6. In an embodiment, after sterilization, the log reduction of 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 pouch. In an embodiment, a lyophilized composition or a semi-gel composition is contained within a pouch. In an embodiment, the pouch includes a moisture-proof layer. In an embodiment, the pouch includes a transparent portion. In an embodiment, the pouch includes a moisture-proof layer and a transparent portion. In an embodiment, the pouch includes a Luer lock connector. In an embodiment, the pouch is about 9 cm 2 to about 30 cm 2 in area, which can be, for example, about 9 cm 2 , about 10 cm 2 , about 11 cm 2 , about 12 cm 2 , about 13 cm 2 , about 14 cm 2 , about 15 cm 2 , about 16 cm 2 , about 17 cm 2 , about 18 cm 2 , about 19 cm 2 , about 20 cm 2 , about 21 cm 2 , about 22 cm 2 , about 23 cm 2 , about 24 cm 2 , about 25 cm 2 , about 26 cm 2 , about 27 cm 2 , about 28 cm 2 , about 29 cm 2 , or about 30 cm 2 (including all ranges and values therebetween).

[0098] In an embodiment, the pouch contains from about 0.2 mL to about 10 mL of the composition, which can be, for example, 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 embodiments, the composition is contained within a mixing vessel (e.g., a syringe, pouch, vial, or tube). In embodiments, the composition is contained within a mixing syringe.

[0100] In embodiments, the storage period of the kit exceeds about 1 year. In embodiments, the storage period of the kit is about 1 year.

[0101] In embodiments, the present disclosure provides a kit comprising a lyophilized composition, a gel dispensing tray, and a light guide. In embodiments, the lyophilized composition is contained within a pouch. In embodiments, the kit is enclosed in a sealed tray.

[0102] The methods of the present disclosure The present disclosure provides compositions and methods for use in the treatment of tympanic membrane perforations. In embodiments, the perforation is a chronic perforation of the tympanic membrane. In embodiments, the otological material comprises a semi-gel composition that cures in situ to form a polymeric matrix (e.g., a hydrogel scaffold). In embodiments, the method comprises administering the semi-gel to a desired site of the patient's ear and curing the material in situ. In embodiments, the method comprises administering the semi-gel composition to an otological packing material and curing the semi-gel composition after administration to the packing material.

[0103] In embodiments, the present disclosure provides a method of treating a tympanic membrane perforation, which comprises (i) applying the semi-gel composition of the present disclosure to the tympanic membrane, (ii) inserting a light guide into the external auditory canal, (iii) curing the composition by light irradiation.

[0104] In embodiments, the light irradiation comprises irradiating the 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 irradiation intensity of about 100 mW / cm 2 ~ about 100,000 mW / cm 2 of the composition.

[0105] In an embodiment, a composition of about 50 μL to about 500 μL is applied to the eardrum. In an embodiment, a composition of about 10 μL to about 500 μL is applied to the eardrum, which can be, 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 an embodiment, a composition of about 200 μL is applied to the eardrum.

[0106] In an embodiment, the light irradiation includes 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 light irradiation intensity of about 100 mW / cm 2 ~ about 1000 mW / cm 2 In an embodiment, the light irradiation includes 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 light irradiation intensity of about 100 mW / cm 2 ~ about 10000 mW / cm 2 In an embodiment, the light irradiation includes 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 light irradiation intensity of about 100 mW / cm 2 ~ about 100000 mW / cm 2 In an embodiment, the light irradiation includes irradiating the composition (e.g., 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 light irradiation power of about 2 mW to about 1 W. In an embodiment, the light irradiation includes irradiating the composition (e.g., 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 light irradiation 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 340 nm, about 345 nm, about 350 nm, about 355 nm, about 360 nm, about 365 nm, about 370 nm, about 375 nm, about 380 nm, about 385 nm, about 390 nm, about 395 nm, about 400 nm, about 405 nm, about 410 nm, about 415 nm, about 420 nm, about 425 nm, about 430 nm, about 435 nm, about 440 nm, about 445 nm, about 450 nm, about 455 nm, about 460 nm, about 465 nm, about 470 nm, about 475 nm, about 480 nm, about 485 nm, about 490 nm, about 495 nm, about 500 nm, about 505 nm, about 510 nm, about 515 nm, about 520 nm, about 525 nm, about 530 nm, about 535 nm, about 540 nm, about 545 nm, about 550 nm, about 555 nm, about 560 nm, about 565 nm, about 570 nm, about 575 nm, about 580 nm, about 585 nm, about 590 nm, about 595 nm, about 600 nm, about 605 nm, about 610 nm, about 615 nm, about 620 nm, about 625 nm, about 630 nm, about 635 nm, about 640 nm, about 645 nm, about 650 nm, about 655 nm, about 660 nm, about 665 nm, about 670 nm, about 675 nm, about 680 nm, about 685 nm, about 690 nm, about 695 nm, about 700 nm, about 705 nm, about 710 nm, about 715 nm, about 720 nm, about 725 nm, about 730 nm, about 735 nm, about 740 nm, about 745 nm, about 750 nm, about 755 nm, about 760 nm, about 765 nm, about 770 nm, about 775 nm, about 780 nm, about 785 nm, about 790 nm, about 795 nm, or about 800 nm (including all ranges and values therebetween). In an embodiment, the irradiation wavelength is from about 420 nm to about 530 nm. In an embodiment, the irradiation wavelength is about 450 nm.

[0108] In an embodiment, the light irradiation power is from about 2 mW to about 1 W. In an embodiment, the light irradiation power is from 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, about 38 mW, about 40 mW, about 42 mW, about 44 mW, about 46 mW, about 48 mW, about 50 mW, about 60 mW, about 70 mW, about 80 mW, about 90 mW, about 100 mW, about 110 mW, about 120 mW, about 130 mW, about 140 mW, about 150 mW, about 160 mW, about 170 mW, about 180 mW, about 190 mW, about 200 mW, about 300 mW, about 400 mW, about 500 mW, about 600 mW, about 700 mW, about 800 mW, about 900 mW, or about 1000 mW (including all ranges and values therebetween). In an embodiment, the light irradiation power is from about 15 mW to about 50 mW. In an embodiment, the irradiation power is about 25 mW.

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

[0110] In an embodiment, the curing time is from about 1 second to about 300 seconds (sec), 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, about 120 seconds, about 130 seconds, about 140 seconds, about 150 seconds, about 160 seconds, about 170 seconds, about 180 seconds, about 190 seconds, about 200 seconds, 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 an embodiment, the curing time is from about 5 seconds to about 120 seconds. In an embodiment, the curing time is from about 12 seconds to about 300 seconds. In an embodiment, the curing time is 90 seconds.

[0111] In an embodiment, the light irradiation (i.e., the curing time) is continuous. In an embodiment, the light irradiation (i.e., the curing time) is performed as one or more bursts, such as 1 burst, 2 bursts, 3 bursts, 4 bursts or 5 bursts. For example, in an embodiment, the curing time is about 90 seconds and the light irradiation is performed as 3 bursts of about 30 seconds each. In an embodiment, the light irradiation is performed as 1 burst. In an embodiment, the light irradiation is performed as 2 bursts. In an embodiment, the light irradiation is performed as 3 bursts. In an embodiment, the time between bursts is from 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 an embodiment, the time between bursts is about 10 seconds.

[0112] In an embodiment, the present disclosure provides a method for treating a tympanic membrane perforation in a patient in need of treatment, which includes: (a) adding a solvent (such as physiological saline) to any one of the lyophilized compositions of the present disclosure and mixing to form a semi-gel composition; (b) administering the semi-gel composition to the perforated site of the patient's tympanic membrane; and (c) curing the applied semi-gel composition by light irradiation. In an embodiment, the lyophilized composition is mixed with about 0.1 mL to about 2 mL of a solvent (such as physiological saline), which can be, for example, 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 an embodiment, the lyophilized composition is mixed with about 1 mL of physiological saline. In an embodiment, the semi-gel composition is directly applied to the perforated site of the patient's tympanic membrane and cured by any of the light irradiation methods described in the present disclosure. In an embodiment, the curable semi-gel composition provides a scaffold for the migration of endogenous cells and is replaced by newly formed tympanic membrane tissue.

[0113] In an embodiment, the patient in need of treatment is a child. In an embodiment, the patient in need of treatment is an adolescent under 18 years old. In an embodiment, the patient in need of treatment is an adult. In an embodiment, the patient is 5 years old or older, or approximately at least 5 years old or older. In an embodiment, the patient is 5 to 11 years old, such as about 5 years old, about 6 years old, about 7 years old, about 8 years old, about 9 years old, about 10 years old, or about 11 years old (including all ranges and values therebetween).

[0114] In an embodiment, the method of the present disclosure provides a perforation treatment to a patient in need of treatment, and 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% (including all ranges and values therebetween)) of the patient's tympanic membrane is affected by the perforation. In an embodiment, the method of the present disclosure provides a perforation treatment to a patient in need of treatment, and at least about 5% of the patient's tympanic membrane is affected by the perforation. In an embodiment, the method of the present disclosure provides a perforation treatment to a patient in need of treatment, and at least about 10% of the patient's tympanic membrane is affected by the perforation. In an embodiment, at least about 15% of the patient's tympanic membrane is affected by the perforation. In an embodiment, at least about 20% of the patient's tympanic membrane is affected by the perforation. In an embodiment, at least about 25% of the patient's tympanic membrane is affected by the perforation. In an embodiment, at least about 30% of the patient's tympanic membrane is affected by the perforation. In an embodiment, at least about 50% of the patient's tympanic membrane is affected by the perforation. In an embodiment, at least about 80% of the patient's tympanic membrane is affected by the perforation.

[0115] In embodiments, the method of the present disclosure provides a tympanic membrane perforation closure rate of at least about 40%. In embodiments, the method of the present disclosure provides a tympanic membrane perforation closure rate of about ≥50%. In embodiments, the method of the present disclosure provides a tympanic membrane perforation closure rate of about ≥70%. In embodiments, the method of the present disclosure provides a tympanic membrane perforation closure rate of about ≥75%. In embodiments, the method of the present disclosure provides a tympanic membrane perforation closure rate of about >75%. In embodiments, the perforation closure rate is determined by visual evaluation. In embodiments, a microscope or an endoscope is used for visual evaluation.

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

[0117] In embodiments, this method provides a tympanic membrane perforation closure rate or a pure tone average score of >75% compared to the baseline with a single application. In embodiments, this method increases the tympanic membrane perforation closure rate at 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, this treatment increases the tympanic membrane perforation closure rate at about 3 months after administration.

Examples

[0118] Example 1: Sterilization of the lyophilized composition Procedure The kit was sterilized by irradiation at a dose of 25 kGy. The irradiated samples were cultured in a medium at 37 °C for 24 hours and 1 week. Thereafter, the samples were placed in an extraction medium in a biosafety cabinet using sterilized instruments. After extraction, a portion of the extraction medium was placed on a selective agar medium and cultured overnight at 37 °C to identify various microorganisms (both bacteria and fungi) that might be present.

[0119] Results After culturing for 24 hours and 1 week, no microorganisms were seen in any of the selective media used in the tests (Figures 1 and 2). In contrast, hundreds of colonies indicating the presence of bacteria were seen in the non-irradiated samples. This suggested that the irradiation level was sufficient to kill the microorganisms that might have been present before sterilization.

[0120] Conclusions Irradiation seems to be sufficient for sterilization. When compared with the non-irradiated control group, the log reduction between the non-sterilized and sterilized samples was 3.4 - 3.6.

[0121] Example 2: Curing and Characterization of the Semi-Gel Procedure In an office environment, the lyophilized composition was rehydrated with 1 mL of physiological saline and mixed for 20 - 30 seconds to form a semi-gel composition. After preparing the treatment site, the semi-gel composition was applied and the tympanic membrane perforation was covered with a 1-mm overlap. Using an optical guide suitable for curing light, the semi-gel was irradiated for 1.5 minutes.

[0122] Results The flow curve test of the semi-gel was performed on samples reconstituted in pouches with n = 4 at a shear rate of 0.01 s -1 ~10 s -1 at a shear rate of 1 s -1 and viscosity values in the range of 0.01 kPa·s to 0.3 kPa·s were obtained.

[0123] Example 3: Establishment and Testing of an In Vivo Animal Model Procedure The tympanic membranes of adult male chinchillas were perforated with a laser, and perforations of similar size were created. Perforations that remained open for 8 weeks were considered chronic. A semi-gel composition was applied to the tympanic membrane perforations and cured with curing light and a light guide to form a curable composition.

[0124] Results The perforations healed within 3 weeks. Fourteen weeks after treatment with the curable composition, the tympanic membrane and cochlea were evaluated by histopathological analysis. The tympanic membrane perforations treated with the curable composition healed, and the regenerated tissue had the same structure as natural tympanic membrane tissue. The hair cells within the cochlea were intact, indicating that the curable composition was not ototoxic.

[0125] Example 4: Evaluation of the potential ototoxicity of the curable composition Procedure The tympanic membranes of adult guinea pigs were perforated with a scalpel and then immediately treated with the semi-gel composition. Next, the semi-gel composition was cured using a light guide and curing light.

[0126] Results The perforations were monitored over a 4-week period, during which time the perforations healed. Ototoxicity was evaluated by auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAE). The results from the ABR and DPOAE tests showed that hearing and the integrity of the hair cells were not lost, indicating that the curable composition had no potential for ototoxicity.

[0127] Example 5: Otolaryngology gel kit Exemplary otolaryngology gel kits of the present disclosure are shown in FIGS. 1A - C. The components included in the kit are shown in Table 1. [Table 1]

[0128] An otolaryngology gel kit (i.e., an otolaryngology gel patch kit) provides a composition of the present disclosure comprising hyaluronic acid (HA), methacrylated chitosan (MeCS), and riboflavin monophosphate (RM). The composition is supplied in a pouch. The contents of the gel pouch are mixed with physiological saline (1 mL of physiological saline) before application to the patient. After adding the physiological saline, re-cap the pouch and then knead to mix the dry reagent and the physiological saline. After mixing, cut off the end of the pouch and then empty the contents into the attached tray. The uncured composition is a semi-gel that is applied directly to the perforation through the external auditory canal using a curette or other preferred instrument. When exposed to blue light from a curing light, the semi-gel of the composition forms a hard three-dimensional gel patch that fills the perforation site. Animal experiments show that the gel material stays in place for less than 4 weeks.

[0129] Example 6: Clinical study on the safety and efficacy of an otolaryngology gel The following example shows a clinical study to evaluate the safety and efficacy of the composition of the present disclosure in the repair of chronic tympanic membrane perforations. In this study, i) the effect of the composition as a gel patch on promoting the natural healing process of closing chronic tympanic membrane perforations greater than 25% with a single application was evaluated, and ii) the safety of the composition and the application procedure was evaluated.

[0130] This study also evaluated i) the success of the gel application procedure (e.g., variations in anatomical status, perforation site, TMP size, and / or patient age), ii) the time taken to apply the gel once, iii) the time from application of the gel patch to healing of the tympanic membrane, iv) the change in hearing in all patient populations, v) the safety of the intended use of the gel patch in the target population, and vi) pain and discomfort were evaluated using a visual analog scale (VAS).

[0131] The main treatment evaluation item is to measure whether the complete closure rate of tympanic membrane perforation reported by clinical visual evaluation exceeds 75% at the fifth visit (3 months) after the application of the gel composition. The safety evaluation item of the incidence rate of serious adverse events related to the device and procedure during and after administration is also measured. The additional secondary evaluation items include complete tympanic membrane perforation closure visually evaluated at the fifth visit after successful application of the gel composition under sedation, success rate / failure rate of the treatment, median time to complete a single application procedure measured in minutes, comparison of pure tone average score and air-bone gap (ABG) baseline and audiometry data at the fifth visit, comparison of word recognition score data at baseline and the fifth visit, comparison of tympanometry measurement data of tympanic membrane mobility at baseline and the fifth visit, and median time to closure of tympanic membrane perforation measured on a weekly basis after treatment.

[0132] Test subject criteria: Patients are eligible to participate in this study if they meet all of the following criteria at the baseline visit. 1. If of appropriate age, have the willingness and ability to provide informed consent, consent of a legally authorized representative (LAR), and commitment. 2. Females and males at least 5 years old. 3. Criteria for the affected ear: a. The perforation is in the tympanic membrane with >25%, b. The perforation does not close spontaneously after 4 weeks of careful follow-up observation, c. The perforation does not heal actively, d. The perforation can be observed by endoscope or microscope, e. Earwax does not block the perforation.

[0133] Exclusion criteria: Patients are ineligible to participate in this study if they meet any of the following criteria. 1. The perforation is a marginal perforation (a perforation with an area without tympanic membrane between the perforation and the bony canal). 2. Exudative or non-exudative active otitis media. 3. Otorrhea from the middle ear has persisted for more than 3 months. 4. History of cleft palate. 5. Is receiving radiation therapy or is taking corticosteroids, immunosuppressants, or chemotherapy. 6. Is currently taking systemic antibiotics, antibiotic ear drops, and / or steroid ear drops. 7. Is currently infected with bacteria or a virus. 8. Fever (body temperature > 100°F) at the time of the first treatment. 9. Has been diagnosed with a cholesteatoma tumor in the middle ear. 10. Has a history of malignant external ear tumors within 3 years of the eligibility screening. 11. Abrasion / laceration of the external ear canal. 12. Severe medical conditions that may prevent the patient from participating in all procedures required for the study. 13. The principal investigator of the clinical trial feels that the patient is unable to cooperate with the application procedures. 14. The parent / LAR feels that the patient is unable to cooperate with the application procedures. 15. Crustacean allergy. 16. Is pregnant or has been found to have a possibility of being pregnant. 17. Adults lacking the capacity to consent.

[0134] Study design: Patients undergo screening at the first visit. Eligible patients receive treatment by applying otolaryngology gel at the second visit (day 0) and submit a pain score after the treatment. To determine the perforation closure rate, the perforation is monitored 1 week (third visit) and 3 weeks (fourth visit) after the treatment. At 12 weeks after the treatment (fifth visit), the closure of the perforation and hearing are evaluated. Audiometry and tympanometry are performed at the fifth visit (end of the study visit). Adverse events are observed at each visit.

[0135] Procedure for applying the otolaryngology gel: The patient is laid supine on the examination chair, and the ear is placed under a microscope or endoscope. The otolaryngologist administers the uncured semi-gel composition using a clean microsurgical instrument. The semi-gel composition is applied to the perforation such that the uncured semi-gel composition and the TM tissue around the perforation overlap by approximately 1 mm. Since this procedure is performed under a microscope or endoscope, the otolaryngologist can accurately cover the perforation with the gel. This procedure is a clean procedure and does not require sterilization. Before applying the gel composition, do not manipulate or abrade the edge of the perforation. After applying the uncured gel composition material to cover the perforation site, the otolaryngologist hardens the material using the provided reusable curing light and disposable light guide. The provided safety glasses are worn by the patient, clinician, and any other person in the room. The curing light emits intermittent beeping sounds during the 90-second curing time. The entire procedure takes approximately 10 minutes for ear treatment.

[0136] Incorporation by reference All references, articles, publications, patents, patent publications, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes. However, the descriptions of the references, articles, publications, patents, patent publications, and patent applications cited herein should not be construed 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 - drying composition comprising: (i) a base 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 photo - initiator, and 0 wt% to about 20 wt% of arginine; and (ii) a diluent.

2. The pre - freeze - drying composition according to claim 1, wherein the base comprises about 60 wt% of methacrylated chitosan.

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

4. The pre - freeze - drying composition according to any one of claims 1 - 3, wherein the base comprises about 0.12 wt% of a photo - initiator, and the photo - initiator is riboflavin.

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

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

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

8. A freeze - drying composition comprising: about 0 wt% to about 10 wt% of 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 photo - initiator; and about 0 wt% to about 20 wt% of arginine.

9. The freeze - drying composition according to claim 8, wherein the composition comprises about 1 wt% of water.

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

11. The freeze - drying composition according to claim 8 or 10, wherein the methacrylated chitosan is about 25% to about 40% methacrylated.

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

13. The freeze - drying composition according to any one of claims 8 - 12, wherein the composition comprises about 0.12 wt% of a photo - initiator, and the photo - initiator is riboflavin.

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

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

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

17. The semi-gel composition according to claim 16, wherein the composition comprises about 2% by weight of methacrylated chitosan.

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

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

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

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

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

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

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

26. A curable composition comprising: from about 80% to about 99% by weight of a diluent; from about 1% to about 3% by weight of methacrylated chitosan; from about 0.5% to about 2% by weight of hyaluronic acid; from about 0.0010% to about 0.01% by weight of a photoinitiator; from about 0% to about 0.5% by weight of arginine.

27. The curable composition according to claim 26, wherein the composition comprises about 2% by weight of methacrylated chitosan.

28. The curable composition according to claim 26 or 27, wherein the methacrylated chitosan is methacrylated by about 25% to about 40%.

29. The curable composition according to any one of claims 26 to 28, wherein the composition contains about 1% by weight of hyaluronic acid.

30. The curable composition according to any one of claims 26 to 29, wherein the composition contains about 0.004% by weight of a photoinitiator, and the photoinitiator is riboflavin.

31. The curable composition according to any one of claims 26 to 30, wherein the composition contains about 0.3% by weight of arginine.

32. The curable composition according to any one of claims 26 to 31, wherein the composition contains about 97% by weight of a diluent.

33. The curable composition according to claim 26 or 32, wherein the diluent is physiological saline.

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

35. The composition is irradiated with light at a wavelength of about 200 nm to about 800 nm, with a curing time of about 1 second to about 300 seconds, and at a light irradiation intensity of about 100 mW / cm ２ to about 100,000 mW / cm ２ The curable composition according to any one of claims 26 to 34, which is prepared by irradiating the semi-gel composition with light having the above light irradiation intensity.

36. The curable composition according to claim 35, wherein the light irradiation includes a curing time of about 90 seconds.

37. The composition according to any one of claims 8 to 25, wherein the composition is sterilized by ionizing radiation.

38. The sterilized composition according to claim 37, wherein the ionizing radiation is electron beam radiation.

39. The sterilized composition according to claim 37 or 38, wherein the logarithmic reduction of colony forming units is 3.4 to 3.6 after sterilization.

40. A kit comprising the composition according to any one of claims 8 to 25, a gel dispensing tray, and an optical guide.

41. The kit according to claim 40, wherein the sterilized composition is contained in a pouch.

42. The kit according to claim 40 or 41, wherein the kit is enclosed in a sealed tray.

43. The kit according to any one of claims 40 to 42, wherein the pouch includes a moisture-proof layer and a transparent portion.

44. A method for treating a tympanic membrane perforation, comprising: (a). applying the semi-gel composition according to any one of claims 16 to 25 to the tympanic membrane; (b). inserting an optical guide into the external auditory canal; (c). curing the composition by light irradiation. The light irradiation includes 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 light irradiation intensity of about 100 mW / cm ２ to about 100000 mW / cm ２ The method as described above.

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

46. A method for treating a tympanic membrane perforation of a patient in need of treatment, (a) Adding physiological saline to the lyophilized composition according to any one of claims 8 to 15 and mixing to form a semi-gel composition; (b) Administering the semi-gel composition to the tympanic membrane perforation site of the patient in need of treatment; (c) Curing the applied semi-gel composition by light irradiation. The method as described above. (Claim 47) (Claim 44) The method according to any one of claims 44 to 46, wherein the patient is 5 years old or older. (Claim 48) (Claim 47) The method according to claim 47, wherein the patient is 5 to 11 years old. (Claim 49) (Claim 48) The method according to any one of claims 44 to 48, wherein at least about 25% of the tympanic membrane of the patient is affected by the perforation. (Claim 50) (Claim 49) The method according to any one of claims 44 to 49, wherein the patient has a tympanic membrane perforation for at least 4 weeks. (Claim 51) (Claim 50) The method according to any one of claims 44 to 50, wherein the lyophilized composition is mixed with about 0.1 mL to about 2 mL of physiological saline. (Claim 52) (Claim 51) The method according to claim 51, wherein the lyophilized composition is mixed with about 1 mL of physiological saline. (Claim 53) (Claim 52) The method according to any one of claims 44 to 52, wherein the curing time lasts for about 90 seconds. (Claim 54) (Claim 53) The method according to any one of claims 44 to 53, wherein the cured semi-gel is biodegradable. (Claim 55) (Claim 54) The method according to any one of claims 44 to 54, wherein the cured semi-gel remains in the applied tympanic membrane for less than about 4 weeks. (Claim 56) (Claim 55) The method according to any one of claims 44 to 55, wherein the tympanic membrane perforation closure rate becomes about ≧ 75% by the treatment. (Claim 57) (Claim 56) The method according to any one of claims 44 to 56, wherein the tympanic membrane perforation closure rate becomes > 75% with one application by the treatment. (Claim 58) (Claim 57) The method according to any one of claims 44 to 57, wherein the tympanic membrane perforation closure rate increases about 3 months after administration by the treatment.

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