A composite hemostatic fabric and method of making same

Abstract

The present application relates to the technical field of hemostatic fabric, in particular to a composite hemostatic fabric and a preparation method thereof, the composite hemostatic fabric comprises a long-chain polysaccharide solution, glycerol, bovine thrombin, bovine serum albumin and an inorganic hemostatic agent, wherein the content of the long-chain polysaccharide solution ranges from 0.01% to 0.5%, the mass concentration of glycerol in the solution ranges from 0.5% to 15%, and the base fabric material includes but is not limited to defatted cotton gauze, all-cotton non-woven fabric, polyester non-woven fabric, polyester viscose fiber blended non-woven fabric, polyurethane non-woven fabric, polyurethane sponge sheet, melamine resin sponge sheet, sponge strip and defatted cotton flower. The present application has the advantages of mild reaction conditions, low energy consumption, easy continuous production, high production efficiency, low cost, low shedding rate of the inorganic hemostatic agent, avoidance of adverse events, simple debridement after hemostasis, no tissue burning during the hemostasis process, successful hemostasis for patients with blood diseases, good enzyme activity of the composite hemostatic fabric after accelerated aging, and suitability for normal temperature storage.

Description

Technical Field

[0001] This invention relates to the field of hemostatic fabric technology, specifically to a composite hemostatic fabric and its preparation method. Background Technology

[0002] Uncontrollable massive bleeding is a leading cause of death in traffic accidents, war, and major natural disasters. Because the human or animal coagulation system requires a relatively slow activation process, it cannot handle large, rapid bleeding like arterial hemorrhage, resulting in nearly half of all deaths occurring before hospitalization. In some cases, the trauma and bleeding are minor, requiring only basic first aid and standard coagulation function to stop the bleeding. It is noteworthy that patients with coagulation disorders may not be able to stop bleeding even from small wounds using conventional methods, leading to life-threatening uncontrollable bleeding. In modern warfare, with the widespread use of tourniquets, the probability of death from massive bleeding in the limbs has greatly decreased. However, bleeding in areas such as the chest, abdomen, head, neck, and junctions that cannot be controlled by tourniquets often results in massive hemorrhage due to a lack of effective treatment. Injuries to these areas are often accompanied by rupture of major arteries, easily leading to death due to delayed or ineffective hemostasis. This type of battlefield hemorrhage has become the leading cause of fatal massive bleeding deaths in modern warfare. Furthermore, because bleeding sites in the torso and surrounding areas are deep, it is often difficult to use forceps for emergency hemostasis on the battlefield, making hemostasis a challenging aspect of combat wound care. If massive hemorrhage is not treated promptly, excessive blood loss can lead to death, a very serious global problem; the World Health Organization reports that 1.9 million people die from excessive bleeding worldwide each year. Therefore, effective and rapid hemostasis is crucial in emergency treatment of sudden accidents in daily life, in wound hemostasis during hospital surgeries, and especially in the care of wounded soldiers in war. Thus, developing safe, effective, rapid, portable, and low-cost pre-hospital hemostatic agents, while also meeting the hemostatic needs of patients with impaired coagulation function, has always been an urgent need for emergency hemostasis.

[0003] Currently, commercially available topical hemostatic products can be broadly categorized into the following six types based on their raw materials:

[0004] 1. Fibrin glue: The main functional components of this type of product are fibrinogen and thrombin. Fibrin glue hemostatic materials are expensive and require strict storage conditions, making them unsuitable for use outside of hospitals.

[0005] 2. Gelatin and collagen; these products are also relatively expensive.

[0006] 3. Polysaccharide hemostatic agents, such as alginate, chitosan, and starch;

[0007] 4. Artificially synthesized polymer materials, such as cyanoacrylate, polyethylene glycol, polyglycolic acid, and polyacrylic acid, are all polymeric hemostatic materials;

[0008] 5. Inorganic hemostatic agents, including natural or synthetic zeolite, kaolin, montmorillonite, etc.

[0009] 6. Cellulose materials: As the earliest used wound dressing and hemostatic materials, cotton bandages or gauze are accepted by people due to their convenience, ease of use and good biocompatibility, but their hemostatic effect is not good.

[0010] Numerous animal and clinical trials have demonstrated that among the six hemostatic materials mentioned above, inorganic and polysaccharide hemostatic agents are the most effective for stopping massive bleeding from battlefield wounds, and are also convenient to store and transport. However, these hemostatic agents have traditionally been in powder form, leaving a large amount of powder adhering to the wound after hemostasis, making subsequent debridement difficult and introducing various safety issues. Furthermore, the use of inorganic hemostatic agents such as zeolite molecular sieves for hemostasis often results in massive exothermic burns to the wound surface, which is another challenge in their application. Studies have shown that conventional inorganic hemostatic agents are ineffective in stopping bleeding in patients with impaired coagulation function, failing to achieve successful hemostasis. Therefore, further technical improvements are needed for the use of inorganic hemostatic agents in stopping bleeding in patients with impaired coagulation function.

[0011] The coagulation process is generally considered to involve two relatively independent pathways converging into a common pathway: thrombin formation as the endpoint of the reaction. Thrombin is the catalyst for the final target reaction of the entire coagulation system—fibrinogen hydrolysis—and is the most crucial protease in the coagulation catalysis reaction. It has a strong fibrinogen-hydrolyzing ability, and controlling thrombin synthesis and increasing its activity can be considered the fundamental way for the coagulation system to achieve its hemostatic function. The coagulation reaction process is a multi-step cascade reaction process (i.e., the coagulation cascade model), mainly including the extrinsic coagulation pathway and the intrinsic coagulation pathway, and the convergence of the extrinsic and intrinsic coagulation pathways at factor Xa, entering the common coagulation pathway. Coagulation factor II (Prothrombin) is catalyzed by factors Xa, Va, and Ca. 2+The resulting prothrombinase complex is activated into thrombin, which cleaves fibrinogen to form fibrin monomers. With the help of coagulation factor XIIIa, these fibrin monomers cross-link to form a fibrin network, leading to thrombosis. Inorganic hemostatic agents have abundant pores, and their surfaces or the inner surfaces of these pores are negatively charged. The intrinsic coagulation pathway begins with the activation of coagulation factor XII by the negatively charged inorganic hemostatic agent, followed by the cascade propagation and activation of coagulation factors XI, IX, X, and VIII. Therefore, inorganic hemostatic agents can quickly stop bleeding in patients with normal coagulation function. However, in patients with severe hemophilia, the use of inorganic hemostatic agents for hemostasis is ineffective due to the absence of coagulation factors VIII and IX, which impairs the intrinsic coagulation pathway.

[0012] Quikclot COMBAT GAUZE, developed by Z-Medica in the United States in 2006, uses non-woven fabric fibers adhered with a large number of tiny kaolinite hemostatic agents. This combat gauze is widely used for stopping massive bleeding from combat wounds. The non-woven fabric fibers are obtained by soaking, drying, slitting, vacuum packaging, and sterilizing in a solution containing kaolinite, glycerin, and water. However, this invention has a drawback: when the combat gauze is placed in a solution system (or comes into contact with blood), the kaolinite adhering to the surface of the non-woven fibers can detach freely. During contact with the wound, these tiny kaolinite particles may enter human tissue, posing a potential clinical risk. Furthermore, it is ineffective in stopping bleeding in patients with coagulation disorders.

[0013] The invention, titled "A Hemostatic Complex and Its Preparation Method" (CN201810625854.7), discloses a method for preparing a hemostatic complex and its applications. This invention eliminates the need for adhesives in the hemostatic complex. The invention utilizes a chemically matched coupling between a molecular sieve and fibers. The molecular sieve is synthesized chemically, and the chemical synthesis reaction conditions for molecular sieves generally require high temperature and high pressure (the temperature in the implementation case is required to be 100 degrees Celsius). The synthesis of zeolite molecular sieves is time-consuming, has low production capacity, and high energy consumption. The high-temperature reaction process causes certain damage to the substrate fibers, and the production process generates a large amount of wastewater and solid waste, which is environmentally unfriendly, inefficient, and costly.

[0014] The invention, "A hemostatic fabric containing trypsin and its preparation method" (CN111249516A), still uses the chemical matching coupling between molecular sieve and fiber (the temperature is required to be 100 degrees Celsius in all implementation cases). The molecular sieve is grown on the fiber surface by chemical synthesis and then compounded with trypsin to obtain a composite hemostatic material. The invention of synthesizing molecular sieve on the fiber surface still has shortcomings such as harsh reaction conditions, high energy consumption and long production cycle.

[0015] The invention described in CN115990284A, titled "A slurry for coating hemostatic gauze and hemostatic gauze prepared therefrom", uses a suspending agent to prepare the coating slurry. However, an adhesive is still added to the slurry. The hemostatic gauze prepared by this invention has a soft feel, but it is not effective in stopping bleeding for patients with coagulation disorders.

[0016] Therefore, manufacturing a composite hemostatic fabric that does not use any adhesives, has a low rate of inorganic hemostatic agent shedding due to the substrate fiber being loaded, has a soft feel, is suitable for use in various harsh environments, has high production efficiency, low energy consumption, is environmentally friendly, and has high procoagulant activity, enabling successful hemostasis in hemophilic mice after tail amputation (and can also be used by patients with coagulation disorders) remains a challenging technical problem. Summary of the Invention

[0017] The purpose of this invention is to provide a composite hemostatic fabric and its preparation method, in order to solve the problem mentioned in the background art that the existing technology cannot manufacture a composite hemostatic fabric that does not use any adhesive, has a mild production process, low energy consumption, low shedding rate of inorganic hemostatic agent loaded on the substrate fiber, is suitable for use in harsh environments, and has high procoagulant activity that can achieve successful hemostasis of tail amputation in hemophilic mice (and can also be used by patients with coagulation disorders).

[0018] To achieve the above objectives, the present invention provides the following technical solution: a composite hemostatic fabric, wherein the composite hemostatic fabric comprises a long-chain polysaccharide solution, glycerol, bovine thrombin, bovine serum albumin and an inorganic hemostatic agent, wherein the content of the long-chain polysaccharide solution ranges from 0.01% to 0.5%, and the mass concentration of glycerol in the solution ranges from 0.5% to 15%.

[0019] As a further step of the present invention, the substrate fiber material includes, but is not limited to, degreased cotton gauze, all-cotton nonwoven fabric, polyester nonwoven fabric, nonwoven fabric blended with polyester viscose fiber, polyurethane nonwoven fabric, polyurethane sponge sheet, melamine resin sponge sheet, sponge strip, and degreased cotton.

[0020] As a further step of the present invention, the mixed solution system includes one or more long-chain polysaccharides or polysaccharides such as xanthan gum, konjac gum, camu camu gum, guar gum, carrageenan, and locust bean gum. The content of long-chain polysaccharides ranges from 0.01% to 0.5%, and the content of long-chain polysaccharides with a molecular length greater than 3 μm is greater than 1%. Preferably, the long-chain polysaccharide is a mixture of xanthan gum and konjac gum, and the mass ratio of xanthan gum to konjac gum is 8:2 to 4:6.

[0021] As a further step of the present invention, the inorganic hemostatic agent is not limited to synthetic molecular sieves or natural inorganic hemostatic agents. Synthetic molecular sieves include any one or more of the following: X-type molecular sieve, Y-type molecular sieve, A-type molecular sieve, ZSM-5 molecular sieve, chalcogenide, β-type molecular sieve, mordenite, L-type molecular sieve, P-type molecular sieve, AlPO4-5 type molecular sieve, and AlPO4-11 type molecular sieve.

[0022] As a further step of the present invention, the natural inorganic hemostatic agent is such as: kaolin, clinoptilolite, diaspore, montmorillonite, sodium bentonite, calcium bentonite, polyphosphate and other oxides and other inorganic hemostatic materials; before use, the inorganic hemostatic agent is soaked in a 5M concentration of strontium chloride, calcium chloride and magnesium chloride solution at room temperature for 12 hours once or multiple times to obtain an ion exchange inorganic hemostatic agent.

[0023] A method for preparing a composite hemostatic fabric includes the following steps:

[0024] S1: Stir the mixed solution of long-chain polysaccharide, glycerol and hemostatic agent at high speed, place the substrate fiber in the solution, take it out and dry it to prepare hemostatic fabric for later use;

[0025] S2: Weigh out appropriate amounts of bovine thrombin and bovine serum albumin, and dissolve them in water to prepare a bovine thrombin and bovine serum albumin solution.

[0026] S3: Add an appropriate amount of HEPES and adjust the pH to 6-8 using 0.1M saline or sodium hydroxide.

[0027] S4: Immerse the above hemostatic fabric in a solution of bovine thrombin and bovine serum albumin, react for a period of time, remove and vacuum dry to a certain extent, cut, vacuum package and sterilize to obtain the composite hemostatic fabric.

[0028] As a further step of the present invention, the inorganic hemostatic agent is kaolin, zeolite, or bentonite, with a content ranging from 0.01% to 20%; the glycerol mass concentration in the solution is preferably in the range of 1% to 5%; the D50 range of the inorganic hemostatic agent is 1nm to 1000nm, and the D50 range is preferably 20nm to 500nm; preferably, it is a mixture of kaolin and zeolite or a mixture of kaolin and bentonite, with a mass ratio of kaolin to zeolite of 9:1 to 2:8; when a mixture of kaolin and bentonite is selected, the mass ratio of kaolin to bentonite is 9:1 to 2:8. The resulting solution is prepared by high-speed stirring at a ratio of 3:7. The substrate fiber is then immersed in the solution or passed through the solution under mechanical traction. It is then dried at 105°C or in a drying tunnel to obtain the hemostatic fabric. Appropriate amounts of bovine thrombin and bovine serum albumin are weighed in a ratio of 1:100 to 1:1000, preferably 1:100 to 1:500. The concentration of the bovine thrombin and bovine serum albumin solution is 1–50 g / L; the glycerol concentration in the solution ranges from 0.5% to 10%. The hemostatic fabric is then immersed in the bovine thrombin and bovine serum albumin solution or passed through the solution under mechanical traction. After vacuum drying to a certain degree, it is slit, vacuum-packed, and sterilized to obtain the composite hemostatic fabric.

[0029] As a further step of the present invention, the mass ratio of the long-chain polysaccharide xanthan gum and konjac gum in the solution system is 7:3 to 4:6. The long-chain polysaccharide preferably has a molecular length of more than 3 μm and a content of more than 1%, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent type A molecular sieve with a D50 of 100 nm is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the immersion is repeated twice in a 5M calcium chloride solution at room temperature for 12 hours. Then, it is filtered and cleaned. The mass concentration of the inorganic hemostatic agent calcium-based type A molecular sieve is 3%. The mass concentration of glycerol in the solution ranges from 1% to 5%. The remainder is deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is immersed in the solution and dried at 105°C to prepare a hemostatic fabric.

[0030] The mass ratio of bovine thrombin to bovine serum albumin is 1:100 to 1:500, and the concentration of the bovine thrombin and bovine serum albumin solution is 1 to 10 g / L. An appropriate amount of HEPES is added, and the pH is adjusted to 6 to 8 using 0.1M saline or sodium hydroxide. The remainder is deionized water. The above hemostatic fabric is immersed in the bovine thrombin and bovine serum albumin solution, then vacuum dried to a certain extent, cut, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0031] Compared with the prior art, the beneficial effects of the present invention are:

[0032] The solution system of this invention contains one or more long-chain polysaccharides or polysaccharides such as xanthan gum, konjac gum, camu camu gum, guar gum, carrageenan, and locust bean gum, one or more inorganic hemostatic agents, and a certain amount of glycerol. After high-speed stirring, the solution contains long-chain polysaccharides, inorganic hemostatic agents, and glycerol uniformly and stably. When the substrate fiber is immersed in the solution, a large amount of solution will be adsorbed on the surface of the substrate fiber. After drying, the long-chain polysaccharides and inorganic hemostatic agents will entangle to form a net structure and be loaded on the surface of the substrate fiber. Water or solution can quickly penetrate into the interior of the net structure.

[0033] Inorganic hemostatic agents have abundant pores, allowing some long-chain polysaccharide molecules or side chains to insert into the inner surface of these pores while simultaneously being encapsulated by the long-chain polysaccharides. The resulting entanglement of the long-chain polysaccharides and the inorganic hemostatic agent forms a net-like structure, which, after drying on the fiber surface, becomes the hemostatic fabric. When the hemostatic fabric is impregnated with a solution of bovine thrombin and bovine serum albumin, or sprayed with such a solution, the solution rapidly penetrates the net-like structure. When the bovine thrombin and bovine serum albumin in the solution encounter the inorganic hemostatic agent within the net-like structure, the positively charged amino acid residues of the proteins (bovine thrombin and bovine serum albumin) can interact with the negatively charged surface (Zeta). The inorganic hemostatic agent (potential) attracts itself on its surface or within its pores. The pores of the inorganic hemostatic agent have a diameter of approximately 0.4–1.3 nm, which generally cannot accommodate intact protein molecules. Therefore, proteins such as bovine thrombin and bovine serum albumin are rapidly captured and adsorbed onto the surface or pores of the inorganic hemostatic agent. Since the surface of the hemostatic fabric (i.e., the substrate fiber) is already loaded with the inorganic hemostatic agent and long-chain polysaccharides, the long-chain polysaccharides and inorganic hemostatic agent become entangled. Some long-chain polysaccharide molecules or side chains insert into the inner surface of the inorganic hemostatic agent pores or are encapsulated by long-chain polysaccharide molecules. When the hemostatic fabric adsorbs bovine thrombin and bovine serum albumin, which have multidimensional spatial structures, the strong adsorption between the inorganic hemostatic agent and biological proteins such as bovine thrombin and bovine serum albumin, along with the synergistic effect of the long-chain polysaccharides, makes the net-like structure on the surface of the substrate fiber more stable, forming a long-chain polysaccharide inorganic-biological protein complex, i.e., a composite hemostatic fabric. The inorganic hemostatic agent is firmly loaded onto the surface of the substrate fiber and is not easily detached, avoiding the risk of it entering the human body.

[0034] Experimental results show that the long-chain polysaccharide inorganic-biological protein complex with a net-like structure possesses protein corona properties, exhibiting excellent procoagulant activity. When blood rapidly enters the net-like structure composed of the long-chain polysaccharide inorganic-biological protein complex, the highly procoagulant protein corona initiates a coagulation reaction, significantly shortening hemostasis time and reducing blood loss. Due to the high procoagulant protein corona formed on the surface of the substrate fibers, which exhibits high thrombin activity, the composite hemostatic fabric successfully achieved hemostasis in hemophilic mice after tail amputation, unlike conventional combat gauze which failed to stop the bleeding. This greatly expands the product's application areas. The composite hemostatic fabric not only maintains the softness of the substrate fibers but also makes it suitable for use in various complex and harsh environments.

[0035] The paper "A Hemostatic Complex and Its Preparation Method" (CN201810625854.7) describes the in-situ chemical synthesis of zeolite molecular sieves on the surface of substrate fiber. Generally, this requires high temperature and high pressure (the temperature in the implementation case is required to be 100 degrees Celsius). The reaction temperature is high and the energy consumption is high. The production process generates a large amount of waste liquid and solid waste, which is not environmentally friendly. The chemical synthesis of molecular sieves takes a long time and is costly.

[0036] Early sandwich structures adhered to tiny particles, but the amount of adhesive used was small and the adhesion effect was poor. Inorganic hemostatic agents were unstable and easily detached. With a larger amount of adhesive, inorganic hemostatic agents were less likely to detach, but they were prone to blocking the molecular sieve channels. Once the molecular sieve channels were blocked, their hemostatic performance was affected.

[0037] This invention employs a two-step method. The first step involves loading an inorganic hemostatic agent, long-chain polysaccharides, and glycerol onto the surface of a substrate fiber to prepare a hemostatic fabric. The second step involves the specific adsorption and binding of bovine thrombin or trypsin and bovine serum albumin to the inorganic hemostatic agent and long-chain polysaccharides loaded on the substrate fiber, resulting in a synergistic effect and the preparation of a structurally stable long-chain polysaccharide-inorganic-biological protein complex, i.e., a composite hemostatic fabric. In the first step, the long-chain polysaccharides and inorganic hemostatic agents in the net-like structure hemostatic fabric are intertwined, preventing rapid flocculation and precipitation due to the interaction between the inorganic hemostatic agent and the biological protein. The second step, preparing the long-chain polysaccharide-inorganic-biological protein composite hemostatic fabric, creatively utilizes the strong adsorption characteristics of inorganic hemostatic agents and proteins. The synergistic effect of the long-chain polysaccharides, inorganic hemostatic agents, and proteins yields a highly procoagulant protein corona complex, which rapidly promotes coagulation and reduces blood loss upon contact with blood. Thrombin activity tests on the composite hemostatic fabric demonstrate its high procoagulant activity. Compared to the failure of conventional combat gauze (COMBAT GAUZE) to achieve hemostasis in hemophilic mice after tail amputation, the composite hemostatic fabric successfully achieved hemostasis in hemophilic mice after tail amputation, greatly expanding the product's application areas. The low shedding rate of the inorganic hemostatic agent significantly reduces the risk of adverse clinical events, simplifies wound cleaning after hemostasis, and avoids tissue burns during the hemostasis process.

[0038] The production process of this invention features mild reaction conditions, low energy consumption, easy continuous production, high production efficiency, and low cost. The inorganic hemostatic agent has a low shedding rate, avoiding adverse events. Post-hemostasis wound cleaning is simple, and the hemostasis process does not burn tissue. It can successfully stop bleeding in patients with blood disorders. The composite hemostatic fabric maintains good enzyme activity even after accelerated aging and is suitable for room temperature storage. Attached Figure Description

[0039] Figure 1 This is a schematic diagram of the preparation steps of the structure of the present invention;

[0040] Figure 2 This is a schematic diagram of the coagulation waterfall model of the present invention;

[0041] Figure 3 This is a schematic diagram of the xanthan gum molecular structure of the present invention;

[0042] Figure 4 This is a schematic diagram of the molecular structure of konjac gum according to the present invention;

[0043] Figure 5 This is a schematic diagram of the composite hemostatic gauze structure of the present invention. Detailed Implementation

[0044] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0045] Please see Figures 1 to 5 One embodiment provided by the present invention:

[0046] A composite hemostatic fabric, embodiment 1:

[0047] The solution system of this invention contains one or more long-chain polysaccharides (or polysaccharides) such as xanthan gum, konjac gum, camu camu camu gum, guar gum, carrageenan, and locust bean gum. Preferably, the long-chain polysaccharides have a molecular length of 3 μm or more and a content of 1% or more. The content range of the long-chain polysaccharides is 0.01% to 0.5%, with a preferred range of 0.02% to 0.05%. The mass concentration of glycerol in the solution ranges from 0.5% to 15%. The solution system also contains one or more small inorganic hemostatic agents such as kaolin, zeolite, and bentonite. The inorganic hemostatic agents are not limited to natural or synthetic agents, and the D50 range of the inorganic hemostatic agent is 1 nm to 500 nm, with a solid content range of 0.01% to 20%. After the obtained solution is stirred at high speed, the substrate fiber is placed in the solution, removed, and dried to prepare a hemostatic fabric for later use.

[0048] Weigh appropriate amounts of bovine thrombin and bovine serum albumin in a ratio of (1:100) to (1:1000), dissolve them in water to prepare a bovine thrombin and bovine serum albumin solution with a concentration of 1–50 g / L; the glycerol concentration in the solution should be 0.5%–10%; add an appropriate amount of HEPES, and adjust the pH to 6–8 using 0.1M saline or sodium hydroxide. Immerse the hemostatic fabric in the bovine thrombin and bovine serum albumin solution, react for a period of time, remove it, vacuum dry it to a certain degree, cut it, vacuum package it, and sterilize it to obtain the composite hemostatic fabric.

[0049] The solution system contains one or more long-chain polysaccharides (or polysaccharides) such as xanthan gum, konjac gum, camu ...

[0050] The inorganic hemostatic agent is not limited to natural or synthetic, and may be one or more types, with a content ranging from 0.01% to 20%; the mass concentration of glycerol in the solution ranges from 0.5% to 15%, preferably from 1% to 5%; the D50 of the inorganic hemostatic agent ranges from 1 nm to 1000 nm, and preferably from 20 nm to 500 nm; kaolin and zeolite or a mixture of kaolin and bentonite are preferred, with a mass ratio of kaolin to zeolite of (9:1) to (2:8); when a mixture of kaolin and bentonite is selected, the mass ratio of kaolin to bentonite is (9:1) to (3:7); after the obtained solution is stirred at high speed, the substrate fiber is immersed in the solution or the substrate fiber is passed through the solution under the traction of a mechanical device, and dried at 105°C or in a drying tunnel to obtain the hemostatic fabric.

[0051] Inorganic hemostatic agents are not limited to synthetic molecular sieves or natural inorganic hemostatic agents. Synthetic molecular sieves include any one or more of the following: X-type molecular sieve, Y-type molecular sieve, A-type molecular sieve, ZSM-5 molecular sieve, chalcogenide, β-type molecular sieve, mordenite, L-type molecular sieve, P-type molecular sieve, AlPO4-5 type molecular sieve, and AlPO4-11 type molecular sieve. Natural inorganic hemostatic agents include: kaolin, clinoptilolite, diaspore, montmorillonite, sodium bentonite, calcium bentonite, polyphosphates, and other oxides and other inorganic hemostatic materials. Before use, inorganic hemostatic agents are soaked in a 5M concentration of strontium chloride, calcium chloride, and magnesium chloride solution at room temperature for 12 hours once or multiple times to obtain ion-exchange inorganic hemostatic agents.

[0052] Weigh out appropriate amounts of bovine thrombin and bovine serum albumin in a ratio of (1:100) to (1:1000), preferably (1:100) to (1:500). The concentration of the bovine thrombin and bovine serum albumin solution is 1 to 50 g / L; the glycerol concentration in the solution ranges from 0.5% to 10%. The hemostatic fabric is then impregnated in the bovine thrombin and bovine serum albumin solution, or passed through the solution under mechanical traction. Afterward, it is vacuum dried to a certain degree, cut, vacuum-packed, and sterilized to obtain the composite hemostatic fabric.

[0053] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 3μm or longer and contains more than 1% of it, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100nm), is impregnated in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the impregnation is repeated twice with the same 5M calcium chloride solution at room temperature for 12 hours. The mixture is then filtered and cleaned. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 3%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The hemostatic fabric was impregnated in the bovine thrombin and bovine serum albumin solution, then vacuum-dried to a certain extent, slit, vacuum-packed, and sterilized to prepare the composite hemostatic fabric.

[0054] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.13 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 0.14%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 0.16%. The in vitro coagulation time is 45 s. The thrombin activity of the composite hemostatic fabric is 1.45 NIH U. The composite hemostatic fabric successfully stopped hemostasis in the tail of hemophilia mice, and the survival rate of the mice was 100%.

[0055] Implementation 2:

[0056] The solution system contained xanthan gum (long-chain polysaccharide) and konjac gum in a mass ratio of (7:3) to (4:6). The preferred long-chain polysaccharide had a molecular length of 3 μm or more and a content of 1% or more, with a concentration of 0.02% to 0.05%. Inorganic hemostatic agent type A molecular sieve (D50 of 100 nm) was impregnated in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the impregnation was repeated twice with the same 5M calcium chloride solution at room temperature for 12 hours, followed by thorough cleaning. The mass concentration of the inorganic hemostatic agent calcium-based type A molecular sieve was 3%. The mass concentration of glycerol in the solution ranged from 1% to 5%, with the remainder being deionized water. The resulting solution was stirred at 4000 rpm for 10 minutes. The substrate fiber was impregnated in the solution, dried at 105℃ to a certain degree, slit, vacuum-packed, and sterilized to prepare the hemostatic fabric.

[0057] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.35 cm. Under the condition of ultrasonic cleaning for 1 minute, the inorganic hemostatic agent shedding rate in water is 84.21%. Under the condition of ultrasonic cleaning for 5 minutes, the inorganic hemostatic agent shedding rate in water is 88.26%. The in vitro coagulation time is about 124 s. The thrombin activity of the composite hemostatic fabric is 0 NIH U. Hemostasis of tail amputation of hemophilia mice failed and all mice died.

[0058] The difference between Example 2 and Example 1 is that the prepared hemostatic fabric was not impregnated with bovine thrombin and bovine serum albumin solution. The composite hemostatic fabric could not form substances with high prothrombin activity, therefore, the use of the composite hemostatic fabric for tail amputation in hemophilic mice failed to achieve hemostasis, and all mice died.

[0059] Implementation 3:

[0060] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 3μm or longer and contains more than 1% of it, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100nm), is impregnated in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the impregnation is repeated twice with the same 5M calcium chloride solution at room temperature for 12 hours, followed by thorough cleaning. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105℃ to prepare a hemostatic fabric. The concentration of bovine serum albumin solution is 1–10 g / L. An appropriate amount of HEPES is added, and the pH is adjusted to 6–8 using 0.1M saline or sodium hydroxide. The remainder is deionized water. The above-mentioned hemostatic fabric was impregnated in bovine serum albumin solution, vacuum dried to a certain extent, cut, vacuum packaged, and sterilized to prepare composite hemostatic fabric.

[0061] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.41 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 0.18%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 0.20%. The in vitro coagulation time is 119 s. The thrombin activity of the composite hemostatic fabric is 0 NIH U. Hemostasis of tail amputation of hemophilia mice using the composite hemostatic fabric failed, and all mice died.

[0062] The difference between Example 3 and Example 1 is that the prepared composite hemostatic fabric, when immersed in bovine serum albumin solution, does not contain bovine thrombin. Since the composite hemostatic fabric lacks bovine thrombin, it cannot form substances with high prothrombin activity. Therefore, the use of the composite hemostatic fabric to stop hemostasis in tail amputation of hemophilia mice failed, and all mice died.

[0063] Implementation 4:

[0064] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6), with the long-chain polysaccharide preferably having a molecular length of 3 μm or more and a content of more than 1%, and a concentration of 0.02% to 0.05%. The mass ratio of the inorganic hemostatic agent kaolin to Y-type molecular sieve is (9:1) to (6:4), with D50 values ​​of 500 nm and 400 nm, respectively. The solution is soaked in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the soaking is repeated twice with a 5M calcium chloride solution at room temperature for 12 hours, followed by thorough cleaning. The mass concentration of the inorganic hemostatic agents calcium-based kaolin and calcium-based Y-type molecular sieve is 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The above hemostatic fabric was impregnated in the bovine thrombin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0065] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.43 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 0.12%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 0.13%. The in vitro coagulation time is 41 s. The thrombin activity of the composite hemostatic fabric is 1.52 NIH U. The composite hemostatic fabric successfully stopped hemostasis in the tail of hemophilia mice, and the survival rate of the mice was 100%.

[0066] The difference between Example 4 and Example 1 is that two different inorganic calcium-based hemostatic agents were used.

[0067] Implementation 5:

[0068] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6), with the long-chain polysaccharide preferably having a molecular length of 3 μm or more and a content of more than 1%, and a concentration of 0.02% to 0.05%. The mass ratio of the inorganic hemostatic agent kaolin to Y-type molecular sieve is (9:1) to (6:4), with D50 values ​​of 500 nm and 400 nm, respectively, and a mass concentration of 2% for both kaolin and Y-type molecular sieve. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The above hemostatic fabric was impregnated in the bovine thrombin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0069] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.60 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 1.01%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 1.26%. The in vitro coagulation time is 76 s. The thrombin activity of the composite hemostatic fabric is 1.13 NIH U. The composite hemostatic fabric successfully stopped hemostasis in the tail of hemophilia mice, and the survival rate of the mice was 100%.

[0070] The difference between Example 5 and Example 1 is that the inorganic hemostatic agent was used directly without ion exchange. The experimental results show that the inorganic hemostatic agent after calcium, magnesium and strontium exchange is more conducive to the adsorption of bovine thrombin, reducing in vitro clotting time and increasing thrombin activity.

[0071] Implementation 6:

[0072] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 3μm or longer and contains more than 1% of it, with a concentration of 0.02% to 0.05%. The mass ratio of the inorganic hemostatic agents kaolin and bentonite is (9:1) to (6:4), with D50 values ​​of 2μm and 3μm respectively. The mixture is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the immersion is repeated twice with the same 5M calcium chloride solution at room temperature for 12 hours. The mixture is then thoroughly cleaned. The mass concentrations of the inorganic hemostatic agents calcium-based A kaolin and calcium-based bentonite are 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The above hemostatic fabric was impregnated in the bovine thrombin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0073] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.51 cm. Under the condition of ultrasonic cleaning for 1 minute, the inorganic hemostatic agent shedding rate in water is 4.24%. Under the condition of ultrasonic cleaning for 5 minutes, the inorganic hemostatic agent shedding rate in water is 5.26%. The in vitro coagulation time is 79 s. The thrombin activity of the composite hemostatic fabric is 1.46 NIH U. The composite hemostatic fabric can still successfully stop bleeding when the tail of hemophilic mice is cut off, and the survival rate of mice is 100%.

[0074] The difference between Example 6 and Example 1 is that the D50 values ​​of the inorganic hemostatic agents kaolin and bentonite are 2μm and 3μm, respectively. Compared with the 100nm D50 value of the inorganic hemostatic agent type A molecular sieve in Example 1, the particle size of the inorganic hemostatic agent is larger. The larger particle size of the inorganic hemostatic agent increases the difficulty of encapsulating long-chain polysaccharide molecules, thus increasing the detachment rate in water.

[0075] Implementation 7:

[0076] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 3μm or longer and contains more than 1% of it, with a concentration of 0.02% to 0.05%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin is (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution is 1 to 10 g / L. An appropriate amount of HEPES is added, and the pH is adjusted to 6 to 8 using 0.1M saline or sodium hydroxide; the remainder is deionized water. The above hemostatic fabric is impregnated in the bovine thrombin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare a composite hemostatic fabric.

[0077] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.37 cm, the in vitro clotting time is 354 s, the hemostatic fabric does not detach when not loaded with inorganic hemostatic agent, and the thrombin activity of the composite hemostatic fabric is 0.03 NIH U. The hemostasis of tail amputation of hemophilia mice failed when the composite hemostatic fabric was used, most of the mice died, and the survival rate was 33.3%.

[0078] The difference between Example 7 and Example 1 is that no inorganic hemostatic agent was added to the solution in the first step. The experiment showed that some bovine thrombin was inactivated and did not have the ability to promote coagulation. Therefore, the in vitro coagulation time was longer and the thrombin activity was lower.

[0079] Implementation 8:

[0080] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The preferred long-chain polysaccharide has a molecular length of 3 μm or more and a content of 1% or more, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100 nm), is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, this process is repeated twice, immersing the sieve in the 5M calcium chloride solution at room temperature for 12 hours each time, followed by thorough cleaning. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%. An additional 0.3% of the binder component from the polymer colloidal emulsion is added to the solution, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is immersed in the solution, dried at 105°C to a certain degree, slit, vacuum-packed, and sterilized to prepare the hemostatic fabric.

[0081] The hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 3.56 cm, the in vitro clotting time is 164 s, and the inorganic hemostatic agent shedding rate in water is 5.29% under ultrasonic cleaning for 1 min and 7.26% under ultrasonic cleaning for 5 min. The thrombin activity of the hemostatic fabric is 0 NIH U. However, tail cutting hemostasis failed in hemophilia mice, and all mice died.

[0082] The difference between Example 8 and Example 1 is that an appropriate amount of adhesive is added to the solution in the first step, and the solution is not impregnated with bovine thrombin and bovine serum albumin solution in the second step. Therefore, the bovine thrombin activity of the hemostatic fabric is 0 NIH U, and the longer in vitro clotting time may be due to the adhesive blocking the inorganic hemostatic agent.

[0083] Implementation 9:

[0084] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The preferred long-chain polysaccharide has a molecular length of 3 μm or more and a content of 1% or more, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100 nm), is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, this process is repeated twice, immersing the sieve in the 5M calcium chloride solution at room temperature for 12 hours. The sieve is then thoroughly cleaned. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%. An additional 5% of the binder component from the polymer colloidal emulsion is added to the solution, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is immersed in the solution, dried at 105°C to a certain degree, slit, vacuum-packed, and sterilized to prepare the hemostatic fabric.

[0085] The hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 6.33 cm, the in vitro clotting time is 215 s, and the inorganic hemostatic agent shedding rate in water is 0.28% under the condition of ultrasonic cleaning for 1 min and 0.34% under the condition of ultrasonic cleaning for 5 min. The thrombin activity of the hemostatic fabric is 0 NIH U. However, the tail cutting hemostasis failed in hemophilia mice, and all mice died.

[0086] The difference between Example 9 and Example 8 is that more adhesive was added. The increase in adhesive content effectively reduced the ultrasonic detachment rate, and the increased adhesive content further prolonged the in vitro coagulation time.

[0087] Implementation 10:

[0088] In this embodiment, the concentration of the long-chain polysaccharide xanthan gum in the solution system is 0.02%–0.05%; the long-chain polysaccharide preferably has a molecular length of 3 μm or more and a content of 1% or more; the inorganic hemostatic agent type A molecular sieve (D50 of 100 nm) is impregnated in a 5M calcium chloride solution at room temperature for 12 hours, separated from the solution, and then impregnated twice more in a 5M calcium chloride solution at room temperature for 12 hours, and then cleaned; the mass concentration of the inorganic hemostatic agent calcium-based type A molecular sieve is 2%; the mass concentration of glycerol in the solution ranges from 1% to 5%; the balance is deionized water, and the resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The above hemostatic fabric was impregnated in the bovine thrombin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0089] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.37 cm, the in vitro clotting time is 115 s, and the inorganic hemostatic agent shedding rate in water is 1.71% under ultrasonic cleaning for 1 min and 1.86% under ultrasonic cleaning for 5 min. The thrombin activity of the composite hemostatic fabric is 1.42 NIH U. It can successfully stop bleeding when the tail is cut off in hemophilic mice, and the survival rate of the mice is 100%.

[0090] The difference between Example 10 and Example 1 is that only one long-chain polysaccharide, xanthan gum, was added to the solution in the first step. Experiments show that the combined use of long-chain polysaccharides has a better effect, and the combined use of long-chain polysaccharides can appropriately reduce the shedding of inorganic hemostatic agents.

[0091] Implementation 11:

[0092] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 3μm or longer and contains more than 1% of it, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100nm), is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, this process is repeated twice, immersing the sieve in the 5M calcium chloride solution at room temperature for 12 hours each time. The sieve is then thoroughly cleaned. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The hemostatic fabric was immersed in the bovine thrombin and bovine serum albumin solution for 20 seconds, then removed, vacuum-dried to a certain extent, slit, vacuum-packed, and sterilized to prepare the composite hemostatic fabric.

[0093] The difference between Example 11 and Example 1 is that the second step of immersing the hemostatic fabric in the solution is 20 seconds.

[0094] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.55 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 0.18%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 0.22%. The in vitro coagulation time is 47 s. The thrombin activity of the composite hemostatic fabric is 1.39 NIH U. The composite hemostatic fabric successfully stopped hemostasis in the tail of hemophilia mice, and the survival rate of the mice was 100%.

[0095] Implementation 12:

[0096] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 3μm or longer and contains more than 1% of it, with a concentration of 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100nm), is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, this process is repeated twice, immersing the sieve in the 5M calcium chloride solution at room temperature for 12 hours each time. The sieve is then thoroughly cleaned. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 2%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The hemostatic fabric was immersed in the bovine thrombin and bovine serum albumin solution for 10 seconds, then removed, vacuum-dried to a certain extent, slit, vacuum-packed, and sterilized to prepare the composite hemostatic fabric.

[0097] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.36 cm. Under ultrasonic cleaning conditions of 1 min, the inorganic hemostatic agent shedding rate in water is 0.21%; under ultrasonic cleaning conditions of 5 min, the shedding rate is 0.26%. The external coagulation time is 49 s, and the thrombin activity of the composite hemostatic fabric is 1.38 NIH U. Hemostasis was successfully achieved in hemophilic mice after tail amputation using the composite hemostatic fabric, with a 100% survival rate. The experimental results of this embodiment demonstrate that the present invention is suitable for continuous industrial production.

[0098] The difference between Example 12 and Example 11 is that the hemostatic fabric is immersed in the solution for 10 seconds in the second step. The difference is not significant, possibly because the immersion time is shorter than the vacuum drying time, so the length of the immersion time has little impact.

[0099] Implementation 13:

[0100] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 1% or more with a molecular length of 5 μm or more, and its concentration is 0.01% to 0.03%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100 nm), is impregnated in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, the impregnation is repeated twice with the same 5M calcium chloride solution at room temperature for 12 hours, followed by thorough cleaning. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 3%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin was (1:100) to (1:500), and the concentration of the bovine thrombin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The above hemostatic fabric was impregnated in the bovine thrombin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0101] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.01 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 0.14%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 0.15%. The in vitro coagulation time is 46 s. The thrombin activity of the composite hemostatic fabric is 1.44 NIH U. The composite hemostatic fabric successfully stopped hemostasis in the tail of hemophilia mice, and the survival rate of the mice was 100%.

[0102] The difference between Example 13 and Example 1 is that the content of long-chain polysaccharide molecules is different.

[0103] Implementation 14:

[0104] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 1% or more with a molecular length of 3 μm or more, and its concentration is 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100 nm), is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, this process is repeated twice, immersing the sieve in the 5M calcium chloride solution at room temperature for 12 hours each time. The sieve is then thoroughly cleaned. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 3%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The ratio of trypsin to bovine serum albumin was (1:100) to (1:500), and the concentration of the trypsin and bovine serum albumin solution was 1 to 10 g / L. An appropriate amount of HEPES was added, and the pH was adjusted to 6 to 8 using 0.1 M saline or sodium hydroxide. The remainder was deionized water. The hemostatic fabric was impregnated in the trypsin and bovine serum albumin solution, vacuum dried to a certain extent, slit, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

[0105] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.53 cm. Under the condition of ultrasonic cleaning for 1 minute, the inorganic hemostatic agent shedding rate in water is 0.15%. Under the condition of ultrasonic cleaning for 5 minutes, the inorganic hemostatic agent shedding rate in water is 0.17%. The in vitro coagulation time is 50 s. The thrombin activity of the composite hemostatic fabric is 1.38 NIH U. The composite hemostatic fabric successfully stopped hemostasis in the tail of hemophilia mice, and the survival rate of the mice was 100%.

[0106] The difference between Example 14 and Example 1 is that trypsin was used. The experimental results show that trypsin and bovine thrombin have no significant effect on the thrombin activity of the composite hemostatic fabric.

[0107] Implementation 15:

[0108] In this embodiment, the mass ratio of the long-chain polysaccharide xanthan gum to konjac gum in the solution system is (7:3) to (4:6). The long-chain polysaccharide is preferably 1% or more with a molecular length of 3 μm or more, and its concentration is 0.02% to 0.05%. The inorganic hemostatic agent, type A molecular sieve (D50 of 100 nm), is immersed in a 5M calcium chloride solution at room temperature for 12 hours. After separation from the solution, this process is repeated twice with a 5M calcium chloride solution at room temperature for 12 hours, followed by thorough cleaning. The mass concentration of the inorganic hemostatic agent, calcium-based type A molecular sieve, is 3%. The mass concentration of glycerol in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is impregnated in the solution and dried at 105°C to prepare a hemostatic fabric. The trypsin solution concentration is 0.1–5 g / L; an appropriate amount of HEPES is added, and the pH is adjusted to 6–8 using 0.1M saline or sodium hydroxide; the remainder is deionized water. The above-mentioned hemostatic fabric was impregnated with trypsin solution, vacuum dried to a certain extent, cut, vacuum packaged, and sterilized to prepare composite hemostatic fabric.

[0109] The composite hemostatic fabric prepared in this embodiment has a soft hand feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.51 cm. Under the condition of ultrasonic cleaning for 1 min, the inorganic hemostatic agent shedding rate in water is 6.34%. Under the condition of ultrasonic cleaning for 5 min, the inorganic hemostatic agent shedding rate in water is 7.31%. The in vitro coagulation time is 98 s. The thrombin activity of the composite hemostatic fabric is 0.04 NIH U. Hemostasis of tail amputation in hemophilia mice failed using the composite hemostatic fabric, and the survival rate of the mice was 33.3%.

[0110] The difference between Example 15 and Example 14 is that bovine serum albumin was not added to the protein solution. The bovine thrombin activity of the composite hemostatic fabric in this example is 0.04 NIH U. The test results show that bovine thrombin (or trypsin) needs to be used in conjunction with bovine serum albumin to ensure its thrombin activity.

[0111] Implementation 16:

[0112] Commercially available Combat Gauze consists of inorganic hemostatic materials (clay, kaolin) attached to the fiber surface. It has a soft feel and is suitable for hemostasis of various complex wounds. The average bending length is 2.04 cm. Under ultrasonic cleaning conditions of 1 min, the clay detachment rate from the gauze fibers is 91.2%; under ultrasonic cleaning conditions of 5 min, the clay detachment rate from the gauze fibers is 96.8%. The in vitro coagulation time is 117 s. The thrombin activity of the composite hemostatic fabric is 0 NIHU. Hemostasis of tail amputated hemophilic mice using the composite hemostatic fabric failed, and all mice died.

[0113] The performance testing conducted on this invention is as follows:

[0114] Sodium citrate (109 mmol / L) solution was mixed with fresh pig whole blood at a volume ratio of 1:9 and shaken well. An appropriate amount (X mL) of 0.2 M calcium chloride solution was added to a 5 mL centrifuge tube containing a certain amount of sodium citrate-anticoagulated pig whole blood. The blood clotting time was adjusted to 8–10 minutes, which is the natural clotting time. An appropriate amount (X mL) of 0.2 M calcium chloride solution and a certain amount of composite hemostatic fabric were added to a 5 mL centrifuge tube, along with a certain amount of sodium citrate-anticoagulated pig whole blood. The mixture was shaken well, and the clotting time was recorded as the in vitro clotting time.

[0115] Water shedding rate:

[0116] A certain amount of composite hemostatic fabric was soaked in water and sonicated for 1 minute and 5 minutes respectively. The composite hemostatic fabric or gauze was then removed and suspended above the soaking solution until no liquid dripped down. The soaking solution was then filtered to collect the detached particles, which were dried, weighed, and the detachment rate was calculated. This indicator assesses how many particles might enter the liquid when the product comes into contact with it.

[0117] The softness of a fabric is characterized by its bend length.

[0118] The bending length of a fabric refers to the length of a strip of fabric that extends downwards to 7.1° under its own weight when one end is suspended in the air. GB / T18318.1-2009 "Determination of bending properties of textiles - Part 1: Inclined plane method".

[0119] In this application, the test method for bending length is in accordance with GB / T 18318.1-2009 "Determination of bending properties of textiles - Part 1: Inclined plane method".

[0120] The average bending length is obtained by dividing the sum of the warp bending length and the weft bending length by 2.

[0121] The calculation method for the average bending length referenced the data processing method adopted in HG / T5254-2017 "Determination of Stiffening Effect of Stiffening Agents in Textile Dyeing and Finishing Auxiliaries".

[0122] The smaller the bending length value obtained from the test, the better the softness of the fabric, which means the worse the stiffness of the fabric. The larger the bending length, the less soft the fabric, which means the higher the stiffness of the fabric.

[0123] The sample was cut into strips 2 cm wide and stored at room temperature for 20 hours in an environment with a humidity of 60-70%. The average bending length was then measured on a 702-type stiffness tester according to the test method of national standard GB / T18318.1-2009.

[0124] Hemostasis time after tail amputation in hemophilic mice:

[0125] The tail of the hemophilia mouse was cut 1 cm from the end. The tail of the hemophilia mouse was bandaged with 4 layers of composite hemostatic fabric. Hemostasis was achieved when no more blood seeped out of the composite hemostatic fabric. The hemostasis time was recorded. Each sample was tested in triplicate and the average value was calculated. The survival rate of the hemophilia mice within 8 hours was also recorded and the survival rate was calculated.

[0126] Thrombin activity assay using chromogenic substrate method:

[0127] Thrombin undergoes enzymatic hydrolysis with its specific chromogenic substrate (CS-01(38), HYPHEN BioMed), which releases a substance with a chromogenic group (pNA). The absorbance of the chromogenic substrate at 405 nm is positively correlated with the activity of bovine thrombin.

[0128] A certain amount of composite hemostatic fabric sample, 1 mL of thrombin chromogenic substrate (4 mg / mL), and 1 mL of HEPES buffer (20 mM HEPES, 150 mM NaCl, pH 7.4) were thoroughly mixed in a centrifuge tube. The mixture was incubated at 37°C with shaking for 2 min, and then 500 μL of glacial acetic acid was added to terminate the reaction. The supernatant was collected by centrifugation, and the process was repeated twice. Finally, the absorbance at 405 nm was measured using 2 mL of the total supernatant.

[0129] Standard bovine thrombin solutions of different concentrations were prepared, and standard curves were plotted after measuring absorbance. Thrombin activity in the samples was calculated based on the standard curves. The stability method for the composite hemostatic fabric was as follows: the composite hemostatic fabric was cut into aluminum foil bags, sealed in vacuum packaging, sterilized, and then placed in a 60℃ oven for accelerated aging for 97 days (60℃*97D). Storage at room temperature was equivalent to 3 years.

[0130] Table 1 below shows the performance of composite hemostatic fabrics:

[0131]

[0132]

[0133] Table 2 below shows the experimental results of the hemostatic performance of composite hemostatic fabrics:

[0134]

[0135]

[0136] Table 3 below shows the Zeta potential of inorganic hemostatic agents.

[0137] 1 talc -16.67 2 clinoptilolite -21.51 3 Diaspore -15.39 4 Magnesium-rich montmorillonite -16.87 5 Calcium-based bentonite -14.12 6 Siliceous zeolite -14.24 7 diatomite -16.31 8 Calcium montmorillonite -17.56 9 Ca-Y molecular sieve -22.48 10 Ca-A molecular sieve -21.64 11 X-type molecular sieve -19.95 12 ZSM-5 molecular sieve -18.27 13 chabazite -16.98 14 β-molecular sieve -17.39 15 L-type molecular sieve -19.02 16 P-type molecular sieve -20.06 17 Kaolin -22.34

[0138] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. A method for preparing a composite hemostatic fabric, characterized in that, Includes the following steps: S1: A mixed solution of long-chain polysaccharides, glycerol, and an inorganic hemostatic agent is stirred at high speed. The substrate fiber is then immersed in the stirred mixed solution, removed, and dried to obtain a hemostatic fabric. The long-chain polysaccharide contains one or more of xanthan gum, konjac gum, camu ... S2: Weigh bovine thrombin and bovine serum albumin, dissolve them in water to prepare a bovine thrombin and bovine serum albumin solution; S3: Add an appropriate amount of HEPES and adjust the pH to 6-8 using 0.1M sodium hydroxide; S4: The above hemostatic fabric is immersed in a solution of bovine thrombin and bovine serum albumin. After reacting for a period of time, it is removed, vacuum dried to a certain extent, cut, vacuum packaged, and sterilized to obtain the composite hemostatic fabric.

2. The method for preparing a composite hemostatic fabric according to claim 1, characterized in that: The inorganic hemostatic agent is kaolin, zeolite, or bentonite, with a content ranging from 0.01% to 20%; the mass concentration of glycerol in the solution ranges from 1% to 5%.

3. The method for preparing a composite hemostatic fabric according to claim 2, characterized in that: The D50 range of the inorganic hemostatic agent is 1 nm to 1000 nm.

4. The method for preparing a composite hemostatic fabric according to claim 3, characterized in that: The inorganic hemostatic agent is a mixture of kaolin and zeolite or kaolin and bentonite, with a mass ratio of kaolin to zeolite of 9:1 to 2:

8.

5. The method for preparing a composite hemostatic fabric according to claim 4, characterized in that: When the inorganic hemostatic agent is a mixture of kaolin and bentonite, the mass ratio of kaolin to bentonite is 9:1 to 3:

7. After the resulting mixed solution is stirred at high speed, the substrate fiber is immersed in the mixed solution and dried at 105°C to obtain the hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin is 1:100 to 1:1000. The above hemostatic fabric is immersed in the bovine thrombin and bovine serum albumin solution, reacted for a period of time, removed, vacuum dried to a certain extent, cut, vacuum packaged, and sterilized to obtain the composite hemostatic fabric.

6. The method for preparing a composite hemostatic fabric according to claim 5, characterized in that: The bovine thrombin to bovine serum albumin mass ratio is 1:100 to 1:500, and the concentration of the bovine thrombin to bovine serum albumin solution is 1 to 50 g / L.

7. The method for preparing a composite hemostatic fabric according to claim 6, characterized in that: The long-chain polysaccharide is xanthan gum and konjac gum in a mass ratio of 7:3 to 4:6, with a molecular length greater than 3 μm, a content greater than 1%, and a concentration of 0.02% to 0.05%.

8. The method for preparing a composite hemostatic fabric according to claim 1, characterized in that: The inorganic hemostatic agent is a type A molecular sieve with a D50 of 100 nm. It is immersed in a 5M calcium chloride solution at room temperature for 12 hours, separated from the solution, and then immersed twice more in the same 5M calcium chloride solution at room temperature for 12 hours. After filtration and cleaning, the inorganic hemostatic agent (calcium-based type A molecular sieve) has a mass concentration of 3%. The glycerol mass concentration in the solution ranges from 1% to 5%, with the remainder being deionized water. The resulting mixed solution is stirred at 4000 rpm for 10 minutes. The substrate fiber is then immersed in the mixed solution and dried at 105°C to prepare a hemostatic fabric. The mass ratio of bovine thrombin to bovine serum albumin is 1:100 to 1:500, and the concentration of the bovine thrombin and bovine serum albumin solution is 1 to 10 g / L. An appropriate amount of HEPES is added, and the pH is adjusted to 6 to 8 using 0.1 M sodium hydroxide. The remainder is deionized water. The above hemostatic fabric is immersed in the bovine thrombin and bovine serum albumin solution, then vacuum dried to a certain extent, cut, vacuum packaged, and sterilized to prepare the composite hemostatic fabric.

9. A composite hemostatic fabric prepared by the method of claim 1, characterized in that: The substrate fiber materials include degreased cotton gauze, all-cotton nonwoven fabric, polyester nonwoven fabric, nonwoven fabric blended with polyester viscose fiber, polyurethane nonwoven fabric, polyurethane sponge sheet, melamine resin sponge sheet, sponge strip, and degreased cotton.

10. The composite hemostatic fabric according to claim 9, characterized in that: The long-chain polysaccharide is a mixture of xanthan gum and konjac gum, with a mass ratio of xanthan gum to konjac gum of 8:2 to 4:

6.

11. The composite hemostatic fabric according to claim 9, characterized in that: The inorganic hemostatic agent includes synthetic molecular sieves or natural inorganic hemostatic agents. The synthetic molecular sieves include any one or more of the following: X-type molecular sieve, Y-type molecular sieve, A-type molecular sieve, ZSM-5 molecular sieve, chalcogenide, β-type molecular sieve, mordenite, L-type molecular sieve, P-type molecular sieve, AlPO4-5 type molecular sieve, and AlPO4-11 type molecular sieve.

12. The composite hemostatic fabric according to claim 9, characterized in that: The inorganic hemostatic agent includes one or more of the following: kaolin, clinoptilolite, sodium bentonite, and calcium bentonite. Before use, the inorganic hemostatic agent is soaked in a 5M concentration of strontium chloride, calcium chloride, and magnesium chloride solution at room temperature for 12 hours once or multiple times to obtain an ion-exchange inorganic hemostatic agent.

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