Preparation method and use method of hemostatic micro powder

Abstract

The invention discloses a preparation method and a use method of hemostatic micro-powder, and the preparation method comprises the following steps: S20, adding a primary amine polymer, an auxiliary A and an auxiliary B into a polyaldehyde polysaccharide biomolecule solution, and stirring at room temperature until the mixture is stable and viscous to obtain a viscous substance; s30, freeze-drying the viscous substance, and grinding to obtain hemostatic micro powder; the use method comprises the following steps: spraying the hemostatic micro-powder onto a wound or dissolving the hemostatic micro-powder in water to form wound colloid, covering the wound with the wound colloid, and instantly adsorbing moisture on a wet wound surface through capillary action force, so that a gel layer is quickly formed, the wound is quickly blocked, and hemostasis is completed; in addition, due to the structural properties of the micro-hemostasis micro powder, the micro-hemostasis micro powder can be stored at room temperature, is convenient to spray and operate, is suitable for irregular wounds, has high wet adhesive force, high mechanical strength and high biological safety, can be independently used, and can be used for quickly stopping bleeding of tension wounds with irregular wet interfaces and large bleeding amount.

Description

Technical Field

[0001] The present invention relates to the field of wound treatment products, and in particular to a preparation method and a use method of hemostatic micropowder. Background Art

[0002] Hemostatic materials based on thrombin, gelatin, chitosan, collagen, fibrin, α-cyanoacrylate, etc., or new hemostatic technologies based on the instant in situ formation of an isolation gel layer, generally have one or more problems such as slow wound sealing, low temperature storage requirements, inconvenient operation, the need for a flat wound surface, low wet adhesion, low mechanical strength, biosafety risks, inability to use alone, and tissue adhesion.

[0003] Therefore, in complex and uncertain wounds, especially tension wounds with irregular wet interfaces and heavy bleeding, it is difficult to achieve rapid hemostasis.

[0004] The present invention aims to solve the problem of how to prepare a rapid hemostatic material that has the characteristics of being able to be stored at room temperature, easy to spray, suitable for irregular wound surfaces, high wet adhesion, high mechanical strength, high biosafety, and can be used alone, so as to be able to quickly stop hemorrhage in tension wounds with irregular wet interfaces and large bleeding volumes. Summary of the Invention

[0005] In order to overcome the deficiencies of the prior art, the present invention provides a method for preparing and using hemostatic micropowder.

[0006] The technical solution adopted by the present invention to solve its technical problem is: A method for preparing hemostatic micropowder, comprising: S20, adding a primary amine polymer, an additive A, and an additive B to the polyaldehyde-polysaccharide biomolecule solution, and stirring at room temperature until a stable viscous state is obtained to obtain a viscous substance; S30. Freeze-drying the viscous substance and grinding it to obtain hemostatic micropowder.

[0007] The polyaldehyde-polysaccharide biomolecules and primary amine polymers form a viscous substance with a network cross-linked structure through a nucleophilic addition-dehydration reaction between the aldehyde and the primary amine (forming a Schiff base bond), as well as hydrogen bonding between the hydroxyl group and the primary amine. This method is highly feasible and can easily produce a hemostatic micropowder that is easy to store.

[0008] Furthermore, the preparation of the polyaldehyde-polysaccharide biomolecule solution comprises the following steps: S10, slowly dripping the strong oxidant solution into the polysaccharide biomolecule solution while heating; Using strong oxidants to oxidize polysaccharide biomolecules, the C2-C3 bond of the glucose unit is broken to produce two aldehyde groups, which lay the foundation for subsequent reactions with other substances; S11, centrifuge and wash 4-7 times at room temperature, and collect the precipitate; The function of centrifugal washing is to separate the unreacted strong oxidant, reaction by-products and other impurities in the reaction system from the product, effectively improve the purity of polyaldehyde polysaccharide biomolecules, reduce the impact of impurities on subsequent reactions and product performance, and ensure the quality and safety of the final prepared hemostatic micropowder.

[0009] S12, drying of the precipitate to obtain polyaldehyde-polysaccharide biomolecules; The drying process can remove moisture from the precipitate to obtain dry polyaldehyde-polysaccharide biomolecules, which helps to improve the stability of the product and prevent the polyaldehyde-polysaccharide biomolecules from hydrolysis or other chemical reactions during storage. It also facilitates subsequent weighing, dissolution, and storage operations.

[0010] S13. Dissolve the polyaldehyde polysaccharide biomolecules in water to obtain a polyaldehyde polysaccharide biomolecule solution.

[0011] Dissolving the dried polyaldehyde polysaccharide biomolecules in water allows them to be evenly dispersed in the water, forming a polyaldehyde polysaccharide biomolecule solution. This uniform polyaldehyde polysaccharide biomolecule solution facilitates uniform mixing and reaction with primary amine polymers, additives, and other substances, ensuring reaction consistency and product performance stability. Furthermore, by controlling the amount of dissolved polyaldehyde polysaccharide biomolecules and the amount of water, the concentration of the polyaldehyde polysaccharide biomolecule solution can be adjusted to reach an appropriate mass concentration range to meet the requirements for subsequent preparation of hemostatic micropowders and ensure the good performance of the final product.

[0012] Furthermore, the polysaccharide biomolecule is one of chondroitin sulfate, hyaluronic acid, heparin, glucomannan, chitosan, pullulan, starch, and sodium alginate; Among them, chondroitin sulfate has good biocompatibility and can combine well with human tissues. The hydrophilic groups in its structure can bind a large amount of water, which helps to keep the wound moist and promote wound healing. Hyaluronic acid has the function of protecting wounds and assisting in hemostasis. It can form a lubricating protective film on the wound surface, reducing friction between the wound and the outside world, preventing wound adhesion, and preventing the invasion of bacteria and other microorganisms. Heparin can promote rapid hemostasis at the wound site and has anti-inflammatory properties. It can reduce the inflammatory response of the wound, facilitate wound recovery, and reduce tissue damage and bleeding caused by inflammation. Glucomannan can form a stable gel structure, which is beneficial for the subsequent molding and performance control of hemostatic materials; Chitosan has natural antibacterial properties, which can inhibit the growth of bacteria in wounds, activate coagulation factors, adsorb red blood cells, etc., and accelerate the hemostasis process; Pullulan can thicken the solution, giving the resulting hemostatic material suitable viscosity and rheological properties, making it easier to process and use. Furthermore, it can be gradually metabolized in the body without producing long-term residues, thus reducing biosafety risks. Starch can form a supporting structure, and its gel properties help absorb components in the blood and promote coagulation; Sodium alginate can form a stable covering layer on the wound surface, playing the role of stopping bleeding and protecting the wound; The average molecular weight of the above polysaccharide biomolecules is 10,000 to 150,000.

[0013] If the molecular weight is too low, the chain length of the polyaldehyde polysaccharide biomolecules formed is short and cannot form an effective three-dimensional network structure. The gel layer forms slowly, stops bleeding slowly, and has insufficient mechanical strength. If the molecular weight is too high, the viscosity of the reaction system is high, the oxidation efficiency is low, the solubility of the product is reduced, and even tissue adhesion may be caused.

[0014] Furthermore, the mass concentration of the polyaldehyde polysaccharide biomolecule solution is 0.05 to 0.60 g / mL.

[0015] This concentration range ensures that the polyaldehyde-polysaccharide biomolecules provide sufficient active aldehyde sites when subsequently reacting with substances such as primary amine polymers. For example, if the concentration is too low, the number of aldehyde groups will be insufficient, resulting in insufficient cross-linking reaction with primary amine polymers and an inability to form sufficient chemical bonds to build a stable three-dimensional network structure. On the other hand, if the concentration is too high, the aldehyde groups will be too dense, the reaction will be too intense, and the reaction will be difficult to control. This may lead to localized excessive cross-linking, affecting the uniformity and performance of the product.

[0016] Furthermore, the sodium perhalate is one of sodium perchlorate, sodium perbromate and sodium periodate, and its mass concentration in water is 0.03-0.5 g / mL. Furthermore, in step S10, the reaction volume ratio of the polysaccharide biomolecule solution to the strong oxidant solution is 1:0.5-2.5, and / or the heating reaction temperature is 25-60° C., and / or the heating reaction time is 3-18 hours.

[0017] The reaction volume ratio of the polysaccharide biomolecule solution to the strong oxidant solution, the heating reaction temperature, and the heating reaction time directly control the degree of oxidation of the polysaccharide biomolecules. If the oxidant dosage is too low, the temperature is too low, or the heating reaction time is too short, the groups on the polysaccharide biomolecules will not be fully oxidized, and sufficient reactive groups such as aldehyde groups will not be generated. This will affect the subsequent cross-linking reaction with the primary amine polymer, resulting in poor final product performance, such as insufficient wet adhesion and unsatisfactory hemostatic effects. Conversely, if the oxidant dosage is too high, the temperature is too high, or the heating reaction time is too long, the polysaccharide molecules may be over-oxidized, destroying their original structure and causing polysaccharide molecular chain breakage, which will also affect product performance. For example, it may reduce the mechanical strength of the product and prevent it from forming a stable covering layer on the wound surface.

[0018] Furthermore, the strong oxidant solution is prepared by dissolving sodium perhalate in deionized water.

[0019] Specifically, the sodium perhalate is one of sodium perchlorate, sodium perbromate and sodium periodate, and its mass concentration in water is 0.03-0.5 g / mL.

[0020] Furthermore, the primary amine polymer is one of polyacrylamide, polyacrylamine, polyetheramine, and polylysine, with an average molecular weight of 100,000 to 5,000,000, and / or a mass concentration of 0.05 to 0.6 g / mL.

[0021] Different primary amine polymers have distinct molecular structures and chemical properties, forming different cross-linking structures with polysaccharide biomolecules. For example, the amide groups on the polyacrylamide molecular chain can cross-link with the aldehyde groups produced by polysaccharide oxidation, forming a cross-linked network with a certain degree of flexibility and stability. Polylysine, on the other hand, contains multiple amino groups, which are more reactive with aldehyde groups and may form a denser cross-linking structure.

[0022] When it comes to molecular weight selection for primary amine polymers, a low molecular weight results in shorter polymer chains, fewer cross-linking sites, and an imperfect cross-linked network. This can lead to insufficient mechanical strength, wet adhesion, and other properties of the gel layer formed during use. As the molecular weight increases, the chain length increases, providing more primary amine groups for cross-linking, making the cross-linked network denser and more stable, thereby improving the product's performance. However, if the molecular weight is too high, the chains may become too bulky and entangled, which can affect their movement and diffusion, hindering the uniformity of the cross-linking reaction and potentially reducing the product's solubility, leading to problems in practical applications.

[0023] Furthermore, the auxiliary agent A is one of lauryl glucoside, rhamnolipid, sophoroyl ester, sodium lauroyl glutamate, and sucrose palmitate, and / or has a mass concentration of 0.02 to 0.15 mg / mL.

[0024] The above substances have a certain effect of regulating the surface properties of the gel layer. When the hemostatic micropowder is applied to the wound, it can make the micropowder better adhere to the wound surface, increase the contact area with the wound, and adapt to the irregular wet interface wound.

[0025] Furthermore, the auxiliary agent B is polyethylene glycol, and its molecular weight is 200-1000, and / or its mass concentration is 0.1-0.5 mg / mL.

[0026] Polyethylene glycol has a thickening effect. During room temperature stirring, it increases the viscosity of the solution and, in conjunction with other ingredients, forms a stable, viscous substance. This viscous substance facilitates subsequent freeze-drying and grinding, facilitating the formation of a fine, uniform hemostatic micropowder. Furthermore, during the subsequent freeze-drying process, polyethylene glycol reduces the formation of ice crystals, preventing components such as polyaldehydes and polysaccharides from losing their activity or structural damage due to ice crystal disruption during the freeze-drying process. It acts as a protective agent, ensuring that the freeze-dried product maintains optimal morphology and properties, thereby improving the quality and stability of the hemostatic micropowder.

[0027] A method for using hemostatic micropowder comprises spraying the hemostatic micropowder prepared by the above-mentioned hemostatic micropowder preparation method onto a wound or dissolving the hemostatic micropowder in water to form a wound colloid, which covers the wound.

[0028] The prepared hemostatic micropowder is sprayed onto the wound or dissolved in water to form a wound colloid that covers the wound. The hemostatic micropowder instantly absorbs moisture through capillary action on the wet wound surface, promoting the rapid participation of amino groups in the wound tissue in rebuilding the synergistic force of Schiff base bonds and hydrogen bonds, thereby quickly forming a gel layer and achieving rapid wound closure and hemostasis. After gelation, the free amino groups are almost completely depleted, and no Schiff base bonds and hydrogen bonds are formed, thus avoiding adhesion to surrounding tissue. In addition, due to the structural properties of the microhemostatic micropowder, it can be stored at room temperature, is easy to spray, and is suitable for irregular wound surfaces. It has high wet adhesion, high mechanical strength, and high biosafety. It can be used alone to quickly stop bleeding in tension wounds with irregular wet interfaces and heavy bleeding.

[0029] Furthermore, the spraying amount of the hemostatic micropowder is 0.09-0.8 g / cm 2 .

[0030] Experiments have shown that applying hemostatic micropowder within this range helps form a uniform gel network on the wound surface with moderate mechanical strength. However, using too much hemostatic micropowder can result in an excessively thick gel layer, which can compress tissue and hinder wound recovery.

[0031] The beneficial effects of the present invention are: In the present application, the raw material polyaldehyde polysaccharide biomolecules are prepared with high biosafety and cross-linking activity. Primary amine polymers are added to the polyaldehyde polysaccharide biomolecule solution, so that the polyaldehyde polysaccharide biomolecule solution and the primary amine polymer generate Schiff base bonds / hydrogen bonds to form a viscous substance with a network cross-linking structure. The viscous substance is freeze-dried and ground to obtain a hemostatic micropowder that is easy to store. The prepared hemostatic micropowder is sprayed onto the wound or dissolved in water to form a wound colloid, which covers the wound. The hemostatic micropowder instantly absorbs moisture on the wet wound surface through capillary action, promotes the amino groups in the wound tissue to quickly participate in the reconstruction of the synergistic force of Schiff base bonds / hydrogen bonds, thereby quickly forming a gel layer, achieving rapid sealing of the wound and completing hemostasis. After gelation, the free amino groups are almost exhausted, and no Schiff base bonds / hydrogen bonds are formed, thereby avoiding adhesion to surrounding tissues. Experiments have shown that this solution can produce hemostatic micropowder with a hemostasis time of 5 seconds and does not cause tissue adhesion during use. It can be stored at room temperature, is easy to spray, is suitable for irregular wound surfaces, has high wet adhesion, high mechanical strength, high biosafety, and can be used alone. It can quickly stop bleeding in tension wounds with irregular wet interfaces and heavy bleeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The present invention will be further described below with reference to the accompanying drawings and examples.

[0033] Figure 1 This is a state diagram of the hemostatic micropowder in this embodiment; Figure 2 1. The diagrams are of the states before, during and after the application of the hemostatic micropowder of this embodiment to the kidney wound of an experimental rabbit; Figure 3 This is a section of pathological tissue of the cecum of an experimental rabbit after the hemostatic micropowder of this embodiment was used on the kidney wound. DETAILED DESCRIPTION

[0034] The reagents and instruments used in the embodiments of the present invention are all commercially available products.

[0035] In the existing technology, continuous bleeding caused by accidental or surgical trauma can easily cause adhesion of surrounding tissues or organs, posing a serious threat to the patient's life safety. The choice of dressing and how to manually intervene to achieve rapid hemostasis are particularly important.

[0036] At present, hemostatic materials based on thrombin, gelatin, chitosan, collagen, fibrin, α-cyanoacrylate, etc. have been clinically used, but they generally have one or more problems such as slow wound sealing, low temperature storage requirement, inconvenient operation, need for a smooth wound surface, low wet adhesion, low mechanical strength, biosafety risks, and cannot be used alone.

[0037] Therefore, rapid hemostasis is difficult to achieve in complex and uncertain wounds, especially tension wounds with irregular wet interfaces and heavy bleeding. Several improvements have been proposed, such as a research paper (DOI: 10.1002 / adma.201905761) that synthesizes a hyperbranched polymer with a hydrophobic backbone and hydrophilic binder (catechol) branches via a Michael addition reaction. This hydrophilic binder then forms a gel layer, triggering the reaction with water, to seal and stop hemorrhage in wet wounds. However, while this gel layer exhibits high wet adhesion, mechanical strength, biosafety, and ease of operation, it indiscriminately adheres to surrounding tissues, causing adhesion.

[0038] The following is a further detailed description of the present invention through specific embodiments in the form of examples, but it should not be understood that the scope of the present invention is limited to the following examples.

[0039] Among them, the reagents and instruments used in the embodiments of the present invention are all common products available on the market.

[0040] Example 1 1. Synthesis of polyaldehyde cassava starch: Add cassava starch (average molecular weight of 60,000) into a round-bottom flask containing deionized water and stir at room temperature until the concentration is a homogeneous solution or dispersion state, with a concentration of 0.1 g / mL; protect from light, dissolve sodium periodate in deionized water to form a sodium periodate solution of 0.08 g / mL; protect from light, slowly add sodium perchlorate solution dropwise to the stirred cassava starch solution, with the volume ratio of cassava starch solution to sodium perchlorate solution being 1:1, and heat at 60°C for 12 hours. After the reaction is completed, centrifuge and wash 5 times at room temperature. After collecting the precipitate, vacuum dry it at -1 Bar pressure at room temperature for 24 hours to obtain polyaldehyde cassava starch.

[0041] 2. Preparation of hemostatic micropowder: 1 L of polyaldehyde cassava starch obtained in step (1) was dissolved in deionized water to prepare a polyaldehyde cassava starch solution with a concentration of 0.2 g / mL. 22 g of polyacrylamide (average molecular weight 4 million), 3 mg of rhamnolipid with a concentration of 0.02 mg / mL, and 10 mg of polyethylene glycol (molecular weight 200) with a concentration of 0.1 mg / mL were added to the polyaldehyde cassava starch solution, and stirred at room temperature until a stable viscous state was obtained. The viscous substance was freeze-dried for 48 h and ground to obtain hemostatic micropowder A. The state of hemostatic micropowder A is referred to Figure 1 .

[0042] 3. Use of Hemostatic Micropowder A: Spray hemostatic micropowder A onto the wound of the rabbit's kidney and cecal mucosa. Figure 2 and Figure 3 , the spraying amount is 0.3g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0043] Example 2 1. Synthesis of polyaldehyde pullulan: Add pullulan (average molecular weight of 100,000) to a round-bottom flask containing deionized water and stir at room temperature until the concentration is a homogeneous solution or dispersion state, with a concentration of 0.09 g / mL; protect from light, dissolve sodium perchlorate in deionized water to form a sodium perchlorate solution of 0.07 g / mL; protect from light, slowly add sodium perchlorate solution dropwise to the stirred pullulan solution, with the volume ratio of pullulan solution to sodium perchlorate solution being 1:1, heat at 60°C for 12 h, and after the reaction is completed, centrifuge and wash 5 times at room temperature. After collecting the precipitate, vacuum dry it at -1 Bar pressure at room temperature for 24 h to obtain polyaldehyde pullulan.

[0044] 2. Preparation of hemostatic micropowder: The polyaldehyde pullulan obtained in step (1) was dissolved in deionized water to prepare a polyaldehyde pullulan solution with a concentration of 0.02 g / mL. 20 g of polyetheramine (average molecular weight 3 million), 2.5 mg of lauryl glucoside and 10 mg of polyethylene glycol (molecular weight 500) were added to the polyaldehyde pullulan solution, and the mixture was stirred at room temperature until it became a stable viscous state. The viscous substance was freeze-dried for 96 hours and ground to obtain hemostatic micropowder B.

[0045] 3. Use of Hemostatic Micropowder B: Spray hemostatic micropowder B onto the wet substrate or wound at a spraying rate of 0.3g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0046] Example 3 1. Synthesis of polyaldehyde hyaluronic acid: Add hyaluronic acid (average molecular weight of 100,000) into a round-bottom flask containing deionized water and stir at room temperature until the concentration reaches a homogeneous solution or dispersion state of 0.1 g / mL. Dissolve sodium periodate in deionized water to form a 0.1 g / mL sodium periodate solution in the dark. Slowly drop the strong sodium periodate solution into the stirred hyaluronic acid solution in the dark, with the volume ratio of hyaluronic acid solution to sodium periodate solution being 1:1. Heat at 60°C for 12 h. After the reaction is completed, centrifuge and wash 5 times at room temperature. Collect the precipitate and vacuum dry it at -1 Bar pressure at room temperature for 24 h to obtain polyaldehyde hyaluronic acid.

[0047] 2. Preparation of hemostatic micropowder: The polyaldehyde hyaluronic acid obtained in step (1) is dissolved in deionized water to prepare a polyaldehyde hyaluronic acid solution with a concentration of 0.02 g / mL; 22 g of concentrated polyacrylamide (average molecular weight 4 million), 2.5 mg of lauryl glucoside, and 10 mg of polyethylene glycol with a molecular weight of 200 and a concentration of 0.1 mg / mL are added to the polyaldehyde hyaluronic acid solution; the mixture is stirred at room temperature until it becomes a stable viscous state; the viscous substance is freeze-dried for 48 h, and then ground to obtain hemostatic micropowder C.

[0048] 3. Use of Hemostatic Micropowder C: Spray hemostatic micropowder C onto the wet substrate or wound at a spraying rate of 0.2g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0049] Example 4 1. Synthesis of polyaldehyde chondroitin sulfate: Add chondroitin sulfate (average molecular weight of 10,000) to a round-bottom flask containing deionized water and stir at room temperature until the concentration is a homogeneous solution or dispersion at 0.03 g / mL. Protect from light, dissolve sodium perchlorate in deionized water to form a sodium perchlorate solution at 0.03 g / mL. Protect from light, slowly add sodium perchlorate solution dropwise to the stirred chondroitin sulfate solution at a volume ratio of 1:0.5. Heat at 25°C for 3 hours. After the reaction is completed, centrifuge and wash four times at room temperature. Collect the precipitate and vacuum dry it at -0.7 Bar for 24 hours at room temperature to obtain polyaldehyde chondroitin sulfate.

[0050] 2. Preparation of hemostatic micropowder: The polyaldehyde chondroitin sulfate obtained in step (1) is dissolved in deionized water to prepare a polyaldehyde chondroitin sulfate solution with a concentration of 0.05 g / mL; a polyacrylamine solution with a concentration of 0.05 g / mL and an average molecular weight of 5 million, a rhamnolipid with a concentration of 0.1 mg / mL, and polyethylene glycol with a molecular weight of 1000 and a concentration of 0.1 mg / mL are added to the polyaldehyde chondroitin sulfate solution; the mixture is stirred at room temperature until it becomes a stable viscous state; the viscous substance is freeze-dried for 12 hours and ground to obtain hemostatic micropowder D.

[0051] 3. Use of Hemostatic Powder D: Spray hemostatic powder D onto the wet substrate or wound at a spraying rate of 0.09g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0052] Example 5 1. Synthesis of polyaldehyde glucomannan: Add glucomannan (average molecular weight of 150,000) to a round-bottom flask containing deionized water and stir at room temperature until the concentration reaches 0.6 g / mL, which is a homogeneous solution or dispersion. Dissolve sodium perbromate in deionized water to form a 0.1 g / mL sodium perbromate solution in the dark. Slowly add sodium perchlorate solution dropwise to the stirred glucomannan solution in a volume ratio of 1:2, and heat at 25°C for 18 h. After the reaction is complete, centrifuge and wash five times at room temperature. Collect the precipitate and vacuum dry it at -0.7 Bar for 24 h at room temperature to obtain polyaldehyde glucomannan.

[0053] 2. Preparation of hemostatic micropowder: The polyaldehyde glucomannan obtained in step (1) was dissolved in deionized water to prepare a polyaldehyde glucomannan solution with a concentration of 0.05 g / mL, and a polyetheramine solution with a concentration of 0.5 g / mL and an average molecular weight of 5 million, a sophorol ester with a concentration of 0.15 mg / mL, and polyethylene glycol with a molecular weight of 200 and a concentration of 0.5 mg / mL were added to the polyaldehyde glucomannan solution, and stirred at room temperature until a stable viscous state was obtained; the viscous substance was freeze-dried for 12 hours and ground to obtain hemostatic micropowder E.

[0054] 3. Use of Hemostatic Micropowder E: Spray hemostatic micropowder E onto the wet substrate or wound at a spraying rate of 0.09 g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0055] Example 6 1. Synthesis of polyaldehyde heparin: Add heparin (average molecular weight of 100,000) to a round-bottom flask containing deionized water and stir at room temperature until the concentration reaches a homogeneous solution or dispersion state of 0.20 g / mL. Dissolve sodium periodate in deionized water in the dark to form a sodium periodate solution of 0.5 g / mL. Slowly add sodium perchlorate solution dropwise to the stirred heparin solution in the dark, with the volume ratio of heparin solution to sodium periodate solution being 1:0.5. Heat at 25°C for 18 h. After the reaction is complete, centrifuge and wash seven times at room temperature. Collect the precipitate and vacuum dry it at -0.7 Bar for 24 h at room temperature to obtain polyaldehyde heparin.

[0056] 2. Preparation of hemostatic micropowder: The polyaldehyde heparin obtained in step (1) is dissolved in deionized water to prepare a polyaldehyde heparin solution with a concentration of 0.05 g / mL, polyacrylamine with a concentration of 0.08 g / mL and an average molecular weight of 1 million, sodium lauroyl glutamate with a concentration of 0.15 mg / mL, and polyethylene glycol with a molecular weight of 400 and a concentration of 0.2 mg / mL are added to the polyaldehyde heparin solution, and stirred at room temperature until a stable viscous state is obtained; the viscous substance is freeze-dried for 12 h and ground to obtain hemostatic micropowder F.

[0057] 3. Use of Hemostatic Micropowder F: Spray hemostatic micropowder F onto the wet substrate or wound at a spraying rate of 0.7g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0058] Example 7 1. Synthesis of polyaldehyde chitosan: Add chitosan (average molecular weight of 50,000) into a round-bottom flask with deionized water and stir at room temperature until the concentration is a homogeneous solution or dispersion state, with a concentration of 0.03 g / mL; protect from light, dissolve sodium periodate in deionized water to form a sodium periodate solution of 0.2 g / mL; protect from light, slowly add sodium perchlorate solution dropwise to the stirred chitosan solution, with the volume ratio of chitosan solution to sodium periodate solution being 1:2.5, heat at 60°C for 3 h, and after the reaction is completed, centrifuge and wash 5 times at room temperature. After collecting the precipitate, vacuum dry it at -0.7 Bar pressure at room temperature for 24 h to obtain polyaldehyde chitosan.

[0059] 2. Preparation of hemostatic micropowder: The polyaldehyde chitosan obtained in step (1) is dissolved in deionized water to prepare a polyaldehyde chitosan solution with a concentration of 0.05 g / mL, and a polylysine solution with a concentration of 0.6 g / mL and an average molecular weight of 2 million, a sucrose palmitate with a concentration of 0.1 mg / mL, and polyethylene glycol with a molecular weight of 500 and a concentration of 0.5 mg / mL are added to the polyaldehyde chitosan solution, and stirred at room temperature until a stable viscous state is obtained; the viscous substance is freeze-dried for 12 hours and ground to obtain hemostatic micropowder G.

[0060] 3. Use of Hemostatic Micropowder G: Spray hemostatic micropowder G onto the wet substrate or wound at a spraying rate of 0.2g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0061] Example 8 1. Synthesis of polyaldehyde sodium alginate: Sodium alginate (average molecular weight of 100,000) was added to a round-bottom flask containing deionized water and stirred at room temperature until the concentration was a homogeneous solution or dispersion of 0.05 g / mL. Sodium perbromate was dissolved in deionized water in the dark to form a sodium perbromate solution of 0.03 g / mL. Sodium perbromate solution was slowly added dropwise to the stirred sodium alginate solution in the dark, with a volume ratio of sodium alginate solution to sodium perbromate solution of 1:1.5. The mixture was heated at 50°C for 6 h. After the reaction was completed, the mixture was centrifuged and washed 5 times at room temperature. The precipitate was collected and vacuum dried at -0.7 Bar for 24 h at room temperature to obtain polyaldehyde sodium alginate.

[0062] 2. Preparation of hemostatic micropowder: dissolve the polyaldehyde sodium alginate obtained in step (1) in deionized water to prepare a polyaldehyde sodium alginate solution with a concentration of 0.05 g / mL; add a polylysine solution with a concentration of 0.05 g / mL and an average molecular weight of 100,000; sodium lauroyl glutamate with a concentration of 0.02 mg / mL; and polyethylene glycol with a molecular weight of 1000 and a concentration of 0.1 mg / mL to the polyaldehyde sodium alginate solution; stir at room temperature until the mixture becomes stable and viscous; freeze-dry the viscous substance for 12 h, and grind it to obtain hemostatic micropowder H.

[0063] 3. Use of Hemostatic Micropowder H: Spray Hemostatic Micropowder H onto the wet substrate or wound at a spraying rate of 0.8g / cm 2 The hemostatic time, wet adhesion, tissue adhesion strength, and mechanical strength obtained are recorded in Table 1.

[0064] The data obtained from the tests on the hemostatic micropowders AH in Examples 1-8 are summarized in the following table, see Table 1 for details:

[0065] As can be seen from Table 1, the hemostatic micropowders AH prepared in this embodiment can quickly gel and seal the wound when sprinkled on the wound, with a gelation time of approximately 5.0s-5.3s; they have high wet adhesion to the wound, which can reduce the probability of the gel falling off the wound, help to improve the degree of close adhesion between the gel and the wound, and fully exert the hemostatic effect; the tissue adhesion strength of hemostatic micropowders AH is 0 kPa, indicating that the hemostatic micropowders do not indiscriminately adhere to surrounding tissues while achieving hemostasis, which is advantageous over some improved methods; the rupture resistance pressure is between 17.6kPa-18.1kPa, with high mechanical strength, suitable for matching the elastic modulus of treating internal organs such as the heart, kidneys, and cecum, and is beneficial for maintaining the normal physiological function of internal organs while treating wounds.

[0066] The above is a specific description of the preferred implementation of the present invention, but the invention is not limited to the embodiments. Those skilled in the art can make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A method for preparing hemostatic micropowder, characterized in that: include: S20, adding a primary amine polymer, an additive A, and an additive B to the polyaldehyde-polysaccharide biomolecule solution, and stirring at room temperature until a stable viscous state is obtained to obtain a viscous substance; S30. Freeze-drying the viscous substance and grinding it to obtain hemostatic micropowder.

2. The preparation method according to claim 1, wherein The preparation of the polyaldehyde polysaccharide biomolecule solution comprises the following steps: S10, slowly dripping the strong oxidant solution into the polysaccharide biomolecule solution while heating; S11, centrifuge and wash 4-7 times at room temperature, and collect the precipitate; S12, drying of the precipitate to obtain polyaldehyde-polysaccharide biomolecules; S13. Dissolve the polyaldehyde polysaccharide biomolecules in water to obtain a polyaldehyde polysaccharide biomolecule solution.

3. The preparation method according to claim 2, wherein: The polysaccharide biomolecule is one of chondroitin sulfate, hyaluronic acid, heparin, glucomannan, chitosan, pullulan, starch, and sodium alginate, and has an average molecular weight of 10,000 to 150,000.

4. The preparation method according to claim 1, wherein: The mass concentration of the polyaldehyde polysaccharide biomolecule solution is 0.05-0.60 g / mL.

5. The preparation method according to claim 2, wherein In step S10, the reaction volume ratio of the polysaccharide biomolecule solution to the strong oxidant solution is 1:0.5-2.5, and / or the heating reaction temperature is 25-60° C., and / or the heating reaction time is 3-18 hours.

6. The preparation method according to claim 1, wherein: The primary amine polymer is one of polyacrylamide, polyacrylamine, polyetheramine and polylysine, with an average molecular weight of 100,000 to 5,000,000 and / or a mass concentration of 0.05 to 0.6 g / mL.

7. The preparation method according to claim 1, wherein: The auxiliary agent A is one of lauryl glucoside, rhamnolipid, sophoroyl ester, sodium lauroyl glutamate, and sucrose palmitate, and / or has a mass concentration of 0.02 to 0.15 mg / mL.

8. The preparation method according to claim 1, wherein: The auxiliary agent B is polyethylene glycol, and its molecular weight is 200-1000, and / or its mass concentration is 0.1-0.5 mg / mL.

9. A method for using hemostatic micropowder, characterized by: The hemostatic micropowder prepared by the method for preparing hemostatic micropowder according to any one of claims 1 to 8 is sprayed onto the wound or dissolved in water to form a wound colloid, which covers the wound.

10. The method of use as described above is characterized in that: The spraying amount of the hemostatic micropowder is 0.09-0.8 g / cm 2 .

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