Preparation method of antibacterial hemostatic medical sponge

Abstract

The invention discloses a preparation method of antibacterial and hemostatic medical sponge, and relates to the technical field of preparation of medical sponge. The preparation method of the antibacterial hemostatic medical sponge comprises the following steps: cleaning silkworm cocoons after removing impurities, and immersing the cleaned silkworm cocoons in a neutral protease solution to complete degumming so as to obtain fibroin fibers; the preparation method comprises the following steps: dissolving chitosan in an acetic acid solution, dissolving polyvinyl alcohol in deionized water, adding sodium alginate, adding a silk fibroin solution to form a composite solution, dropwise adding an AgNO3 solution, introducing CO2, maintaining the pressure, quickly releasing the pressure, foaming, injecting into a mold, carrying out gradient freezing to form a multistage-aperture sponge matrix, immersing in a genipin ethanol solution, and transferring into a 5% CaC 12 solution for soaking. The silk fibroin provides a biocompatible substrate and promotes cell adhesion, the sponge is constructed into a hierarchical pore structure with a compact surface and a loose interior, and liquid absorption and blood coagulation factor enrichment are accelerated.

Description

Technical Field

[0001] The invention relates to the technical field of preparation of medical sponges, in particular to a method for preparing an antibacterial and hemostatic medical sponge. Background Art

[0002] Fabric damage can easily cause bleeding and breed bacteria. Effective hemostatic and antibacterial methods can reduce unnecessary casualties. Therefore, the development of a hemostatic material that can both stop bleeding and be antibacterial is of great significance in trauma treatment and surgery.

[0003] Chitosan is a natural cationic polysaccharide that is non-toxic, biocompatible, non-immunogenic, and can be degraded by lysozyme to produce glucosamine. Chitosan also has antibacterial, hemostatic, immune-regulating, wound-healing promoting, wound exudate absorption, and scar reduction properties.

[0004] For example, an antibacterial hemostatic sponge and its preparation method and application disclosed in Chinese Invention Publication No. CN113425889B achieves hemostatic and antibacterial effects by adding the hemostatic drug 6-aminocaproic acid and the antibacterial drug nano-zinc oxide. Furthermore, because chitosan quaternary ammonium salt has good bactericidal and antibacterial properties, and the added nano-zinc oxide also has good bactericidal and antibacterial properties, the antibacterial and hemostatic properties of the chitosan quaternary ammonium salt itself, combined with the hemostatic drugs 6-aminocaproic acid and the antibacterial drugs nano-zinc oxide, improve the overall antibacterial and hemostatic properties of the material. Carboxymethyl chitosan has certain hemostatic properties, and by adding the hemostatic drugs 6-aminocaproic acid and the antibacterial drugs nano-zinc oxide, even better hemostatic and antibacterial effects can be achieved.

[0005] Currently, sponges in existing technologies often use drugs loaded with them, such as metronidazole and polyhexamethylene hydrochloride, to achieve antibacterial properties, but there is a risk of sudden drug release. Although the above-mentioned sponges improve their antibacterial properties by adding nano-zinc oxide, they do not optimize the pore structure and cannot achieve the enrichment of hemostatic factors. Summary of the Invention

[0006] In view of the deficiencies of the prior art, the present invention provides a method for preparing an antibacterial hemostatic medical sponge, which solves the problems raised in the above background technology.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions: a method for preparing an antibacterial hemostatic medical sponge, comprising removing impurities from silk cocoons, cleaning them, immersing them in a neutral protease solution to complete degumming, obtaining silk fibroin fibers, and then dissolving them in a LiBr solution for dialyzation and purification to obtain a silk fibroin solution;

[0008] Chitosan was dissolved in 2% acetic acid solution, polyvinyl alcohol was dissolved in 90°C deionized water, sodium alginate was added and stirred, and the obtained silk fibroin solution was added to form a chitosan-polyvinyl alcohol-sodium alginate-silk fibroin composite solution;

[0009] Adding AgNO3 solution dropwise to the obtained composite solution to form a silver-containing composite solution colloid after reaction;

[0010] The colloid is placed in a high-pressure reactor, CO2 is introduced, the pressure is maintained and then the pressure is quickly released to foam, and then injected into a mold for gradient freezing to form a multi-level pore size sponge matrix;

[0011] The sponge matrix was immersed in 1% genipin ethanol solution, then in 5% CaCl2 solution, and rinsed with deionized water to obtain a calcified antibacterial sponge.

[0012] Medical zeolite powder and 0.5% chitosan acetate solution are mixed into a suspension, and the suspension is evenly loaded on the surface of the sponge by a spraying method to obtain an antibacterial hemostatic sponge.

[0013] A further improvement of the technical solution of the present invention is that the silk cocoons are cleaned after being decontaminated, immersed in a neutral protease solution to complete degumming, and the silk fibroin fibers are obtained, and then dissolved in a LiBr solution for dialysis purification to obtain a silk fibroin solution, comprising:

[0014] The cocoons were chopped and placed in a titanium alloy tank of an ultrasonic cleaning machine. They were cleaned with 0.5% NaHCO3 solution and ultrasonically treated at 60°C and 280W for 35 minutes to remove lipids and impurities. They were then rinsed with deionized water until neutral and drained through a 300-mesh filter.

[0015] The pretreated cocoons were transferred to a conical glass reactor equipped with a constant temperature oscillating water bath, 2% neutral protease solution was added, and the water bath parameters were set to a constant temperature of 60°C and an oscillation frequency of 140 rpm. The reaction was carried out for 5 hours. After the reaction, the degummed silk fibers were separated using a centrifuge and washed three times with deionized water.

[0016] Degummed silk fibroin fibers were added to a high-temperature resistant glass reactor, and a 9.3 mol / L LiBr solution was injected at a ratio of 1:9 w / v. After stirring at 145 rpm, the solution was dissolved in a 65°C water bath for 1 hour to form a light yellow viscous liquid. The solution was transferred to a dialysis bag and placed in a dialysis tank with stirring at 40 rpm. The water was changed every 3 hours and the dialysis was continued for 72 hours. After dialysis, the undissolved impurities were removed by centrifugation to obtain a silk fibroin solution.

[0017] Further improvements to the present invention's technical solution include: the added 0.5% NaHCO3 solution has a liquid-to-solid ratio of 20:1; and the added 2% neutral protease solution has a pH of 7.0 and a liquid-to-solid ratio of 15:1. Neutral protease specifically degrades sericin at a pH of 7.0, avoiding the chain breakage of silk fibroin caused by traditional alkaline methods. Furthermore, silk fibroin exhibits excellent biocompatibility, mechanical properties, degradation resistance, and easy plasticity. Therefore, this application uses silk fibroin as the substrate for the preparation of medical sponges.

[0018] A further improvement of the technical solution of the present invention is that: chitosan is dissolved in a 2% acetic acid solution, polyvinyl alcohol is dissolved in 90°C deionized water, sodium alginate is added and stirred, and the obtained silk fibroin solution is added to form a composite solution, comprising:

[0019] Chitosan powder was added to a polytetrafluoroethylene beaker, and a 2% acetic acid solution was added at a concentration of 3% w / v. The mixture was placed in a constant temperature water bath with a magnetic stirrer. The reaction parameters were set at 40°C and 500 rpm. Stirring was continued for 4 hours until the mixture was completely dissolved. The undissolved particles were removed by filtering with a 200-mesh nylon filter cloth to obtain a transparent chitosan colloid.

[0020] PVA particles were added to a laminated glass reactor, and deionized water was added at a concentration of 5% w / v. After stirring, the temperature was raised to 90°C and maintained for 1 hour to form a homogeneous transparent solution. The solution was then sterilized in a high-temperature autoclave at 121°C for 20 minutes and cooled to room temperature to obtain a PVA solution.

[0021] The transparent chitosan colloid and polyvinyl alcohol solution were transferred to a sandwich reactor in a volume ratio of 1:2. After stirring, they were mixed at a constant temperature of 40°C for 30 minutes. SA powder was slowly added to a final concentration of 1.5% w / v. The mixture was stirred for 1 hour until no visible particles were found. The silk fibroin solution was added and the mixture was transferred to a titanium alloy tank of an ultrasonic cleaner for ultrasonic treatment at 40°C for 20 minutes to remove bubbles to obtain a composite solution.

[0022] A further improvement of the technical solution of the present invention is that the added silk fibroin solution: transparent chitosan colloid: polyvinyl alcohol solution = 1:1:2, and the viscosity of the final composite solution is ≥5000mPa·s and pH = 5.2-5.5.

[0023] A further improvement of the technical solution of the present invention is that: the AgNO3 solution is added dropwise to the obtained composite solution to form a silver-containing composite solution colloid after reaction, comprising:

[0024] AgNO3 solution was slowly added dropwise at a rate of 0.5 mL / min to a final concentration of 0.5 mmol / L. Nitrogen protection was turned on, and the composite solution was filtered through a 0.22 μm microporous filter membrane to remove large particle aggregates and stored in a light-shielding sealed container.

[0025] A further improvement to the technical solution of the present invention is that the colloid is placed in an autoclave, CO2 is introduced, the pressure is maintained, and then the pressure is rapidly released to foam, and then injected into a mold for gradient freezing to form a multi-level pore sponge matrix, which includes: connecting the composite solution colloid to a CO2 gas source, pressurizing it at a rate of 1.5MPa / min to 8MPa, maintaining the pressure for 2 hours to allow the CO2 to fully penetrate the interior of the colloid, triggering rapid pressure release at a rate of >15MPa / s, so that the composite solution colloid forms a multi-level pore size, injecting the foamed colloid into a polytetrafluoroethylene mold pre-cooled to 4°C, cooling the temperature from 4°C to -20°C at a rate of -1°C / min for 4 hours to directionally freeze the micropores on the surface, and cooling the temperature from -20°C to -80°C at a rate of -5°C / min for 2 hours to fix the internal macroporous structure, thereby forming a multi-level pore sponge matrix. The rapid pressure release of >15MPa / s induces CO2 gas-liquid phase separation, forming a gradient structure with dense surface and loose interior, and the dual pore formation improves the water absorption rate.

[0026] A further improvement of the technical solution of the present invention is that: after immersing the sponge base in a 1% genipin ethanol solution, it is transferred to a 5% CaCl2 solution for immersion, and rinsed with deionized water to obtain a calcified antibacterial sponge, comprising:

[0027] The sponge substrate was immersed in a 1% genipin ethanol solution with a solution volume to sponge mass ratio of 10:1 v / w, and shaken at 60 rpm at 25°C for 12 hours. The sponge was transferred to a 5% CaCl2 solution and allowed to stand for 1 hour. After that, the residual liquid on the surface was removed by centrifugation at 800 rpm in a centrifugal dehydrator and rinsed three times with deionized water.

[0028] The further improvement of the technical solution of the present invention is that the added 1% genipin ethanol solution has a pH of 6.0, the sponge matrix is adjusted with 0.1 mol / L PBS buffer when immersed, and the added 5% CaCl2 solution has a liquid-to-solid ratio of 15:1 to the sponge.

[0029] A further improvement of the technical solution of the present invention is that: the medical zeolite powder is mixed with 0.5% chitosan acetate solution to form a suspension, and the suspension is evenly loaded on the surface of the sponge by a spraying method to obtain an antibacterial hemostatic sponge, comprising:

[0030] The sponge sprayed with the suspension was vacuumed to 10 Pa at -20°C for 4 hours to make the moisture content ≤3% to prevent ice crystals from damaging the pore structure. After drying, the dry sponge was placed in a double-layer polypropylene sterilization bag and placed on an irradiation conveyor belt for sterilization.

[0031] Beneficial effects

[0032] Compared with the existing technology, the beneficial effects of the present invention are: providing a biocompatible substrate and promoting cell adhesion through silk fibroin, and constructing a multi-level porous structure with a dense surface and loose interior of the sponge, accelerating liquid absorption and coagulation factor enrichment; in terms of antibacterial, the introduction of silver ions and chitosan produces a dual antibacterial mechanism, and improves the chelation of calcium ions with sodium alginate to enhance mechanical strength, while activating the platelet coagulation pathway, increasing the liquid absorption rate and liquid absorption capacity, and the silk fibroin and chitosan matrix can be completely enzymatically hydrolyzed within 28 days, and the degradation products are non-cytotoxic. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The figure is a schematic flow chart of the preparation method of the antibacterial hemostatic medical sponge. DETAILED DESCRIPTION

[0034] Various exemplary embodiments, features, and aspects of the present application will be described in detail below with reference to the accompanying drawings. The same reference numerals in the accompanying drawings represent elements with the same or similar functions. Although various aspects of the embodiments are shown in the accompanying drawings, the drawings are not necessarily drawn to scale unless otherwise indicated.

[0035] The word “exemplary” is used exclusively herein to mean “serving as an example, example, or illustration.” Any embodiment described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0036] In addition, numerous specific details are provided in the following specific examples to better illustrate the present application. Those skilled in the art will appreciate that the present application can be practiced without certain specific details. In some instances, methods, means, and components well known to those skilled in the art are not described in detail in order to highlight the main purpose of the present application.

[0037] The present invention provides a method for preparing an antibacterial hemostatic medical sponge, comprising:

[0038] S100, cleaning the silk cocoons after removing impurities, immersing them in a neutral protease solution to complete degumming, obtaining silk fibroin fibers, and then dissolving them in a LiBr solution for dialysis purification to obtain a silk fibroin solution; comprising:

[0039] S101, after the cocoons are chopped, they are placed in a titanium alloy tank of an ultrasonic cleaning machine and cleaned with a 0.5% NaHCO3 solution. They are ultrasonically treated at 60°C and 280W for 35 minutes to remove lipids and impurities. The cocoons are then rinsed with deionized water until neutral and filtered through a 300-mesh filter to drain.

[0040] S102, transferring the pretreated cocoons to a conical glass reactor equipped with a constant temperature oscillating water bath, adding 2% neutral protease solution, setting the water bath parameters to a constant temperature of 60°C and an oscillation frequency of 140 rpm, and reacting for 5 hours. After the reaction, the degummed silk fibroin fibers were separated using a centrifuge and washed three times with deionized water;

[0041] S103. Add the degummed silk fibroin fibers into a high-temperature resistant glass reactor, inject 9.3 mol / L LiBr solution at a ratio of 1:9 w / v, stir at 145 rpm, and dissolve in a 65°C water bath for 1 hour to form a light yellow viscous liquid. Transfer the solution into a dialysis bag, place it in a dialysis tank and stir at 40 rpm. Change the water every 3 hours and continue dialysis for 72 hours. After dialysis, centrifuge to remove undissolved impurities to obtain a silk fibroin solution.

[0042] The liquid-to-solid ratio of the added 0.5% NaHCO3 solution was 20:1;

[0043] The pH of the added 2% neutral protease solution was 7.0, and the liquid-to-solid ratio was 15:1.

[0044] Neutral protease specifically degrades sericin at pH 7.0, avoiding the breakage of silk fibroin chains caused by traditional alkaline methods. Furthermore, silk fibroin has excellent biocompatibility, mechanical properties, degradation properties, and plasticity. Therefore, this application uses silk fibroin as a substrate in the preparation of medical sponges.

[0045] S200, dissolving chitosan in 2% acetic acid solution, dissolving polyvinyl alcohol in 90° C. deionized water, adding sodium alginate and stirring, adding the obtained silk fibroin solution to form a chitosan-polyvinyl alcohol-sodium alginate-silk fibroin composite solution; comprising:

[0046] S201, chitosan powder was added to a polytetrafluoroethylene beaker, 2% acetic acid solution was injected at a concentration of 3% w / v, and the mixture was placed in a constant temperature water bath with a magnetic stirrer. The reaction parameters were set at 40°C and 500 rpm, and stirring was continued for 4 hours until completely dissolved. Undissolved particles were removed by filtering with a 200-mesh nylon filter cloth to obtain a transparent chitosan colloid;

[0047] S202, adding polyvinyl alcohol particles to a laminated glass reactor, adding deionized water at a concentration of 5% w / v, stirring, heating to 90° C. and maintaining for 1 hour to form a homogeneous transparent solution, placing in a high-temperature sterilizer at 121° C. for 20 minutes, and cooling to room temperature to obtain a polyvinyl alcohol solution;

[0048] S203. Transfer the transparent chitosan colloid and polyvinyl alcohol solution to a sandwich reactor in a volume ratio of 1:2. After stirring, mix at a constant temperature of 40°C for 30 minutes, slowly add SA powder to a final concentration of 1.5% w / v, continue stirring for 1 hour until no visible particles are present, add silk fibroin solution, and transfer the mixture to a titanium alloy tank of an ultrasonic cleaner for ultrasonic treatment at 40°C for 20 minutes to remove bubbles, thereby obtaining a composite solution.

[0049] The added silk fibroin solution: transparent chitosan colloid: polyvinyl alcohol solution = 1:1:2, and the final composite solution has a viscosity of ≥5000 mPa·s and a pH of 5.2-5.5.

[0050] S300, adding AgNO3 solution dropwise to the obtained composite solution to form a silver-containing composite solution colloid after reaction;

[0051] Adding AgNO3 solution dropwise to the obtained composite solution, the reaction forms a silver-containing composite solution colloid, comprising:

[0052] S301. Slowly add AgNO3 solution dropwise at a rate of 0.5 mL / min to a final concentration of 0.5 mmol / L. Turn on nitrogen protection, filter the composite solution through a 0.22 μm microporous filter membrane to remove large particle aggregates, and store in a light-shielding sealed container.

[0053] S400, placing the colloid in a high-pressure reactor, introducing CO2, maintaining the pressure and then rapidly releasing the pressure to foam, injecting the colloid into a mold for gradient freezing to form a multi-level pore size sponge matrix; including:

[0054] S401. Connect the composite solution colloid to a CO2 gas source, pressurize it to 8MPa at a rate of 1.5MPa / min, and maintain the pressure for 2 hours to allow CO2 to fully penetrate into the interior of the colloid, triggering rapid pressure relief. The pressure relief rate is greater than 15MPa / s, and the composite solution colloid forms a multi-level pore size. Inject the foamed colloid into a polytetrafluoroethylene mold pre-cooled to 4°C, and reduce the temperature from 4°C to -20°C at a rate of -1°C / min, and maintain it for 4 hours to directionally freeze the micropores on its surface. Reduce the temperature from -20°C to -80°C at a rate of -5°C / min, and maintain it for 2 hours to fix the internal macroporous structure, thereby obtaining a multi-level pore sponge matrix.

[0055] The rapid pressure relief of >15MPa / s induces the separation of CO2 gas-liquid phases, forming a gradient structure with dense surface and loose interior, and the double pore formation increases its water absorption rate.

[0056] S500, immersing the sponge base in a 1% genipin ethanol solution, then in a 5% CaCl2 solution, and rinsing with deionized water to obtain a calcified antibacterial sponge;

[0057] S501. Immerse the sponge substrate in a 1% genipin ethanol solution with a solution volume to sponge mass ratio of 10:1 v / w, and shake at 60 rpm at 25°C for 12 hours. Transfer the sponge to a 5% CaCl2 solution, let it stand for 1 hour, then use a centrifugal dehydrator to centrifuge at 800 rpm to remove the residual liquid on the surface, and rinse with deionized water three times.

[0058] The added 1% genipin ethanol solution has a pH of 6.0. The sponge matrix is adjusted with 0.1 mol / L PBS buffer when immersed. The added 5% CaCl2 solution has a liquid-to-solid ratio of 15:1 to the sponge.

[0059] S600, mixing medical zeolite powder and 0.5% chitosan acetate solution into a suspension, and uniformly loading the suspension onto the surface of a sponge by spraying to obtain an antibacterial hemostatic sponge, comprising:

[0060] S601. The sponge sprayed with the suspension is vacuumed to 10 Pa at -20°C and maintained for 4 hours to make the moisture content ≤3% to prevent ice crystals from damaging the pore structure. After drying, the dried sponge is placed in a double-layer polypropylene sterilization bag and placed on an irradiation conveyor belt for sterilization.

[0061] It should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus.

[0062] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing an antibacterial hemostatic medical sponge, characterized in that: include: The silk cocoons are cleaned after being decontaminated, immersed in a neutral protease solution to complete degumming, and the silk fibroin fibers are obtained, which are then dissolved in a LiBr solution and dialyzed to obtain a silk fibroin solution; Chitosan was dissolved in 2% acetic acid solution, polyvinyl alcohol was dissolved in 90°C deionized water, sodium alginate was added and stirred, and the obtained silk fibroin solution was added to form a composite solution; Adding AgNO3 solution dropwise to the obtained composite solution to form a silver-containing composite solution colloid after reaction; The colloid is placed in a high-pressure reactor, CO2 is introduced, the pressure is maintained and then the pressure is quickly released to foam, and then injected into a mold for gradient freezing to form a multi-level pore size sponge matrix; The sponge matrix was immersed in 1% genipin ethanol solution, then in 5% CaCl2 solution, and rinsed with deionized water to obtain a calcified antibacterial sponge. Medical zeolite powder and 0.5% chitosan acetate solution are mixed into a suspension, and the suspension is evenly loaded on the surface of the sponge by a spraying method to obtain an antibacterial hemostatic sponge.

2. The method for preparing an antibacterial hemostatic medical sponge according to claim 1, wherein: The silk cocoons are cleaned after being decontaminated, immersed in a neutral protease solution to complete degumming, and the silk fibroin fibers are obtained, and then dissolved in a LiBr solution for dialysis purification to obtain a silk fibroin solution, comprising: The cocoons were chopped and placed in a titanium alloy tank of an ultrasonic cleaning machine. They were cleaned with 0.5% NaHCO3 solution and ultrasonically treated at 60°C and 280W for 35 minutes to remove lipids and impurities. They were then rinsed with deionized water until neutral and drained through a 300-mesh filter. The pretreated cocoons were transferred to a conical glass reactor, and 2% neutral protease solution was added. The reaction parameters were set at a constant temperature of 60°C and an oscillation frequency of 140 rpm. The reaction was carried out for 5 hours. After the reaction, the degummed silk fibers were separated and washed with deionized water three times. Degummed silk fibroin fibers were injected with 9.3 mol / L LiBr solution at a ratio of 1:9 w / v, stirred at 145 rpm, and dissolved in a 65°C water bath for 1 hour to form a light yellow viscous liquid. The solution was transferred to a dialysis bag and placed in a dialysis tank with stirring at 40 rpm. The water was changed every 3 hours and the dialysis was continued for 72 hours. After dialysis, the undissolved impurities were removed by centrifugation to obtain a silk fibroin solution.

3. The method for preparing an antibacterial hemostatic medical sponge according to claim 2, wherein: The liquid-to-solid ratio of the added 0.5% NaHCO3 solution was 20:1; The pH of the added 2% neutral protease solution was 7.0, and the liquid-to-solid ratio was 15:

1.

4. The method for preparing an antibacterial hemostatic medical sponge according to claim 1, wherein: The method comprises dissolving chitosan in a 2% acetic acid solution, dissolving polyvinyl alcohol in 90° C. deionized water, adding sodium alginate and stirring, and adding the obtained silk fibroin solution to form a composite solution, comprising: Chitosan powder was injected into 2% acetic acid solution at a concentration of 3% w / v, and the reaction parameters were set at 40°C and 500 rpm. Stirring was continued for 4 hours until it was completely dissolved. Undissolved particles were removed by filtering with 200-mesh nylon filter cloth to obtain a transparent chitosan colloid. Add polyvinyl alcohol particles to deionized water at a concentration of 5% w / v, stir, heat to 90°C and maintain for 1 hour to form a homogeneous transparent solution, sterilize at 121°C for 20 minutes, and cool to room temperature to obtain a polyvinyl alcohol solution; The transparent chitosan colloid and polyvinyl alcohol solution were transferred to a sandwich reactor in a volume ratio of 1:

2. After stirring, they were mixed at a constant temperature of 40°C for 30 minutes. SA powder was slowly added to a final concentration of 1.5% w / v, and the silk fibroin solution was added. The mixture was ultrasonically treated at 40°C for 20 minutes to remove bubbles to obtain a composite solution.

5. The method for preparing an antibacterial hemostatic medical sponge according to claim 4, characterized in that: The added silk fibroin solution: transparent chitosan colloid: polyvinyl alcohol solution = 1:1:2, and the final composite solution has a viscosity of ≥5000 mPa·s and a pH of 5.2-5.

5.

6. The method for preparing an antibacterial hemostatic medical sponge according to claim 1, characterized in that: The step of adding AgNO3 solution dropwise to the obtained composite solution to form a silver-containing composite solution colloid after reaction comprises: AgNO3 solution was slowly added dropwise at a rate of 0.5 mL / min to a final concentration of 0.5 mmol / L. Nitrogen protection was turned on, and the composite solution was filtered through a 0.22 μm microporous filter membrane to remove large particle aggregates and stored in a light-shielding sealed container.

7. The method for preparing an antibacterial hemostatic medical sponge according to claim 1, characterized in that: The colloid is placed in a high-pressure reactor, CO2 is introduced, the pressure is maintained and then the pressure is quickly released to foam, and the colloid is injected into a mold for gradient freezing to form a multi-level pore size sponge matrix, including: The composite solution colloid was connected to a CO2 gas source, pressurized to 8 MPa at a rate of 1.5 MPa / min, maintained at pressure for 2 hours, and then rapid pressure relief was triggered. The foaming colloid was injected into a mold pre-cooled to 4°C, and the temperature was lowered from 4°C to -20°C at a rate of -1°C / min, maintained for 4 hours, and then lowered from -20°C to -80°C at a rate of -5°C / min, maintained for 2 hours to obtain a multi-level pore size sponge matrix.

8. The method for preparing an antibacterial hemostatic medical sponge according to claim 1, characterized in that: The method comprises: immersing the sponge base in a 1% genipin ethanol solution, then immersing the sponge base in a 5% CaCl2 solution, and then rinsing the sponge base with ionized water to obtain a calcified antibacterial sponge. The sponge substrate was immersed in 1% genipin ethanol solution with a solution volume to sponge mass ratio of 10:1 v / w, and shaken at 60 rpm at 25°C for 12 hours. The sponge was transferred to 5% CaCl2 solution, allowed to stand for 1 hour, and then centrifuged to remove the residual liquid on the surface, and rinsed with deionized water three times.

9. The method for preparing an antibacterial hemostatic medical sponge according to claim 8, characterized in that: The pH of the added 1% genipin ethanol solution was 6.0, and the sponge matrix was adjusted with 0.1 mol / L PBS buffer when immersed. The liquid-to-solid ratio of the added 5% CaCl2 solution to the sponge was 15:

1.

10. The method for preparing an antibacterial hemostatic medical sponge according to claim 1, characterized in that: The medical zeolite powder and 0.5% chitosan acetate solution are mixed into a suspension, and the suspension is evenly loaded on the surface of the sponge by a spraying method to obtain an antibacterial hemostatic sponge, comprising: The sponge sprayed with the suspension was vacuumed to 10 Pa at -20°C for 4 hours to make the moisture content ≤3%. After drying, the dried sponge was placed in a double-layer polypropylene sterilization bag and placed on an irradiation conveyor belt for sterilization.

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