Antibacterial cross-linked nanofiber membrane based on rabbit skin and hair proteins and method for preparing the same

Abstract

The application provides an antibacterial crosslinked nanofiber film based on rabbit skin and hair protein and a preparation method thereof, and the preparation method comprises the following steps: rabbit skin gelatin extraction; rabbit hair keratin extraction; rabbit hair keratin solution dialysis purification; rabbit hair keratin powder preparation; nanosilver dispersion liquid preparation; ice acetic acid-nanosilver dispersion liquid preparation; nanospinning liquid preparation; NHS / EDC composite crosslinking agent preparation; and silver-doped gelatin / protein crosslinked composite nanofiber film preparation. The application constructs a three-phase composite nanofiber film material of rabbit hair keratin-rabbit skin gelatin-silver by using a crosslinking reaction. Not only is the bottleneck of poor antibacterial performance of the protein matrix nanofiber film broken, but also the problem of too strong hydrophilicity of the protein matrix nanofiber film is solved, and a neutral quasi-two-dimensional material between hydrophilicity and hydrophobicity is obtained. The air permeability of the material is ensured, and the nanofiber is more easily adapted to the matrix due to the similar fiber structure of the nanofiber and human skin.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of new materials, in particular to an antibacterial cross-linked nanofiber membrane based on rabbit skin and hair protein and a preparation method thereof. BACKGROUND

[0002] Wounds are defined as the phenomenon of skin or mucosal epidermis layer continuity interruption caused by physical or thermal damage, and the complexity of the healing process brings challenges to the clinical treatment of acute and chronic skin damage. Therefore, it is of great significance to develop innovative wound dressings with skin repair function to promote the wound healing process.

[0003] Nanofiber membranes have excellent biocompatibility and physical properties due to their unique three-dimensional structure and high porosity, making them advanced biomaterials in the field of medical dressings. Small pore size design helps gas exchange and limits microbial penetration, effectively reducing the risk of infection.

[0004] When selecting nanofiber membrane materials that can be used as dressings, attention should be paid to the mechanical strength, plasticity of the material, and whether it has natural cell binding sites to ensure that cells can adhere to it and accelerate cell growth and repair process. Today, nanofiber membranes prepared using traditional polymer materials such as polycaprolactone (PCL), polyvinyl alcohol (PVA), polyethylene glycol (PEG), and polylactic acid (PLA) already have good mechanical strength and plasticity, but they often lack natural cell binding sites, which can affect cell adhesion and growth. One of the research hotspots is to find a natural polymer material with good mechanical strength and plasticity, as well as a sufficient number of natural cell binding sites, as the base material for nanofiber membranes. SUMMARY

[0005] The present application aims to provide an antibacterial cross-linked nanofiber membrane based on rabbit skin and hair protein and a preparation method thereof to solve at least one of the above technical problems in the prior art.

[0006] To solve the above technical problems, the present application provides a preparation method of an antibacterial cross-linked nanofiber membrane based on rabbit skin and hair protein, which comprises the following steps:

[0007] S10. Extracting rabbit skin gelatin;

[0008] S20. Extracting rabbit hair keratin;

[0009] Mixing the rabbit hair with the cysteine reagent at a bath ratio of 1:30 to obtain a mixed solution;

[0010] Adjusting the pH value of the mixed solution to the range of 8-12 using NaOH solution;

[0011] In 85 0 C, nitrogen was introduced into the mixed solution under air-isolated condition, and the reaction was carried out for 1-10 hours, and then the rabbit hair keratin solution was obtained by centrifugal filtration;

[0012] S30. Dialysis purification of the rabbit hair keratin solution

[0013] The rabbit hair keratin solution was placed in a dialysis bag with a cut-off of 8000-14000, and dialysis treatment was carried out at 20 0 C to 40 0 C until the rabbit hair keratin solution became white;

[0014] S40. Preparation of rabbit hair keratin powder

[0015] The rabbit hair keratin solution purified in step S30 was mixed with polyethylene glycol 20000, and dehydration was carried out at 4°C, and the dehydrated product was freeze-dried using a freeze-drying machine to obtain keratin powder;

[0016] S50. Preparation of silver nano-dispersion

[0017] The pH of the carboxymethyl chitosan solution was adjusted to 12 using NaOH, and silver nitrate solution was added dropwise under continuous uniform stirring at 60-80°C to obtain silver nano-dispersion;

[0018] S60. Preparation of silver nano-dispersion

[0019] Ice acetic acid was dissolved in the silver nano-dispersion prepared in step S50, and the mass ratio of ice acetic acid in the solution was 80%, and then ice acetic acid-silver nano-dispersion was obtained;

[0020] S70. Preparation of silver-gelatin-protein three-phase nano-spinning solution

[0021] Keratin powder and gelatin were dissolved in the ice acetic acid-silver nano-dispersion, and the concentration of keratin in the ice acetic acid-silver nano-dispersion was 12-22%; the mass ratio of gelatin to keratin powder was 1:1 to 6:1;

[0022] Finally, silver-gelatin-protein three-phase nano-spinning solution was obtained;

[0023] S80. Preparation of NHS / EDC composite crosslinking agent:

[0024] EDC and NHS were dissolved in anhydrous ethanol respectively to prepare NHS / EDC composite crosslinking agent; the mass ratio of EDC to NHS in the NHS / EDC composite crosslinking agent was 1:1 to 1:5, and the content of EDC in the NHS / EDC composite crosslinking agent was 50-400 millimoles per liter;

[0025] S90. Preparation of silver-doped gelatin / protein cross-linked composite nanofiber membrane

[0026] The silver-gelatin-protein three-phase nanospinning solution and the NHS / EDC composite cross-linking agent are respectively placed in two syringes, and electrospinning is performed by using an electrospinning device to obtain a silver-doped gelatin / protein cross-linked composite nanofiber membrane.

[0027] Further, in step S20, the concentration of the cysteine reagent is 3-10% / L.

[0028] Preferably, the concentration of the cysteine reagent is 5%.

[0029] Further, in step S20, the concentration of the NaOH solution is 8-15 mol / L; preferably, 10 mol / L.

[0030] Further, in step S50, the concentration of the carboxymethyl chitosan solution is 0.1-0.2%. Preferably, 0.15%.

[0031] Further, in step S50, the carboxymethyl chitosan solution is continuously stirred at a uniform speed at 70°C.

[0032] Further, in step S50, the mass ratio of the silver nitrate solution is 1.7%, and the pH value is 12.

[0033] Further, the following steps are further included:

[0034] S11. The fur-bearing rabbit skin is soaked in a 0.05-0.3 g / mL aqueous inorganic salt solution at a solid-liquid ratio of 1:10 to 1:50 for 12-48 h.

[0035] After the subcutaneous fat is scraped off, the water is drained;

[0036] A paste is obtained by mixing a 3%-9% aqueous inorganic sulfide solution with Ca(OH)2 solid powder, the paste is uniformly coated on the fur-bearing rabbit skin, and the skin is placed at 10-40°C for 2-5 h, and then washed with water to separate the skin from the fur.

[0037] Further, the following steps of rabbit skin pretreatment are further included:

[0038] S12. The rabbit skin after the fur is removed is soaked in a 0.005-0.05 g / mL aqueous inorganic salt solution at a solid-liquid ratio of 1:2 to 1:7, soaked for 1-10 h, then washed with pure water to remove the residual inorganic salt on the surface of the rabbit skin, and then dried at 25°C.

[0039] Further, step S10 further includes the following steps:

[0040] S13. The pretreated rabbit skin is added into the hydrochloric acid solution with a material-to-liquid ratio of 1:2 to 1:7, soaked for 6 to 15 minutes and then taken out; the mass concentration of the hydrochloric acid solution is 0.5% to 2%;

[0041] The rabbit skin is washed with distilled water until the pH of the surface of the rabbit skin is 7.

[0042] Further, the step S10 further comprises the following steps:

[0043] S14. The rabbit skin treated in the step S13 is soaked in deionized water, and then hydrochloric acid is poured into the deionized water until the pH of the deionized water is adjusted to 3-5;

[0044] The water bath oscillation is performed at 60℃ for 4 to 7 hours.

[0045] The supernatant is collected after centrifugation, and the supernatant is subjected to water removal at 60℃ to obtain the solid rabbit skin gelatin.

[0046] Further, the step S20 comprises the following steps:

[0047] S21. Rabbit hair pretreatment

[0048] The rabbit hair is soaked in anhydrous ethanol for 20-60 minutes to remove soluble impurities such as fat, and then washed with distilled water and dried.

[0049] Further, the inorganic salt aqueous solution is one or more of sodium chloride, magnesium chloride and potassium chloride.

[0050] Further, in the step S90, the working parameters during electrospinning are as follows: voltage 15-30 kV; distance between the receiving device and the spinning port 10-30 cm; extrusion rate 0.2-2 mL / h; temperature 20-30℃; and relative humidity 5-50%.

[0051] Further, the inorganic sulfide aqueous solution is one or more of sodium sulfide, potassium sulfide and magnesium sulfide.

[0052] The second aspect of the present application discloses an antibacterial cross-linked nanofiber membrane prepared by the above preparation method.

[0053] By adopting the above technical solution, the present application has the following beneficial effects:

[0054] The application provides an antibacterial cross-linked nanofiber membrane based on rabbit skin and hair protein and a preparation method thereof.

[0055] Compared with existing protein-based nanofiber membrane materials, the application has the following beneficial effects:

[0056] 1) The application uses discarded rabbit hair and rabbit skin, thereby reducing the generation of industrial waste and providing an environmentally friendly and sustainable medical dressing design scheme, which meets the current global trend of environmental protection and sustainable development.

[0057] 2) The application forms a three-phase system by using nanosilver ions, rabbit skin gelatin and rabbit hair keratin, thereby increasing the antibacterial performance of the material and further increasing the "natural cell binding site". Meanwhile, the doping of nanoparticles increases the roughness of the surface of the nanofiber to some extent and increases the adsorption performance of the overall material.

[0058] 3) The application adds a cross-linking agent, thereby overcoming the problem that pure protein nanofibers are too hydrophilic and have a large solubility in water. The nanofiber membrane has certain hydrophobic properties, can still maintain a skin-like structure with gaps and fiber morphology in a wet state, and has excellent mechanical properties.

[0059] 4) The application greatly improves the added value of waste, creates new economic benefits, reduces the production cost of natural protein nanofiber membrane dressings, and has great market application potential. BRIEF DESCRIPTION OF DRAWINGS

[0060] In order to more clearly illustrate the specific embodiments of the application or the technical solutions in the prior art, the following will briefly introduce the drawings needed to be used in the specific embodiments or prior art description. Obviously, the drawings in the following description are some embodiments of the application, and those skilled in the art can also obtain other drawings according to these drawings without creative labor.

[0061] Figure 1 SEM picture of the application example 1;

[0062] Figure 2 SEM picture of the application example 2;

[0063] Figure 3 SEM picture of Example 3 of the present application;

[0064] Figure 4 SEM picture of Example 4 of the present application;

[0065] Figure 5 SEM picture of Example 5 of the present application;

[0066] Figure 6 SEM picture of Comparative Example 1;

[0067] Figure 7 Comparison chart of antibacterial test results of Example 1 and Comparative Example 1. DETAILED DESCRIPTION

[0068] The present application will be further explained in conjunction with specific embodiments.

[0069] Example 1:

[0070] 1) After the fur of the meat rabbit skin was washed to remove dirt, the fur of the meat rabbit skin was placed in a 0.1 g / mL NaCl aqueous solution at a material-liquid ratio of 1:20 and soaked for 24 h. After the subcutaneous fat was scraped off, the water was drained. A 5% inorganic sulfide aqueous solution was mixed with an appropriate amount of solid Ca(OH)2 powder to form a paste, which was then uniformly coated on the fur of the meat rabbit skin. The coated fur of the meat rabbit skin was placed in a 25 0 C for 4 h, and then water-washed to separate the skin and fur of the meat rabbit.

[0071] 2) Rabbit skin pretreatment: the rabbit skin was soaked in a 0.01 g / mL NaCl aqueous solution at a material-liquid ratio of 1:5, and then washed with pure water to remove the residual inorganic salt on the surface. The rabbit skin was dried in a 25 0 C oven.

[0072] 3) Rabbit skin gelatin extraction: the pretreated rabbit skin was soaked in a 1% hydrochloric acid solution at a material-liquid ratio of 1:6 for 10 min, and then taken out and washed with distilled water until the surface pH was 7.

[0073] The washed sample was soaked in deionized water at a mass ratio of 1:3, and the pH was adjusted to 4 using hydrochloric acid. The sample was subjected to water bath oscillation at 60 0 C for 6 h. After the reaction was completed, the sol system was subjected to high-speed centrifugation, and the supernatant was collected. The supernatant was dried at 60 0 C, and the solid gelatin sample was obtained.

[0074] 4) Rabbit fur pretreatment: the rabbit fur was soaked in anhydrous ethanol for 30 min to remove fat and other soluble impurities, and then washed with distilled water and dried in a drying machine.

[0075] 5) Rabbit hair keratin extraction: the pretreated rabbit hair was mixed with a mixed solution of 5% L-cysteine and 8 mol / L urea at a bath ratio of 1:30, the pH value was adjusted to the range of 10.5 using a 10 mol / L NaOH solution, and the mixture was reacted at 60 0 C under the condition of air isolation and nitrogen introduction for 5 hours, and finally the rabbit hair keratin solution was obtained by centrifugal filtration.

[0076] 6) Dialysis purification of rabbit hair keratin solution: the rabbit hair keratin solution was placed in a dialysis bag with a cut-off of 8000-14000, and the water was changed every 6 hours under the condition of 25 0 C until the solution became white.

[0077] 7) Preparation of rabbit hair keratin powder: the purified keratin solution was mixed with polyethylene glycol 20000, and then dehydrated at 4°C, and the dehydrated product was freeze-dried using a freeze-drying machine to obtain keratin powder. The amount of polyethylene glycol 20000 was about 15-18% of the keratin solution.

[0078] 8) Preparation of silver nanodispersion: a 0.15% carboxymethyl chitosan solution was prepared, the pH of the solution was adjusted to 12 with NaOH, and the solution was continuously stirred at 70°C. Slowly add a 1.7% silver nitrate solution with a mass ratio of 1.7% and pH=12, and continuously stir at constant temperature for 1 hour to complete the preparation of the silver nanodispersion.

[0079] 9) Preparation of glacial acetic acid-silver nanodispersion: dissolve glacial acetic acid in the silver nanodispersion, and the mass ratio of glacial acetic acid in the solution is 80%.

[0080] 10) Preparation of silver-gelatin-protein three-phase nanospinning solution: dissolve gelatin and keratin in the glacial acetic acid-silver nanodispersion prepared in step 9 at a mass ratio of 3:1 to prepare a spinning solution with a total protein concentration of 16%.

[0081] 11) Preparation of NHS / EDC composite crosslinking agent: dissolve EDC and NHS in anhydrous ethanol respectively, prepare a composite crosslinking agent solution with a mass ratio of EDC to NHS of 1:2, and at the same time keep the content of EDC in the composite crosslinking agent solution at 200 millimoles per liter.

[0082] 12) Preparation of silver-doped gelatin / protein cross-linked composite nanofiber membrane: the spinning solution and the composite crosslinking agent were placed in two adjacent syringes of the same height, and the silver-doped gelatin / protein cross-linked composite nanofiber membrane was obtained by electrospinning.

[0083] The hydrophilic and hydrophobic properties of the nanofiber membrane were evaluated according to the national standard "GB / T 14210" using a JC2000DM contact angle measuring instrument. According to the test standard GB4789.3-2016, the antibacterial properties of different concentrations of silver-containing nanofiber membranes and the blank control group against gram-negative (Escherichia coli) and gram-positive (Staphylococcus aureus) were explored by plate counting method.

[0084] At the same time, the mechanical properties of the nanofiber membrane were evaluated using an Instron 5569. The sample size was 10 mm wide, 30 mm long, and 2 mm thick. The tensile test was performed at a strain rate of 10 mm / min.

[0085] Example 2

[0086] This example is basically the same as Example 1, except that:

[0087] The content of EDC in the composite crosslinking agent solution in step 11 of this example is 50 millimoles per liter.

[0088] Example 3

[0089] This example is basically the same as Example 1, except that:

[0090] The content of EDC in the composite crosslinking agent solution in step 11 of this example is 400 millimoles per liter.

[0091] Example 4

[0092] This example is basically the same as Example 1, except that:

[0093] The mass ratio of EDC to NHS in step 11 of this example is 1:1.

[0094] Example 5

[0095] This example is basically the same as Example 1, except that:

[0096] The mass ratio of EDC to NHS in step 11 of this example is 1:3.

[0097] Comparative Example 1:

[0098] The main difference between this example and Example 1 is that it is not treated with a crosslinking agent. That is, step 11 is removed, and the remaining parameters remain unchanged.

[0099] The test results show that the performance of the samples treated in the examples of the present application is shown in the following table.

[0100]

[0101] The results of the above table show that the tensile strength and elongation at break of Example 1 described in the present application are both more than one time the value of Comparative Example 1. The initial contact angle has also been significantly improved, especially the contact angle after 1 second, with a significant difference. At the same time, as a hydrophilic or even water-soluble protein nanofiber membrane, the finished product prepared by the preparation method described in the present application has a significant change in the contact angle before and after 1 second. It can be seen that the example described in the present application has good mechanical properties and certain hydrophobic properties.

[0102] By comparing Figure 1 , Figure 4 , Figure 5 and Figure 6 , it can be seen that when the crosslinking agent concentration is less than 50 mmol / L, the crosslinking degree is not high, and the electron microscope has no big difference with Comparative Example 1 without using crosslinking agent. When the crosslinking agent concentration is 200 mmol / L, the crosslinking effect appears, the fibers form a network with obvious pores between the fibers, and the fibers have obvious fiber morphology. When the crosslinking agent continues to exceed 400 mmol / L, the surface pores disappear, forming a plane.

[0103] By comparing Figure 1 , Figure 2 , Figure 3 and Figure 6 , it can be found that in Comparative Example 1, the nanofibers in the sample are independent of each other and have no correlation between each other. In Example 2, when NHS:EDC is 1:1, partial crosslinking occurs in the nanofiber membrane, and part still has the characteristics of individual fibers, which is due to the low molar ratio of crosslinking agent, which limits the mobility of crosslinking agent and gelatin chain in the fiber, thereby resulting in low crosslinking efficiency. In Example 1, when EDC:NHS is 1:2, crosslinking occurs between the fibers of the nanofiber membrane, while the morphology of the nanofiber is maintained, forming a fibrous reticular structure similar to skin fibers. In Example 5, when NHS:EDC is 3:1, the morphology of the nanofiber has been destroyed, and the pore structure cannot be seen.

[0104] Comparative Example 2:

[0105] The main difference compared with Example 1 is that there is no silver nanoparticle doped in the fiber membrane. Step 8 is cancelled, and the same amount of distilled water is used instead of the original silver nanoparticle dispersion in step 9.

[0106] Figure 7 It can be clearly seen that the antibacterial performance of the sample of Comparative Example 2 is much lower than that of Example 1 described in the present application for both Staphylococcus aureus and Escherichia coli. It shows that the nanofiber membrane described in the present application not only does not weaken the antibacterial performance due to the addition of protein, but also has obvious improvement in antibacterial performance.

[0107] The above conclusion shows that the silver-doped gelatin / protein antibacterial cross-linked nanofiber membrane based on natural meat rabbit fur and the preparation method thereof can greatly improve the strength, hydrophobicity and antibacterial performance of the protein-based nanofiber membrane on the basis of reducing waste of meat rabbit fur in a pure green and environmentally friendly way, and has a good application prospect.

[0108] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present application, but not to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for preparing a rabbit skin, hair protein-based antibacterial cross-linked nanofiber membrane, characterized by, The method comprises the following steps: S10. Extracting rabbit skin gelatin; S20. Extracting rabbit hair keratin; Mixing the rabbit hair with the cysteine reagent at a bath ratio of 1:30 to obtain a mixed solution; Adjusting the pH value of the mixed solution to 10.5 by using a NaOH solution; In 60 0 Under the condition of C, under the condition of air isolation, nitrogen was introduced into the mixed solution, and the reaction was carried out for 5 hours, and then rabbit hair keratin solution was obtained by centrifugal filtration. S30. Dialysis purification of the rabbit hair keratin solution Placing the rabbit hair keratin solution in a dialysis bag with a cut-off of 8000-14000, and performing dialysis treatment at 25℃ until the rabbit hair keratin solution becomes white; S40. Preparing rabbit hair keratin powder Mixing polyethylene glycol 20000 with the rabbit hair keratin solution purified in step S30, and then dehydrating the mixture at 4℃, and then freeze-drying the dehydrated product by using a freeze dryer to obtain keratin powder; S50. Preparing a nano-silver dispersion Adjusting the pH value of a carboxymethyl chitosan solution to 12 by using NaOH, and then adding a silver nitrate solution drop by drop under the condition of uniform stirring at 70℃ to obtain a nano-silver dispersion; S60. Preparing an acetic acid-nano-silver dispersion Dissolving acetic acid in the nano-silver dispersion prepared in step S50, so that the mass ratio of acetic acid in the solution is 80%, and then obtaining an acetic acid-nano-silver dispersion; S70. Preparing a silver-gelatin-protein three-phase nano spinning solution Dissolving keratin powder and gelatin in the acetic acid-nano-silver dispersion, and the concentration of keratin in the acetic acid-nano-silver dispersion is 16%; the mass ratio of gelatin to keratin powder is 3:1; Finally, a silver-gelatin-protein three-phase nano spinning solution is obtained; S80. Preparing an NHS / EDC composite crosslinking agent Dissolving EDC and NHS in anhydrous ethanol respectively to obtain an NHS / EDC composite crosslinking agent; the mass ratio of EDC to NHS in the NHS / EDC composite crosslinking agent is 1:2, and the content of EDC in the NHS / EDC composite crosslinking agent is 200 millimoles per liter; S90. Preparing a silver-doped gelatin / protein crosslinked composite nanofiber membrane Placing the silver-gelatin-protein three-phase nano spinning solution and the NHS / EDC composite crosslinking agent in two syringes respectively, and then performing spinning by using an electrospinning device to obtain a silver-doped gelatin / protein crosslinked composite nanofiber membrane.

2. The production method according to claim 1, characterized by, In step S20, the concentration of the cysteine reagent is 5%.

3. The preparation method according to claim 1, characterized in that, In step S20, the concentration of the NaOH solution is 10 mol / L.

4. The method of claim 1, wherein, In step S50, the concentration of the carboxymethyl chitosan solution is 0.15%.

5. The preparation method according to claim 1, characterized in that, In step S50, the mass ratio of the silver nitrate solution is 1.7%, and the pH value is 12.

6. The method of claim 1, wherein, The method further comprises the following steps: S11. Placing the rabbit skin with hair in a 0.1 g / mL NaCl aqueous solution at a material-liquid ratio of 1:20, and soaking for 24 h; After removing the subcutaneous fat, draining the water; Mixing a 5% inorganic sulfide aqueous solution with Ca(OH)2 solid powder to obtain a paste, uniformly applying the paste on the rabbit skin with hair, and then placing the rabbit skin with hair at 25℃ for 4 h, and then washing the rabbit skin with water to separate the skin from the hair; S12. The rabbit skin after the rabbit hair is stripped is soaked in the NaCl aqueous solution with a concentration of 0.01 g / mL according to a material-to-liquid ratio of 1:5, and after soaking for 1-10 hours, the residual inorganic salt on the surface of the rabbit skin is removed by washing with pure water, and then the rabbit skin is dried at 25°C; S13. The pretreated rabbit skin is added into the hydrochloric acid solution according to a material-to-liquid ratio of 1:6, and after soaking for 10 minutes, the rabbit skin is taken out; the mass concentration of the hydrochloric acid solution is 1%; The rabbit skin is washed with distilled water until the pH of the surface of the rabbit skin is 7.

7. The preparation method according to claim 6, characterized in that, Step S10 further comprises the following steps: S14. The rabbit skin treated in step S13 is soaked in the deionized water, and then the hydrochloric acid is poured into the deionized water until the pH of the deionized water is adjusted to 4; The water bath is shaken at 60°C for 6 hours; After centrifugal treatment, the supernatant is collected; after removing the water at 60°C, the solid rabbit skin gelatin is obtained.

8. The method of claim 1, wherein, Step S20 comprises the following steps: S21. Rabbit hair pretreatment The rabbit hair is soaked in anhydrous ethanol for 30 minutes to remove soluble impurities, and then washed clean with distilled water, and then dried.

9. An antibacterial cross-linked nanofiber membrane prepared by the preparation method of any one of claims 1-8.

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