CN113209365B

CN113209365B - Multifunctional closed hemostatic wound dressing and preparation method thereof

Info

Publication number: CN113209365B
Application number: CN202110599634.3A
Authority: CN
Inventors: 李飞翰; 杨黄浩; 张进; 潘高星; 林倩; 潘佳铭; 刘泽群
Original assignee: Fuzhou University
Current assignee: Fuzhou Bomei Biotechnology Co.,Ltd.
Priority date: 2021-05-31
Filing date: 2021-05-31
Publication date: 2022-04-01
Anticipated expiration: 2041-05-31
Also published as: CN113209365A

Abstract

The invention discloses a multifunctional closed hemostatic wound dressing which is prepared by uniformly mixing chitosan, blood coagulation factor-loaded nanoparticles, Pluronic F-127 and catechol at room temperature. The multifunctional closed hemostatic wound dressing has the advantages of high adhesion speed in a humid environment, high hemostatic efficiency, strong antibacterial activity, safety, no toxicity, good biocompatibility, low price, convenience in operation and the like, is suitable for aortic hemostasis, is simple in preparation method, and can be produced in a large scale.

Description

Multifunctional closed hemostatic wound dressing and preparation method thereof

Technical Field

The invention belongs to the field of biomaterial preparation and biomedical application, and particularly relates to a composite hemostatic material prepared from blood coagulation factor-loaded nanoparticles, Pluronic F-127, chitosan and catechol, and a preparation method thereof.

Background

The research and development of a new generation of high-efficiency hemostatic material to meet the requirements of military treatment on stepness and high hemostasis efficiency is always an urgent research hotspot of military war guarantee and clinical medicine at present. The hemostatic materials in the market at present mainly comprise fibrin glue, oxidized regenerated cellulose, cyanoacrylate, zeolite, chitosan and the like. For example, the Quikclot hemostatic material developed by Z-medical company is a composite zeolite hemostatic material based on 75% synthetic zeolite, which has a remarkable hemostatic effect but has obvious side effects of water absorption and heat release, animal experiments show that the highest temperature of water absorption and heat release of the material can reach about 100 ℃, which can cause serious thermal injury to injured tissues, and the powdery hemostatic material is easy to remain in a vessel cavity, thus causing inconvenience for subsequent treatment. Fibrin glue can form fibrin clot to adhere to wound surface, can reduce the formation of hematoma on wound surface, and is suitable for stopping bleeding after peritoneum and abdominal cavity viscera bleeding. The oxidized regenerated cellulose has the functions of inhibiting bacteria and preventing postoperative adhesion, is suitable for hemostasis of organs in abdominal cavity, but is avoided in neurosurgery operation. The cyanoacrylate also has strong adhesion effect, can fill the tissue defect part and stimulate the growth of granulation tissue, so the cyanoacrylate is suitable for skin injury patients. Most of the hemostatic products are only used in non-war time, and are effective for partial small bleeding of wounds, and no quick and effective hemostatic material is available in the market at present in the aspect of large-scale acute bleeding occurring in war, acute burst explosion and other events. Therefore, it is necessary to develop a medical biological adhesive with fast hemostasis, good biocompatibility and high adhesive strength in a humid environment in vivo.

Chitosan is a natural mucopolysaccharide having good biocompatibility, and has been widely used as a biomedical material. The chitosan can promote hemostasis by adsorbing platelets in a solid state; in the liquid state, chitosan also has the function of aggregating erythrocytes. Based on the advantages, the hemostatic material taking chitosan as the base material shows good application effect in the aspect of wound hemostasis. In addition, chitosan also has the effect of inhibiting the growth of various bacteria and fungi. The wound dressing made of chitosan has water and oxygen absorption and oxygen permeability, and wound tissues under the dressing can obtain enough oxygen, thereby being beneficial to epithelial cells to gather from the periphery to cover the wound. Patent CN106822986B discloses a preparation method of a chitosan-agar oligosaccharide porous bead hemostatic material, which comprises S1, preparing a chitosan-agar oligosaccharide composite solution; s2, adding a cross-linking agent 1, 3-diglycidyl ether glycerol into the composite alcohol solution, dropwise adding the chitosan-agar oligosaccharide composite solution into the composite alcohol solution containing 1, 3-diglycidyl ether glycerol under the condition of ultrasonic power of 130-150W, stopping ultrasonic after dropwise adding, and stirring at 600-800 rpm for 30 minutes; and S3, filtering the beads, washing and freeze-drying to obtain the microsphere. The hemostatic material prepared by the technical scheme has good biocompatibility, safety and no toxicity; can promote blood coagulation rapidly, can finish rapid hemostasis in a certain application range, but has no adhesion to wet tissues and is not suitable for aortic hemorrhage and visceral hemorrhage. And the preparation steps are complicated, so that the method is not suitable for batch production.

Disclosure of Invention

In order to solve the technical problems, the invention provides a composite hemostatic material prepared from blood coagulation factor-loaded nanoparticles, Pluronic F-127 (nonionic triblock copolymer formed by polyoxyethylene-polyoxypropylene-polyoxyethylene), chitosan and catechol, wherein the blood coagulation factor-loaded PVA (polyvinyl alcohol) nanoparticles are used as a matrix material, and the Pluronic F-127 subjected to terminal thiolation is used as an adhesive to form a double network inside by virtue of self-assembly and chelation between the catechol and chitosan biomacromolecules. The prepared composite hemostatic material has the advantages of high bonding strength in a humid environment, high hemostatic efficiency, strong antibacterial activity, safety, no toxicity, good biocompatibility, low price, convenience in operation and the like, and is suitable for large-range acute hemorrhage.

The technical scheme provided by the invention is as follows:

a preparation method of the multifunctional closed hemostatic wound dressing comprises the following steps:

(1) dissolving a chitosan solution in an aluminum chloride hexahydrate solution, and stirring at a constant temperature to obtain a solution S1;

(2) adding bovine serum albumin and a blood coagulation factor into the polyvinyl alcohol solution, and stirring to obtain a solution S2;

(3) adding polycaprolactone into dichloromethane, and stirring to obtain a solution S3;

(4) emulsifying S2 in S3, and evaporating the organic solvent to obtain a solution S4;

(5) diluting 10wt% Pluronic F-127 in Hepes buffer, and stirring at room temperature to obtain solution S5;

(6) dissolving catechol in a mixed solution of deionized water and ethanol, and stirring at constant temperature to obtain a solution S6;

(7) and uniformly mixing the solutions S1, S4, S5 and S6 at room temperature to obtain the multifunctional closed hemostatic wound dressing.

Preferably, in the solution S1 in the step (1), the concentration of chitosan is 0.001-10 g/mL; the concentration of the aluminum chloride hexahydrate is 0.001-10 g/mL; the stirring temperature is 10-60 ℃; the stirring time is 0.5-6 h.

Preferably, in the solution S2 in the step (2), the mass fraction of the polyvinyl alcohol is 0.2-5%; the mass fraction of the bovine serum albumin is 0.01-0.5%; the mass fraction of the blood coagulation factor is 0.01-0.5%; the stirring temperature is 30-50 ℃; the stirring time is 1-4 h.

Preferably, in the solution S3 in the step (3), the mass fraction of polycaprolactone is 0.5-5%; the stirring temperature is 30-50 ℃; the stirring time is 1-4 h.

Preferably, in the step (4), S2 is emulsified in S3, stirred and evaporated for 0.5-3 h by using a magnetic stirrer.

Preferably, in the solution S5 in the step (5), the mass fraction of the Pluronic Pluronic F-127 is 0.01-0.1%.

Preferably, in the solution S6 in the step (6), the concentration of the catechol is 0.001-10 g/mL; the stirring temperature is 20-35 ℃; the stirring time is 15-30 min.

Preferably, in the step (7), the mass ratio of the solution S1 to the solution S6 to the solution S5 to the solution S4 is 1: 500: 0.1: 500-500: 1: 2.5: 1; the mixing speed is 0.01-1 mL/min; the stirring time is 1-10 min.

The invention also discloses a composite hemostatic material prepared by any one of the preparation methods.

Compared with the prior art, the invention has the following technical advantages:

(1) the composite hemostatic material can self-assemble in molecules to form a double-layer supramolecular network, wherein a protonated amino group on a chitosan side chain and a catechol group of catechol generate hydrogen bond action, and a first layer of cross-linked network is formed through self-assembly and chelation among macromolecules; the Pluronic F-127 wraps the blood coagulation factor nano-particles with negative charges, the whole body is negatively charged, and the chitosan side chain protonated amino group generates electrostatic interaction to form a second layer of cross-linked network inside. In addition, the cationic effect of chitosan can rapidly capture and activate red blood cells and platelets; pluronic is a surfactant and forms micelles in water to diffuse out nanoparticles, so that Pluronic F-127 can improve the diffusivity of the nanoparticles, and the blood coagulation factor nanoparticles can rapidly permeate skin mucosa, thereby accelerating the release of the blood coagulation factor. By means of covalent crosslinking of catechol and chitosan and electrostatic interaction between Pluronic F-127 and chitosan, a double network is formed inside the prepared composite hemostatic material, so that gaps among hydrogel stable molecules are very small, wounds can be sealed, the effect of rapid hemostasis is achieved, and the use of large-dose hemorrhage is met.

(2) The double-layer supermolecule network formed among different materials in the invention greatly enhances the adhesion effect and good tensile property of the composite hemostatic material in a wet environment, and the added blood coagulation factor nanoparticles can specifically adsorb red blood cells, improve the success rate of underwater hemostasis and have very wide application prospect.

(3) On one hand, the composite hemostatic material prepared by the invention promotes blood coagulation based on the function of the chitosan/blood coagulation factor nano-particles for aggregating erythrocytes; on the other hand, the rapid sealing of injured tissues can be realized based on the ultra-strong wet tissue bonding capacity of a large amount of phenolic hydroxyl contained in the catechol solution, the prepared hemostatic material has the advantages of high bonding speed in a wet environment, high hemostatic efficiency, strong antibacterial activity, safety, no toxicity, good biocompatibility, low price, convenience in operation and the like, is suitable for aortic hemostasis, is simple in preparation method, and can be produced in a large scale.

Drawings

FIG. 1 is the result of cytotoxicity test of the novel multifunctional closed hemostatic wound dressing prepared in example 6 of the present invention;

FIG. 2 is a graph showing the results of a wet adhesion strength test of the novel multifunctional occlusive hemostatic wound dressing prepared in example 6 of the present invention;

FIG. 3 is the test result of the mechanical strength of the novel multifunctional closed hemostatic wound dressing prepared in example 6 of the present invention;

fig. 4 is a scanning electron microscope image of the novel multifunctional closed hemostatic wound dressing prepared in example 6 of the present invention.

Detailed Description

For a further understanding of the invention, reference will now be made to the preferred embodiments of the invention by way of example, and it is to be understood that the description is intended to further illustrate features and advantages of the invention, and not to limit the scope of the claims.

Example 1

A preparation method of a novel multifunctional closed hemostatic wound dressing comprises the following steps:

(1) 15 g of aluminum hexachloride was dissolved in water, and 15 g of chitosan was added thereto, followed by stirring at 40 ℃ for 6 hours, to obtain a solution S1 in which the concentration of chitosan was 0.67 g/mL and the concentration of aluminum chloride hexahydrate was 0.67 g/mL.

(2) Adding 2 g of PVA into deionized water, stirring for 4h at 40 ℃, adding 25 mg of BSA and 10 mg of blood coagulation factor, and dissolving to obtain a blood coagulation factor-loaded nanoparticle solution S2, wherein the mass fraction of the polyvinyl alcohol is 2%, the mass fraction of the bovine serum albumin is 0.025%, and the mass fraction of the blood coagulation factor is 0.01%;

(3) dissolving 250 mg of PCL (polycaprolactone) in dichloromethane, stirring at 30 ℃ for 4h to obtain a solution S3, wherein the mass fraction of the polycaprolactone is 3%;

(4) emulsifying the solution S2 in the solution S3 by using a probe type ultrasonic homogenizer, and stirring for 2 hours by using a magnetic stirrer to completely evaporate the organic solvent to obtain a solution S4;

(5) diluting Pluronic F-127 with 10% by mass in Hepes buffer solution to 0.02% to obtain solution S5;

(6) dissolving 15 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 30 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 1 g/mL;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 20: 10: 100: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Example 2

(1) dissolving 20 g of aluminum chloride hexahydrate in water, adding 20 g of chitosan, and stirring at 45 ℃ for 4 hours to obtain a solution S1, wherein the concentration of the chitosan is 1 g/mL; the concentration of the aluminum chloride hexahydrate is 1 g/mL;

(2) adding 4 g of PVA into deionized water, stirring for 4h at 35 ℃, adding 25 mg of BSA and 10 mg of blood coagulation factor, and dissolving to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 4%, the mass fraction of the bovine serum albumin is 0.025%, and the mass fraction of the blood coagulation factor is 0.01%;

(3) 100 mg of PCL was dissolved in dichloromethane; stirring at 30 ℃ for 4h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 1%;

(5) diluting Pluronic F-127 with 10% by mass in Hepes buffer solution to 0.03% to obtain solution S5;

(6) dissolving 20 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 35 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 0.67 g/mL;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 20: 20: 80: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Example 3

(1) dissolving 20 g of aluminum chloride hexahydrate in water, adding 20 g of chitosan, and stirring at 45 ℃ for 4 hours to obtain a solution S1, wherein the concentration of the chitosan is 1 g/mL, and the concentration of the aluminum chloride hexahydrate is 1 g/mL;

(2) adding 2 g of PVA into deionized water, stirring for 4h at 45 ℃, adding 30mg of BSA and 15 mg of blood coagulation factor, and dissolving to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 4%, the mass fraction of the bovine serum albumin is 0.03%, and the mass fraction of the blood coagulation factor is 0.015%;

(3) 200 mg of PCL was dissolved in dichloromethane; stirring at 35 ℃ for 3.5 h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 2%;

(4) emulsifying the solution S2 in the solution S3 by using a probe type ultrasonic homogenizer, and stirring for 3 hours by using a magnetic stirrer to completely evaporate the organic solvent to obtain a solution S4;

(6) dissolving 40 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 35 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 1.5 g/mL;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 20: 20: 100: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Example 4

(1) dissolving 9 g of aluminum chloride hexahydrate in water, adding 9 g of chitosan, and stirring at 45 ℃ for 4 hours to obtain a solution S1, wherein the concentration of the chitosan is 0.43 g/mL, and the concentration of the aluminum chloride hexahydrate is 0.43 g/mL;

(2) adding 1 g of PVA into deionized water, stirring for 4h at 30 ℃, and then adding 20mg of BSA and 20mg of blood coagulation factor to dissolve to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 1%, the mass fraction of the bovine serum albumin is 0.02%, and the mass fraction of the blood coagulation factor is 0.02%;

(3) 300 mg of PCL was dissolved in dichloromethane; stirring at 35 ℃ for 3.5 h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 3.5%;

(5) diluting Pluronic F-127 with 10% by mass in Hepes buffer solution to 0.04% to obtain solution S5;

(6) dissolving 9 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 35 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 0.43 g/mL;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 15: 50: 150: 1 for 10 min to obtain the hemostatic composite material with adhesive performance.

Example 5

(2) adding 2 g of PVA into deionized water, stirring for 64h at 30 ℃, and then adding 20mg of BSA and 20mg of blood coagulation factor to dissolve to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 2%, the mass fraction of the bovine serum albumin is 0.02%, and the mass fraction of the blood coagulation factor is 0.02%;

(3) 300 mg of PCL was dissolved in dichloromethane; stirring at 30 ℃ for 4h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 3.5%;

(6) dissolving 10 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 35 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 0.54 g/mL;

Example 6

(2) adding 2 g of PVA into deionized water, stirring for 4h at 40 ℃, and then adding 30mg of BSA and 30mg of blood coagulation factor to dissolve to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 2%, the mass fraction of the bovine serum albumin is 0.03%, and the mass fraction of the blood coagulation factor is 0.03%;

(3) 150 mg of PCL was dissolved in dichloromethane; stirring at 30 ℃ for 4h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 1.5%;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 50: 50: 150: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Example 7

(2) adding 2 g of PVA into deionized water, stirring for 4h at 40 ℃, and then adding 15 mg of BSA and 15 mg of blood coagulation factor to dissolve to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 2%, the mass fraction of the bovine serum albumin is 0.015%, and the mass fraction of the blood coagulation factor is 0.015%;

(3) 300 mg of PCL was dissolved in dichloromethane; stirring at 40 ℃ for 3 h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 3.5%;

(6) dissolving 9 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 35 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 0.67 g/mL;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 50: 50: 200: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Example 8

(1) dissolving 9 g of aluminum chloride hexahydrate in water, adding 15 g of chitosan, and stirring at 45 ℃ for 4 hours to obtain a solution S1, wherein the concentration of the chitosan is 0.71 g/mL, and the concentration of the aluminum chloride hexahydrate is 0.71 g/mL;

(2) adding 1 g of PVA into deionized water, stirring for 4h at 45 ℃, and then adding 20mg of BSA and 10 mg of blood coagulation factor to dissolve to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 1%, the mass fraction of the bovine serum albumin is 0.02%, and the mass fraction of the blood coagulation factor is 0.01%;

(3) 250 mg of PCL was dissolved in dichloromethane; stirring at 50 ℃ for 2.5 h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 2%;

(6) dissolving 15 g of catechol in deionized water and ethanol (1: 1) solution, and stirring at 35 ℃ for 20 min to obtain solution S6, wherein the concentration of the catechol is 1 g/mL;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 25: 25: 150: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Example 9

(1) dissolving 15 g of aluminum hexachloride in water, adding 15 g of chitosan, and stirring at 50 ℃ for 5 hours to obtain a solution S1, wherein the concentration of the chitosan is 0.67 g/mL, and the concentration of the aluminum hexachloride hexahydrate is 0.67 g/mL;

(2) adding 3 g of PVA into deionized water, stirring for 4h at 50 ℃, adding 25 mg of BSA and 25 mg of blood coagulation factor, and dissolving to obtain a solution S2, wherein the mass fraction of the polyvinyl alcohol is 3%, the mass fraction of the bovine serum albumin is 0.025%, and the mass fraction of the blood coagulation factor is 0.025%;

(3) 200 mg of PCL was dissolved in dichloromethane; stirring at 30 ℃ for 4h to obtain a solution S3, wherein the mass fraction of polycaprolactone is 2%;

(7) mixing the solution S1, the solution S4, the solution S6 and the solution S5 according to the weight ratio of 50: 50: 250: 1 for 10 min to obtain the composite hemostatic material with adhesive property.

Measurement of Performance

The hemostatic materials prepared in examples 1-9 were used in rat heart hemostasis experiments, as shown in table 1: the composite hemostatic material prepared by the invention can rapidly stop bleeding within 5s, the medical gauze can stop bleeding within about 20s, and meanwhile, the prepared material can be adhered to wet environments such as heart and the like, is expected to replace the traditional operation suture line and becomes a new generation of hemostatic medical material.

TABLE 1 hemostatic material prepared according to the present invention used for rat heart hemostasis data

TABLE 2 test results of medical Properties of hemostatic materials prepared according to the present invention

As can be seen from the medical performance test results in Table 2, the hemostatic material prepared by the invention has good non-toxicity, no allergy and no irritation. The cytotoxicity test results shown in fig. 1 show that the composite hemostatic material of the present invention has a cell activity of more than 90% after co-culturing with cells for 1,3, and 7 days, and thus the hemostatic material has good biological safety.

Cytotoxicity test:

in a 96-well plate, the material was placed in the wells and 100.0 μ L of LO2 cell suspension (density = 5000 cells mL) was added^-1) And at 5% CO₂Incubate at 37 ℃ for 1,3, and 7 days in a humid atmosphere, and replace the medium every other day. At predetermined time intervals, the original medium in the wells of the cell culture plate was removed, 100.0 μ L of fresh serum-free medium (containing 10% CCK-8 reagent) was added per well, and cultured at 5% for 45 minutes at 37 ℃. Cells seeded in 96-well plates without extraction medium served as positive controls. Absorbance was measured at 450 nm using a microplate reader (model 550, Bio-Rad, Hercules, Calif., USA). The specific results are shown in FIG. 1.

Wet adhesion test:

the adhesion properties of the hydrogels were tested on a 100N load cell using an Instron machine 1185 (Instron, Boston, MA, USA) according to lap shear test (ASTM F2255-05) standard. To prepare the test samples, fresh pig skin obtained from a supermarket was cut into 4.0cm rectangular sections x 1.0cm, with a sheet thickness of 0.2cm, and soaked with PBS buffer before use. For better application in the hydrogel, the sections were removed and the remaining PBS buffer was ground with filter paper. The hydrogel was then coated directly onto the opposite sides of the two pigskin sheets with a contact area of 1.0 x 1.0 square centimeters. After contacting the two pigskin pieces with a load of 250.0g for 40min, they were placed in a tester to evaluate the adhesive strength of the hydrogel at room temperature. Finally, the adhesive strength of the hydrogel was measured to be 70.93. + -. 5.63 kPa.

Mechanical strength test:

tensile strength measurements were performed by texture analyzer (SMS, ltd. hamilton, MA, USA) at room temperature to measure the mechanical properties of the composite hydrogels. Each sample was cut into a 30.0 mm rectangular block × 10.0 mm × 2.0 mm, and the stretching rate was set to 10.0 mm min-1 for the stretching test until breakage. The stress intensity was finally measured to be 194.54. + -. 0.91 kPa, and the Young's modulus was 10.67. + -. 1.18 kPa.

And (3) scanning electron microscope test:

the morphology of the hydrogel was studied using a Scanning Electron Microscope (SEM). Prior to observation, the lyophilized samples were coated with a thin gold conductive layer using a sputter coater. Then, the surface structure of the hydrogel was examined using Nova NanoSEM 230 (FEI corporation, Hillsboro, OR, USA). The hydrogel was found to exhibit a porous structure.

It will be understood by those skilled in the art that the foregoing is merely a preferred embodiment of the invention, and is not intended to limit the invention, and that any modification, equivalent replacement or improvement made within the spirit and principle of the invention should be included within the scope of protection of the invention.

Claims

1. A preparation method of the multifunctional closed hemostatic wound dressing is characterized by comprising the following steps:

2. The method of claim 1, wherein: in the solution S1 in the step (1), the concentration of chitosan is 0.001-10 g/mL; the concentration of the aluminum chloride hexahydrate is 0.001-10 g/mL; the stirring temperature is 10-60 ℃; the stirring time is 0.5-6 h.

3. The method of claim 1, wherein: in the solution S2 in the step (2), the mass fraction of polyvinyl alcohol is 0.2-5%; the mass fraction of the bovine serum albumin is 0.01-0.5%; the mass fraction of the blood coagulation factor is 0.01-0.5%; the stirring temperature is 30-50 ℃; the stirring time is 1-4 h.

4. The method of claim 1, wherein: in the solution S3 in the step (3), the mass fraction of polycaprolactone is 0.5-5%; the stirring temperature is 30-50 ℃; the stirring time is 1-4 h.

5. The method of claim 1, wherein: in the step (4), S2 is emulsified in S3, stirred and evaporated for 0.5-3 h by a magnetic stirrer.

6. The method of claim 1, wherein: in the solution S5 in the step (5), the mass fraction of Pluronic F-127 is 0.01-0.1%.

7. The method of claim 1, wherein: in the solution S6 in the step (6), the concentration of catechol is 0.001-10 g/mL; the stirring temperature is 20-35 ℃; the stirring time is 15-30 min.

8. The method of claim 1, wherein: in the step (7), the mass ratio of the solution S1, the solution S6, the solution S5 and the solution S4 is 1: 500: 0.1: 500-500: 1: 2.5: 1; the stirring time is 1-10 min.

9. The multifunctional closed hemostatic wound dressing prepared by the preparation method of any one of claims 1 to 8.