Medical collagen solution for wound repair and preparation process thereof

Abstract

This invention provides a medical collagen solution for wound repair and its preparation process, belonging to the field of collagen solution preparation technology. The preparation process includes the following steps: preparation of a zinc tea polyphenol complex; preparation of zinc tea polyphenol complex liposomes; preparation of thermosensitive composite microspheres; and preparation of the collagen solution. The collagen solution of this invention contains thermosensitive composite microspheres prepared by first encapsulating the zinc tea polyphenol complex with liposomes and then compounding it with recombinant human collagen type III. The thermosensitive composite microspheres are thermosensitive and can achieve intelligent release. Under human body temperature, the microspheres quickly release recombinant human collagen type III and zinc tea polyphenol complex liposomes, continuously providing nutrition and support for wound repair, which is beneficial for long-term wound repair and healing.

Description

Technical Field

[0001] This invention relates to the field of collagen liquid preparation technology, specifically to a medical collagen liquid for wound repair and its preparation process. Background Technology

[0002] In the field of trauma treatment, wound repair is a crucial step. Effective wound repair not only affects the patient's physical recovery but also directly impacts their quality of life. Currently, there are various products available for wound repair, such as traditional petroleum jelly gauze, various antibacterial ointments, and some biological agents containing growth factors.

[0003] However, these existing wound care products still have many shortcomings. While traditional petroleum jelly gauze can protect the wound to some extent, its poor breathability makes it prone to infection and hinders wound healing. Antibacterial ointments, although able to inhibit the growth of some bacteria, often fail to effectively eliminate excess free radicals generated during wound repair. Excessive oxidative stress can damage wound tissue, exacerbating inflammation and hindering normal wound healing. Biological agents containing growth factors can promote cell proliferation, but their stability is poor, they are expensive, and their absorption and utilization efficiency in the body is limited, making it difficult to provide long-term support for wound repair.

[0004] In the process of wound repair, it is necessary to effectively inhibit pathogens and reduce the risk of infection, provide a good growth environment for cells to promote cell proliferation and tissue regeneration, and at the same time solve the problem of drug absorption and continuous release in wound tissue in order to achieve long-term effective wound repair.

[0005] Therefore, developing a safe, efficient, multifunctional, and intelligently released medical collagen solution for wound repair, along with its preparation process, has significant practical implications and clinical application value. Summary of the Invention

[0006] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a medical collagen liquid for wound repair and its preparation process.

[0007] A medical collagen solution for wound repair and its preparation process, comprising the following steps:

[0008] S1: Preparation of Zinc Complex of Tea Polyphenols

[0009] Tea polyphenol extract was obtained by desorption of tea powder with a desorbent and then extraction. The tea polyphenol solution and zinc chloride solution were then mixed, and NaHCO3 was added to adjust the pH of the mixture to 6. The reaction was then carried out to prepare the tea polyphenol zinc complex.

[0010] S2: Preparation of Zinc Tea Polyphenol Complex Liposomes

[0011] Tea polyphenol zinc complex liposomes were prepared by encapsulating tea polyphenol zinc complex using soybean phospholipids and cholesterol as raw materials.

[0012] S3: Preparation of thermosensitive composite microspheres

[0013] N-isopropylacrylamide was used to graft sodium alginate to prepare grafted modified sodium alginate. Then, the grafted modified sodium alginate and chitosan were used as raw materials to encapsulate tea polyphenol zinc complex liposomes and recombinant human collagen type III to prepare thermosensitive composite microspheres.

[0014] S4: Preparation of Collagen Solution

[0015] Add 1-3 parts by weight of sodium hyaluronate, 2-3 parts by weight of trehalose, 2-3 parts by weight of glycerol, 3-5 parts by weight of glyceryl caprylate, 1-2 parts by weight of capryloyl hydroxamic acid, 1-2 parts by weight of hexanediol and 10-12 parts by weight of thermosensitive composite microspheres to 80-100 parts by weight of deionized water, stir and mix, then sterilize to obtain collagen solution.

[0016] Furthermore, the preparation of the tea polyphenol zinc complex in step S1 specifically includes the following steps:

[0017] S1.1: Wet 3-5 parts by weight of tea powder evenly with 15-20 parts by weight of desorbent, desorb for 8-10 minutes to allow the desorbent to fully penetrate the tea powder, then add 80-100 parts by weight of pure water at 90-100℃ and extract for 10-12 minutes. After extraction, centrifuge at 4000-4200r / min for 10-12 minutes, take the supernatant to obtain tea polyphenol extract;

[0018] S1.2: Mix the tea polyphenol solution and zinc chloride solution, then adjust the pH of the mixture to 6 with 0.1 mol / L NaHCO3, stir at a constant speed for 20-30 min, then centrifuge to collect the precipitate. Wash the precipitate with distilled water and anhydrous ethanol 2-3 times in sequence, and freeze-dry under vacuum to obtain a solid powder, namely the tea polyphenol zinc complex.

[0019] Furthermore, the preparation of tea polyphenol zinc complex liposomes in step S2 specifically includes the following steps:

[0020] S2.1: Dissolve 0.7-1 parts by weight of soybean lecithin and 0.25-0.3 parts by weight of cholesterol in 20-30 parts by weight of chloroform-ethanol mixed solvent. After mixing evenly, add 0.05-0.08 parts by weight of tea polyphenol zinc complex and stir thoroughly for 20-30 minutes. Then, use a rotary evaporator to rotary evaporate at 40-60℃ for 1-2 hours to obtain lipid membrane.

[0021] S2.2: Add 1-2 parts by weight of lipid membrane to 10-12 parts by weight of PBS buffer preheated to 55-60℃ and pH 6.5, and then stir magnetically at 300-500 r / min for 1-2 h to obtain the hydrated suspension.

[0022] S2.3: The hydrated suspension was sonicated in an ice bath for 5-8 minutes. The sonicated suspension was then filtered through a 0.45 μm microporous membrane and transferred to a high-pressure homogenizer for homogenization at 800 bar for 4-5 cycles to obtain a homogenized suspension.

[0023] S2.4: The homogenized suspension was centrifuged in an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa at a centrifugal force of 3000-4000 for 10-15 min, and then filtered through a 0.22 μm sterile filter membrane to obtain tea polyphenol zinc complex liposomes.

[0024] Furthermore, the preparation of the temperature-sensitive composite microspheres in step S3 specifically includes the following steps:

[0025] S3.1: Stir 1-2 parts by weight of 1 wt% sodium alginate solution magnetically at 70-80℃ and heat for 20-30 min, then add 0.23-0.4 parts by weight of initiator and maintain at 40-50℃ for 20-30 min. After that, add 1-2 parts by weight of N-isopropylacrylamide and continue the reaction for 3-4 h. At the end of the reaction, pour the reactants into 200-230 parts by weight of ethanol to terminate the chemical reaction, then wash 2-3 times and dry to obtain grafted modified sodium alginate.

[0026] S3.2: Disperse 1.5-2 parts by weight of grafted modified sodium alginate in 100-120 parts by weight of distilled water, stir magnetically for 20-30 minutes, then add 5-10 parts by weight of calcium carbonate, sonicate for 5-10 minutes, and mix to form the aqueous phase. Add 3-4 parts by weight of emulsifier Span-80 to 200-220 parts by weight of liquid paraffin oil, stir and mix evenly to form the oil phase.

[0027] S3.3: Disperse 2-3 parts by weight of chitosan in 100-120 parts by weight of 0.5 wt% acetic acid solution and stir magnetically for 20-30 min to obtain a crosslinking agent. Add the aqueous phase to the oil phase and emulsify by mechanical stirring for 5-8 min. Then add 2-3 parts by weight of zinc tea polyphenol complex liposomes and 2-3 parts by weight of recombinant human collagen type III and continue emulsifying for 10-12 min. Then add 1-2 parts by weight of acetic acid and 2-3 parts by weight of calcium carbonate and react for 2-3 h to obtain an emulsion. Add the emulsion dropwise to the crosslinking agent and crosslink for 20-30 min. Then centrifuge, filter, wash and dry to obtain thermosensitive composite microspheres.

[0028] Further, the desorbent in step S1.1 is a 40-60 wt% aqueous ethanol solution.

[0029] Further, in step S1.2, the tea polyphenol solution and zinc chloride solution are mixed in a molar ratio of 1:2-3.

[0030] Furthermore, in step S2.1, the volume ratio of chloroform to ethanol is 4:1.

[0031] Furthermore, in step S2.3, the power of the ultrasonic treatment is 200-220W, with a working time of 2 seconds and an interval of 3 seconds.

[0032] Furthermore, the initiator in step S3.1 is potassium persulfate.

[0033] A medical collagen liquid for wound repair, prepared by the preparation process of a medical collagen liquid for wound repair as described in any one of claims 1-9.

[0034] Compared with the prior art, the present invention has at least the following beneficial effects:

[0035] 1. The collagen solution of this invention contains a tea polyphenol zinc complex. Tea polyphenols themselves have strong antioxidant capabilities, and their catechins and other components can eliminate excess free radicals generated during wound repair. After forming a complex with zinc, it helps reduce oxidative stress damage to wound tissues, prevents excessive inflammatory response, creates a favorable microenvironment for wound repair, promotes cell proliferation and tissue regeneration, and the complex formed by the two can produce a synergistic effect, enhancing the inhibitory ability against common wound pathogens such as Staphylococcus aureus and Escherichia coli, reducing the risk of wound infection, thereby alleviating inflammatory response and promoting normal wound healing. At the same time, the zinc ions in the tea polyphenol zinc complex can participate in intracellular signal transduction pathways, regulate cell growth and proliferation, promote fibroblast proliferation and collagen synthesis, and help wound healing and tissue repair.

[0036] 2. This invention combines a zinc polyphenol complex with recombinant human collagen type III. As an important component of the extracellular matrix, recombinant human collagen type III provides a good growth scaffold for cells, promoting cell adhesion, proliferation, and differentiation. In particular, it can accelerate the proliferation of cells closely related to wound repair, such as fibroblasts, laying the foundation for wound repair. Zinc ions participate in the synthesis and activation of various enzymes and can regulate intracellular signal transduction pathways, further promoting cell mitosis and proliferation. Working together with collagen type III, it can significantly increase the cell proliferation rate and accelerate the wound healing process.

[0037] 3. In this invention, the zinc polyphenol complex is first encapsulated in liposomes and then compounded with recombinant human type III collagen before being used to prepare thermosensitive composite microspheres. The structure of the liposomes is similar to that of a biological membrane, exhibiting good biocompatibility. Liposomes encapsulating the zinc polyphenol complex can more easily fuse with cells or be taken up by cells, thereby promoting the entry of the zinc polyphenol complex into the cells, improving its absorption and utilization efficiency in wound tissue, and better exerting its role in wound repair. Furthermore, the liposomes can act as a sustained-release carrier, slowly releasing the zinc polyphenol complex. During wound repair, this product can continuously provide the wound with zinc polyphenols, maintaining an effective drug concentration and prolonging its duration of action. Subsequently, it is combined with recombinant human collagen type III and encapsulated to prepare thermosensitive composite microspheres. The thermosensitive composite microspheres prepared using grafted modified sodium alginate and chitosan as raw materials have thermosensitivity and can achieve intelligent release. Under human body temperature, the microspheres will quickly release recombinant human collagen type III and zinc polyphenols liposomes, continuously providing nutrition and support for wound repair, which is conducive to long-term wound repair and healing. Attached Figure Description

[0038] The accompanying drawings, which are incorporated herein and form part of the specification, illustrate embodiments of the invention and, together with the specification, further serve to explain the principles of the invention and enable those skilled in the art to practice and use the invention.

[0039] Figure 1 This is a flowchart illustrating the preparation process of a medical collagen solution for wound repair used in an embodiment of the present invention. Detailed Implementation

[0040] The preparation process of a medical collagen solution for wound repair provided by the present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. It should also be noted that, to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art can use other alternative methods to implement some known technologies; moreover, the accompanying drawings are only for more specific description of the embodiments and are not intended to specifically limit the present invention.

[0041] Example 1

[0042] A medical collagen solution for wound repair and its preparation process, such as Figure 1 As shown, it includes the following steps:

[0043] S1: Preparation of Zinc Complex of Tea Polyphenols

[0044] S1.1: Wet 3 parts by weight of tea powder evenly with 15 parts by weight of 40 wt% ethanol aqueous solution as a desorbent, desorb for 8 min to allow the desorbent to fully penetrate the tea powder, then add 80 parts by weight of 90℃ pure water and extract for 10 min. After extraction, centrifuge at 4000 r / min for 10 min, take the supernatant to obtain tea polyphenol extract.

[0045] S1.2: Mix tea polyphenol solution and zinc chloride solution in a molar ratio of 1:2, then adjust the pH of the mixture to 6 with 0.1 mol / L NaHCO3, stir at a constant speed for 20 min, then centrifuge to collect the precipitate, wash the precipitate twice with distilled water and anhydrous ethanol, and freeze-dry under vacuum to obtain a solid powder, namely tea polyphenol zinc complex.

[0046] S2: Preparation of Zinc Tea Polyphenol Complex Liposomes

[0047] S2.1: Dissolve 0.7 parts by weight of soybean lecithin and 0.25 parts by weight of cholesterol in 20 parts by weight of chloroform-ethanol mixed solvent, with a volume ratio of chloroform to ethanol of 4:1. After mixing evenly, add 0.05 parts by weight of tea polyphenol zinc complex, stir thoroughly for 20 min, and then use a rotary evaporator to rotary evaporate at 40℃ for 1 h to obtain lipid membrane.

[0048] S2.2: Add 1 part by weight of lipid membrane to 10 parts by weight of PBS buffer preheated to 55°C and pH 6.5, and then stir magnetically at 300 r / min for 1 h to obtain the hydrated suspension.

[0049] S2.3: The hydrated suspension was subjected to ultrasonic treatment for 5 minutes in an ice bath at 200W for 2 seconds followed by 3 seconds of intermittent operation. The ultrasonically treated suspension was then passed through a 0.45μm microporous membrane and transferred to a high-pressure homogenizer for cyclic homogenization at 800 bar for 4 cycles to obtain a homogenized suspension.

[0050] S2.4: The homogenized suspension was centrifuged at 3000 centrifugation force for 10 min using an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa, and then filtered through a 0.22 μm sterile filter membrane to obtain tea polyphenol zinc complex liposomes.

[0051] S3: Preparation of thermosensitive composite microspheres

[0052] S3.1: 1 part by weight of 1 wt% sodium alginate solution was magnetically stirred and heated at 70°C for 20 min, then 0.23 parts by weight of potassium persulfate initiator was added, and the mixture was continuously kept at 40°C for 20 min. After that, 1 part by weight of N-isopropylacrylamide was added, and the reaction was continued for 3 h. At the end of the reaction, the reactants were poured into 200 parts by weight of ethanol to terminate the chemical reaction. The mixture was then washed twice and dried to obtain grafted modified sodium alginate.

[0053] S3.2: Disperse 1.5 parts by weight of grafted modified sodium alginate in 100 parts by weight of distilled water, stir magnetically for 20 min, then add 5 parts by weight of calcium carbonate, sonicate for 5 min, and mix to form the aqueous phase. Add 3 parts by weight of emulsifier Span-80 to 200 parts by weight of liquid paraffin oil, stir and mix evenly to form the oil phase.

[0054] S3.3: Disperse 2 parts by weight of chitosan in 100 parts by weight of 0.5 wt% acetic acid solution and stir magnetically for 20 min to obtain a crosslinking agent. Add the aqueous phase to the oil phase and emulsify mechanically for 5 min. Then add 2 parts by weight of zinc tea polyphenol complex liposomes and 2 parts by weight of recombinant human collagen type III and continue emulsifying for 10 min. Then add 1 part by weight of acetic acid and 2 parts by weight of calcium carbonate and react for 2 h to obtain an emulsion. Drop the emulsion into the crosslinking agent and crosslink for 20 min. Then centrifuge, filter, wash and dry to obtain thermosensitive composite microspheres.

[0055] S4: Preparation of Collagen Solution

[0056] Add 1 part by weight of sodium hyaluronate, 2 parts by weight of trehalose, 2 parts by weight of glycerol, 3 parts by weight of glyceryl caprylate, 1 part by weight of capryloyl hydroxamic acid, 1 part by weight of hexanediol and 10 parts by weight of thermosensitive composite microspheres to 80 parts by weight of deionized water, stir and mix, then sterilize to obtain collagen solution.

[0057] Example 2

[0058] A preparation process for a medical collagen solution used for wound repair, such as... Figure 1 As shown, it includes the following steps:

[0059] S1: Preparation of Zinc Complex of Tea Polyphenols

[0060] S1.1: Wet 5 parts by weight of tea powder evenly with 20 parts by weight of 60 wt% ethanol aqueous solution as a desorbent, desorb for 8 min to allow the desorbent to fully penetrate the tea powder, then add 100 parts by weight of 90℃ pure water and extract for 10 min. After extraction, centrifuge at 4000 r / min for 10 min, take the supernatant to obtain tea polyphenol extract.

[0061] S1.2: Mix tea polyphenol solution and zinc chloride solution in a molar ratio of 1:3, then adjust the pH of the mixture to 6 with 0.1 mol / L NaHCO3, stir at a constant speed for 20 min, then centrifuge to collect the precipitate, wash the precipitate with distilled water and anhydrous ethanol 2-3 times in sequence, and freeze-dry under vacuum to obtain solid powder, namely tea polyphenol zinc complex.

[0062] S2: Preparation of Zinc Tea Polyphenol Complex Liposomes

[0063] S2.1: Dissolve 1 part by weight of soybean lecithin and 0.3 parts by weight of cholesterol in 30 parts by weight of chloroform-ethanol mixed solvent, with a volume ratio of chloroform to ethanol of 4:1. After mixing evenly, add 0.08 parts by weight of tea polyphenol zinc complex, stir thoroughly for 20 min, and then use a rotary evaporator to rotary evaporate at 40℃ for 1 h to obtain lipid membrane.

[0064] S2.2: Add 2 parts by weight of lipid membrane to 12 parts by weight of PBS buffer preheated to 55°C and pH 6.5, and then stir magnetically at 300 r / min for 1 h to obtain the hydrated suspension.

[0065] S2.3: The hydrated suspension was subjected to ultrasonic treatment for 5 minutes in an ice bath at 200W for 2 seconds followed by 3 seconds of intermittent operation. The ultrasonically treated suspension was then passed through a 0.45μm microporous membrane and transferred to a high-pressure homogenizer for cyclic homogenization at 800 bar for 4 cycles to obtain a homogenized suspension.

[0066] S2.4: The homogenized suspension was centrifuged at 3000 centrifugation force for 10 min using an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa, and then filtered through a 0.22 μm sterile filter membrane to obtain tea polyphenol zinc complex liposomes.

[0067] S3: Preparation of thermosensitive composite microspheres

[0068] S3.1: 2 parts by weight of 1 wt% sodium alginate solution were magnetically stirred and heated at 70°C for 20 min, then 0.4 parts by weight of potassium persulfate initiator were added, and the mixture was kept at 40°C for 20 min. After that, 2 parts by weight of N-isopropylacrylamide were added, and the reaction was continued for 3 h. At the end of the reaction, the reactants were poured into 230 parts by weight of ethanol to terminate the chemical reaction. The mixture was then washed twice and dried to obtain grafted modified sodium alginate.

[0069] S3.2: Disperse 2 parts by weight of grafted modified sodium alginate in 120 parts by weight of distilled water, stir magnetically for 20 min, then add 10 parts by weight of calcium carbonate, sonicate for 5 min, and mix to form the aqueous phase. Add 4 parts by weight of emulsifier Span-80 to 220 parts by weight of liquid paraffin oil, stir and mix evenly to form the oil phase.

[0070] S3.3: Disperse 3 parts by weight of chitosan in 120 parts by weight of 0.5 wt% acetic acid solution and stir magnetically for 20 min to obtain a crosslinking agent. Add the aqueous phase to the oil phase and emulsify mechanically for 5 min. Then add 3 parts by weight of zinc tea polyphenol complex liposomes and 3 parts by weight of recombinant human collagen type III and continue emulsifying for 10 min. Then add 2 parts by weight of acetic acid and 3 parts by weight of calcium carbonate and react for 2 h to obtain an emulsion. Drop the emulsion into the crosslinking agent and crosslink for 20 min. Then centrifuge, filter, wash and dry to obtain temperature-sensitive composite microspheres.

[0071] S4: Preparation of Collagen Solution

[0072] Add 3 parts by weight of sodium hyaluronate, 3 parts by weight of trehalose, 3 parts by weight of glycerol, 5 parts by weight of glyceryl caprylate, 2 parts by weight of capryloyl hydroxamic acid, 2 parts by weight of hexanediol and 12 parts by weight of thermosensitive composite microspheres to 100 parts by weight of deionized water, stir and mix, then sterilize to obtain collagen solution.

[0073] Example 3

[0074] A preparation process for a medical collagen solution used for wound repair, such as... Figure 1 As shown, it includes the following steps:

[0075] S1: Preparation of Zinc Complex of Tea Polyphenols

[0076] S1.1: Wet 3 parts by weight of tea powder evenly with 15 parts by weight of 40-60 wt% ethanol aqueous solution as a desorbent, desorb for 10 min to allow the desorbent to fully penetrate the tea powder, then add 80 parts by weight of 100℃ pure water and extract for 12 min. After extraction, centrifuge at 4200 r / min for 12 min, take the supernatant to obtain tea polyphenol extract;

[0077] S1.2: Mix tea polyphenol solution and zinc chloride solution in a molar ratio of 1:2, then adjust the pH of the mixture to 6 with 0.1 mol / L NaHCO3, stir at a constant speed for 30 min, then centrifuge to collect the precipitate, wash the precipitate three times with distilled water and anhydrous ethanol, and freeze-dry under vacuum to obtain a solid powder, namely tea polyphenol zinc complex.

[0078] S2: Preparation of Zinc Tea Polyphenol Complex Liposomes

[0079] S2.1: Dissolve 0.7 parts by weight of soybean lecithin and 0.25 parts by weight of cholesterol in 20 parts by weight of chloroform-ethanol mixed solvent, with a volume ratio of chloroform to ethanol of 4:1. After mixing evenly, add 0.05 parts by weight of tea polyphenol zinc complex, stir thoroughly for 30 min, and then use a rotary evaporator to rotary evaporate at 60℃ for 2 h to obtain lipid membrane.

[0080] S2.2: Add 1 part by weight of lipid membrane to 10 parts by weight of PBS buffer preheated to 60°C and pH 6.5, and then stir magnetically at 500 r / min for 2 h to obtain the hydrated suspension.

[0081] S2.3: The hydrated suspension was subjected to ultrasonic treatment for 8 minutes in an ice bath at 220W for 2 seconds followed by 3 seconds of intermittent operation. The ultrasonically treated suspension was then passed through a 0.45μm microporous membrane and transferred to a high-pressure homogenizer for 5 cycles of homogenization at 800 bar to obtain the homogenized suspension.

[0082] S2.4: The homogenized suspension was centrifuged at 4000 centrifugation force for 15 min using an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa, and then filtered through a 0.22 μm sterile filter membrane to obtain tea polyphenol zinc complex liposomes.

[0083] S3: Preparation of thermosensitive composite microspheres

[0084] S3.1: 1 part by weight of 1 wt% sodium alginate solution was magnetically stirred and heated at 80°C for 30 min, then 0.23 parts by weight of potassium persulfate initiator was added, and the mixture was continuously kept at 50°C for 30 min. After that, 1 part by weight of N-isopropylacrylamide was added, and the reaction was continued for 4 h. At the end of the reaction, the reactants were poured into 200 parts by weight of ethanol to terminate the chemical reaction. The mixture was then washed 3 times and dried to obtain grafted modified sodium alginate.

[0085] S3.2: Disperse 1.5 parts by weight of grafted modified sodium alginate in 100 parts by weight of distilled water, stir magnetically for 30 min, then add 5 parts by weight of calcium carbonate, sonicate for 10 min, and mix to form the aqueous phase. Add 3 parts by weight of emulsifier Span-80 to 200 parts by weight of liquid paraffin oil, stir and mix evenly to form the oil phase.

[0086] S3.3: Disperse 2 parts by weight of chitosan in 100 parts by weight of 0.5 wt% acetic acid solution and stir magnetically for 30 min to obtain a crosslinking agent. Add the aqueous phase to the oil phase and emulsify mechanically for 8 min. Then add 2 parts by weight of zinc tea polyphenol complex liposomes and 2 parts by weight of recombinant human collagen type III and continue emulsifying for 12 min. Then add 1 part by weight of acetic acid and 2 parts by weight of calcium carbonate and react for 3 h to obtain an emulsion. Drop the emulsion into the crosslinking agent and crosslink for 30 min. Then centrifuge, filter, wash and dry to obtain temperature-sensitive composite microspheres.

[0087] S4: Preparation of Collagen Solution

[0088] Add 1 part by weight of sodium hyaluronate, 2 parts by weight of trehalose, 2 parts by weight of glycerol, 3 parts by weight of glyceryl caprylate, 1 part by weight of capryloyl hydroxamic acid, 1 part by weight of hexanediol and 10 parts by weight of thermosensitive composite microspheres to 80 parts by weight of deionized water, stir and mix, then sterilize to obtain collagen solution.

[0089] Comparative Example 1

[0090] Compared with Example 1, Comparative Example 1 differs in that the zinc polyphenol complex liposomes in steps S1-S2 and S3.3 are removed, while the remaining steps remain unchanged to prepare collagen solution, and it is referred to as Comparative Example 1.

[0091] Comparative Example 2

[0092] Compared with Example 1, Comparative Example 2 differs in that the recombinant human collagen type III in step S3.3 is removed, while the remaining steps are unchanged to prepare collagen solution, and it is referred to as Comparative Example 2.

[0093] Comparative Example 3

[0094] Compared with Example 1, Comparative Example 3 differs in that step S3.1 is removed, and the grafted modified sodium alginate in step S3.2 is replaced with sodium alginate, while the remaining steps remain unchanged to prepare collagen solution. This is referred to as Comparative Example 3.

[0095] Antibacterial test results:

[0096] Table 1. Results of antibacterial performance testing in Examples and Comparative Example 1

[0097]

[0098] As can be seen from the data in Table 1, the zinc polyphenol complex added in this invention can enhance the inhibitory effect on common pathogens of wounds, such as Staphylococcus aureus and Escherichia coli, thereby giving the collagen liquid an antibacterial effect.

[0099] Determination of healing rate in a rat burn model:

[0100] Healthy adult rats, typically weighing between 200-250g, were selected. They were acclimatized for one week prior to the experiment to ensure good health. The rats were anesthetized with an anesthetic, and their backs were shaved to expose the skin. Standardized deep second-degree burns were caused by applying 90°C hot water to the skin for a specified period. The rats were randomly divided into seven groups of 10 each. The first six groups were treated with the collagen solutions prepared in Examples 1-3 and Comparative Examples 1-3. The seventh group served as a blank control group, receiving only physiological saline treatment. The healing rate was measured.

[0101] Healing rate (%) = [(initial wound area - current wound area) / initial wound area] × 100%. See Table 3 for the measurement results.

[0102] Table 2. Results of healing rate determination in Examples 1-3 and Comparative Examples 1-3

[0103]

[0104] As can be seen from the data in Comparative Examples 1-2 in Table 2, the combination of zinc tea polyphenol complex and recombinant human collagen type III can significantly improve the cell proliferation rate and accelerate the wound healing process.

[0105] The data from Comparative Example 3 show that the healing rate of Comparative Example 3 is lower than that of the Example. This is because the composite microspheres prepared from sodium alginate and chitosan are not thermosensitive and cannot be released quickly after contact with the human body. Instead, they can only be released slowly. As a result, some recombinant human collagen type III and zinc tea polyphenol complex liposomes are not released and therefore cannot play a role, resulting in a low healing rate.

[0106] The above embodiments are merely illustrative of the principles and effects of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

Claims

1. A preparation process for a medical collagen solution used for wound repair, characterized in that, Includes the following steps: S1: Preparation of Zinc Complex of Tea Polyphenols Tea polyphenol extract was obtained by desorption of tea powder with a desorbent and then extraction. The tea polyphenol solution and zinc chloride solution were then mixed, and NaHCO3 was added to adjust the pH of the mixture to 6. The reaction was then carried out to prepare the tea polyphenol zinc complex. S2: Preparation of Zinc Tea Polyphenol Complex Liposomes Tea polyphenol zinc complex liposomes were prepared by encapsulating tea polyphenol zinc complex using soybean phospholipids and cholesterol as raw materials. S3: Preparation of thermosensitive composite microspheres N-isopropylacrylamide was used to graft sodium alginate to prepare grafted modified sodium alginate. Then, the grafted modified sodium alginate and chitosan were used as raw materials to encapsulate tea polyphenol zinc complex liposomes and recombinant human collagen type III to prepare thermosensitive composite microspheres. S4: Preparation of Collagen Solution Add 1-3 parts by weight of sodium hyaluronate, 2-3 parts by weight of trehalose, 2-3 parts by weight of glycerol, 3-5 parts by weight of glyceryl caprylate, 1-2 parts by weight of capryloyl hydroxamic acid, 1-2 parts by weight of hexanediol and 10-12 parts by weight of thermosensitive composite microspheres to 80-100 parts by weight of deionized water, stir and mix, then sterilize to obtain collagen solution.

2. The preparation process of a medical collagen solution for wound repair according to claim 1, characterized in that, Step S1, the preparation of the zinc polyphenol complex, specifically includes the following steps: S1.1: Wet 3-5 parts by weight of tea powder evenly with 15-20 parts by weight of desorbent, desorb for 8-10 minutes to allow the desorbent to fully penetrate the tea powder, then add 80-100 parts by weight of pure water at 90-100℃ and extract for 10-12 minutes. After extraction, centrifuge at 4000-4200r / min for 10-12 minutes, take the supernatant to obtain tea polyphenol extract; S1.2: Mix the tea polyphenol solution and zinc chloride solution, then adjust the pH of the mixture to 6 with 0.1 mol / L NaHCO3, stir at a constant speed for 20-30 min, then centrifuge to collect the precipitate. Wash the precipitate with distilled water and anhydrous ethanol 2-3 times in sequence, and freeze-dry under vacuum to obtain a solid powder, namely the tea polyphenol zinc complex.

3. The preparation process of a medical collagen solution for wound repair according to claim 2, characterized in that, Step S2, the preparation of tea polyphenol zinc complex liposomes, specifically includes the following steps: S2.1: Dissolve 0.7-1 parts by weight of soybean lecithin and 0.25-0.3 parts by weight of cholesterol in 20-30 parts by weight of chloroform-ethanol mixed solvent. After mixing evenly, add 0.05-0.08 parts by weight of tea polyphenol zinc complex and stir thoroughly for 20-30 minutes. Then, use a rotary evaporator to rotary evaporate at 40-60℃ for 1-2 hours to obtain lipid membrane. S2.2: Add 1-2 parts by weight of lipid membrane to 10-12 parts by weight of PBS buffer preheated to 55-60℃ and pH 6.5, and then stir magnetically at 300-500 r / min for 1-2 h to obtain the hydrated suspension. S2.3: The hydrated suspension was sonicated in an ice bath for 5-8 minutes. The sonicated suspension was then filtered through a 0.45 μm microporous membrane and transferred to a high-pressure homogenizer for homogenization at 800 bar for 4-5 cycles to obtain a homogenized suspension. S2.4: The homogenized suspension was centrifuged in an ultrafiltration centrifuge tube with a molecular weight cutoff of 10 kDa at a centrifugal force of 3000-4000 for 10-15 min, and then filtered through a 0.22 μm sterile filter membrane to obtain tea polyphenol zinc complex liposomes.

4. The preparation process of a medical collagen solution for wound repair according to claim 3, characterized in that, Step S3, the preparation of the thermosensitive composite microspheres, specifically includes the following steps: S3.1: Stir 1-2 parts by weight of 1 wt% sodium alginate solution magnetically at 70-80℃ and heat for 20-30 min, then add 0.23-0.4 parts by weight of initiator and maintain at 40-50℃ for 20-30 min. After that, add 1-2 parts by weight of N-isopropylacrylamide and continue the reaction for 3-4 h. At the end of the reaction, pour the reactants into 200-230 parts by weight of ethanol to terminate the chemical reaction, then wash 2-3 times and dry to obtain grafted modified sodium alginate. S3.2: Disperse 1.5-2 parts by weight of grafted modified sodium alginate in 100-120 parts by weight of distilled water, stir magnetically for 20-30 minutes, then add 5-10 parts by weight of calcium carbonate, sonicate for 5-10 minutes, and mix to form the aqueous phase. Add 3-4 parts by weight of emulsifier Span-80 to 200-220 parts by weight of liquid paraffin oil, stir and mix evenly to form the oil phase. S3.3: Disperse 2-3 parts by weight of chitosan in 100-120 parts by weight of 0.5 wt% acetic acid solution and stir magnetically for 20-30 min to obtain a crosslinking agent. Add the aqueous phase to the oil phase and emulsify by mechanical stirring for 5-8 min. Then add 2-3 parts by weight of zinc tea polyphenol complex liposomes and 2-3 parts by weight of recombinant human collagen type III and continue emulsifying for 10-12 min. Then add 1-2 parts by weight of acetic acid and 2-3 parts by weight of calcium carbonate and react for 2-3 h to obtain an emulsion. Add the emulsion dropwise to the crosslinking agent and crosslink for 20-30 min. Then centrifuge, filter, wash and dry to obtain thermosensitive composite microspheres.

5. The preparation process of a medical collagen solution for wound repair according to claim 2, characterized in that, The desorbent in step S1.1 is a 40-60 wt% aqueous ethanol solution.

6. The preparation process of a medical collagen solution for wound repair according to claim 2, characterized in that, In step S1.2, the tea polyphenol solution and zinc chloride solution are mixed in a molar ratio of 1:2-3.

7. The preparation process of a medical collagen solution for wound repair according to claim 3, characterized in that, In step S2.1, the volume ratio of chloroform to ethanol is 4:

1.

8. The preparation process of a medical collagen solution for wound repair according to claim 3, characterized in that, In step S2.3, the ultrasonic treatment power is 200-220W, with a working time of 2s and an interval of 3s.

9. The preparation process of a medical collagen solution for wound repair according to claim 4, characterized in that, In step S3.1, the initiator is potassium persulfate.

10. A medical collagen solution for wound repair, characterized in that, It is prepared by the preparation process of a medical collagen liquid for wound repair as described in any one of claims 1-9.

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