Zwitterionic modified aminopolysaccharide-based material, preparation method and application thereof, and hemostatic powder

Abstract

The application belongs to the technical field of biomedical materials, and particularly relates to a zwitterionic modified amino polysaccharide-based material, a preparation method and application thereof, and a hemostatic powder. The application provides a zwitterionic modified amino polysaccharide-based material, which comprises a zwitterionic modified amino polysaccharide-based polymer and oxidized hyaluronic acid; the zwitterionic modified amino polysaccharide-based polymer is obtained by reaction of an amino polysaccharide-based polymer and a zwitterionic monomer; and the mass ratio of the zwitterionic modified amino polysaccharide-based polymer to the oxidized hyaluronic acid is 3 / 1-1 / 3. The zwitterionic modified amino polysaccharide-based material provided by the application has excellent tissue adhesion performance, can effectively stay in a damaged area and block a wound, and has excellent water absorption / blood absorption performance, portability and a sprayable characteristic.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biomedical materials, and particularly relates to a zwitterion-modified amino polysaccharide-based material, a preparation method and application thereof, and a hemostatic powder. BACKGROUND

[0002] Uncontrolled bleeding caused by severe trauma causes more than 2 million deaths worldwide each year, and is one of the important reasons for the death of civilians and military personnel. The body's inherent coagulation cascade process is difficult to effectively stop massive bleeding, and rapid and effective hemostatic intervention using hemostatic materials is essential to save lives.

[0003] At present, a variety of hemostatic materials have been reported, such as gauze, sponge, hydrogel and powder. For most hemostatic materials, it is still difficult to handle the rapid closure of deep, narrow and irregularly shaped non-compressible wounds. The powdered hemostatic material has excellent portability, can be directly sprayed and filled in the damaged area, and is suitable for hemostasis of non-compressible wounds. However, the existing marketed hemostatic powders such as QuickClot based on zeolite, Celox based on chitosan and Arista based on starch all lack sufficient tissue adhesion. This makes these hemostatic powders easily washed away under rapid or high pressure blood flow, which seriously hinders their application in hemostasis. SUMMARY

[0004] The purpose of the present application is to provide a zwitterion-modified amino polysaccharide-based material, a preparation method and application thereof, and a hemostatic powder. The zwitterion-modified amino polysaccharide-based material provided by the present application has excellent tissue adhesion performance, can effectively stay in the damaged area and block the wound; at the same time, has excellent water / blood absorption performance, portability and sprayability.

[0005] In order to achieve the above purpose, the present application provides the following technical scheme:

[0006] The present application provides a zwitterion-modified amino polysaccharide-based material, comprising a zwitterion-modified amino polysaccharide-based polymer and oxidized hyaluronic acid.

[0007] The zwitterion-modified amino polysaccharide-based polymer is obtained by reaction of an amino polysaccharide-based polymer and a zwitterion monomer.

[0008] The mass ratio of the zwitterion-modified amino polysaccharide-based polymer to the oxidized hyaluronic acid is 3 / 1 to 1 / 3.

[0009] Preferably, the preparation method of the zwitterion-modified amino polysaccharide-based polymer comprises the following steps:

[0010] Mixing the aminopolysaccharide-based polymer, the zwitterionic monomer, the initiator and the solvent to obtain a mixed solution; performing graft copolymerization reaction on the mixed solution to obtain a reaction product;

[0011] Performing dialysis and drying on the reaction product in sequence to obtain the zwitterionic modified aminopolysaccharide-based polymer.

[0012] Preferably, the aminopolysaccharide-based polymer comprises any one or more of chitosan, O-carboxymethyl chitosan, chitosan quaternary ammonium salt and hydroxybutyl chitosan.

[0013] Preferably, the zwitterionic monomer is methacrylate sulfobetaine.

[0014] Preferably, the mass ratio of the aminopolysaccharide-based polymer to the zwitterionic monomer is 5 / 1-1 / 20.

[0015] Preferably, the mixing comprises the following steps:

[0016] Dissolving the aminopolysaccharide-based polymer in the solvent to obtain an aminopolysaccharide-based polymer solution, the mass percentage content of the aminopolysaccharide-based polymer solution being 0.5-5%;

[0017] Then mixing the aminopolysaccharide-based polymer solution, the zwitterionic monomer and the initiator.

[0018] Preferably, the temperature of the graft copolymerization reaction is 50-60℃, and the time is 12-24h; the graft copolymerization reaction is performed in a protective gas atmosphere.

[0019] The drying mode comprises one or more of freeze drying, room temperature drying, vacuum drying and heating drying, and the temperature of the heating drying is 30-100℃.

[0020] The present application provides a preparation method of the zwitterionic modified aminopolysaccharide-based material as described in the above technical solution, comprising the following steps:

[0021] Mixing the zwitterionic modified aminopolysaccharide-based polymer and the oxidized hyaluronic acid to obtain the zwitterionic modified aminopolysaccharide-based material.

[0022] The present application provides an application of the zwitterionic modified aminopolysaccharide-based material as described in the above technical solution or the zwitterionic modified aminopolysaccharide-based material prepared by the preparation method as described in the above technical solution in preparing a hemostatic material.

[0023] The present application provides a hemostatic powder, comprising the zwitterionic modified aminopolysaccharide-based material as described in the above technical solution or the zwitterionic modified aminopolysaccharide-based material prepared by the preparation method as described in the above technical solution.

[0024] The present application provides a zwitterionic modified amino polysaccharide-based material, comprising a zwitterionic modified amino polysaccharide-based polymer and oxidized hyaluronic acid; the zwitterionic modified amino polysaccharide-based polymer is obtained by reaction of an amino polysaccharide-based polymer and a zwitterionic monomer; and the mass ratio of the zwitterionic modified amino polysaccharide-based polymer to the oxidized hyaluronic acid is 3 / 1-1 / 3. Compared with the prior art, the present application has the following beneficial effects: the zwitterionic modified amino polysaccharide-based material provided by the present application selects a zwitterionic modified amino polysaccharide-based polymer and oxidized hyaluronic acid as key components, and has good hydrophilic performance, so that the zwitterionic modified amino polysaccharide-based material can quickly absorb tissue fluid. Under water / blood triggering, the zwitterionic modified amino polysaccharide-based material provided by the present application can quickly gel under the synergistic effect of non-covalent interaction (electrostatic interaction and hydrogen bond) and covalent interaction (Schiff base reaction). The electrostatic interaction is provided by the zwitterionic polymer chains in the zwitterionic modified amino polysaccharide-based polymer structure with negative charge and positive charge; at the same time, the zwitterionic modified amino polysaccharide-based polymer can provide a large number of intermolecular and intramolecular hydrogen bonds; in addition, the Schiff base reaction between the aldehyde group in the oxidized hyaluronic acid and the amino group on the amino polysaccharide-based polymer molecular chain can form a dynamic covalent bond in the gel network. These interactions enable the zwitterionic modified amino polysaccharide-based material provided by the present application to construct a hydrogel network structure in situ after absorbing liquid and swelling, so as to form a porous structure of an adhesive hydrogel network, which can effectively stay in the injury area and block the wound, and is conducive to maintaining the sealing and moist environment of the wound surface, while reducing potential tissue damage and promoting wound healing.

[0025] The present application provides a preparation method of the zwitterionic modified amino polysaccharide-based material described in the above technical solution. The preparation method provided by the present application has easy-to-obtain raw materials, simple preparation process, and does not use any chemical crosslinking agent, and the raw materials all have good biocompatibility and biodegradability. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 Nuclear magnetic resonance hydrogen spectrum of the poly-methacrylic acid sulfobetaine grafted modified chitosan prepared for Example 1;

[0027] Figure 2 Property change of the poly-methacrylic acid sulfobetaine grafted modified chitosan-based adhesive hemostatic powder prepared for Example 2 in deionized water;

[0028] Figure 3 Water absorption / blood absorption performance of the poly-methacrylic acid sulfobetaine grafted modified chitosan-based adhesive hemostatic powder prepared for Example 2;

[0029] Figure 4Tissue adhesion of the poly-methacrylic acid sulfobetaine grafted modified chitosan-based adhesive hemostatic powder prepared in Example 2;

[0030] Figure 5 Biocompatibility of the poly-methacrylic acid sulfobetaine grafted modified chitosan-based adhesive hemostatic powder prepared in Example 2;

[0031] Figure 6 In-vitro blood coagulation time of the poly-methacrylic acid sulfobetaine grafted modified chitosan-based adhesive hemostatic powder prepared in Example 2;

[0032] Figure 7 Red blood cell and platelet adhesion of the poly-methacrylic acid sulfobetaine grafted modified chitosan-based adhesive hemostatic powder prepared in Example 2. DETAILED DESCRIPTION

[0033] The present application provides a zwitterionic modified amino polysaccharide-based material, comprising a zwitterionic modified amino polysaccharide-based polymer and oxidized hyaluronic acid.

[0034] The zwitterionic modified amino polysaccharide-based polymer is obtained by reacting an amino polysaccharide-based polymer and a zwitterionic monomer.

[0035] The mass ratio of the zwitterionic modified amino polysaccharide-based polymer to the oxidized hyaluronic acid is 3 / 1 to 1 / 3.

[0036] In the present application, all the raw materials / components are commercially available products well known to those skilled in the art, unless otherwise specified.

[0037] The zwitterionic modified amino polysaccharide-based material provided by the present application comprises a zwitterionic modified amino polysaccharide-based polymer. In the present application, the zwitterionic modified amino polysaccharide-based polymer is obtained by reacting an amino polysaccharide-based polymer and a zwitterionic monomer. In the present application, the zwitterionic modified amino polysaccharide-based polymer is preferably a powder material.

[0038] In the present application, the preparation method of the zwitterionic modified amino polysaccharide-based polymer preferably comprises the following steps:

[0039] The amino polysaccharide-based polymer, the zwitterionic monomer, the initiator and the solvent are mixed to obtain a mixed solution; the mixed solution is subjected to graft copolymerization to obtain a reaction product;

[0040] The reaction product is subjected to dialysis and drying in sequence to obtain the zwitterionic modified amino polysaccharide-based polymer.

[0041] The present application mixes the aminopolysaccharide-based polymer, the zwitterionic monomer, the initiator and the solvent to obtain a mixed solution; and performs graft copolymerization reaction on the mixed solution to obtain a reaction product. In the present application, the aminopolysaccharide-based polymer is preferably a natural aminopolysaccharide-based polymer. The aminopolysaccharide-based polymer preferably includes any one or more of chitosan, O-carboxymethyl chitosan (O-CMCS), chitosan quaternary ammonium salt and hydroxybutyl chitosan, and in the embodiments, can be chitosan or O-CMCS. The zwitterionic monomer is preferably sulfobetaine methacrylate (SBMA). The initiator is preferably ammonium persulfate. The solvent is preferably an acetic acid aqueous solution or water. The mass percentage content of the acetic acid aqueous solution is preferably 0.5-1%. The water is preferably deionized water. The mass ratio of the aminopolysaccharide-based polymer to the zwitterionic monomer is preferably 5 / 1-1 / 20, and more preferably 2 / 1-1 / 10, and in the embodiments, can be 1 / 5, 2 / 1, 1 / 1, 1 / 2 or 1 / 10. The mass ratio of the zwitterionic monomer to the initiator is preferably 20 / 1-10 / 1. The present application can ensure that the zwitterionic monomer and the aminopolysaccharide-based polymer fully perform graft copolymerization reaction by controlling the mass of the aminopolysaccharide-based polymer and the zwitterionic monomer. In the present application, the mixing preferably includes the following steps: dissolving the aminopolysaccharide-based polymer in a solvent to obtain an aminopolysaccharide-based polymer solution. The mass percentage content of the aminopolysaccharide-based polymer solution is preferably 0.5-5%, and more preferably 0.5-2%. After the dissolving, an initial aminopolysaccharide-based polymer solution is obtained, and the present application preferably degasses the initial aminopolysaccharide-based polymer solution with N2 to obtain an aminopolysaccharide-based polymer solution. The temperature of the N2 degassing is preferably 55-60°C, and the time of the N2 degassing is preferably 30-60 min. After obtaining the aminopolysaccharide-based polymer solution, the present application mixes the aminopolysaccharide-based polymer solution, the zwitterionic monomer and the initiator. In the present application, the temperature of the graft copolymerization reaction is preferably 50-60°C, and the time is preferably 12-24 h; the graft copolymerization reaction is preferably performed in a protective gas atmosphere, and the protective gas is preferably nitrogen. The graft copolymerization reaction is preferably performed under stirring. In the present application, the dialysis is preferably performed in deionized water, and the dialysis bag used in the dialysis has a molecular weight cut-off of 14000 Da. The drying mode preferably includes one or more of freeze drying, room temperature drying, vacuum drying and heating drying, and the temperature of the heating drying is preferably 30-100°C. In the present application, the zwitterionically modified aminopolysaccharide-based polymer obtained by drying is a powder material.

[0042] In this invention, one of the raw materials for preparing the zwitterionic modified aminopolysaccharide polymer is that the aminopolysaccharide polymer has good water absorption and mechanical strength, and can promote hemostasis by concentrating blood. Furthermore, under the action of naturally occurring enzymes in body fluids (such as lysozyme and N-acetylglucosidase), the aminopolysaccharide polymer gradually degrades, releasing N-acetyl-β-D-glucosamine, promoting the proliferation and migration of fibroblasts, facilitating the orderly deposition of collagen, and stimulating an increase in the synthesis level of natural hyaluronic acid at the wound site, thus promoting soft tissue re-epithelialization.

[0043] This invention improves the water absorption and crosslinking properties of the resulting zwitterionic modified aminopolysaccharide polymer by grafting zwitterionic monomers onto the polymer, thereby enhancing the tissue adhesion of the zwitterionic modified aminopolysaccharide material.

[0044] The zwitterionic modified aminopolysaccharide-based material provided by this invention includes oxidized hyaluronic acid (OHA). In this invention, the oxidized hyaluronic acid is preferably a powder material.

[0045] In this invention, the oxidized hyaluronic acid exhibits good biocompatibility and biodegradability. Furthermore, it promotes cell migration and proliferation, thereby promoting tissue regeneration. In this invention, the mass ratio of the zwitterionic modified aminopolysaccharide polymer to the oxidized hyaluronic acid is 3 / 1 to 1 / 3, preferably 1 / 1 to 1 / 3, and in the examples, it can be 1 / 1 or 1 / 3. This invention, by compounding the zwitterionic modified aminopolysaccharide polymer and the oxidized hyaluronic acid according to the above mass ratio, yields a zwitterionic modified aminopolysaccharide material that can rapidly form a hydrosol upon absorbing water. Then, through the synergistic effect of non-covalent interactions (electrostatic interactions and hydrogen bonding) and covalent interactions (Schiff base reaction), it triggers in-situ self-gelation, forming a porous adhesive hydrogel, which can effectively remain in the damaged area and seal the wound.

[0046] In this invention, the zwitterionic modified aminopolysaccharide-based material is preferably a powder material. The zwitterionic modified aminopolysaccharide-based powder material provided by this invention can achieve tissue adhesion on a wet tissue surface through a dry crosslinking mechanism; rapid liquid absorption and dehydration of the tissue surface allow polymer chains to penetrate into the tissue, thereby forming entanglements with tissue fibers. Simultaneously, the removal of water at the wound interface allows the reaction between the aldehyde groups in the hydrogel and the amine groups on the tissue to form stable covalent bonds. Ultimately, the zwitterionic modified aminopolysaccharide-based material provided by this invention is transformed into a viscous hydrogel with a strong adhesive interface, which is beneficial for maintaining wound sealing, keeping the environment moist, reducing potential tissue damage, and promoting wound healing.

[0047] This invention provides a method for preparing the zwitterionic modified aminopolysaccharide-based material described above, comprising the following steps:

[0048] The zwitterionic modified aminopolysaccharide polymer and the oxidized hyaluronic acid are mixed to obtain the zwitterionic modified aminopolysaccharide material.

[0049] The present invention does not have any special requirements for the specific implementation of the mixture of the zwitterionic modified aminopolysaccharide polymer and the oxidized hyaluronic acid.

[0050] This invention provides the application of the zwitterionic modified aminopolysaccharide-based material described in the above technical solution or the zwitterionic modified aminopolysaccharide-based material prepared by the preparation method described in the above technical solution in the preparation of hemostatic materials.

[0051] This invention uses zwitterionic modified aminopolysaccharide polymers (i.e., zwitterionic grafted modified aminopolysaccharide polymers) and oxidized hyaluronic acid as key components to obtain zwitterionic modified aminopolysaccharide powder materials. The zwitterionic modified aminopolysaccharide powder materials provided by this invention can rapidly absorb water at the wound site, quickly adhere and seal the wound through electrostatic interaction and dynamic chemical cross-linking, and concentrate blood cells, thereby achieving efficient hemostasis. The zwitterionic modified aminopolysaccharide powder materials provided by this invention have a simple preparation process, good biocompatibility and biodegradability, can be sprayed, and have excellent water absorption and blood absorption properties, making them particularly suitable as hemostatic materials for hemostasis and tissue repair of non-pressure wounds.

[0052] This invention provides a hemostatic powder, comprising the zwitterionic modified aminopolysaccharide material described in the above-described technical solution or the zwitterionic modified aminopolysaccharide material prepared by the preparation method described in the above-described technical solution. The hemostatic powder provided by this invention can rapidly absorb water at the wound site, quickly adhere and seal the wound through electrostatic interaction and dynamic chemical cross-linking, and concentrate blood cells, thereby achieving efficient hemostasis. The hemostatic powder provided by this invention has a simple preparation process, good biocompatibility and biodegradability, can be sprayed, and has excellent water absorption and blood absorption properties, making it particularly suitable as a hemostatic material for hemostasis and tissue repair of non-pressure wounds.

[0053] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0054] Example 1: Preparation of chitosan powder grafted with polymethacrylic acid sulfobetaine

[0055] Chitosan was dissolved in a 1% acetic acid aqueous solution to prepare a 0.5 wt% chitosan aqueous solution, which was then degassed under N2 at 60°C for 30 minutes. Ammonium persulfate and zwitterionic sulfonyl betaine monomer (SBMA) were added sequentially to the chitosan aqueous solution, with a chitosan to SBMA mass ratio of 1 / 5 and an SBMA to ammonium persulfate mass ratio of 20 / 1. The mixture was then stirred at 60°C under N2 atmosphere for 12 hours, and after dialyzing and drying, polymethyl methacrylate sulfonyl betaine grafted modified chitosan (CS-g-5PSBMA, abbreviated as CSB5) powder was obtained. The polymethyl methacrylate sulfonyl betaine grafted modified chitosan (CS-g-5PSBMA, abbreviated as CSB5) was characterized by proton nuclear magnetic resonance spectroscopy. Figure 1 Compared to the chitosan NMR spectrum, the CSB5 NMR spectrum shows a methyl proton signal (-CH2-C(CH3)-) on the SBMA backbone at around 1 ppm, a methylene proton signal at around 2 ppm, and a methyl proton signal bonded to N at 3.25 ppm. + The presence of N(CH3)2 indicates that PSBMA was successfully grafted onto chitosan. This powder exhibits some tissue adhesion properties, but it cannot form a stable gel under excessive body fluid; the powder flows off the tissue surface and fails to adhere to the tissue. Adjusting the mass ratio of chitosan to SBMA to 2 / 1, 1 / 1, 1 / 2, and 1 / 10 yields CSB0.5, CSB1, CSB2, and CSB10 powders, respectively. All of these powders possess some tissue adhesion properties. With increasing SBMA content, the adhesion properties of the resulting CSB powders improve, but none of the powders can form a stable gel under excessive body fluid.

[0056] Example 2: Preparation of chitosan-based adhesive hemostatic powder grafted with polymethacrylic acid sulfobetaine

[0057] The polymethyl methacrylate sulfobetaine-grafted modified chitosan (CSB5) powder prepared in Example 1 was mixed with oxidized hyaluronic acid (OHA) powder at a 1:1 mass ratio. After shaking and mixing, a polymethyl methacrylate sulfobetaine-grafted modified chitosan-based adhesive hemostatic powder (CSB5-OHA) was obtained. When deionized water was dropped onto the surface of this powder, it rapidly absorbed water, swelled, and gelled. Figure 2 CSB0.5, CSB1, CSB2, and CSB10 powders were mixed with oxidized hyaluronic acid (OHA) powder at a 1:1 mass ratio to obtain CSB0.5-OHA, CSB1-OHA, CSB2-OHA, and CSB10-OHA powders, respectively. All prepared CSB-OHA powders exhibited excellent water absorption / blood absorption properties (e.g., ...). Figure 3 As shown in the image, it can quickly absorb water, gel, and adhere to the tissue surface, sealing the wound, quickly stopping bleeding, and providing a moist environment for the wound, thereby promoting wound healing.

[0058] Example 3: Preparation of O-carboxymethyl chitosan powder grafted with polymethacrylic acid sulfobetaine

[0059] O-Carboxymethyl Chitosan (O-CMCS) was dissolved in deionized water to prepare a 1 wt% O-CMCS aqueous solution, which was then degassed under N2 at 60°C for 30 minutes. Ammonium persulfate and zwitterionic sulfobetaine methacrylate monomer (SBMA) were added sequentially to the O-CMCS aqueous solution, with a mass ratio of O-CMCS to SBMA of 1 / 5 and a mass ratio of SBMA to ammonium persulfate of 10 / 1. The mixture was then stirred at 60°C under N2 atmosphere for 12 hours, and after dialyzing and drying, poly(sulfobetaine methacrylate) grafted modified O-carboxymethyl chitosan powder was obtained. This powder could not form a stable gel structure under water / blood conditions.

[0060] Example 4: Preparation of O-carboxymethyl chitosan-based adhesive hemostatic powder grafted with polymethacrylate sulfobetaine

[0061] The polymethacryloyloxyethylphosphorylcholine grafted modified O-carboxymethyl chitosan powder prepared in Example 3 was mixed with oxidized hyaluronic acid powder at a mass ratio of 1 / 3. After shaking and mixing, polymethacrylic acid sulfobetaine grafted modified O-carboxymethyl chitosan-based adhesive hemostatic powder was obtained. This powder has excellent water absorption and water / blood triggered self-gelling properties, as well as excellent tissue adhesion.

[0062] Example 5: Performance Testing

[0063] 1. Water absorption / blood absorption properties

[0064] The water / blood absorption properties of the polymethacrylate sulfobetaine-grafted chitosan-based adhesive hemostatic powders (CSB5-OHA, CSB0.5-OHA, CSB1-OHA, CSB2-OHA, and CSB10-OHA) prepared in Example 2 were tested within 30 seconds using PBS solution (phosphate buffered solution) at pH 7.4 or anticoagulated rabbit blood. Figure 3 It is known that the adhesive hemostatic powder material CSB-OHA has good water / blood absorption capacity. When used for wound hemostasis, it can quickly absorb liquid, adhere to the wound, concentrate blood, and effectively promote hemostasis.

[0065] 2. Tissue adhesion properties

[0066] Tissue adhesion strength tests were conducted using a universal testing machine to evaluate the adhesion of the CSB-OHA (CSB2-OHA, CSB5-OHA, and CSB10-OHA) adhesive hemostatic powder materials prepared in Example 2 to tissues. The tissue adhesion strength was tested using an overlap shear test. A 1 cm wide fresh pigskin piece was used, with the fat layer removed using a scalpel, and rinsed with PBS (pH=7.4) for 30 minutes before the test. 15 mg of hemostatic powder was evenly sprayed onto the surface of a wet pigskin piece, and then another pigskin piece of the same size was used to bond a 10 × 10 mm area. 2 Cover. After gently pressing the two pieces of pigskin at room temperature for 30 seconds, the tensile shear strength of the specimens was tested. The results showed that the CSB-OHA adhesive hemostatic powder material has good tissue adhesion. Figure 4 ).

[0067] 3. Biocompatibility

[0068] The toxicity of the CSB-OHA (CSB2-OHA, CSB5-OHA, and CSB10-OHA) adhesive hemostatic powders prepared in Example 2 to NIH 3T3 cells was tested using the extract method. Before testing, all samples were sterilized by UV irradiation for 2 hours on a clean laboratory bench. All sterilized samples were then immersed in DMEM medium at a specific ratio. After obtaining the extract, the extract was used to replace the culture medium for NIH 3T3 cells. After culturing for 24 hours, the proliferation rate of NIH 3T3 cells in the wells was tested using a CCK-8 assay kit. 0.1 mL of DMEM medium served as a negative control. Cell experiments showed that the CSB-OHA adhesive hemostatic powder had good cell compatibility, and its extract did not affect the growth and proliferation of NIH 3T3 cells.

[0069] Red blood cell suspension was prepared by centrifuging anticoagulated rabbit whole blood at 3000 rpm for 10 min. After washing the red blood cells three times with PBS, a 5% (v / v) red blood cell suspension was prepared. Then, CSB-OHA was added to 500 μL of the suspension at 0.5 g / mL. The centrifuge tubes were incubated at 37°C for 1 hour, then centrifuged at 3000 rpm for 5 min, and the absorbance of the supernatant was measured. PBS and deionized water were used as negative and positive controls, respectively. The hemolysis experiment showed that the CSB-OHA adhesive hemostatic powder did not cause red blood cell rupture, with a relative hemolysis rate of less than 5%, indicating good blood compatibility. Figure 5 The above results demonstrate that the CSB-OHA adhesive hemostatic powder prepared in Example 2 has good biocompatibility.

[0070] 4. In vitro coagulation properties

[0071] 50 μL of calcified whole blood was added to a well plate containing 5 mg of sample to evaluate the clotting time of the adhesive hemostatic powder. The blood was incubated at 37°C. The blood was washed with PBS at predetermined times until a stable clot formed, and this time was defined as the clotting time. Calcified whole blood served as a blank control group.

[0072] Dynamic whole blood coagulation index tests were performed on the adhesive hemostatic powder materials (CSB2-OHA, CSB5-OHA, and CSB10-OHA) prepared in this invention. 50 μL of calcified whole blood was added to 3 mg of hemostatic powder and incubated in a 37°C water bath for 30 seconds. Then, 10 mL of deionized water was added, and the supernatant was collected to measure the absorbance and calculate the coagulation index. 50 μL of recalcified whole blood was added to 10 mL of deionized water as a blank control group. Longer coagulation time and higher coagulation index indicate poorer coagulation performance. Figure 6 It can be seen that the coagulation time and coagulation index of CSB-OHA adhesive hemostatic powder material are both lower than those of gauze, indicating that CSB-OHA adhesive hemostatic powder material has good in vitro coagulation performance.

[0073] 5. Platelet and erythrocyte adhesion properties

[0074] Platelet adhesion properties of the adhesive hemostatic powder materials prepared in this invention were tested. Platelet-rich plasma (50 μL) was added dropwise to the dried CSB-OHA (CSB2-OHA, CSB5-OHA, and CSB10-OHA) adhesive hemostatic powder materials, and then incubated at 37°C for 1 hour. Unadhesive platelets were eluted with PBS, and the number of platelets adhering to the CSB-OHA adhesive hemostatic powder materials was tested using a lactate dehydrogenase (LDH) kit.

[0075] The erythrocyte adhesion properties of the adhesive hemostatic powder material prepared in this invention were tested. 50 μL of erythrocyte suspension was added to 5 mg of hemostatic powder, and then incubated at 37°C for 1 hour. Unadhered erythrocytes were washed off with PBS. Then, the sample was immersed in 2 mL of deionized water for 1 hour to lyse the adhered erythrocytes, and the absorbance of the supernatant was measured to determine the erythrocyte adhesion rate. Figure 7 It is known that CSB-OHA adhesive hemostatic powder has good adhesion properties to platelets and red blood cells, which allows CSB-OHA adhesive hemostatic powder to quickly gather platelets and red blood cells when used for wound hemostasis, thus achieving efficient hemostasis.

[0076] Comparative Example 1: Polymethacrylic acid sulfobetaine grafted with chitosan-based mixed hyaluronic acid powder

[0077] The polymethacrylate sulfobetaine-grafted modified chitosan (CSB5) powder prepared in Example 1 was mixed with hyaluronic acid (HA) powder in a 1:1 mass ratio. After shaking and mixing, a CSB5 / HA mixed powder was obtained. This powder has certain tissue adhesion properties, but under the action of excessive body fluid, the powder is lost on the tissue surface and cannot form a gel.

[0078] As can be seen from the above embodiments, the zwitterionic modified aminopolysaccharide-based material provided by the present invention uses zwitterionic modified aminopolysaccharide-based polymer and oxidized hyaluronic acid as key components. It can rapidly absorb water at the wound site, and through electrostatic interaction and dynamic chemical cross-linking, quickly adhere to and seal the wound, concentrating blood cells, thereby achieving efficient hemostasis. Simultaneously, the zwitterionic modified aminopolysaccharide-based material provided by the present invention has a simple preparation process, good biocompatibility and biodegradability, can be sprayed, and possesses excellent water absorption and blood absorption properties, making it particularly suitable as a hemostatic material for hemostasis and tissue repair of non-pressure wounds.

[0079] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. Other embodiments can be obtained based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A zwitterionic modified aminopolysaccharide-based material, characterized in that, Including zwitterionic modified aminopolysaccharide polymers and oxidized hyaluronic acid; The zwitterionic modified aminopolysaccharide polymer is obtained by reacting an aminopolysaccharide polymer with a zwitterionic monomer. The aminopolysaccharide polymer includes any one or more of chitosan, O-carboxymethyl chitosan, chitosan quaternary ammonium salt and hydroxybutyl chitosan. The zwitterionic monomer is methacrylate sulfobetaine. The mass ratio of the zwitterionic modified aminopolysaccharide polymer to oxidized hyaluronic acid is 3 / 1 to 1 / 3.

2. The zwitterionic modified aminopolysaccharide-based material according to claim 1, characterized in that, The preparation method of the zwitterionic modified aminopolysaccharide polymer includes the following steps: The aminopolysaccharide polymer, zwitterionic monomer, initiator, and solvent are mixed to obtain a mixed solution; the mixed solution is then subjected to a graft copolymerization reaction to obtain the reaction product. The reaction products were sequentially dialyzed and dried to obtain the zwitterionic modified aminopolysaccharide polymer.

3. The zwitterionic modified aminopolysaccharide-based material according to claim 1 or 2, characterized in that, The mass ratio of the aminopolysaccharide polymer to the zwitterionic monomer is 5 / 1 to 1 / 20.

4. The zwitterionic modified aminopolysaccharide-based material according to claim 2, characterized in that, The mixing process includes the following steps: The aminopolysaccharide polymer is dissolved in a solvent to obtain an aminopolysaccharide polymer solution, wherein the mass percentage of the aminopolysaccharide polymer solution is 0.5-5%. Then the aminopolysaccharide polymer solution, zwitterionic monomer, and initiator are mixed.

5. The zwitterionic modified aminopolysaccharide-based material according to claim 2, characterized in that, The graft copolymerization reaction is carried out at a temperature of 50-60°C for 12-24 hours; the graft copolymerization reaction is carried out in a protective gas atmosphere. The drying method includes one or more of freeze drying, room temperature drying, vacuum drying and heat drying, wherein the temperature of heat drying is 30~100℃.

6. The method for preparing the zwitterionic modified aminopolysaccharide-based material according to any one of claims 1 to 5, characterized in that, Includes the following steps: The zwitterionic modified aminopolysaccharide polymer and the oxidized hyaluronic acid are mixed to obtain the zwitterionic modified aminopolysaccharide material.

7. The application of the zwitterionic modified aminopolysaccharide material according to any one of claims 1 to 5 or the zwitterionic modified aminopolysaccharide material prepared by the preparation method according to claim 6 in the preparation of hemostatic materials.

8. A hemostatic powder, characterized in that, Includes the zwitterionic modified aminopolysaccharide-based material according to any one of claims 1 to 5 or the zwitterionic modified aminopolysaccharide-based material prepared by the preparation method according to claim 6.

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