Liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment and preparation method and application thereof

Abstract

The present application relates to the technical field of biological hydrogel for tympanic membrane treatment, and specifically discloses a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, a preparation method and application thereof, the liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment comprises the following raw materials in percentage by weight: 8-12% of polyvinyl alcohol, 2-4% of oxidized dextran, 15-25% of functional complex, and the balance is solvent; the liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared by the present application utilizes the film-forming property of polyvinyl alcohol, realizes the functional recovery of tympanic membrane vibration through the transformation of the thin film from low modulus to high modulus by rapid cross-linking / solvent volatilization, and forms a tympanic membrane perforation treatment mode, which has the advantages of simple preparation method, mild preparation condition, high process feasibility, and the obtained hydrogel can be simply added to the tympanic membrane perforation site by using a simple needle, and there is no requirement for the operation level of the operator.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological hydrogel for tympanic membrane treatment, more particularly, it relates to a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment and a preparation method and application thereof. BACKGROUND

[0002] Tympanic membrane perforation caused by external force or inflammation is a common clinical problem leading to hearing loss worldwide. Tympanic membrane perforation reduces the transmission efficiency and frequency range of sound, thereby causing hearing loss, speech communication disorders and decreased quality of life. Perforation area that does not reach 50% of the total tympanic membrane area and does not involve the malleus can usually heal by itself, but during natural healing, patients have a certain degree of hearing loss and infection risk, which brings inconvenience to life.

[0003] A degradable biological tympanic membrane repair material is disclosed in Chinese patent with application publication number CN118718094A, which is made of gelatin spinning solution and crosslinking solution, and can replace temporal fascia and other autologous tissues to achieve tympanic membrane tissue repair. Its advantage lies in that it can be absorbed and degraded, has good support and repair properties. The patent with application publication number CN110302425A uses various biomaterials to obtain a crystal glue material that can be used for tympanic membrane perforation, which has the advantages of safety, non-toxicity and low immunogenicity. The patent with application publication number CN109069699A uses solvents to prepare silk fibroin biocompatible polyurethane membranes for repairing tympanic membrane perforation, which has the advantage of good biodegradation and acoustic vibration properties. However, the manufacturing process of these materials is complex, and they still need to be implanted under anesthesia with delicate surgery, which requires a high level of operation from the surgeon and has the disadvantage of easy occurrence of ear leakage. Therefore, it is of great clinical significance and application value to develop a tympanic membrane repair material with simple manufacturing process and convenient use. SUMMARY

[0004] To solve the problem of complex manufacturing process of existing tympanic membrane perforation repair materials, and still need to be implanted under anesthesia with delicate surgery, which requires a high level of operation from the surgeon and has the disadvantage of easy occurrence of ear leakage, the present application provides a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment and a preparation method and application thereof.

[0005] The present application provides a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, which adopts the following technical scheme:

[0006] A liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following raw materials by weight percentage: polyvinyl alcohol 8-12%, oxidized dextran 2-4%, functional complex 15-25%, and the balance is solvent.

[0007] Preferably, the liquid fast-drying antibacterial hydrogel for treating tympanic membrane perforation comprises the following raw materials by weight percentage: polyvinyl alcohol 10%, oxidized dextran 3%, functional complex 20%, and the balance is solvent.

[0008] Preferably, the solvent comprises anhydrous ethanol and deionized water in a mass ratio of 4-6:3-5.

[0009] Preferably, the oxidized dextran is prepared by the following method:

[0010] (1) Dissolve dextran in deionized water to prepare a dextran solution with a mass concentration of 3-6%;

[0011] (2) Under light-proof conditions, add sodium periodate solution to the dextran solution, adjust the pH value to 4.5-5, and stir at 20-25°C for 5-6h, then add ethylene glycol and continue stirring for 1-2h to terminate the reaction, obtaining a reaction liquid;

[0012] (3) After removing sodium periodate from the reaction liquid by dialysis, concentrate by rotary evaporation and freeze-dry to obtain the oxidized dextran.

[0013] Preferably, the oxidation degree of the oxidized dextran is 20-30%.

[0014] Preferably, the functional complex comprises tannic acid, carboxymethyl chitosan and ε-polylysine in a mass ratio of 7-9:2-4:1.

[0015] A preparation method of a liquid fast-drying antibacterial hydrogel for treating tympanic membrane perforation, comprising the following steps:

[0016] At room temperature, polyvinyl alcohol, oxidized dextran and functional complex are sequentially added to the solvent, and the solute is uniformly distributed in the solvent by magnetic stirring under heating, and defoaming is performed by standing, to form a liquid fast-drying antibacterial hydrogel for treating tympanic membrane perforation.

[0017] Preferably, the heating specifically refers to heating to 65-75°C.

[0018] Preferably, the magnetic stirring time is 8-15min.

[0019] A liquid fast-drying antibacterial hydrogel for treating tympanic membrane perforation is applied in the repair of tympanic membrane perforation.

[0020] Preferably, the application method of the liquid fast-drying antibacterial hydrogel for treating tympanic membrane perforation comprises the following steps: at room temperature, 0.5-2μL of the liquid fast-drying antibacterial hydrogel for treating tympanic membrane perforation is added to the tympanic membrane perforation through a needle, and after waiting for 30-60s, the effect of rapidly restoring hearing is achieved.

[0021] In summary, the present application has the following beneficial effects:

[0022] The liquid rapid-drying antibacterial hydrogel prepared by the present application for tympanic membrane perforation treatment utilizes the film-forming property of polyvinyl alcohol, and through rapid cross-linking / solvent volatilization, the transformation of the film from low modulus to high modulus is completed, the function recovery of tympanic membrane vibration is realized, and a new type of tympanic membrane perforation treatment mode is formed.

[0023] In the preparation process of the hydrogel, oxidized dextran and a functional complex are introduced, wherein the oxidized dextran and the polyvinyl alcohol main chain form an interpenetrating network through a dynamic Schiff base bond, so that the mechanical strength of the hydrogel is significantly improved, and the ear pressure fluctuation caused by sneezing and the like can be tolerated; the functional complex is obtained by compounding tannic acid, carboxymethyl chitosan and epsilon-polylysine, and both tannic acid and epsilon-polylysine have inhibitory effect on a plurality of bacteria and fungi, so that the antibacterial rate can be significantly improved, the antibacterial spectrum can be expanded, infection and inflammation can be prevented, and the cochlear toxicity risk of traditional silver ion antibacterial agents can be effectively avoided; the introduction of carboxymethyl chitosan, on the one hand, forms a weak hydrogen bond with the phenolic hydroxyl group of tannic acid in the normal ear canal environment, so that the gel remains in a pre-crosslinked liquid state, and on the other hand, forms a pH-sensitive hydrogen bond network with tannic acid in the inflammation area after tympanic membrane perforation, so that the carboxymethyl chitosan is protonated and rapid gelation is triggered, the film-forming time is shortened to 30 seconds, and the cross-linking effect is enhanced.

[0024] The liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared by the present application has simple preparation method, mild preparation conditions, high process feasibility, and the obtained hydrogel can be simply added dropwise to the tympanic membrane perforation site, so that the operation level of the operator is not required, the hydrogel added dropwise only triggers strong cross-linking in the perforation area, and the functions of precise in-situ film formation, rapid sealing, mechanical adaptation and long-acting antibiosis are integrated. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 The modulus characterization diagram of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 of the present application.

[0026] Figure 2 The schematic diagram of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 of the present application for repairing tympanic membrane perforation.

[0027] Figure 3 The hearing characterization diagram of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 of the present application for repairing tympanic membrane perforation.

[0028] Figure 4 The in-vivo antibacterial effect characterization diagram of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 and Comparative Example 1 of the present application.

[0029] Figure 5 Figure for in vitro antibacterial effect characterization of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared for Example 1 and Comparative Example 1 of the present application.

[0030] Figure 6 Figure for degradation performance characterization of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared for Example 1 of the present application.

[0031] Figure 7 Figure for adhesion characterization of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared for Example 1 of the present application. DETAILED DESCRIPTION

[0032] The present application is further described in detail below in conjunction with examples.

[0033] The raw materials used in the examples and comparative examples of the present application are as follows:

[0034] Polyvinyl alcohol, CP Pharmacopoeia Standard, CAS No. 9002-89-5, active ingredient content 99.6%, purchased from Rui Cheng Kang Pharmaceutical Technology (Shaanxi) Co., Ltd.;

[0035] Dextran is β-dextran, CAS No. 1439905-58-4, active ingredient content 99%, purchased from Xi'an Tianzheng Pharmaceutical Auxiliary Material Co., Ltd.;

[0036] Tannic acid, CAS No. 1401-55-4, active ingredient content 90%, brand Xinxinjie, purchased from Shandong Aicai Biological Technology Co., Ltd.;

[0037] Carboxymethyl chitosan, CAS No. 83512-85-0, active ingredient content 99%, model CP2020, purchased from Xi'an Tianzheng Pharmaceutical Auxiliary Material Co., Ltd.;

[0038] Epsilon-polylysine, CAS No. 25104-18-1, active ingredient content 99%, particle size 100 mesh, brand Xinxinjie, purchased from Shandong Aicai Biological Technology Co., Ltd.;

[0039] Preparation Example 1 and Comparative Preparation Example 1 provide an oxidized dextran.

[0040] Preparation Example 1

[0041] The oxidized dextran is prepared by the following method:

[0042] (1) Add dextran into deionized water, stir at 40°C until completely dissolved, and prepare a dextran solution with a mass concentration of 4%;

[0043] (2) Under the light-avoiding condition, sodium periodate solution was added dropwise into the dextran solution, 0.1M hydrochloric acid was used to adjust the pH value to 4.8, the reaction was stirred at 25℃ for 6h, then ethylene glycol was added to continue stirring for 2h to terminate the reaction, and a reaction liquid was obtained, wherein the mass concentration of the sodium periodate solution was 5%, the mass ratio of the dextran solution to the sodium periodate solution was 1:0.36, and the addition amount of the ethylene glycol was 40% of the mass of the sodium periodate solution;

[0044] The reaction liquid was loaded into a dialysis bag with a molecular weight cut-off of 6kDa, and dialysis was performed in deionized water at 4℃ for 72h, and the water was changed every 8h to completely remove the sodium periodate, and a dialysate was obtained;

[0045] The dialysate was concentrated to 45% of the original volume under the condition of 50℃ and 0.1MPa by rotary evaporation, and then freeze-dried at-20℃ for 48h to obtain the oxidized dextran.

[0046] The degree of oxidation was 25% determined by the hydroxylamine hydrochloride method, and the dialysate was verified to be free of free IO4 by potassium iodide-starch test paper. - .

[0047] Comparative Preparation Example 1

[0048] The oxidized dextran was prepared by the following method:

[0049] (1) Dextran was added into deionized water, and stirred at 40℃ until completely dissolved to prepare a dextran solution with a mass concentration of 4%;

[0050] (2) Under the light-avoiding condition, sodium periodate solution was added dropwise into the dextran solution, 0.1M hydrochloric acid was used to adjust the pH value to 4.8, the reaction was stirred at 25℃ for 24h, then ethylene glycol was added to continue stirring for 2h to terminate the reaction, and a reaction liquid was obtained, wherein the mass concentration of the sodium periodate solution was 5%, the mass ratio of the dextran solution to the sodium periodate solution was 1:0.72, and the addition amount of the ethylene glycol was 50% of the mass of the sodium periodate solution;

[0051] The reaction liquid was loaded into a dialysis bag with a molecular weight cut-off of 6kDa, and dialysis was performed in deionized water at 4℃ for 72h, and the water was changed every 8h to completely remove the sodium periodate, and a dialysate was obtained;

[0052] The dialysate was concentrated to 45% of the original volume under the condition of 50℃ and 0.1MPa by rotary evaporation, and then freeze-dried at-20℃ for 48h to obtain the oxidized dextran.

[0053] The degree of oxidation was 60% determined by the hydroxylamine hydrochloride method, and the dialysate was verified to be free of free IO4 by potassium iodide-starch test paper. - .

[0054] Examples 1-3 provide a liquid fast-drying antibacterial hydrogel for eardrum perforation treatment.

[0055] Example 1

[0056] A liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following raw materials by weight percentage: polyvinyl alcohol 8%, oxidized dextran in Preparation Example 1 2%, functional complex 15%, and the balance being solvent;

[0057] The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 4:3.

[0058] The functional complex comprises tannic acid, carboxymethyl chitosan and ε-polylysine in a mass ratio of 7:2:1.

[0059] A preparation method of a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following steps:

[0060] At room temperature, polyvinyl alcohol, oxidized dextran and functional complex are sequentially added to the solvent, and the temperature is raised to 65℃. The magnetic stirring time is controlled to be 15 min. The solute is uniformly distributed in the solvent by magnetic stirring. After standing for 1 h to remove bubbles, a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment is formed.

[0061] Example 2

[0062] A liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following raw materials by weight percentage: polyvinyl alcohol 10%, oxidized dextran in Preparation Example 1 3%, functional complex 20%, and the balance being solvent;

[0063] The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 5:4.

[0064] The functional complex comprises tannic acid, carboxymethyl chitosan and ε-polylysine in a mass ratio of 8:3:1.

[0065] A preparation method of a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following steps:

[0066] At room temperature, polyvinyl alcohol, oxidized dextran and functional complex are sequentially added to the solvent, and the temperature is raised to 70℃. The magnetic stirring time is controlled to be 12 min. The solute is uniformly distributed in the solvent by magnetic stirring. After standing for 1.5 h to remove bubbles, a liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment is formed.

[0067] Example 3

[0068] A liquid fast-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following raw materials by weight percentage: polyvinyl alcohol 12%, oxidized dextran in Preparation Example 1 4%, functional complex 25%, and the balance being solvent;

[0069] The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 6:5.

[0070] The functional complex comprises tannic acid, carboxymethyl chitosan and epsilon-polylysine in a mass ratio of 9:4:1.

[0071] A preparation method of a liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following steps:

[0072] At room temperature, polyvinyl alcohol, oxidized dextran and a functional complex are sequentially added to the solvent, the temperature is raised to 75 DEG C, the magnetic stirring time is controlled to be 8 min, the solute is uniformly distributed in the solvent by using magnetic stirring, and the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment is formed by standing for 2 h to remove bubbles.

[0073] In order to verify the comprehensive performance of the antibacterial hydrogel prepared in examples 1-3, the applicant sets up comparative examples 1-3, as follows:

[0074] Comparative example 1 is the same as example 1, except that the functional complex comprises tannic acid and epsilon-polylysine in a mass ratio of 7:3, as follows:

[0075] A liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following raw materials in weight percentage: polyvinyl alcohol 8%, oxidized dextran 2% in comparative preparation example 1, functional complex 15%, and the balance being solvent;

[0076] The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 4:3.

[0077] The functional complex comprises tannic acid and epsilon-polylysine in a mass ratio of 7:3.

[0078] A preparation method of a liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following steps:

[0079] At room temperature, polyvinyl alcohol, oxidized dextran and a functional complex are sequentially added to the solvent, the temperature is raised to 65 DEG C, the magnetic stirring time is controlled to be 15 min, the solute is uniformly distributed in the solvent by using magnetic stirring, and the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment is formed by standing for 1 h to remove bubbles.

[0080] Comparative example 2

[0081] Comparative example 2 is the same as example 1, except that the functional complex comprises tannic acid and carboxymethyl chitosan in a mass ratio of 7:3, as follows:

[0082] A liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment, comprising the following raw materials in weight percentage: polyvinyl alcohol 8%, oxidized dextran 2%, functional complex 15%, and the balance being solvent;

[0083] The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 4:3.

[0084] The functional complex comprises tannic acid and carboxymethyl chitosan in a mass ratio of 7:3.

[0085] A preparation method of the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment comprises the following steps:

[0086] At room temperature, polyvinyl alcohol, oxidized dextran and a functional complex are sequentially added to the solvent, and the temperature is raised to 65 DEG C. The magnetic stirring time is controlled to be 15 min. The solute is uniformly distributed in the solvent by using magnetic stirring. After standing for 1 h to remove bubbles, the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment is formed.

[0087] Comparative Example 3

[0088] Comparative Example 3 is the same as Example 1, except that the oxidized dextran in Preparation Example 1 is replaced by the oxidized dextran in Comparative Preparation Example 1 in equal mass, as follows:

[0089] A liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment comprises the following raw materials by weight percentage: polyvinyl alcohol 8%, oxidized dextran 2%, functional complex 15%, and the balance is solvent.

[0090] The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 4:3.

[0091] The functional complex comprises tannic acid, carboxymethyl chitosan and epsilon-polylysine in a mass ratio of 7:2:1.

[0092] A preparation method of the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment comprises the following steps:

[0093] At room temperature, polyvinyl alcohol, oxidized dextran and a functional complex are sequentially added to the solvent, and the temperature is raised to 65 DEG C. The magnetic stirring time is controlled to be 15 min. The solute is uniformly distributed in the solvent by using magnetic stirring. After standing for 1 h to remove bubbles, the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment is formed.

[0094] Performance detection

[0095] The comprehensive performance of the liquid quick-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Examples 1-3 and Comparative Examples 1-3 of the application is tested respectively.

[0096] Test 1, tensile strength and elongation at break test of hydrogel

[0097] The liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in the present application examples 1-3 and comparative examples 1-3 was respectively placed in a polytetrafluoroethylene mold, and a film was formed in a pH 6.8 buffer at 37°C for 30 seconds to obtain a sample strip of 20 mm x 10 mm x 100 μm. The sample strip was placed in a stretching device, and a tensile test was performed at a stretching rate of 60 mm / min until the sample was broken. The tensile strength and elongation at break were counted, and the results are shown in Table 1 below and the following Figure 1 .

[0098] Table 1:

[0099]

[0100] Table 1 and the following Figure 1 The results show that the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in the present application examples 1-3 has excellent tensile strength and elongation at break, and the comprehensive mechanical properties are significantly better than those of the comparative examples 1-3, Figure 1 The horizontal coordinate Strain represents the elongation at break, and the vertical coordinate Stress represents the tensile strength.

[0101] Test two, hearing recovery effect test of hydrogel for repairing tympanic membrane perforation

[0102] The tympanic membrane perforation of rats was modeled as follows: 48 male Sprague-Dawley rats (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.) weighing 180-220 g and 6-8 weeks old were adaptively fed for one week. The 48 male Sprague-Dawley rats were randomly divided into 8 groups, 6 rats in each group, and were named blank control group (normal rats), model group (untreated), application examples 1-3, and comparative examples 1-3. Except for the blank control group (normal rats), the rats in the model group (untreated), application examples 1-3, and comparative examples 1-3 were intraperitoneally injected with 2% sodium pentobarbital (0.2 mL / 100 g) to ensure that the rats had no pain reflex. The auricles and external auditory canals of the rats were sterilized with 75% ethanol, the tympanic membrane was positioned under a microscope, and a 1.5 mm sterile needle was used to vertically pierce the tympanic membrane to form a perforation, thereby obtaining a rat tympanic membrane perforation model.

[0103] After modeling, the blank control group (normal rats) and the model group (untreated) were not treated, and the application examples 1-3 and comparative examples 1-3 were prepared using the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in the present application examples 1-3 and comparative examples 1-3. At room temperature, 1 μL was added to the tympanic membrane perforation of the rats through the needle, and the hearing recovery effect at different frequencies was tested by auditory brainstem response (ABR) after the film was formed for 30 seconds. The results are shown in Table 2 below and the followingFigures 2-3 .

[0104] Table 2:

[0105]

[0106] Combine Table 2 and Appendix Figures 2-3 It can be seen that the liquid quick-drying antibacterial hydrogel prepared in Examples 1-3 of the present invention for the treatment of tympanic membrane perforation can achieve the effect of rapid hearing restoration by simply dripping it onto the tympanic membrane perforation with a needle, and the hearing restoration effect is far superior to that of comparative examples 1-3. Figure 3 middle, This is expressed as p<0.05 under the Wilcoxon signed-rank test, meaning that the hearing thresholds in each frequency band after repair are significantly different from those after piercing.

[0107] Experiment 3: In vivo antibacterial effect test of hydrogel

[0108] After establishing a rat tympanic membrane perforation model, 50 μL of a solution containing 10... was injected into the perforation site of the rat tympanic membrane. 8 A bacterial suspension of CFU / mL Staphylococcus aureus was used to simulate infectious perforation. Liquid, quick-drying antibacterial hydrogels prepared in Examples 1-3 and Comparative Examples 1-3 for the treatment of tympanic membrane perforation were applied at room temperature and flow dynamics, with 1 μL dripped into the tympanic membrane perforation site in rats via a needle. After 40 seconds of film formation, the healing of the tympanic membrane was observed using an otoscope. Changes in bacterial count were observed by sampling after application. The results are shown in Table 3 and Appendix. Figure 4 .

[0109] Table 3:

[0110]

[0111] Combine Table 3 and Appendix Figure 4 It can be seen that the liquid quick-drying antibacterial hydrogels prepared in Examples 1-3 of this invention for the treatment of tympanic membrane perforation have significant in vivo antibacterial effects and tympanic membrane perforation treatment effects, and their overall effect is far superior to that of comparative examples 1-3. Figure 4 The left side of the image shows the coating sampling results of Comparative Example 1, and the right side shows the coating sampling results of Example 1.

[0112] In addition to the above tests, the applicant also tested the following properties of the hydrogel:

[0113] I. Test of the antibacterial effect of hydrogel

[0114] The liquid quick-drying antibacterial hydrogels for tympanic membrane perforation prepared in Example 1 and Comparative Example 1 of this invention were respectively added to 10 8The bacterial liquid of CFU / mL of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa was cultured at 37℃ for 6 hours, and then bacterial live and dead staining (DMAO / PI) was performed on the bacterial liquid, and sampling and culture observation were performed, and the specific results are shown in the following table 1. Figure 5 .

[0115] As shown in the following table 2, the bacterial liquid of CFU / mL of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa was cultured at 37℃ for 6 hours, and then bacterial live and dead staining (DMAO / PI) was performed on the bacterial liquid, and sampling and culture observation were performed, and the specific results are shown in the following table 2. Figure 5 It can be known from the following table 2 that in Example 1, almost no bacteria survive, and the in-vitro antibacterial effect is obviously better than that in Comparative Example 1, Figure 5 The left side of the table in Comparative Example 1 is the sampling and culture observation result of bacteria, and the right side is the sampling and culture observation result of bacteria in Example 1.

[0116] II. Degradation performance test of hydrogel

[0117] 400 μL of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 was taken, and after film formation at 37℃ in a pH 6.8 buffer for 30 seconds, a sample to be tested was obtained. The sample to be tested was placed in 5 mL of a PBS solution at 37℃, and the sample was taken out at 3d, 5d and 7d, respectively, and the degradation morphology at different times was observed. The specific results are shown in the following table 3. Figure 6 .

[0118] As shown in the following table 4, the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 was taken, and after film formation at 37℃ in a pH 6.8 buffer for 30 seconds, a sample to be tested was obtained. The sample to be tested was placed in 5 mL of a PBS solution at 37℃, and the sample was taken out at 3d, 5d and 7d, respectively, and the degradation morphology at different times was observed. The specific results are shown in the following table 4. Figure 6 It can be known from the following table 4 that the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in the application has a high degradation rate and a short degradation time.

[0119] III. Adhesion test of hydrogel

[0120] The 90° peeling force of the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 on different substrates was tested according to the national standard GB / T 2792-2014 “Test method for peeling strength of adhesive tape”. The specific results are shown in the following table 5. Figure 7 .

[0121] As shown in the following table 6, the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in Example 1 was taken, and after film formation at 37℃ in a pH 6.8 buffer for 30 seconds, a sample to be tested was obtained. The sample to be tested was placed in 5 mL of a PBS solution at 37℃, and the sample was taken out at 3d, 5d and 7d, respectively, and the degradation morphology at different times was observed. The specific results are shown in the following table 4. Figure 7 It can be known from the following table 6 that the liquid rapid-drying antibacterial hydrogel for tympanic membrane perforation treatment prepared in the application has strong adhesion and can be applied to different substrate materials, has a wide application range, and the selected substrate materials include steel plate substrate, pigskin substrate, acrylic substrate and rubber substrate (silicone rubber is specifically used in this test).

[0122] This specific embodiment is only an explanation of the application and is not a limitation of the application. Those skilled in the art can make modifications to this embodiment without creative contribution after reading this specification, but as long as it is within the scope of the claims of the application, it is protected by the patent law.

Claims

1. A liquid, quick-drying antibacterial hydrogel for the treatment of tympanic membrane perforation, characterized in that, It is made from the following raw materials by weight percentage: 8% polyvinyl alcohol, 2% oxidized dextran, 15% functional complex, and the balance being solvent; The oxidized dextran is prepared by the following method: (1) Dissolve dextran in deionized water to prepare a dextran solution with a mass concentration of 3-6%; (2) Under light-protected conditions, sodium periodate solution was added dropwise to the dextran solution to adjust the pH value to 4.5-5. After stirring at 20-25℃ for 5-6 hours, ethylene glycol was added and stirring was continued for 1-2 hours to terminate the reaction and obtain the reaction solution. (3) After the reaction solution is dialyzed to remove sodium periodate, it is concentrated by rotary evaporation and freeze-dried to obtain oxidized dextran; The degree of oxidation of the oxidized dextran is 20-30%; The functional complex comprises tannic acid, carboxymethyl chitosan, and ε-polylysine in a mass ratio of 7:2:

1. The solvent comprises anhydrous ethanol and deionized water in a mass ratio of 4:3; The liquid, quick-drying antibacterial hydrogel used for the treatment of tympanic membrane perforation is prepared by the following method: At room temperature, polyvinyl alcohol, oxidized dextran, and functional complex were added to the solvent in sequence, the temperature was raised to 65°C, and the magnetic stirring time was controlled to be 15 min. The solute was evenly distributed in the solvent by magnetic stirring, and the mixture was allowed to stand for 1 h to defoam, forming a liquid quick-drying antibacterial hydrogel for the treatment of tympanic membrane perforation.

2. The liquid quick-drying antibacterial hydrogel for treating tympanic membrane perforation according to claim 1, characterized in that, The oxidized dextran is prepared by the following method: (1) Add dextran to deionized water and stir at 40°C until completely dissolved to prepare a 4% (w / w) dextran solution; (2) Under light-protected conditions, sodium periodate solution was added dropwise to the dextran solution, and the pH value was adjusted to 4.8 with 0.1M hydrochloric acid. After stirring at 25°C for 6 hours, ethylene glycol was added and stirring was continued for 2 hours to terminate the reaction, and the reaction solution was obtained. The mass concentration of sodium periodate solution was 5%, the mass ratio of dextran solution to sodium periodate solution was 1:0.36, and the amount of ethylene glycol added was 40% of the mass of sodium periodate solution. The reaction solution was placed in a dialysis bag with a molecular weight cutoff of 6 kDa and dialyzed in deionized water at 4°C for 72 hours, with the water changed every 8 hours to completely remove sodium periodate and obtain dialysate. The dialysate was concentrated to 45% of its original volume by rotary evaporation at 50℃ and 0.1MPa, and then freeze-dried at -20℃ for 48 hours to obtain oxidized dextran. The degree of oxidation of the oxidized dextran is 25%.

3. The application of the liquid quick-drying antibacterial hydrogel of claim 1 or claim 2 for the treatment of tympanic membrane perforation in the repair of tympanic membrane perforation.

4. The application of the liquid quick-drying antibacterial hydrogel for the treatment of tympanic membrane perforation according to claim 3, characterized in that, The application method includes the following steps: At room temperature flow, 0.5-2 μL of the liquid quick-drying antibacterial hydrogel used for tympanic membrane perforation treatment is dripped into the tympanic membrane perforation through a needle, and then waited for 30-60 seconds.

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