Application and preparation method of carboxymethyl chitosan-modified activated zeolite

Abstract

The invention discloses application of carboxymethyl chitosan-modified activated zeolite, a hydrogel biological material and a preparation method, and relates to the technical field of hemostatic and antibacterial materials. The hydrogel biological material is prepared from 1%-30% of a carboxymethyl chitosan-modified activated zeolite composite material, 0.1%-2.5% of carrageenan, 0.05%-2% of konjac glucomannan, 0.02%-2% of calcium chloride, 0.02%-2% of edible glucose, 0.02%-2% of maltodextrin, 0.01%-1% of potassium citrate, 0.01%-2% of xanthan gum, 0.01%-1% of sodium chloride and the balance of water. Wherein the carboxymethyl chitosan-modified activated zeolite endows the hydrogel biological material with excellent hemostatic performance, high adhesiveness, antibacterial property and wound healing promotion performance. When the hydrogel biomaterial is prepared from the hydrogel biomaterial, polysaccharide xanthan gum, konjac gum, carrageenan and other substances, the hydrogel biomaterial can be adsorbed and self-adhered to human skin on the premise of not adding any adhesive, and meanwhile, the hydrogel biomaterial has excellent functions of stopping bleeding, resisting bacteria and promoting wound healing.

Description

Technical Field

[0001] This invention belongs to the field of hemostatic and antibacterial materials technology, and more specifically relates to the use and preparation method of carboxymethyl chitosan-modified activated zeolite. Background Technology

[0002] Quikclot COMBAT GAUZE, developed by Z-Medica in the United States in 2006, is a nonwoven fabric substrate with a large number of tiny kaolinite hemostatic agents adhering to it. This combat gauze is widely used for stopping massive bleeding from combat wounds. The nonwoven fabric substrate fibers are obtained by soaking, drying, folding, vacuum packaging, and sterilization in a solution containing kaolinite, glycerin, and water. However, a drawback of this invention is that when the combat gauze is placed in a solution system (or comes into contact with blood), the kaolinite adhering to the surface of the nonwoven fibers can detach freely. During contact with the wound, these tiny detached kaolinite particles may enter human tissue, posing a potential clinical risk.

[0003] Patent CN201810625854.7 discloses a method for preparing a hemostatic compound and its application. This invention eliminates the need for adhesives in the hemostatic compound. It utilizes a chemically matched coupling between a molecular sieve and fibers. The molecular sieve is chemically synthesized, and the chemical synthesis reaction conditions generally require high temperature and pressure (100℃ in the implementation examples). Zeolite molecular sieve synthesis is time-consuming, has low production capacity, and high energy consumption. The high-temperature reaction process causes damage to the substrate fibers, and the production process generates large amounts of wastewater and solid waste, which is environmentally unfriendly, inefficient, and costly. CN111249516A also employs a chemically matched coupling between a molecular sieve and fibers, where the molecular sieve is chemically synthesized and grown on the fiber surface, then combined with trypsin to obtain a composite hemostatic material. However, this invention still suffers from drawbacks such as harsh reaction conditions, high energy consumption, and long production cycles. CN115990284A uses a suspending agent to prepare a coating slurry, but an adhesive is still added to the slurry to prevent the inorganic hemostatic agent from detaching during clinical use. The hemostatic materials described in CN201810625854.7, CN111249516A, and CN115990284A do not have antibacterial, wound barrier (physical isolation), moisturizing, or wound healing promotion functions.

[0004] Studies have shown that zeolite, being negatively charged, can activate coagulation factors and possesses excellent hemostatic properties, while also exhibiting deodorizing and adsorption functions. Chitosan, being positively charged, also possesses excellent hemostatic and wound-healing properties, along with deodorizing and adsorption capabilities. If the functional groups of carboxymethyl chitosan (CMCS) could be exposed at the interface, enhancing hydrogen bonds, ionic bonds, or hydrophobic interactions with contact surfaces (such as skin and mucous membranes), the adhesiveness of hydrogel hemostatic materials could be improved, thereby preventing them from detaching from the skin. However, this technology has not yet been studied. If chitosan could be modified to enhance its adhesiveness, a novel approach could be developed that achieves excellent hemostasis and self-adhesion to human skin without the addition of any binder.

[0005] COOK's Hemospray uses carbon dioxide, compressed air, and endoscopic tubing to deliver hemostatic powder to the bleeding site in the gastrointestinal tract. However, Hemospray can sometimes cause the hemostatic powder to clog the machine or equipment, or the powder can cover the endoscope lens, obstructing the doctor's view. Summary of the Invention

[0006] Based on this, the present invention provides the uses and preparation method of carboxymethyl chitosan-modified activated zeolite, overcoming the defects of poor adhesion and poor antibacterial properties of hemostatic materials in the prior art.

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

[0008] The first aspect of the present invention provides the use of a carboxymethyl chitosan-modified activated zeolite composite material, the use being in the preparation of a highly adhesive biomaterial that promotes wound healing and has high antibacterial properties;

[0009] Preferably, the adhesiveness is such that it is not easily separated from or detached from the skin.

[0010] While existing technologies do include the combined application of chitosan and modified zeolite in hemostatic materials, these methods typically involve simple mixing without any modification or reaction. This invention, however, mixes carboxymethyl chitosan with modified activated zeolite and induces a reaction. The inventors discovered, through accidental observation, that the reaction of carboxymethyl chitosan and modified activated zeolite under specific conditions creatively enhances the adhesiveness of the biomaterial, preventing it from detaching from the skin. Furthermore, the synergistic effect of the positive charge of CMCS and the negative charge of the modified activated zeolite in the carboxymethyl chitosan-modified activated zeolite composite material, along with the presence of Zn in the zeolite channels, further contributes to this effect. 2+ and Cu 2+ The synergistic effect achieves antibacterial properties while greatly reducing the cytotoxicity of Cu-based zeolites and mitigating the adverse effects of copper in activated zeolites on cytotoxicity.

[0011] A second aspect of the present invention provides a biomaterial comprising a carboxymethyl chitosan-modified activated zeolite composite material and a medically acceptable excipient.

[0012] Preferably, the biomaterial is used for hemostasis, antibacterial properties, and promoting wound healing.

[0013] Preferably, the biomaterial includes hydrogel production materials and hemostatic particles obtained by drying hydrogel biomaterials;

[0014] Preferably, the medically acceptable excipients include carrageenan, sodium chloride, konjac gum, maltodextrin, potassium citrate, calcium chloride, glucose, and xanthan gum.

[0015] Compared with the prior art, the hydrogel production material prepared by the present invention has good adhesion, and the prepared hemostatic agent microparticles will not form a mist-like powder that covers the endoscope lens and affects the doctor's visual effect.

[0016] Preferably, the total raw materials comprise, by mass percentage, the following: 1%-30% carboxymethyl chitosan-modified activated zeolite composite material, 0.1%-2.5% carrageenan, 0.05%-2% konjac gum, 0.02%-2% calcium chloride, 0.02%-2% edible glucose, 0.02%-2% maltodextrin, 0.01%-1% potassium citrate, 0.01%-2% xanthan gum, 0.01%-1% sodium chloride, and the balance being water.

[0017] A third aspect of the present invention provides a method for preparing the biomaterial, applicable to the preparation of the biomaterial, comprising the following steps:

[0018] (1) Prepare carboxymethyl chitosan-modified activated zeolite composite material and weigh each component according to the mass ratio;

[0019] (2) Add carboxymethyl chitosan-modified activated zeolite composite material, carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride to water and stir evenly. Heat to 85°C until boiling to obtain a gel solution.

[0020] (3) Place the gel solution into a specific mold and allow it to cool naturally to obtain the hydrogel biomaterial; or prepare hydrogel biomaterial particles by spraying and cooling, and prepare hemostatic particles after drying the biomaterial particles.

[0021] Compared with existing technologies, this invention mixes carboxymethyl chitosan-modified activated zeolite composite material with other excipients such as carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, and xanthan gum, and forms a stable three-dimensional network structure through cross-linking. This network encapsulates zeolite particles, restricting their free fall and further enhancing adhesion. This innovatively achieves self-adhesion between the hydrogel hemostatic material and human skin without adding any binders (for ease of use), and also gives the hydrogel antibacterial, moisturizing, and barrier properties.

[0022] Preferably, the preparation process of the carboxymethyl chitosan-modified activated zeolite composite material is as follows:

[0023] Chitosan is dispersed in a solvent to achieve a chitosan mass concentration of 5%-30%. Then, an alkaline solution is added to alkalize the chitosan to a pH of 12-14. Next, chloroacetic acid solution is added and stirred, and the solution is neutralized with acid to a pH of 7. The mixture is centrifuged to remove the precipitate, yielding a carboxymethyl chitosan solution. The mass ratio of chloroacetic acid to chitosan is 1:1-5:1. The pH of the chitosan solution is adjusted to 4.5-6. Modified activated zeolite is added at a mass ratio of chitosan solution to modified activated zeolite of 3:1-1:1. Glutaraldehyde is then added to achieve a final mass concentration of 0.5%-1%. The mixture is stirred at room temperature or heated to 50°C for 4-8 hours, centrifuged, and the precipitate is cleaned until neutral and dried to obtain a carboxymethyl chitosan-modified activated zeolite composite material.

[0024] Preferably, the solvent is a mixture of isopropanol and water; and the mass ratio of isopropanol to water is 7:3-9:3.

[0025] Preferably, the alkaline solution is a strongly alkaline solution.

[0026] Preferably, the strongly alkaline solution is a NaOH solution, and the mass concentration of the NaOH solution is 5%-30%.

[0027] Preferably, the acid solution is a strongly acidic solution.

[0028] Preferably, the strong acid solution is an HCl solution, and the mass concentration of the HCl solution is 5%-20%.

[0029] Preferably, the preparation process of the activated modified zeolite is as follows: zeolite is added to a mixed solution containing 2M-4M zinc chloride, 1M-2M titanium tetrachloride, and 1M-3M calcium chloride, such that the mass ratio of zeolite to solution is 1:5-1:10. After stirring and filtering, ammonia gas is introduced and reacted at 30-50℃ for 4-10 hours. After calcination, the modified activated zeolite is obtained.

[0030] Preferably, the stirring speed is 500 rpm to 5000 rpm.

[0031] Preferably, the filtration is performed using a 0.2 μm membrane.

[0032] Preferably, the calcination is carried out at 300℃-500℃ for 2h-6h.

[0033] Compared with existing technologies, the preparation of modified and activated zeolites can achieve zeolite surface modification and ion exchange through impregnation with metal ion solutions. Specifically, the aluminosilicate framework of the zeolite adsorbs Zn through ion exchange. 2+ Ti 4+ Ca 2+ Meanwhile, ZnCl2 can etch zeolite channels, increasing the specific surface area; Ti 4+ Subsequent calcination can form TiO2 nanoparticles, enhancing the material's activity. Ammonia gas is introduced because it can react with metal ions, modulating surface chemical properties, forming amino or nitride active sites, increasing the material's surface alkalinity and hydrophilicity, and enhancing its compatibility with carboxymethyl chitosan.

[0034] Preferably, the gel solution can be sprayed onto the surface of a carrier before cooling, and then naturally cooled to obtain the gel; wherein the carrier includes woven fabric, medical gauze, or cotton fiber.

[0035] Compared with existing technologies, when the gel solution is sprayed onto the carrier material, it is more conducive to the gel solution being carried and encapsulated, preventing it from falling off the skin surface.

[0036] The fourth aspect of this invention provides the application of the biomaterials prepared by the above method in the preparation of medical-related products.

[0037] In summary, the technical effects achieved by this invention are: this invention creatively crosslinks carboxymethyl chitosan with modified activated zeolite, which increases adhesion performance, prevents detachment, and endows biomaterials with antibacterial properties.

[0038] Moreover, the composite material of carboxymethyl chitosan and modified activated zeolite, along with other excipients, can form a three-dimensional network structure that encapsulates zeolite particles, restricting their free fall and further enhancing adhesion. This creatively achieves the effect of self-adhesion between hydrogel hemostatic materials and human skin without adding any binders. Detailed Implementation

[0039] The technical solutions in the embodiments of the present invention will be clearly and completely described below. 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.

[0040] Existing hemostatic materials have poor adhesion and antibacterial properties, and require adhesive materials, which can irritate the skin.

[0041] To achieve the above objectives, the present invention adopts the following technical solution:

[0042] The first aspect of the present invention provides the use of carboxymethyl chitosan-modified activated zeolite composite material in the preparation of highly adhesive biomaterials that promote wound healing and have high antibacterial properties.

[0043] It should be understood that the adhesiveness here refers to the fact that it is not easily separated from the skin and detached.

[0044] When the above technical solution is adopted, the adhesion of the material is significantly improved, preventing the biological material from detaching from the human skin surface. Furthermore, the synergistic effect of the positive charge of CMCS and the negative charge of the modified activated zeolite in the carboxymethyl chitosan-modified activated zeolite composite material, as well as the Zn content within the zeolite channels... 2+ and Cu 2+ The synergistic effect achieves antibacterial properties while greatly reducing the cytotoxicity of Cu-based zeolites and mitigating the adverse effects of copper in activated zeolites on cytotoxicity.

[0045] A second aspect of the present invention provides a biomaterial comprising a carboxymethyl chitosan-modified activated zeolite composite material and a medically acceptable excipient.

[0046] It should be understood that the aforementioned biomaterials are used for hemostasis, antibacterial purposes, and promoting wound healing.

[0047] In some embodiments, the biomaterial includes hydrogel biomaterial and hemostatic granules prepared by drying the hydrogel biomaterial; more preferably, it is a hydrogel biomaterial.

[0048] In some embodiments, medically acceptable excipients include carrageenan, sodium chloride, konjac gum, maltodextrin, potassium citrate, calcium chloride, glucose, and xanthan gum.

[0049] In some embodiments, the total raw materials include the following by mass percentage: 1%-30% carboxymethyl chitosan-modified activated zeolite composite material, 0.1%-2.5% carrageenan, 0.05%-2% konjac gum, 0.02%-2% calcium chloride, 0.02%-2% edible glucose, 0.02%-2% maltodextrin, 0.01%-1% potassium citrate, 0.01%-2% xanthan gum, 0.01%-1% sodium chloride, and the balance being water.

[0050] Examples include: 1% carboxymethyl chitosan-modified activated zeolite composite, 2% carrageenan, 0.06% konjac gum, 0.1% calcium chloride, 0.1% edible glucose, 0.5% maltodextrin, 0.02% potassium citrate, 0.05% xanthan gum, 0.01% sodium chloride, and the balance being water.

[0051] Example composition includes: 30% carboxymethyl chitosan-modified activated zeolite composite, 0.1% carrageenan, 0.5% sodium chloride, 2% konjac gum, 2% maltodextrin, 1% potassium citrate, 0.02% calcium chloride, 0.02% edible glucose, 0.01% xanthan gum, and the balance being water.

[0052] Example composition includes: 20% carboxymethyl chitosan-modified activated zeolite composite, 2.5% carrageenan, 1% sodium chloride, 1% konjac gum, 2% maltodextrin, 1% potassium citrate, 2% calcium chloride, 2% edible glucose, 2% xanthan gum, and the balance being water.

[0053] The third aspect of this invention provides a method for preparing the above-mentioned biomaterial, comprising the following steps:

[0054] (1) Prepare carboxymethyl chitosan-modified activated zeolite composite material and weigh each component according to the mass ratio;

[0055] (2) Add carboxymethyl chitosan-modified activated zeolite composite material, carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride to water and stir evenly. Heat to 85°C until boiling to obtain a gel solution.

[0056] (3) Place the gel solution into a specific mold and allow it to cool naturally to obtain hydrogel biomaterial.

[0057] When the above technical solution is adopted, the carboxymethyl chitosan-modified activated zeolite composite material is mixed with other excipients such as carrageenan, sodium chloride, konjac gum, xanthan gum, maltodextrin, glucose, potassium citrate, and calcium chloride. Through cross-linking, a stable three-dimensional network structure is formed, which encapsulates the zeolite particles, restricts the free fall of zeolite, and further enhances the adhesion. Without adding any binder, the hydrogel hemostatic material is creatively adsorbed and self-adhesed to human skin, and the hydrogel has antibacterial, moisturizing and barrier properties.

[0058] In some embodiments, the preparation process of the carboxymethyl chitosan-modified activated zeolite composite material is as follows:

[0059] Chitosan is dispersed in a solvent to achieve a chitosan mass concentration of 5%-30%; then an alkaline solution is added to alkalize the chitosan to a pH of 12-14; subsequently, chloroacetic acid solution is added and stirred, and the solution is neutralized with an acid solution to a pH of 7. After centrifugation, the precipitate is removed to obtain a carboxymethyl chitosan solution; wherein the mass ratio of chloroacetic acid to chitosan is 1:1-5:1.

[0060] Adjust the pH of the carboxymethyl chitosan solution to 4.5-6, add the modified activated zeolite at a mass ratio of 3:1 to 1:1, and then add glutaraldehyde to make the final mass concentration 0.5%-1%. Stir at room temperature for 4-8 hours, centrifuge, take the precipitate, wash it clean until neutral, and dry it to obtain the carboxymethyl chitosan-modified activated zeolite composite material.

[0061] For example, the alkalization pH value can be selected as a range of values ​​consisting of 12, 13, 14 or any point value, preferably 13-14;

[0062] For example, the centrifugation speed can be selected as 5000 rpm, 6000 rpm, 7000 rpm or any range of values; the centrifugation time can be 10 min, 8 min, 5 min or any range of values.

[0063] For example, the pH value of the carboxymethyl chitosan solution can be selected as a range of values ​​consisting of 4.5, 5.5, 6, or any point value.

[0064] For example, the mass ratio of carboxymethyl chitosan solution to modified activated zeolite can be selected as 3:1, 2:1, 1:1 or a range of values ​​consisting of any points.

[0065] For example, the final mass concentration of glutaraldehyde can be selected as a range of values ​​consisting of 0.5%, 0.75%, 1%, or any point values.

[0066] In some embodiments, the solvent is a polyol, more preferably isopropanol.

[0067] In some embodiments, the alkaline solution is a strongly alkaline solution, more preferably a NaOH solution, and the mass concentration of the NaOH solution is 5%-30%.

[0068] In some embodiments, the acid solution is a strongly acidic solution, more preferably an HCl solution, and the mass concentration of the HCl solution is 5%-20%.

[0069] In some embodiments, the preparation process of activated modified zeolite is as follows: zeolite is added to a mixed solution containing 2M-4M zinc chloride, 1M-2M titanium tetrachloride, and 1M-3M calcium chloride, with a zeolite-to-solution mass ratio of 1:5-1:20. After stirring and filtration, ammonia gas is introduced and reacted at 30-50℃ for 4-10 hours. The mixture is then calcined at high temperature to obtain modified activated zeolite.

[0070] For example, the molar concentration of zinc chloride can be selected as a range of values ​​consisting of 2M, 3M, 4M, or any point values; preferably 2M-3M.

[0071] For example, the molar concentration of titanium tetrachloride can be selected as a range of values ​​consisting of 1M, 2M, 1.5M, or any point values; preferably 1M-1.5M.

[0072] For example, the molar concentration of calcium chloride can be selected as a range of values ​​consisting of 1M, 2M, 3M, or any point values; preferably 1M-2M.

[0073] In some embodiments, after ammonia is introduced, the reaction time at 30-50°C is 4-10 hours; for example, it can be selected as a range of values ​​consisting of 4 hours, 8 hours, 10 hours or any point values; preferably 8 hours to 10 hours.

[0074] In some embodiments, the stirring speed is 500 rpm to 5000 rpm. For example, the stirring speed can be selected as a range of values ​​consisting of 500 rpm, 2000 rpm, 5000 rpm, or any point value; preferably 2000 rpm to 5000 rpm.

[0075] In some embodiments, filtration is performed using a 0.2 μm membrane.

[0076] In some embodiments, calcination is performed at 300°C-500°C for 2-6 hours. For example, the calcination temperature can be selected from a range of 300°C, 400°C, 500°C, or any combination of these values; preferably 400°C-500°C.

[0077] When using the above technical solution, in the process of preparing modified and activated zeolite, the zeolite surface modification and ion exchange are achieved through impregnation with a metal ion solution. Specifically, the aluminosilicate framework of the zeolite adsorbs Zn through ion exchange. 2+ Ti4+ Ca 2+ Meanwhile, ZnCl2 can etch zeolite channels, increasing the specific surface area; Ti 4+ Subsequent calcination can form TiO2 nanoparticles, enhancing the material's activity. Ammonia gas is introduced because it can react with metal ions, modulating surface chemical properties, forming amino or nitride active sites, increasing the material's surface alkalinity and hydrophilicity, and enhancing its compatibility with carboxymethyl chitosan.

[0078] In some embodiments, the gel solution can be thermally sprayed onto the surface of a carrier before cooling, and then naturally cooled to obtain the gel; wherein the carrier includes woven fabric, medical gauze, or cotton fiber.

[0079] When the gel solution is thermally sprayed onto the carrier material, it is more effective in carrying the gel solution, encapsulating it, and preventing it from falling off the skin surface.

[0080] The fourth aspect of this invention provides the application of the biomaterials prepared by the above method in the preparation of medical-related products.

[0081] To better illustrate the technical solution of the present invention, the following embodiments are provided. It should be understood that, unless otherwise stated, all materials used are commercially available.

[0082] Example 1

[0083] The medical hydrogel hemostatic material comprises, by mass percentage, the following components: 1% carboxymethyl chitosan-modified activated zeolite composite material, 2% carrageenan, 0.06% konjac gum, 0.1% calcium chloride, 0.1% edible glucose, 0.5% maltodextrin, 0.02% potassium citrate, 0.05% xanthan gum, 0.01% sodium chloride, and the balance being water.

[0084] The preparation process of the above-mentioned hydrogel hemostatic material is as follows:

[0085] (1) Weigh each component according to the mass ratio;

[0086] (2) Add carboxymethyl chitosan-modified activated zeolite composite material, carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride to water and stir evenly. Heat to 85°C until boiling to obtain a gel solution.

[0087] (3) Hydrogel biomaterial particles are prepared by spraying and cooling, and the biomaterial particles are dried to obtain hemostatic particles. The preparation process of the carboxymethyl chitosan-modified activated zeolite composite material includes:

[0088] Chitosan was dispersed in an isopropanol solution to achieve a chitosan concentration of 5% and an isopropanol to water mass ratio of 7:3. Then, a 10% NaOH solution was added to alkalize the chitosan to a pH of 12. Subsequently, a 10% chloroacetic acid solution was added and stirred, with a chloroacetic acid to chitosan mass ratio of 3:1. The solution was then neutralized to a pH of 7 with a 10% HCl solution. After centrifugation at 5000 rpm for 10 min, the precipitate was removed to obtain a carboxymethyl chitosan solution.

[0089] The pH of the carboxymethyl chitosan solution was adjusted to 4.5. Modified activated zeolite was added at a mass ratio of 1:1 between the carboxymethyl chitosan solution and the modified activated zeolite. Glutaraldehyde was then added to bring the final mass concentration to 0.5%. The mixture was stirred at room temperature for 4 hours, centrifuged at 6000 rpm for 8 minutes, and the precipitate was washed clean until neutral. The precipitate was then dried at 100℃ for 2 hours to obtain the carboxymethyl chitosan-modified activated zeolite composite material.

[0090] The preparation process of modified activated zeolite is as follows: Zeolite is added to a mixed solution containing 2M zinc chloride, 1M titanium tetrachloride and 3M calcium chloride at a mass ratio of 1:5. The mixture is stirred at 500 rpm for 10 h. After filtration through a 0.2 μm membrane, the zeolite is cleaned and then ammonia gas is introduced to react at 30 °C for 10 h. Finally, the mixture is calcined at 300 °C for 4 h to obtain modified activated zeolite.

[0091] Example 2

[0092] The medical hydrogel hemostatic material comprises, by mass percentage, the following components: 30% carboxymethyl chitosan-modified activated zeolite composite material, 0.1% carrageenan, 0.5% sodium chloride, 2% konjac gum, 2% maltodextrin, 1% potassium citrate, 0.02% calcium chloride, 0.02% edible glucose, 0.01% xanthan gum, and the balance being water.

[0093] The preparation process of the above-mentioned hydrogel hemostatic material is as follows:

[0094] (1) Weigh each component according to the mass ratio;

[0095] (2) Add carboxymethyl chitosan-modified activated zeolite composite material, carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride to water and stir evenly. Heat to 85°C until boiling to obtain a gel solution.

[0096] (3) Place the gel solution into a specific mold and allow it to cool naturally to a gel state to obtain the hydrogel biomaterial. The preparation of the carboxymethyl chitosan-modified activated zeolite composite material includes:

[0097] Chitosan was dispersed in isopropanol solution to achieve a chitosan concentration of 30% and an isopropanol to water mass ratio of 8:3. Then, a 10% NaOH solution was added to alkalize the chitosan to a pH of 12. Subsequently, a 10% chloroacetic acid solution was added and stirred, with a chloroacetic acid to chitosan mass ratio of 3:1. The solution was neutralized to pH 7 with a 10% HCl solution. After centrifugation at 5000 rpm for 10 min, the precipitate was removed to obtain a carboxymethyl chitosan solution.

[0098] The pH of the carboxymethyl chitosan solution was adjusted to 4.5. Modified activated zeolite was added at a mass ratio of 2:1 between the carboxymethyl chitosan solution and the modified activated zeolite. Glutaraldehyde was then added to bring the final mass concentration to 0.5%. The mixture was stirred at room temperature for 4 hours, centrifuged at 5000 rpm for 5 minutes, and the precipitate was washed clean until neutral. The precipitate was then dried at 100℃ for 2 hours to obtain the carboxymethyl chitosan-modified activated zeolite composite material.

[0099] The preparation process of modified activated zeolite is as follows: Zeolite is added to a mixed solution containing 4M zinc chloride, 2M titanium tetrachloride and 1M calcium chloride at a mass ratio of 1:8. The mixture is stirred at 2000 rpm for 8 hours, filtered through a 0.2 μm membrane, and the resulting zeolite is cleaned and then reacted with ammonia at 50℃ for 4 hours. Finally, it is calcined at 400℃ for 2 hours to obtain modified activated zeolite.

[0100] Example 3

[0101] The medical hydrogel hemostatic material comprises, by mass percentage, the following: 20% carboxymethyl chitosan-modified activated zeolite composite material, 2.5% carrageenan, 1% sodium chloride, 1% konjac gum, 2% maltodextrin, 1% potassium citrate, 2% calcium chloride, 2% edible glucose, 2% xanthan gum, and the balance being water.

[0102] The preparation process of the above-mentioned hydrogel hemostatic material is as follows:

[0103] (1) Weigh each component according to the mass ratio;

[0104] (2) Add carboxymethyl chitosan-modified activated zeolite composite material, carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride to water and stir evenly. Heat to 85°C until boiling to obtain a gel solution.

[0105] (3) Place the gel solution into a specific mold and allow it to cool naturally to a gel state to obtain the hydrogel biomaterial.

[0106] The preparation process of the carboxymethyl chitosan-modified activated zeolite composite material includes:

[0107] Chitosan was dispersed in isopropanol solution to achieve a chitosan concentration of 15% and an isopropanol to water mass ratio of 9:3. Then, a 10% NaOH solution was added to alkalize the chitosan to a pH of 12. Subsequently, a 10% chloroacetic acid solution was added and stirred, with a chloroacetic acid to chitosan mass ratio of 3:1. The solution was neutralized to pH 7 with a 10% HCl solution. After centrifugation at 7000 rpm for 5 min, the precipitate was removed to obtain a carboxymethyl chitosan solution.

[0108] The pH of the carboxymethyl chitosan solution was adjusted to 4.5. Modified activated zeolite was added at a mass ratio of 3:1 between the carboxymethyl chitosan solution and the modified activated zeolite. Glutaraldehyde was then added to bring the final mass concentration to 0.5%. The mixture was stirred at room temperature for 4 hours, centrifuged at 5000 rpm for 5 minutes, and the precipitate was washed clean until neutral. The precipitate was then dried at 100℃ for 2 hours to obtain the carboxymethyl chitosan-modified activated zeolite composite material.

[0109] The preparation process of modified activated zeolite is as follows: Zeolite is added to a mixed solution containing 3M zinc chloride, 1.5M titanium tetrachloride and 2M calcium chloride at a mass ratio of 1:10. The mixture is stirred at 5000 rpm for 4 hours. After filtration through a 0.2 μm membrane, the zeolite is cleaned and then ammonia gas is introduced to react at 40℃ for 8 hours. Finally, the mixture is calcined at 400℃ for 2 hours to obtain modified activated zeolite.

[0110] Example 4

[0111] The difference between Example 4 and Example 1 is that the preparation process of the modified activated zeolite is as follows: the zeolite is added to a mixed solution containing 2M zinc chloride and 3M calcium chloride at a mass-volume ratio of 1:5. 1M titanium tetrachloride is not added to the solution. Other process steps are the same.

[0112] Example 5

[0113] The difference between Example 5 and Example 1 is that the method for preparing the carboxymethyl chitosan-modified activated zeolite composite material is as follows: chitosan is dispersed in an ethanol solution, while other process steps are the same.

[0114] Example 6

[0115] The difference between Example 6 and Example 1 is that chloroacetic acid was not used.

[0116] Example 7

[0117] The difference between Example 6 and Example 1 is that the preparation process of the modified activated zeolite is as follows: the zeolite is cleaned, dried, and calcined at 300°C for 4 hours to obtain the modified activated zeolite. The remaining process flow is the same as in the example.

[0118] Example 8

[0119] The difference between Example 8 and Example 1 is that the carboxymethyl chitosan-modified activated zeolite composite material was not added, while the other processes were the same.

[0120] Example 9

[0121] Taking Example 3 as an example, the only difference from Example 3 is that the carboxymethyl chitosan-modified activated zeolite composite material is replaced in equal amounts with a mixture of carboxymethyl chitosan and modified zeolite at room temperature, and the mass ratio is 2:1.

[0122] Example 10

[0123] Taking Example 3 as an example, the only difference from Example 3 is that the carboxymethyl chitosan-modified activated zeolite composite material is replaced by an equal amount of chitosan-modified activated zeolite composite material.

[0124] Example 11

[0125] Taking Example 3 as an example, the only difference from Example 3 is that the carboxymethyl chitosan-modified activated zeolite composite material is replaced with an equal amount of carboxymethyl chitosan-zeolite composite material.

[0126] Performance testing

[0127] 1. Cytotoxicity test

[0128] Material preparation

[0129] The hydrogel hemostatic materials of Examples 1-11 were selected and prepared into discs with a diameter of 5 mm and a thickness of 2 mm (or cut according to the actual use shape), with 3 parallel samples for each group.

[0130] Extraction media: serum-free DMEM medium (free of phenol red to avoid interference with absorbance detection), fetal bovine serum (FBS), and phosphate buffered saline (PBS, pH 7.4).

[0131] Cells and reagents

[0132] L929 cell line (ATCC CCL-1) was cryopreserved in liquid nitrogen at -196℃, and after thawing, it was passaged to 3-5 generations to ensure good cell condition (adhesion rate >90%, doubling time 30h).

[0133] Reagents: 0.25% trypsin-EDTA, MTT reagent (5 mg / mL, store protected from light), dimethyl sulfoxide (DMSO), penicillin-streptomycin bispecific antibody (100 U / mL).

[0134] Instruments and equipment

[0135] Carbon dioxide incubator (37℃, 5% CO2), ultra-clean workbench, inverted phase contrast microscope, ELISA reader (detection wavelength 570 nm, reference wavelength 630 nm), 96-well cell culture plate, cell counting plate.

[0136] Experimental procedure:

[0137] 1) L929 cell culture and seeding

[0138] Cell resuscitation and passage:

[0139] Remove L929 cell cryovials from liquid nitrogen, thaw rapidly in a 37°C water bath, transfer to DMEM medium containing 10% FBS, centrifuge at 1000 rpm for 5 min, discard the supernatant, add fresh DMEM medium, and incubate at 37°C with 5% CO2. When cell confluence reaches 90%, digest with 0.25% trypsin for 2 min, add serum-containing DMEM medium to stop digestion, centrifuge at 1000 rpm for 1 min, and count cells (trypan blue staining method, viability > 95%).

[0140] 2) Inoculate into a 96-well plate:

[0141] Adjust the cell concentration to 5×10 4 Add 100 μL of cell suspension to each well and incubate for 24 h until the cells adhere to the wall (under a microscope, they appear spindle-shaped or polygonal, with an adhesion rate >90%).

[0142] 3) Toxicity testing (MTT method)

[0143] Control group: DMEM medium containing 10% FBS (negative control);

[0144] Positive control group: 0.1% Triton X-100 solution (disrupts cell membranes and induces cell death);

[0145] Experimental groups: hydrogels from Examples 5-7 and Comparative Examples 1-5.

[0146] Sample addition and incubation:

[0147] Discard the old culture medium in the 96-well plate, wash once with PBS, add 100 μL of extraction solution to each well of the experimental group, and add 100 μL of the corresponding culture medium to each well of the control group, and continue to culture in an incubator for 24 h, 48 h, and 72 h (to detect cytotoxicity at different time points).

[0148] MTT staining and absorbance measurement:

[0149] Add 20 μL of MTT reagent (5 mg / mL) to each well and continue culturing for 4 hours. Discard the supernatant, add 150 μL of DMSO to each well, and shake for 10 minutes to completely dissolve the Formazan crystals.

[0150] The absorbance (OD value) at 570 nm was measured using an ELISA reader, and the background value was corrected using a reference wavelength of 630 nm.

[0151] Cell viability (%)

[0152] The results are shown in Table 3;

[0153] Table 3. Results of cell viability test

[0154]

[0155] 3. Clotting time test

[0156] In a laboratory environment of 25℃-30℃, take out an appropriate amount of processed frozen pig blood plasma and place it in...

[0157] Place the plasma in a 37°C constant temperature water bath for 30 minutes until it is completely dissolved and ready for use.

[0158] Transfer 2 ml of anticoagulated blood to a 5 mL centrifuge tube using a pipette, and add an appropriate amount of 0.2 mol / L CaCl2 solution.

[0159] After adding the solution, shake well and immediately start timing. React in a 37℃ constant temperature water bath. Observe the state of the solution in the 5mL centrifuge tube. When the solution in the 5mL centrifuge tube becomes a jelly-like colloidal state, the coagulation reaction is considered to be over. The time recorded at this time is considered to be the natural coagulation time. During the experiment, the natural coagulation time is adjusted to between 7-10 min. Record the amount of 0.2 mol / L CaCl2 solution used as X μL. The amount of CaCl2 added at this time is the amount added for the sample coagulation time determination.

[0160] Coagulation time test of samples: 2 mL of pig plasma was transferred to a 5 mL centrifuge tube using a pipette, and X μL of 0.2 mol / L CaCl2 solution was added. Then, 0.300 g of sample (accurate to at least 0.001 g) was quickly poured into the 5 mL centrifuge tube, immediately shaken well, placed in a 37℃ constant temperature water bath, and the timing was started. The mixture was shaken every 3-5 seconds, and the coagulation state of the pig plasma was observed. The coagulation time was recorded when the pig plasma began to coagulate. The results are shown in Table 4.

[0161] Table 4 Bleeding Performance Test

[0162]

[0163] 4. Initial adhesion

[0164] According to the test method under the first method (determination of initial adhesion) of the fourth part of the 2020 edition of the Pharmacopoeia of the People's Republic of China, Volume IV, 0952, the tilt angle was set at 30°, and the model of the hydrogel that adhered to the steel ball for 5 seconds without falling was determined. The larger the model, the better the adhesion performance.

[0165] The experimental procedure is described in Example 4. The results are shown in Table 5.

[0166] Table 5 Adhesion test results

[0167]

[0168] 5. Antibacterial performance test

[0169] The antimicrobial activity of hydrogel biomaterials against Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922), and Candida albicans (ATCC 10231) under dynamic conditions was tested according to the test method of ASTM E2149-13a. The results are shown in Table 6.

[0170] Table 6. Results of Antibacterial Performance Tests

[0171]

[0172] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The application of carboxymethyl chitosan-modified activated zeolite composite material, characterized in that, The intended use is in the preparation of highly adhesive biomaterials that promote wound healing and possess high antibacterial properties; Preferably, the adhesiveness is such that it is not easily separated from or detached from the skin.

2. A biomaterial, characterized in that, The biomaterials include carboxymethyl chitosan-modified activated zeolite composite materials and medically acceptable excipients; Preferably, the biomaterial is used for hemostasis, antibacterial properties, and promoting wound healing; Preferably, the biomaterial includes hydrogel biomaterial or hemostatic particles of dried hydrogel biomaterial; Preferably, the medically acceptable excipients include at least one of carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride.

3. The biomaterial according to claim 2, characterized in that, Based on the total raw materials by mass percentage, the composition includes the following components: 1%-30% carboxymethyl chitosan-modified activated zeolite composite material, 0.1%-2.5% carrageenan, 0.05%-2% konjac gum, 0.02%-2% calcium chloride, 0.02%-2% edible glucose, 0.02%-2% maltodextrin, 0.01%-1% potassium citrate, 0.01%-2% xanthan gum, 0.01%-1% sodium chloride, and the balance being water.

4. A method for preparing a biomaterial, applied to the preparation of the biomaterial according to claim 3, characterized in that, Includes the following steps: (1) Prepare carboxymethyl chitosan-modified activated zeolite composite material and weigh each component according to the mass ratio; (2) Add carboxymethyl chitosan-modified activated zeolite composite material, carrageenan, konjac gum, calcium chloride, edible glucose, maltodextrin, potassium citrate, xanthan gum, and sodium chloride to water and stir evenly. Heat to 85℃-90℃ until boiling to obtain a gel solution. (3) Place the gel solution in a specific mold and allow it to cool naturally to obtain the hydrogel biomaterial; or prepare hydrogel biomaterial particles by spraying and cooling, and prepare hemostatic particles after drying the biomaterial particles.

5. The preparation method according to claim 4, characterized in that, The preparation process of the carboxymethyl chitosan-modified activated zeolite composite material is as follows: Chitosan is dispersed in a solvent to achieve a chitosan concentration of 5%-30% by mass. Then, an alkaline solution is added to alkalize the chitosan to a pH of 12-14. Next, chloroacetic acid solution is added and stirred, and the solution is neutralized with acid to a pH of 7. The mixture is centrifuged to remove the precipitate, yielding a carboxymethyl chitosan solution. The mass ratio of chloroacetic acid to chitosan is 1:1-5:

1. The pH of the carboxymethyl chitosan solution is adjusted to 4.5-6. Modified activated zeolite is added at a mass ratio of 3:1-1:1, followed by the addition of glutaraldehyde to a final concentration of 0.5%-1%. The mixture is stirred at room temperature or heated to 50°C for 4-8 hours. After centrifugation, the precipitate is collected, washed thoroughly until neutral, and dried to obtain a carboxymethyl chitosan-modified activated zeolite composite material.

6. The preparation method according to claim 5, characterized in that, The solvent is a mixture of isopropanol and water; and the mass ratio of isopropanol to water is 7:3-9:

3. The alkaline solution is a strongly alkaline solution; preferably, the strongly alkaline solution is a NaOH solution, and the mass concentration of the NaOH solution is 5%-30%. The acid solution is a strongly acidic solution; preferably, the strongly acidic solution is an HCl solution, and the mass concentration of the HCl solution is 5%-20%.

7. The preparation method according to any one of claims 4 to 6, characterized in that, The preparation process of the activated modified zeolite is as follows: Zeolite is added to a mixed solution containing 2M-4M zinc chloride, 1M-2M titanium tetrachloride and 1M-3M calcium chloride, so that the mass ratio of zeolite to mixed solution is 1:5-1:

10. After stirring and filtering, ammonia gas is introduced and reacted at 30-50℃ for 4-10 hours. After high-temperature calcination, the modified activated zeolite is obtained.

8. The preparation method according to claim 7, characterized in that, The stirring speed is 500 rpm to 5000 rpm; the calcination is carried out at 300℃ to 500℃ for 2 hours to 6 hours.

9. The preparation method according to claim 8, characterized in that, The gel solution is poured into a specific mold before cooling and cooled to form hydrogel biomaterials; it can also be sprayed, sprayed or impregnated on a carrier and cooled to prepare hydrogel biomaterials, or freeze-dried to prepare hemostatic gauze; or sprayed and cooled to prepare hydrogel biomaterial particles, and the particles are freeze-dried to prepare porous hemostatic particles; wherein, the carrier includes woven fabric, medical gauze or cotton fiber, etc.

10. The use of the biomaterial prepared by the method according to any one of claims 4-9 in the preparation of products related to the medical field.

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