A method for preparing a silk fibroin-based hemostatic gel

Abstract

The application discloses a preparation method of a silk fibroin-based hemostatic gel, and comprises the following steps: (1) obtaining small-molecule silk fibroin by hydrolyzing and dialyzing regenerated silk fibroin; (2) modifying the small-molecule silk fibroin by using a modifier to obtain small-molecule modified silk fibroin; and (3) mixing the small-molecule modified silk fibroin with an auxiliary agent, a photoinitiator, modified gelatin and raspberry glycoside in a proportion, and then stirring and reacting in the dark to obtain the silk fibroin-based hemostatic gel. The silk fibroin-based hemostatic gel prepared by the method has the advantages of rapid hemostasis and good adhesion, and can be widely applied in the fields of wound hemostasis and the like.

Description

Technical Field

[0001] This invention relates to the field of hydrogel technology, and more specifically, to a method for preparing a silk fibroin-based hemostatic gel. Background Technology

[0002] Hemostasis is a crucial aspect of clinical emergency care and surgical treatment. Rapid and effective hemostatic materials can significantly reduce blood loss and lower the risk of complications such as infection and shock. Existing hemostatic materials mainly include fibrin glue, gelatin sponge, and chitosan dressings, but they have significant limitations: fibrin glue relies on plasma extraction, is susceptible to viral contamination, and has a long curing time; gelatin sponge easily detaches after absorbing water and swells, and its hemostatic effect is short-lived; while chitosan dressings have good biocompatibility, they have a slow hemostatic speed, and some patients may experience local allergic reactions.

[0003] Silk fibroin, as a natural protein, possesses excellent biocompatibility, biodegradability, and mechanical properties, making it a research hotspot in hemostatic materials in recent years. However, natural silk fibroin has a large molecular weight and poor solubility in aqueous solutions, making it difficult to rapidly diffuse into wound tissue, resulting in low hemostatic efficiency.

[0004] Although existing technologies attempt to reduce the molecular weight of silk fibroin through degradation, acid and alkali hydrolysis easily damage the secondary structure of silk fibroin, leading to a decrease in its biological activity. While enzymatic hydrolysis is gentler, how to precisely control the degree of enzymatic hydrolysis to obtain silk fibroin with a suitable molecular weight remains an unsolved problem. Existing hemostatic gels also have shortcomings in terms of adhesive properties.

[0005] In summary, existing silk fibroin-based hemostatic materials suffer from problems such as slow hemostasis due to excessively large molecular weight, complex cross-linking methods, poor ease of use, and poor adhesion. There is an urgent need to develop a hemostatic gel based on low molecular weight silk fibroin that can achieve rapid hemostasis, is easy to use, and has good adhesive properties. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing a silk fibroin-based hemostatic gel. This invention uses an enzymatic hydrolysis process to prepare low molecular weight silk fibroin, which has the characteristics of rapid hemostasis, convenient use, and good adhesiveness.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] A method for preparing a silk fibroin-based hemostatic gel includes the following steps: (1) regenerated silk fibroin is hydrolyzed and dialyzed to obtain small molecule silk fibroin; (2) the small molecule silk fibroin is modified with a modifier to obtain small molecule modified silk fibroin; (3) the small molecule modified silk fibroin is mixed with an auxiliary agent, a photoinitiator, modified gelatin, and raspberry glycoside in a certain proportion and stirred in the dark to obtain a silk fibroin-based hemostatic gel.

[0009] Further, the specific steps of step (1) are as follows: dissolve the regenerated silk fibroin in purified water to obtain an aqueous solution of silk fibroin, add neutral protease and adjust the pH value, then carry out a hydrolysis reaction, and then dialyze and freeze-dry the solution after the reaction to obtain small molecule silk fibroin.

[0010] Further, the preparation method of regenerated silk fibroin is as follows: Silkworm cocoons are added to a 0.05 mol / L Na₂CO₃ aqueous solution and boiled at 100°C for 25-35 minutes. After boiling, the cocoons are removed and placed in a freshly prepared Na₂CO₃ aqueous solution of equal volume and concentration and boiled at 100°C for 25-35 minutes. The cocoons are then removed, washed, and dried to obtain degummed silk. The degummed silk is then heated with a CaCl₂-C₂H₆O-H₂O solution in a water bath at 65-75°C with stirring for 4-5 minutes. h, after cooling and centrifugation, the supernatant was obtained. The supernatant was placed in a dialysis bag and dialyzed at 4℃ for 3 days to obtain regenerated silk fibroin. The ratio of silkworm cocoon to Na2CO3 aqueous solution was 0.5~2g:50~400mL; the molar ratio of CaCl2, C2H6O, and H2O was 1:2:8; the ratio of degummed silk to CaCl2-C2H6O-H2O solution was 0.1~2g:10~30mL; the molecular weight of regenerated silk fibroin was 10-130 kDa.

[0011] Furthermore, the concentration of the silk fibroin aqueous solution is 20-40 wt%; the neutral protease accounts for 4-6 wt% of the silk fibroin aqueous solution; the pH value is controlled at 4.5-6.5 using 0.1 mol / L NaOH solution; the reaction temperature is 30-50℃, the reaction time is 2-4 hours; the dialysis time is 48-72 hours; the freeze-drying temperature is -80 to -70℃, the vacuum degree is 1-10 Pa, and the time is 48-72 hours; the molecular weight of the small molecule silk fibroin is 1-9 kDa.

[0012] Further, step (2) specifically involves: dissolving small molecule silk fibroin in purified water to obtain a small molecule silk fibroin solution, adding 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide for stirring and reaction, then adding a modifier to react at room temperature, and dialyzing and freeze-drying the resulting solution to obtain small molecule modified silk fibroin.

[0013] Furthermore, the concentration of the small molecule silk fibroin solution is 10-30 wt%; 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide each account for 1-5 wt% of the small molecule silk fibroin solution, and the molar ratio of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to N-hydroxysuccinimide is 1:1; the stirring reaction time is 0.5-1 h; the modifier is one of glycidyl methacrylate, methacrylic anhydride, methacryloyl ethyl isocyanate, and methacryloyl chloride, and the modifier accounts for 5-15 wt% of the small molecule silk fibroin solution; the reaction time at room temperature is 12-24 h; the molecular weight of the dialysis bag is 1000-3500 Da, and the dialysis time is 48-72 h; the freeze-drying temperature is -80 to -70 °C, the vacuum degree is 1-10 Pa, and the drying time is 48-72 h.

[0014] Further, step (3) specifically involves dissolving small molecule modified silk fibroin in purified water to obtain a small molecule modified silk fibroin solution, adding auxiliaries, photoinitiators, modified gelatin, and raspberry glycosides, stirring in the dark to obtain a silk fibroin-based hemostatic gel.

[0015] Furthermore, the concentration of the small molecule modified silk fibroin solution is 10-30 wt%; the auxiliary agent is one or more of sodium hyaluronate, carboxymethyl chitosan, dopamine, polyvinyl alcohol, and carbomer, accounting for 5-10 wt% of the small molecule modified silk fibroin solution; the photoinitiator is one of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylphenylacetone, 2,4,6-trimethylbenzoyl-diphenylphosphine oxide, 2,2-dimethoxy-2-phenylacetophenone, lithium phenyl(2,4,6-trimethylbenzoyl)phosphate, and 2,4,6-trimethylbenzoyl-diphenylphosphine oxide, accounting for 0.1-1 wt% of the small molecule modified silk fibroin solution; the modified gelatin accounts for 1-2 wt% of the small molecule modified silk fibroin solution; the raspberry glycoside accounts for 0.5-1 wt% of the small molecule modified silk fibroin solution; and the reaction time is 12-24 h with stirring in the dark.

[0016] The beneficial effects of this invention are:

[0017] This invention utilizes small-molecule silk fibroin, which can accelerate the wound hemostasis process. This invention can be customized to suit the size and irregular shape of the wound, and is convenient to use with good adhesion. Attached Figure Description

[0018] Figure 1 This is a photograph of a syringe containing the silk fibroin-based hemostatic gel prepared in Example 1. Detailed Implementation

[0019] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0020] This invention discloses a method for preparing a silk fibroin-based hemostatic gel, comprising the following steps:

[0021] Step (1): Add the chopped silkworm cocoons to a 0.05 mol / L Na2CO3 aqueous solution and boil at 100℃ for 25-35 minutes (the ratio of silkworm cocoons to Na2CO3 aqueous solution is 0.5-2 g: 50-400 mL). After boiling, remove the cocoons and place them in a freshly prepared Na2CO3 aqueous solution of equal volume and concentration and boil at 100℃ for 25-35 minutes. Remove the cocoons, wash them with deionized water, and dry them to obtain degummed silk. Then, mix the degummed silk with a CaCl2-C2H6O-H2O solution (CaCl2-C2H6O-H2O solution). The molar ratio of Cl2, C2H6O, and H2O is 1:2:8; the volume ratio of degummed silk to CaCl2-C2H6O-H2O solution is 0.1~2g:10~30mL. The solution is heated in a water bath at 65~75℃ and stirred for 4~5h. After cooling, a silk fibroin solution is obtained. The silk fibroin solution is centrifuged to remove insoluble impurities, and the supernatant is obtained. The supernatant is placed in an MWC3500 dialysis bag and dialyzed in deionized water at 4℃ for 3 days to obtain regenerated silk fibroin (molecular weight 10-130kDa).

[0022] Step (2): Weigh the regenerated silk fibroin, add purified water, stir and dissolve at room temperature to prepare a silk fibroin aqueous solution with a concentration of 20-40 wt%; add neutral protease (neutral protease accounts for 4-6 wt% of the silk fibroin aqueous solution) to the silk fibroin aqueous solution, adjust the pH value to 4.5-6.5 with 0.1 mol / L NaOH solution, and then stir and react in a water bath at 30-50℃ for 2-4 h; transfer the reacted solution to a dialysis bag and dialyze for 48-72 h; after dialysis, freeze dry at -80 to -70℃ and 1-10 Pa for 48-72 h to obtain freeze-dried small molecule silk fibroin (molecular weight of 1-9 kDa).

[0023] Step (3): Weigh the lyophilized small-molecule silk fibroin, add purified water, stir and dissolve at room temperature to prepare a small-molecule silk fibroin solution with a concentration of 10-30 wt%; add 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide sequentially to the small-molecule silk fibroin solution (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide each account for 1-5 wt% of the small-molecule silk fibroin solution; the molar ratio of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide is 1:1) and stir for 0.5-1 hour, add a modifier (one of glycidyl methacrylate, methacrylic anhydride, methacryloyl ethyl isocyanate, or methacryloyl chloride; accounting for 5-15 wt% of the small-molecule silk fibroin solution), react for 12-24 hours, and then load the reacted solution into a 1000-3500 ml container. Dialysis was performed in a Da dialysis bag for 48-72 hours; after dialysis, the fibroin was freeze-dried at -80 to -70°C and 1-10 Pa for 48-72 hours to obtain freeze-dried small molecule modified silk fibroin.

[0024] Step (4): Weigh the lyophilized small-molecule modified silk fibroin, add purified water, stir and dissolve at room temperature to prepare a small-molecule modified silk fibroin solution with a concentration of 10-30 wt%. Then add one or more of the following additives (sodium hyaluronate, carboxymethyl chitosan, dopamine, polyvinyl alcohol, carbomer; accounting for 5-10 wt% of the small-molecule modified silk fibroin solution) and photoinitiators [2-hydroxy-4'-(2-hydroxyethoxy)-2-methylphenylacetone (photoinitiator 2959) and 2,4,6-trimethylbenzoyl-diphenylphosphine oxide (photoinitiator 819)]. One of the following: 2,2-dimethoxy-2-phenylacetophenone (photoinitiator 651), lithium phenyl (2,4,6-trimethylbenzoyl) phosphate (LAP blue light initiator), or 2,4,6-trimethylbenzoyl-diphenylphosphine oxide (photoinitiator TPO-L); comprising 0.1-1 wt% of the small molecule modified silk fibroin solution; modified gelatin (comprising 1-2 wt% of the small molecule modified silk fibroin solution); and raspberry glycoside (comprising 0.5-1 wt% of the small molecule modified silk fibroin solution); reacted with stirring in the dark for 12-24 h to obtain a silk fibroin-based hemostatic gel.

[0025] The prepared silk fibroin-based hemostatic gel can be used in the following two ways:

[0026] (1) It can be applied directly to the wound using a syringe.

[0027] (2) Take medical tape, cut it, peel off the release paper, and place it on a hot table at 80-100℃ for 30-60 minutes; place a hollow mold in the middle of the tape, pour in silk fibroin-based hemostatic gel, and heat at 40-80 W / cm 2Cured and set under ultraviolet light for 20-40 seconds.

[0028] The preferred embodiment is as follows:

[0029] Example 1

[0030] Step (1): Add 30g of chopped silkworm cocoons to 6000mL of 0.05mol / L Na2CO3 aqueous solution and boil at 100℃ for 30 minutes. After removing the cocoons, boil them in a freshly prepared Na2CO3 aqueous solution of equal volume and concentration at 100℃ for 30 minutes. Remove the cocoons, wash them with deionized water, and dry them to obtain degummed silk.

[0031] 21g of degummed silk was mixed with 315mL of CaCl2-C2H6O-H2O solution (the molar ratio of CaCl2, C2H6O, and H2O was 1:2:8) and heated in a water bath at 70℃ for 4 hours with stirring. After cooling, a silk fibroin solution was obtained. The silk fibroin solution was centrifuged to remove insoluble impurities, and the supernatant was obtained. The supernatant was placed in an MWC3500 dialysis bag and dialyzed in deionized water at 4℃ for 3 days to obtain regenerated silk fibroin (molecular weight 30kDa).

[0032] Step (2): In a 100mL flask, add 12g of regenerated silk fibroin and 28g of purified water, stir to dissolve, then add 2g of neutral protease (enzyme activity of 200,000 U / g; purchased from Beijing Solarbio Science & Technology Co., Ltd., catalog number Z8032). Adjust the pH value to 5.0 with 0.1mol / L NaOH solution, then place the flask in a 37℃ water bath and heat and stir for 2h. Transfer the reaction solution to a 10 kDa dialysis bag and dialyze for 72h. After dialysis, freeze-dry at -80℃ and 1Pa for 72h to obtain freeze-dried small molecule silk fibroin (molecular weight of 7kDa).

[0033] Step (3): Add 5g of lyophilized small molecule silk fibroin and 45g of purified water to a 100mL flask and stir to dissolve at room temperature; after the small molecule silk fibroin has dissolved, add 1g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 0.74g of N-hydroxysuccinimide in sequence; after stirring for half an hour, add 2.5g of glycidyl methacrylate, react for 24 hours, and then put the solution into a 3500 Da dialysis bag and dialyze for 72 hours; after dialysis, freeze-dry for 72 hours at -80℃ and 1Pa to obtain lyophilized small molecule modified silk fibroin.

[0034] Step (4): Add 2g of lyophilized small molecule modified silk fibroin and 8g of purified water to a 20mL beaker, stir and dissolve at room temperature, then add 0.5g of sodium hyaluronate, 0.5g of dopamine, 0.05g of LAP blue light initiator, 0.2g of modified gelatin [prepared according to Preparation Example 1 in the patent "A hemostatic gel and its preparation method and application" (CN117414459A)] and 0.1g of raspberry glycoside (purchased from Shaanxi Chenxi Biotechnology Co., Ltd., catalog number CXSWSMG), stir in the dark for 20h to obtain silk fibroin-based hemostatic gel.

[0035] Comparative Example 1

[0036] Steps (1), (2), and (3) are the same as in Example 1. In step (4), 0.5g of sodium hyaluronate and 0.5g of dopamine are removed to form pure silk fibroin gel.

[0037] Comparative Example 2

[0038] Skip steps (1), (2), and (3); change step (4) to: add 1g of sodium hyaluronate and 1g of dopamine, and 8g of purified water to a 20mL beaker, stir and dissolve at room temperature to form a gel.

[0039] Comparative Example 3

[0040] Remove steps (1) and (2); replace the small molecule silk fibroin in step (3) with regenerated silk fibroin [prepared in step (1)], and the other steps are the same as in Example 1.

[0041] Hemostatic performance test:

[0042] Take medical adhesive tape and cut it into four strips of 2cm × 6cm. Peel off the release paper and heat on an 80℃ hot plate for 30 minutes. Place a hollow mold in the middle of each strip and pour the silk fibroin-based hemostatic gel prepared in Example 1 and Comparative Examples 1-3 into the molds. Heat at 60 W / cm². 2 The gel tape was cured and set under ultraviolet light for 20 seconds to obtain gel tapes of Example 1, Comparative Example 1, Comparative Example 2, and Comparative Example 3. The preparation method was the same as that of gel tape of Example 1, except that medical gauze was used instead of medical adhesive tape to obtain gel gauze of Example 1. The hemostatic properties of gel tapes of Example 1, Comparative Example 1, Comparative Example 2, Comparative Example 3, and gel gauze of Example 1 were tested using the following method:

[0043] (1) Thirty rats weighing 250-300g were randomly divided into 5 groups of 6 rats each, namely, experimental group 1, experimental group 2, experimental group 3, experimental group 4 and experimental group 5;

[0044] (2) After anesthetizing the rats in each group, the tails of the rats were docked (three rats in each group were randomly docked 1 cm from the tail tip, and the remaining three rats were docked 2 cm from the tail tip). After 20 seconds of free bleeding, the rats in each group were hemostatically controlled using gel tape from Example 1, gel tape from Comparative Example 1, gel tape from Comparative Example 2, gel tape from Comparative Example 3, and gel gauze from Example 1, and the time was recorded. The timing was stopped after complete hemostasis, and the average hemostasis time of the rats with 1 cm tail docking and the average hemostasis time of the rats with 2 cm tail docking were recorded. The test results are shown in Table 1.

[0045] Table 1

[0046]

[0047] The results of the rat tail amputation hemostasis experiment show that silk fibroin-based hemostatic gel has a rapid hemostatic effect. At the same time, the small molecule silk fibroin can take advantage of its molecular structure to further accelerate the hemostasis of the wound.

[0048] Comparative Example 5

[0049] The steps are the same as in Example 1, except that raspberry glycosides are not added.

[0050] The hemostatic gel adhesion performance of Example 1 and Comparative Example 5 was tested in accordance with YY / T1477.4-2017 "Standard test models for evaluating the performance of contact wound dressings - Part 4: In vitro models for evaluating the potential adhesion of wound dressings".

[0051] The test method is as follows: (1) The hemostatic gel prepared in the examples and comparative examples is evenly applied to two tinplates (10cm×5cm in size), and the two tinplates are attached to both sides of the wound model (prepared according to the wound model in YY / T1477.4-2017) to form a hemostatic gel-wound model combination.

[0052] (2) Place the hemostatic gel-wound model assembly on a horizontal tray, apply a pressure block (the same pressure block as in Appendix A of YY / T1477.4-2017) on the hemostatic gel-wound model assembly, place it at room temperature for 10 min, and then place it in a drying oven at (37±2)℃ for 24 h.

[0053] (3) The hemostatic gel-wound model assembly was mounted on the clamps of the tensile testing machine. The upper and lower clamps held two tinplates respectively, with an initial distance of 50 mm between the two clamps. The assembly was stretched at a rate of 100 mm / min until one piece of hemostatic gel peeled off from the wound model (if the hemostatic gel peeled off from the tinplate, it was considered invalid). The test was repeated 3 times, and the average peeling force was recorded. The test results are shown in Table 2.

[0054] Table 2

[0055]

[0056] The silk fibroin-based hemostatic gel prepared in Example 1 was injected into a syringe, and the resulting product was as follows: Figure 1 As shown.

[0057] The above description is merely a preferred embodiment of the present invention. The scope of protection of the present invention is not limited to the above embodiments. All technical solutions falling within the scope of the present invention's concept are within the scope of protection of the present invention. It should be noted that for those skilled in the art, any improvements and modifications made without departing from the principles of the present invention should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a silk fibroin-based hemostatic gel, characterized by, Includes the following steps: (1) Regenerated silk fibroin was hydrolyzed and dialyzed to obtain small molecule silk fibroin; (2) Small molecule silk fibroin was modified with a modifier to obtain small molecule modified silk fibroin; (3) Small molecule modified silk fibroin was mixed with an auxiliary agent, a photoinitiator, modified gelatin and raspberry glycoside in a certain proportion and stirred in the dark to obtain silk fibroin-based hemostatic gel. The specific steps of step (1) are as follows: dissolve the regenerated silk fibroin in purified water to obtain a silk fibroin aqueous solution, add neutral protease and adjust the pH value, then carry out a hydrolysis reaction, dialyze and freeze-dry the solution after the reaction to obtain small molecule silk fibroin; the concentration of the silk fibroin aqueous solution is 20-40wt%; the neutral protease accounts for 4-6wt% of the silk fibroin aqueous solution; the pH value is controlled at 4.5-6.5 with 0.1mol / L NaOH solution; the reaction temperature is 30-50℃, the reaction time is 2-4 hours; the dialyze time is 48-72h; the freeze-drying temperature is -80 to -70℃, the vacuum degree is 1-10Pa, and the time is 48-72h; the molecular weight of the small molecule silk fibroin is 1-9 kDa; Step (3) specifically involves dissolving small molecule modified silk fibroin in purified water to obtain a small molecule modified silk fibroin solution, adding auxiliaries, photoinitiators, modified gelatin, and raspberry glycosides, stirring in the dark to obtain a silk fibroin-based hemostatic gel.

2. The method of claim 1, wherein the silk fibroin-based hemostatic gel is prepared by the steps of: The method for preparing the regenerated silk fibroin is as follows: Silkworm cocoons are added to a 0.05 mol / L Na₂CO₃ aqueous solution and boiled at 100°C for 25-35 minutes. After boiling, the cocoons are removed and placed in a freshly prepared Na₂CO₃ aqueous solution of equal volume and concentration and boiled at 100°C for 25-35 minutes. The cocoons are then removed, washed, and dried to obtain degummed silk. The degummed silk is then heated in a water bath at 65-75°C with stirring for 4-5 hours in a CaCl₂-C₂H₆O-H₂O solution. After cooling and centrifugation, the supernatant was obtained. The supernatant was placed in a dialysis bag and dialyzed at 4°C for 3 days to obtain regenerated silk fibroin. The ratio of silkworm cocoon to Na2CO3 aqueous solution was 0.5~2g:50~400mL; the molar ratio of CaCl2, C2H6O, and H2O was 1:2:8; the ratio of degummed silk to CaCl2-C2H6O-H2O solution was 0.1~2g:10~30mL; and the molecular weight of regenerated silk fibroin was 10-130 kDa.

3. The method of claim 1, wherein the silk fibroin-based hemostatic gel is prepared by the steps of: Step (2) is as follows: dissolve the small molecule silk fibroin in purified water to obtain a small molecule silk fibroin solution, add 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide and stir to react, then add a modifier and react at room temperature, and then dialyze and freeze dry the solution after reaction to obtain small molecule modified silk fibroin.

4. The method of claim 3, wherein the silk fibroin-based hemostatic gel is prepared by the steps of: The concentration of the small molecule silk fibroin solution is 10-30 wt%; 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide each account for 1-5 wt% of the small molecule silk fibroin solution, and the molar ratio of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide to N-hydroxysuccinimide is 1:1; the stirring reaction time is 0.5-1 h; the modifier is one of glycidyl methacrylate, methacrylic anhydride, methacryloyl ethyl isocyanate, or methacryloyl chloride, and the modifier accounts for 5-15 wt% of the small molecule silk fibroin solution; the reaction time at room temperature is 12-24 h; the molecular weight of the dialysis bag is 1000-3500 Da, and the dialysis time is 48-72 h; the freeze-drying temperature is -80 to -70℃, the vacuum degree is 1-10 Pa, and the drying time is 48-72 h.

5. The method of claim 1, wherein the silk fibroin-based hemostatic gel is prepared by the steps of: The concentration of the small molecule modified silk fibroin solution is 10-30 wt%; the auxiliary agents are one or more of sodium hyaluronate, carboxymethyl chitosan, dopamine, polyvinyl alcohol, and carbomer, accounting for 5-10 wt% of the small molecule modified silk fibroin solution; the photoinitiator is one of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylphenylacetone, 2,4,6-trimethylbenzoyl-diphenylphosphine oxide, 2,2-dimethoxy-2-phenylacetophenone, phenyl(2,4,6-trimethylbenzoyl)lithium phosphate, and 2,4,6-trimethylbenzoyl-diphenylphosphine oxide, accounting for 0.1-1 wt% of the small molecule modified silk fibroin solution; the modified gelatin accounts for 1-2 wt% of the small molecule modified silk fibroin solution; the raspberry glycoside accounts for 0.5-1 wt% of the small molecule modified silk fibroin solution; the reaction time is 12-24 h with stirring in the dark.

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