Gel composition for repairing female genital tract as well as preparation process and application of gel composition

Abstract

The invention discloses a gel composition for repairing female genital tracts as well as a preparation process and application of the gel composition. The composition comprises three functional core systems: a polysaccharide-enzyme compound A which is formed by self-assembly of superoxide dismutase and oxidative degradation modified aloe polysaccharide and has antioxidant and microenvironment regulation functions; a metal ion-polysaccharide slow-release antibacterial body B formed by coordination of zinc ions and low-molecular-weight chitosan provides instant and long-acting antibacterial protection; the plant polyphenol-protein tissue repair body C is formed by non-covalent binding of epigallocatechin gallate and hydrolyzed collagen, and tissue regeneration and functional reconstruction are promoted. The three components are compounded according to a mass ratio of (1-2): (0.5-1): (2-4), and are integrated with a carbomer matrix to form stable gel. The invention further provides a specific preparation process and a synergistic effect mechanism of all the components. The gel has good stability, biocompatibility and repairing efficiency, and is suitable for treating or improving female genital tract mucosa lesion, inflammation or flora imbalance.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of gel composition preparation, in particular to a gel composition for female reproductive tract repair and its preparation process and application. BACKGROUND

[0002] Female reproductive tract health is an important foundation for maintaining women's overall physiological function and quality of life. As the first line of defense against pathogenic microorganisms, the reproductive tract mucosa is prone to damage, inflammation or bacterial imbalance due to infection, surgical trauma, hormonal changes or improper care. Such problems are common in scenarios such as vaginitis, cervical lesions, postpartum repair, and post-radiotherapy mucosa repair. Current clinical treatment methods include antibiotics, antifungal drugs, hormone ointments, and probiotic preparations, which can alleviate symptoms to some extent, but still have obvious limitations.

[0003] Existing gel products mainly focus on single functions, for example: gels containing antibacterial ingredients can inhibit pathogenic bacteria, but long-term use can lead to drug resistance or further imbalance of the microecology; gels containing moisturizing ingredients can relieve dryness, but cannot effectively promote tissue regeneration and repair; and some antioxidant enzyme preparations (such as superoxide dismutase SOD) have theoretical potential, but in actual application, they are difficult to play a sustained and effective antioxidant role due to problems such as poor molecular stability, easy inactivation, and short half-life in the body. In addition, most products lack the ability to regulate the timing of the repair process, and cannot achieve synergistic effects in the antibacterial, antioxidant, and tissue regeneration stages, resulting in low repair efficiency and easy recurrence. SUMMARY

[0004] I) Technical problems solved The purpose of the present application is to make up for the shortcomings of the prior art, and to provide a gel composition for female reproductive tract repair, which comprises: a polysaccharide-enzyme complex A formed by self-assembly of superoxide dismutase and oxidatively degraded modified aloe polysaccharide; a metal ion-polysaccharide sustained-release antibacterial body B formed by coordination bonding of zinc ions and low molecular weight chitosan; a plant polyphenol-protein tissue repair body C composed of epigallocatechin gallate and hydrolyzed collagen through non-covalent bonding; Among them, the mass ratio of polysaccharide-enzyme complex A, metal ion-polysaccharide sustained-release antibacterial body B and plant polyphenol-protein tissue repair body C is (1 ~ 2) : (0.5 ~ 1) : (2 ~ 4).

[0005] Further, the molecular weight of the oxidatively degraded modified aloe polysaccharide is mainly distributed in the range of 50-200 kDa.

[0006] Furthermore, in the polysaccharide-enzyme complex A, the mass ratio of the superoxide dismutase to the modified aloe polysaccharide is 1:5 to 1:10.

[0007] Furthermore, the number-average molecular weight of the low molecular weight chitosan is less than 50 kDa.

[0008] Furthermore, in the metal ion-polysaccharide sustained-release antibacterial compound B, the molar ratio of zinc ions to amino groups on the chitosan molecular chain is 1:2 to 1:4.

[0009] Furthermore, in the plant polyphenol-protein tissue repair body C, the mass ratio of hydrolyzed collagen to epigallocatechin gallate is 20:1.

[0010] The present invention also provides a process for preparing the gel composition, comprising the following steps: (i) Prepare the polysaccharide-enzyme complex A; (ii) Prepare the metal ion-polysaccharide sustained-release antibacterial body B; (iii) Preparation of the plant polyphenol-protein tissue repair body C; (iv) The plant polyphenol-protein tissue repair body C prepared in (iii) is first mixed with the gel matrix, and then the polysaccharide-enzyme complex A and the metal ion-polysaccharide sustained-release antibacterial body B are added and mixed respectively.

[0011] Further, the preparation of the polysaccharide-enzyme complex A in step (i) includes: (i-1) Extraction and modification of aloe polysaccharides: Polysaccharides were extracted and purified from aloe vera leaf pulp gel, and then oxidatively degraded using a hydrogen peroxide-ascorbic acid system to obtain modified aloe polysaccharides; (i-2) Complex self-assembly: Modified aloe polysaccharide and superoxide dismutase were dissolved together in phosphate buffer and allowed to stand at 4°C to form a complex.

[0012] Further, in step (i-1), the reaction conditions for the oxidative degradation are to react at 30-40°C for 1-3 hours.

[0013] Further, the preparation of the metal ion-polysaccharide sustained-release antimicrobial agent B in step (ii) includes: dissolving low molecular weight chitosan in an acetate buffer solution at pH 5.5, stirring and reacting it with zinc sulfate solution at 50°C for 2 hours, then adjusting the pH to neutral to precipitate the complex, and obtaining a powder by centrifugation, washing and freeze-drying.

[0014] Further, the preparation of the plant polyphenol-protein tissue repair body C in step (iii) includes: stirring hydrolyzed collagen and epigallocatechin gallate in deionized water at room temperature in the dark to form a complex.

[0015] Furthermore, the integration described in step (iv) includes: (iv-1) Preparation of primary gel matrix: Carbomer was dispersed in deionized water containing hyaluronic acid, swollen, and then neutralized to pH 5.0 with triethanolamine; (iv-2) Under low-speed stirring, the solutions of the polysaccharide-enzyme complex A, the metal ion-polysaccharide sustained-release antibacterial body B, and the plant polyphenol-protein tissue repair body C are added sequentially to the primary gel matrix; (iv-3) Add glycerol and trehalose, and obtain the final gel product after vacuum degassing.

[0016] The present invention also provides the use of the gel composition in the preparation of a medicament for treating or improving damage, inflammation or dysbiosis of the female reproductive tract mucosa.

[0017] The beneficial effects are as follows: The metal ion-polysaccharide sustained-release antibacterial system (B) exerts its effect immediately after gel application. The complex formed by low molecular weight chitosan and zinc ions through coordination bonds provides rapid and broad-spectrum antibacterial action by disrupting the cell membranes of pathogenic microorganisms through the positive charge of both zinc ions and chitosan itself. Furthermore, this complex slowly degrades under physiological conditions, continuously releasing zinc ions and chitosan oligosaccharides, thus providing long-lasting antibacterial protection and effectively inhibiting the proliferation of pathogenic bacteria, fungi, and other pathogens. More importantly, chitosan can form a transparent, biodegradable physical barrier on the mucosal surface.

[0018] During reproductive tract inflammation and injury, local tissues generate a large number of reactive oxygen species (ROS). These ROS attack cell membranes, proteins, and DNA, exacerbating tissue damage and hindering the repair process. Superoxide dismutase (SOD) in this invention is one of the most important antioxidant enzymes in the body, specifically catalyzing the dismutation reaction of superoxide anion free radicals to generate oxygen and hydrogen peroxide, thereby eliminating free radicals at their source. However, SOD itself is extremely unstable in both in vivo and in vitro environments and is easily inactivated. This invention utilizes the molecular network of aloe polysaccharides to form a complex with SOD through self-assembly, providing a protective effect on SOD and significantly enhancing its stability during storage and in the complex reproductive tract environment, ensuring the sustained release of its biological activity. Continuous removal of excess ROS effectively reduces oxidative stress damage and blocks the vicious cycle of inflammation.

[0019] Epigallocatechin gallate is the most active component of tea polyphenols, while hydrolyzed collagen provides essential nutrient substrates and physical scaffolds for the migration, spread, and proliferation of epithelial cells. This invention combines EGCG with hydrolyzed collagen via non-covalent bonds, not only utilizing collagen as a "cargo carrier" to improve the stability and bioavailability of EGCG, but also achieving synergistic function between the two. EGCG can further scavenge residual free radicals and inhibit inflammatory signaling pathways such as NF-κB, regulating inflammatory responses at the molecular level; simultaneously, it can also stimulate fibroblasts to synthesize collagen and glycosaminoglycans. Detailed Implementation

[0020] 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.

[0021] Example 1: Preparation of polysaccharide-enzyme complex protection system (A) 1. Materials and Methods 1.1 Raw Materials and Reagents Fresh Aloe vera leaves (purchased from a standardized Aloe vera cultivation base in Hainan Province); superoxide dismutase (SOD, activity ≥3000 U / mg, derived from bovine erythrocytes, Sigma-Aldrich); cellulase (derived from Aspergillus niger, activity ≥100 U / mg); pectinase (derived from Aspergillus niger, activity ≥500 U / mg); macroporous adsorption resin AB-8 (Tianjin Bohong Resin Technology Co., Ltd.); hydrogen peroxide (30%), L-ascorbic acid, disodium hydrogen phosphate, sodium dihydrogen phosphate, etc., were all analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd.

[0022] 1.2 Instruments and Equipment High-efficiency tissue homogenizer (F6 / 10, Shanghai Fluke); constant temperature water bath shaker (SHZ-82, Jintan, Jiangsu); low-speed large-capacity centrifuge (LC-4012, Beijing Reiboer); freeze dryer (LGJ-18, Beijing Songyuan Huaxing); UV-Vis spectrophotometer (UV-2600, Shimadzu, Japan); high-performance gel permeation chromatograph (HPGPC, equipped with TSK-GEL G5000PWxl column, Tosoh, Japan).

[0023] 2. Experimental Procedure 2.1 Extraction and purification of aloe polysaccharides Take fresh Aloe vera leaves, wash them with clean water, peel off the outer skin, and accurately weigh 1000g of leaf mesotherapy gel. Place it in a homogenizer, add twice the volume (2000mL) of deionized water, and homogenize at 10000 rpm in an ice bath for 3 minutes to obtain an aloe vera homogenate. Add 0.5% (w / w) of a complex enzyme (cellulase:pectinase = 1:1, w / w) to the homogenate, adjust the pH to 4.5 with 1M citrate-disodium hydrogen phosphate buffer, and enzymatically hydrolyze in a 50°C water bath for 2 hours. After enzymatic hydrolysis, rapidly raise the temperature to 90°C and maintain it for 10 minutes to inactivate the enzyme activity.

[0024] Centrifuge the enzyme-inactivated solution at 8000 rpm for 20 minutes and collect the supernatant. Slowly add 3 volumes of anhydrous ethanol to the supernatant and incubate overnight at 4°C to precipitate the polysaccharide. Centrifuge again at 8000 rpm for 20 minutes and collect the precipitate. Wash the precipitate twice with anhydrous ethanol and acetone, respectively, to obtain crude aloe polysaccharide.

[0025] The crude polysaccharide was redissolved in deionized water to prepare a 50 mg / mL solution. Purification was performed using macroporous adsorption resin AB-8. The resin column had dimensions of φ 5 cm × 50 cm, and the sample loading volume was 1 / 10 of the column volume. Elution was performed sequentially with deionized water and 30% ethanol at a flow rate of 2 BV / h, with the eluent collected using an automatic fraction collector. The polysaccharide content was determined using the phenol-sulfuric acid method, and the polysaccharide fraction eluted in water was collected. The collected polysaccharide solution was concentrated under reduced pressure and then freeze-dried to obtain purified aloe polysaccharide powder for later use.

[0026] 2.2 Oxidative Degradation Modification of Aloe Polysaccharides Accurately weigh 10 g of the purified aloe polysaccharide and dissolve it in 1000 mL of pH 7.0 phosphate buffer to prepare a 1% (w / v) solution. Establish a hydrogen peroxide-ascorbic acid oxidation system: add hydrogen peroxide to a final concentration of 10 mM and ascorbic acid to the polysaccharide solution sequentially. After mixing thoroughly, place the solution in a 35°C constant temperature water bath shaker and react for 2 hours with continuous gentle stirring.

[0027] After the reaction was completed, the reaction solution was immediately placed in ice water to cool and terminate the reaction. The reaction solution was transferred to a dialysis bag with a molecular weight cutoff of 10 kDa and dialyzed against flowing deionized water for 48 hours to thoroughly remove small molecule impurities and reaction reagents. The liquid in the dialysis bag was removed and freeze-dried to obtain modified aloe polysaccharides with a reduced molecular weight.

[0028] The molecular weight distribution of polysaccharides before and after modification was determined by high-performance gel permeation chromatography. The results showed that the molecular weight of unmodified purified aloe polysaccharides was mainly distributed in the range of 400-800 kDa, while after modification with the hydrogen peroxide-ascorbic acid system, the main molecular weight peak shifted significantly forward, and the main distribution range was concentrated in the range of 50-200 kDa, achieving the design target.

[0029] 2.3 Self-assembly of polysaccharide-enzyme complexes Accurately weigh 500 mg of modified aloe polysaccharide and 80 mg of superoxide dismutase (SOD) (mass ratio approximately 6.25:1), and dissolve them together in 100 mL of 0.01 M phosphate buffer, pH 7.4. Stir gently at 200 rpm for 30 minutes at 4°C using a magnetic stirrer to ensure complete dissolution and homogeneity. Then, stop stirring and allow the mixture to stand at 4°C for 12 hours to allow it to complete the self-assembly process.

[0030] The particle size changes before and after self-assembly were measured using a dynamic light scattering nanoparticle size analyzer. It was found that the self-assembled particles formed a complex with a size distribution of 100-300 nm, and the polydispersity index (PDI) was less than 0.2, indicating that the system was homogeneous and stable. The content of free SOD in the supernatant was determined by the BCA method, and the encapsulation efficiency of SOD was calculated to be 92.5%. The obtained polysaccharide-enzyme complex solution was aliquoted and stored at 4°C, thus forming the polysaccharide-enzyme complex protection system (A).

[0031] Example 2: Preparation of a metal ion-polysaccharide sustained-release antibacterial system (B) 1. Materials and Methods 1.1 Raw Materials and Reagents Low molecular weight chitosan (number average molecular weight Mn ≈ 30 kDa, deacetamide ≥ 95%, Sinopharm Chemical Reagent Co., Ltd.); zinc sulfate heptahydrate (analytical grade, Sinopharm Group); glacial acetic acid, sodium hydroxide, etc., were all analytical grade.

[0032] 1.2 Instruments and Equipment Precision pH meter (PHS-3E, Shanghai Instrument & Electronics Scientific Instruments); Heat-collecting constant temperature magnetic stirrer (DF-101S, Zhengzhou Kefeng); Vacuum freeze dryer (same as above).

[0033] 2. Experimental Procedure 2.1 Ionic crosslinking reaction Accurately weigh 5.0 g of low molecular weight chitosan and add it to 500 mL of 1% (v / v) acetate buffer at pH 5.5. Dissolve overnight at room temperature with magnetic stirring to obtain a 1% (w / v) clear chitosan solution.

[0034] Weigh out 4.5 g of zinc sulfate heptahydrate (equivalent to Zn 2+ (Approximately 1.03 g) was dissolved in 50 mL of deionized water to prepare a zinc sulfate solution.

[0035] The chitosan solution was transferred to a 500 mL three-necked flask and placed in a 50°C water bath for constant temperature. While maintaining continuous mechanical stirring (500 rpm), zinc sulfate solution was slowly and dropwise added to the chitosan solution using a constant-pressure dropping funnel, with the addition time controlled within 1 hour. After the addition was complete, the reaction was continued at 50°C and 500 rpm for 2 hours. During this process, the solution gradually changed from clear to slightly turbid, indicating that zinc ions effectively coordinated with the free amino groups on the chitosan molecular chains, forming a zinc-chitosan complex.

[0036] 2.2 Precipitation and Purification After the reaction was complete, the reaction system was allowed to cool naturally to room temperature. While stirring, 1 M sodium hydroxide solution was added dropwise to slowly adjust the pH of the system to 7.0. As the pH increased, the solubility of the zinc-chitosan complex decreased sharply, and it quickly precipitated as a white flocculent precipitate.

[0037] The suspension containing the precipitate was transferred to a centrifuge tube and centrifuged at 10,000 rpm for 15 minutes. The supernatant was carefully discarded. The collected precipitate was washed repeatedly with plenty of deionized water 3-4 times until the washings showed no sulfate ions when tested with barium chloride solution (indicating that the inorganic salts were washed away). Finally, the washed wet precipitate was pre-frozen and then freeze-dried in a freeze dryer for 48 hours to obtain a loose, white powdery metal ion-polysaccharide sustained-release antibacterial system (B).

[0038] The zinc content in the final product was determined by inductively coupled plasma atomic emission spectrometry, and the zinc loading in the complex was calculated to be approximately 8.5% (w / w).

[0039] Example 3: Preparation of Plant Polyphenol-Protein Tissue Repair System (C) 1. Materials and Methods 1.1 Raw Materials and Reagents Hydrolyzed collagen (derived from fish scales, molecular weight approximately 2000 Da, Shaanxi Ciyuan Biotechnology Co., Ltd.); Epigallocatechin gallate (EGCG, purity ≥ 98%, Suzhou Luye Pharmaceutical Co., Ltd.).

[0040] 1.2 Instruments and Equipment Magnetic stirrer (85-2, Shanghai Sile Instruments); Vacuum degassing stirrer (DZ-2, Zhengzhou Saikesi).

[0041] 2. Experimental Procedure Accurately weigh 20.0 g of hydrolyzed collagen and 1.0 g of EGCG (mass ratio 20:1) and add them together to a 1000 mL amber glass reagent bottle. Add 500 mL of nitrogen-deoxygenated deionized water to the bottle and stir at 400 rpm for 1 hour at room temperature in the dark.

[0042] Throughout the stirring process, the system remained clear without any precipitation or turbidity. This indicates that EGCG successfully bound to the peptide chains of hydrolyzed collagen through hydrophobic interactions and intermolecular hydrogen bonds, forming a soluble complex. After stirring, the resulting clear solution was the plant polyphenol-protein tissue repair system (C), which was stored at 4°C in the dark for later use. The EGCG content in the complex solution was determined by the Folin-phenol method and compared with the initial amount added, showing a binding rate exceeding 98%.

[0043] Example 4: Integration of the gel matrix and preparation of the final product 1. Materials and Reagents Carbomer 940 (Lubrizol Specialty Chemicals); Hyaluronic acid (molecular weight 1.5×10⁴) 6 Da (Freda Biochemical); Triethanolamine (analytical grade); Glycerin (pharmaceutical grade); Trehalose (biological reagent).

[0044] 2. Preparation process 2.1 Preparation of Aqueous Phase / Primary Gel Matrix In a 1000 mL beaker, add 800 mL of boiled, deoxygenated, and cooled deionized water. While stirring at high speed (2000 rpm), slowly and evenly sprinkle in 4.0 g of carbomer 940 powder, continuing stirring for 30 minutes to ensure the carbomer is fully dispersed and free of lumps. Then, add 2.0 g of hyaluronic acid and continue stirring until completely dissolved, forming a viscous, translucent dispersion. Let stand for 2 hours to allow the carbomer to fully swell.

[0045] After the carbomer has fully swelled, add a 10% (w / w) aqueous solution of triethanolamine dropwise while stirring at low speed (500 rpm), monitoring the pH of the system as you add, until the pH of the gel matrix is ​​precisely neutralized to 5.0. At this point, the dispersion can be observed to rapidly transform into a clear, transparent primary gel matrix with good viscoelasticity.

[0046] 2.2 Integration of Active Components While stirring at low speed (300 rpm), the active components prepared in the previous steps are added to the primary gel matrix in the following order: First, add the complete plant polyphenol-protein tissue repair system (C) solution (approximately 500 mL, containing 20g collagen and 1g EGCG).

[0047] Then, slowly add all the powder (15.0 g) of the metal ion-polysaccharide sustained-release antibacterial system (B).

[0048] Finally, add the entire solution of the polysaccharide-enzyme complex protection system (A) (approximately 100 mL, containing 0.5 g of modified polysaccharide and 0.08 g of SOD).

[0049] When adding each component, ensure that it is fully dispersed and mixed evenly before adding the next component to avoid excessively high local concentrations or uneven mixing.

[0050] 2.3 Homogenization and Completion After all active ingredients have been added, add 50 mL of glycerol and 10 g of trehalose to the gel system as a humectant and stabilizer. Increase the stirring speed to 800 rpm and stir continuously for 1 hour to ensure the entire system is highly homogeneous.

[0051] The homogenized gel was transferred to a vacuum degassing mixer and degassed for 15 minutes at a vacuum of -0.095 MPa to remove air bubbles introduced during stirring, resulting in a fine, smooth, milky-white, semi-transparent final gel product. It was then dispensed into pharmaceutical-grade aluminum tubes or dedicated dispensing devices, sealed, and stored in a cool, dry place.

[0052] Example 5: Product Properties and Physicochemical Indicators Testing Appearance: This product is a milky white to light yellow translucent gel with a uniform and delicate texture, free of foreign matter.

[0053] pH value: Take 5.0 g of this product, dilute it with 50 mL of freshly boiled and cooled deionized water, stir well, and measure the pH value with a precision pH meter. The pH value is 5.0 ± 0.2.

[0054] Viscosity: Measured using a Brookfield DV-II+ Pro rotational viscometer, rotor #4, at 60 rpm and 25°C, the viscosity was 15,000 ± 2,000 cP.

[0055] SOD activity assay: The SOD activity in the final gel product was determined using the pyrogallol auto-oxidation method. The results showed that each gram of gel contained no less than 50 U of SOD activity, and after one month of accelerated testing at 40°C, the activity retention rate exceeded 85%, demonstrating that the polysaccharide-enzyme complex protection system significantly improved the stability of SOD in the formulation.

[0056] Example 6: In-depth study of the mechanism of action The repair mechanism of this invention is based on the temporal synergistic effect of three active core systems, and its molecular and cellular biological basis is as follows: 6.1 The Preemptive Barrier and Debridement Effect of the Metal Ion-Polysaccharide Sustained-Release Antibacterial System (B) Following genital tract infection or injury, pathogenic microorganisms (such as Escherichia coli, Candida albicans, Gardnerella vaginalis, etc.) proliferate rapidly, disrupting the mucosal barrier and triggering inflammation. Chitosan in System B, through its polycationic properties, adsorbs onto the negatively charged bacterial cell membrane surface, altering membrane permeability and causing leakage of intracellular components, leading to cell death. Zinc ions can penetrate the cell wall, interfering with the metabolic enzyme system of microorganisms and generating reactive oxygen species that kill them. The synergistic effect of these two components provides a rapid and sustained antibacterial effect. In vitro antibacterial tests show that the minimum inhibitory concentration (MIC) of this product against the aforementioned common pathogens is within the range of 32-128 μg / mL, exhibiting broad-spectrum antibacterial activity.

[0057] More importantly, the hydrophilic gel layer formed by chitosan on the mucosal surface not only physically blocks pathogens but also absorbs wound exudate, keeping the wound moist. This aligns with the modern theory of moist healing and is beneficial for the migration of subsequent repair cells.

[0058] 6.2 Oxidative stress regulation and microenvironment improvement of polysaccharide-enzyme complex protective system (A) During infection and inflammation, activated immune cells (such as neutrophils) produce a "respiratory burst," releasing large amounts of reactive oxygen species (ROS) such as superoxide anions. Excessive ROS attack lipids, proteins, and DNA, leading to cell dysfunction and apoptosis, and continuously activating pro-inflammatory signaling pathways such as NF-κB, forming a vicious cycle.

[0059] In System A, SOD is the first and most important line of defense for removing ROS. It catalytically disproportionates superoxide anions into hydrogen peroxide and oxygen. Hydrogen peroxide can then be further broken down into harmless water by catalase or glutathione peroxidase inherent in the reproductive tract. This efficiently removes O2• - SOD inhibits the chain reaction of oxidative stress at its source.

[0060] We validated this using a human vaginal epithelial cell (VK2 / E6E7) model. Oxidative stress injury was induced with hydrogen peroxide (H2O2), followed by the addition of the extract of the gel from this invention (mainly containing System A). Results showed that, compared to the injury group, the treatment group exhibited significantly reduced intracellular ROS levels (approximately 65%), a marked decrease in apoptosis rate, and a substantial downregulation of the mRNA expression levels of pro-inflammatory factors IL-6 and TNF-α. This demonstrates that System A effectively alleviates oxidative stress and secondary inflammatory responses, creating a crucial "anti-inflammatory-repair" microenvironment for tissue repair.

[0061] 6.3 Tissue regeneration and functional reconstruction of the plant polyphenol-protein tissue repair system (C) After infection is controlled and oxidative stress is alleviated, repair enters a phase dominated by cell proliferation and tissue remodeling. System C plays a core role in this phase.

[0062] EGCG has many functions: Antioxidant and anti-inflammatory effects: EGCG itself is a powerful antioxidant that can directly scavenge various free radicals and chelate metal ions to inhibit the Fenton reaction. More importantly, it can inhibit the expression of various pro-inflammatory factors at the transcriptional level by inhibiting IKK phosphorylation, preventing IκB degradation and NF-κB nuclear translocation.

[0063] Promoting fibroblast proliferation and collagen synthesis: Our scratch assay and CCK-8 assay using human dermal fibroblasts (HDF) showed that the EGCG-containing complex significantly promoted cell migration and proliferation. Western blotting results showed that EGCG upregulated the protein expression levels of type I and type III collagen. The mechanism may be related to the activation of pro-survival and proliferative signaling pathways such as MAPK / ERK and PI3K / Akt.

[0064] Promotes angiogenesis: In chicken embryo allantoic membrane experiments, this product demonstrated the ability to promote angiogenesis, which is crucial for delivering nutrients and oxygen to repaired tissues.

[0065] Hydrolyzed collagen directly provides the amino acids (such as proline and glycine) and signal peptides required for cell proliferation. These small peptides can be directly taken up by cells as raw materials for synthesizing new collagen. At the same time, certain collagen peptides with specific sequences also have chemotactic effects, attracting fibroblasts and epithelial cells to migrate to the site of injury.

[0066] The two are bound by non-covalent bonds. The protective effect of EGCG prolongs the biological function of collagen, while collagen acts as a carrier, increasing the effective concentration and retention time of EGCG locally. In an animal (rat) vaginal mucosal injury model, the treatment group using the gel of this invention showed a 40% shorter recovery time of mucosal epithelial integrity compared to the blank gel group, and the thickness of newly formed epithelium and the density of submucosal collagen were significantly better than the control group.

[0067] 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.

Claims

1. A gel composition for the repair of the female reproductive tract, characterized in that, Include: A polysaccharide-enzyme complex A is formed by the self-assembly of superoxide dismutase and oxidatively degraded aloe polysaccharides. Metal ion-polysaccharide sustained-release antimicrobial agent B is formed by the coordination bonding of zinc ions and low molecular weight chitosan. Plant polyphenol-protein tissue repair complex C is composed of epigallocatechin gallate and hydrolyzed collagen through non-covalent bonding; The mass ratio of polysaccharide-enzyme complex A, metal ion-polysaccharide sustained-release antibacterial agent B, and plant polyphenol-protein tissue repair agent C is (1 ~ 2): (0.5 ~ 1): (2 ~ 4).

2. The gel composition according to claim 1, characterized in that, The molecular weight of the oxidatively degraded aloe polysaccharides is mainly distributed in the range of 50-200 kDa. The number average molecular weight of the low molecular weight chitosan is less than 50 kDa.

3. The gel composition according to claim 1, characterized in that, In the polysaccharide-enzyme complex A, the mass ratio of superoxide dismutase to modified aloe polysaccharide is 1:5 to 1:10; In the metal ion-polysaccharide sustained-release antibacterial compound B, the molar ratio of zinc ions to amino groups on the chitosan molecular chain is 1:2 to 1:

4. In the plant polyphenol-protein tissue repair body C, the mass ratio of hydrolyzed collagen to epigallocatechin gallate is 20:

1.

4. A process for preparing the gel composition according to any one of claims 1-3, characterized in that, Includes the following steps: (i) Prepare the polysaccharide-enzyme complex A; (ii) Prepare the metal ion-polysaccharide sustained-release antibacterial body B; (iii) Preparation of the plant polyphenol-protein tissue repair body C; (iv) The plant polyphenol-protein tissue repair body C prepared in (iii) is first mixed with the gel matrix, and then the polysaccharide-enzyme complex A and the metal ion-polysaccharide sustained-release antibacterial body B are added and mixed respectively.

5. The preparation process of the gel composition according to claim 4, characterized in that, The preparation of the polysaccharide-enzyme complex A in step (i) includes: (i-1) Extraction and modification of aloe polysaccharides: Polysaccharides were extracted and purified from aloe vera leaf pulp gel, and then oxidatively degraded using a hydrogen peroxide-ascorbic acid system to obtain modified aloe polysaccharides; (i-2) Complex self-assembly: Modified aloe polysaccharide and superoxide dismutase were dissolved together in phosphate buffer and allowed to stand at 4°C to form a complex.

6. The preparation process of the gel composition according to claim 4, characterized in that, In step (i-1), the reaction conditions for the oxidative degradation are 1-3 hours at 30-40°C.

7. The preparation process of the gel composition according to claim 4, characterized in that, The preparation of the metal ion-polysaccharide sustained-release antimicrobial agent B in step (ii) includes: dissolving low molecular weight chitosan in an acetate buffer solution at pH 5.5, stirring and reacting it with zinc sulfate solution at 50°C for 2 hours, then adjusting the pH to neutral to precipitate the complex, and obtaining a powder by centrifugation, washing and freeze-drying.

8. The preparation process of the gel composition according to claim 4, characterized in that, The preparation of the plant polyphenol-protein tissue repair compound C in step (iii) includes: stirring hydrolyzed collagen and epigallocatechin gallate in deionized water at room temperature in the dark to form a complex.

9. The preparation process of the gel composition according to claim 4, characterized in that, The integration described in step (iv) includes: (iv-1) Preparation of primary gel matrix: Carbomer was dispersed in deionized water containing hyaluronic acid, swollen, and then neutralized to pH 5.0 with triethanolamine; (iv-2) Under low-speed stirring, the solutions of the polysaccharide-enzyme complex A, the metal ion-polysaccharide sustained-release antibacterial body B, and the plant polyphenol-protein tissue repair body C are added sequentially to the primary gel matrix; (iv-3) Add glycerol and trehalose, and obtain the final gel product after vacuum degassing.

10. Use of the gel composition according to any one of claims 1-3 in the preparation of a medicament for treating or improving damage, inflammation or dysbiosis of the female reproductive tract mucosa.