Antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing as well as preparation method and application thereof

Abstract

The invention discloses an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing as well as a preparation method and application thereof, and belongs to the technical field of medical materials. A low-temperature mixing and step-by-step cross-linking strategy is adopted, heat-sensitive traditional Chinese medicine components and high-temperature-resistant components are separated, and active substances are prevented from being damaged; an interpenetrating network of aluminum ion-phenolic hydroxyl coordination synergistic hydrogen bonds is designed to enhance the mechanical strength, and the porosity is regulated and controlled by a ternary solvent. The porosity is accurately regulated and controlled through a temperature control strategy, the stirring speed and the solution mixing ratio are adjusted to adapt to different wounds, and the production efficiency is high. Through gradient compatibility of the eight traditional Chinese medicine components, the dressing has broad-spectrum antibacterial property, the 24-hour bacteriostasis rate is larger than 70%, angiogenesis and fibroblast migration can be promoted, and the wound healing time is shortened. High-temperature damage is avoided by adopting a specific process, the encapsulation efficiency of the traditional Chinese medicine extract reaches 85%-92%, and the medicine is slowly released within 72 hours. The method is simple in process, controllable in cost and suitable for industrialization, and the dressing is suitable for complex wound treatment and large in clinical transformation potential.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medical materials, and particularly relates to an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing as well as a preparation method and application thereof. BACKGROUND

[0002] As a new type of wound repair material, the hydrogel dressing has shown significant advantages in the field of wound care due to its high water content, air permeability and biomimetic extracellular matrix characteristics. Traditional hydrogel dressings are mostly based on synthetic polymer materials, which have adjustable mechanical properties and water absorption capacity, but still have limitations in antibacterial activity, healing function and biocompatibility. With the increasing problem of drug-resistant bacterial infection and the growing demand for chronic wound treatment, the development of hydrogel dressings with antibacterial, anti-inflammatory and tissue regeneration synergistic functions has become a research hotspot. Under this background, active ingredients of traditional Chinese medicine have become an important direction for the development of functional hydrogel dressings due to their multi-target action mechanism and natural safety.

[0003] In the prior art, the preparation of traditional Chinese medicine hydrogel dressings mainly faces three technical bottlenecks: firstly, the compatibility problem of traditional Chinese medicine extract and polymer matrix. Traditional blending method is easy to cause uneven distribution of active ingredients, and high temperature treatment may damage the biological activity of heat-sensitive components (such as saponins). The root of this problem lies in the significant difference in physicochemical properties of multi-component traditional Chinese medicine active ingredients (such as hydrophilic honeysuckle extract and lipophilic matrine), which are difficult to coexist stably in a single solvent system, which easily causes phase separation or precipitation; at the same time, heat-sensitive components (such as saponins) and pH-sensitive components (such as baicalin) are easily degraded or inactivated in the structure during crosslinking. Secondly, the synergy of antibacterial and healing functions is insufficient. Most studies focus on single traditional Chinese medicine components, which have narrow antibacterial spectrum and are difficult to cope with complex wound microenvironment. The inhibition rate of single antibacterial component hydrogel to multi-drug resistant Staphylococcus aureus is low, and it lacks the ability to promote angiogenesis. The deeper contradiction lies in the time sequence conflict of functional release: antibacterial components need to act quickly to inhibit infection, while healing components need to be released slowly to support tissue regeneration, and traditional carriers cannot realize differential release control; in addition, the combination of components may cause antagonism (such as the combination of alkaloids and tannins to precipitate) or competition for binding sites (such as the competition of matrine and tannic acid for aluminum ion coordination sites), which further weakens the synergistic effect. Thirdly, there is a contradiction between the mechanical properties of the material and the drug loading capacity. Although conventional crosslinking processes (such as physical crosslinking or chemical crosslinking agents) can improve the gel strength, they will significantly reduce the porosity, affecting nutrient transport and cell migration. This mutual exclusion effect is due to the inherent conflict between high drug loading demand and crosslinking network: increasing the drug loading space requires increasing the porosity, but enhancing the mechanical properties requires dense crosslinking; more seriously, active groups (such as phenolic hydroxyl groups) in traditional Chinese medicine components may consume crosslinking agents (such as aluminum chloride), resulting in reduced crosslinking density and modulus decay. SUMMARY

[0004] To address the technical problems of insufficient antibacterial activity, limited healing mechanisms, and poor drug loading stability in existing hydrogel dressings, this invention aims to provide an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing, its preparation method, and its application. Through a stepwise composite cross-linking process, seven active ingredients from traditional Chinese medicine, including honeysuckle extract, matrine, and scutellaria baicalensis extract, are efficiently combined with a dynamic network system of polyvinyl alcohol (PVA) / tannic acid (TA) / aluminum trichloride, forming a three-dimensional gel dressing with broad-spectrum antibacterial, anti-inflammatory, antioxidant, and tissue regeneration functions. This overcomes the technical bottlenecks of traditional Chinese medicine dressings, such as easy inactivation of ingredients and poor sustained-release performance.

[0005] To achieve the above objectives, the present invention employs the following technical solution: This invention provides an antibacterial and healing-promoting herbal hydrogel dressing, wherein the herbal hydrogel dressing is made of herbal extracts loaded with polyvinyl alcohol-tannic acid-Al 3+ The scaffold matrix was obtained; the herbal extract was composed of two or more of the following: honeysuckle extract, matrine, scutellaria baicalensis extract, centella asiatica extract, salvia miltiorrhiza extract, onion extract, ginsenoside Rb, and ginsenoside Rg; the polyvinyl alcohol-tannic acid-Al 3+ The scaffold matrix was obtained by crosslinking polyvinyl alcohol, tannic acid and aluminum trichloride.

[0006] Preferably, the herbal extract is composed of honeysuckle extract, matrine, scutellaria baicalensis extract and centella asiatica extract.

[0007] Preferably, the herbal extract is composed of danshen extract, onion extract, ginsenoside Rb1 and ginsenoside Rg1.

[0008] More preferably, the herbal extract is composed of honeysuckle extract, matrine, scutellaria baicalensis extract, centella asiatica extract, salvia miltiorrhiza extract, onion extract, ginsenoside Rb1, and ginsenoside Rg1.

[0009] More preferably, the honeysuckle extract has a mass-volume concentration of 0.1 g / mL to 0.4 g / mL, matrine has a mass-volume concentration of 10-30 mg / mL, scutellaria baicalensis extract has a mass-volume concentration of 0.2-0.7 g / mL, centella asiatica extract has a mass-volume concentration of 0.2-0.7 g / mL, salvia miltiorrhiza extract has a volume concentration of 0.7-1 mL / mL, onion extract has a mass-volume concentration of 0.2-0.5 g / mL, ginsenoside Rb1 has a mass-volume concentration of 3-5 mg / mL, and ginsenoside Rg has a mass-volume concentration of 2-3 mg / mL.

[0010] Preferably, the honeysuckle extract, scutellaria baicalensis extract, salvia miltiorrhiza extract, centella asiatica extract, and onion extract are commercially available products or obtained by the following methods.

[0011] The honeysuckle extract was prepared as follows: honeysuckle was dried and pulverized, then deionized water was added at a ratio of 1:15 g / mL to extract the extract, which was then boiled at 90°C for 2 hours. The extract was then filtered, concentrated, and dried.

[0012] The Scutellaria baicalensis extract was prepared as follows: Scutellaria baicalensis root slices (2-4 mm), dried and pulverized, added to deionized water at a ratio of 1:1515 g / mL, decocted at 100℃ for 3 hours, centrifuged, and purified.

[0013] The Centella asiatica extract was prepared as follows: Centella asiatica was dried and pulverized, and extracted with 70% ethanol at a material-to-liquid ratio of 1:10 g / mL at 75°C for 2 hours. The extract was then concentrated and purified with AB-8 macroporous resin and 60% ethanol.

[0014] The preparation of the danshen extract is as follows: Danshen slices of 2-3 mm were added, and 60% ethanol was added at a material-to-liquid ratio of 1:8 g / mL. The mixture was then refluxed at 70°C for 3 hours, filtered, concentrated, and filtered through 0.45 μm micropores.

[0015] The onion extract was prepared as follows: onion slices with a thickness of 0.5 mm were added to deionized water at a material-to-liquid ratio of 1:5 g / mL and extracted at 90 °C for 2 h. The extract was then filtered, concentrated, and spray-dried.

[0016] This invention provides a method for preparing an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing, comprising: Step 1: Polyvinyl alcohol, tannic acid, and crosslinking agent aluminum trichloride are added to a ternary solvent and mixed. After heating and stirring to react, the mixture is kept at a constant temperature and allowed to stand to obtain polyvinyl alcohol-tannic acid-Al. 3+ Matrix solution; Step 2: Add the Chinese herbal extract to the ternary solvent and stir to obtain a mixed solution of Chinese herbal medicine; Step 3: Add the mixed Chinese medicine solution to polyvinyl alcohol-tannic acid-Al 3+ The matrix solution was mixed to obtain an antibacterial and healing-promoting herbal hydrogel dressing.

[0017] In step 1, the ternary solvent is prepared by mixing glycerol, water and ethanol in a volume ratio of (2~3):(3~4):(4~5).

[0018] Preferably, the ternary solvent is a mixture of glycerol, water and ethanol in a volume ratio of 3:4:5.

[0019] In step 1, the mass ratio of polyvinyl alcohol, tannic acid and aluminum trichloride is (5~6):(5~6):1.

[0020] Preferably, the mass ratio of polyvinyl alcohol, tannic acid and aluminum trichloride is 5:5:1.

[0021] In step 1, the heating and stirring reaction temperature is 60~80℃, the reaction time is 0.5~1h, the stirring speed is 200~400rpm, and the standing time is 1~2h.

[0022] Preferably, the stirring reaction temperature is 70°C, the reaction time is 0.5 h, the stirring speed is 200 rpm, and the standing time at 70°C is 1 h.

[0023] In step 2, the stirring temperature is 20-30℃, the speed is 200-400 rpm, and the time is 10-30 min.

[0024] Preferably, the stirring temperature is 26°C, the stirring speed is 300 rpm, and the stirring time is 10 min.

[0025] In step 3, the traditional Chinese medicine mixture solution is reacted with polyvinyl alcohol-tannic acid-Al 3+ The volume ratio of the skeleton matrix solution is 1:1~2.

[0026] Preferably, the traditional Chinese medicine mixture solution contains polyvinyl alcohol-tannic acid-Al 3+ The volume ratio of the skeleton matrix solution is 1:1.

[0027] The mixing speed is 200-400 rpm, and the time is 10-30 min.

[0028] This invention provides the application of the above-mentioned antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing in the preparation of a treatment for diabetic foot ulcers or burn infected wounds.

[0029] Compared with the prior art, the present invention has the following beneficial effects: The antibacterial and healing-promoting herbal hydrogel dressing provided by this invention achieves functional integration and performance optimization by constructing a synergistic system of a specific cross-linked matrix matrix and multi-component herbal extracts. This herbal hydrogel dressing forms a broad-spectrum antibacterial spectrum covering Gram-positive bacteria (such as Staphylococcus aureus), Gram-negative bacteria (such as Escherichia coli), and fungi, with a 24-hour antibacterial rate >70%. Simultaneously, tanshinone extract and ginsenosides significantly promote angiogenesis and fibroblast migration by activating the VEGF and TGF-β1 signaling pathways. Experiments showed that the wound healing time in rats was shortened by 38.6%, which is superior to single-component dressings (which only shortened the time by 13.2%–24.7%). Polyvinyl alcohol-tannic acid-Al 3+The matrix matrix forms a three-dimensional network through the coordination of polyvinyl alcohol's hydroxyl groups and tannic acid's phenolic hydroxyl groups with aluminum trichloride. Its dynamic cross-linking characteristics ensure structural stability and provide loading space for traditional Chinese medicine components, solving the phase separation problem of traditional blending methods. A combination of eight types of traditional Chinese medicine components, including honeysuckle extract and matrine, is selected. Through the spatiotemporal synergy of multi-target antibacterial mechanisms and healing-promoting components, the narrow antibacterial spectrum of single components is overcome. Aluminum trichloride acts as a cross-linking agent, forming metal-phenol coordination bonds with tannic acid, avoiding the competition of active groups in the traditional Chinese medicine components for cross-linking sites, and maintaining the balance between cross-linking density and drug loading. By limiting the traditional Chinese medicine extracts to two or more combinations, component antagonism is avoided, and a synergistic effect of broad-spectrum antibacterial and multi-pathway healing-promoting is formed through functional complementarity.

[0030] Furthermore, the concentration of honeysuckle extract was controlled within the range of 100-400 mg / mL to ensure the effective concentration of antibacterial components such as chlorogenic acid while avoiding excessive binding of phenolic hydroxyl groups with tannins due to excessively high concentrations; the concentration of matrine at 10-30 mg / mL balanced its interaction with Al. 3+ Coordination competition to prevent tannic acid-Al 3+ The cross-linked network is disrupted; the combination of 200-700 mg / mL of Scutellaria baicalensis extract and 200-700 g / mL of Centella asiatica extract complements the pH sensitivity of baicalin and the collagen-promoting function of Centella asiatica glycoside; the high concentration of 0.7-1 mL / mL of Salvia miltiorrhiza extract ensures the stable dispersion of the lipid-soluble components of tanshinone; the sulfide concentration gradient of 200-500 mg / mL of onion extract exerts both antibacterial effects and avoids precipitation with alkaloids; the specific ratio of ginsenoside Rb1 (3-5 mg / mL) and Rg (2-3 mg / mL) enhances the dispersion stability of saponin components in the cross-linked matrix by regulating the HLB value of saponin components, while reducing structural degradation during thermal processing.

[0031] The present invention provides a method for preparing an antibacterial and healing-promoting herbal hydrogel dressing. This method achieves stable loading and synergistic function of active ingredients by constructing a functionalized matrix and a herbal loading system in a stepwise manner. A framework matrix with a porous network and metal coordination sites is formed through crosslinking of polyvinyl alcohol, tannic acid, and aluminum trichloride. 3+Coordination with the phenolic hydroxyl groups of tannins enhances mechanical strength while preserving the porous structure to accommodate traditional Chinese medicine (TCM) components. A ternary solvent system is used to dissolve TCM extracts with diverse properties, avoiding phase separation of hydrophilic / lipophilic components. Low-temperature stirring reduces degradation of heat-sensitive components, achieving an encapsulation rate of 85%–92%, more than 30% higher than traditional blending methods (<65%). The TCM solution is mixed with a matrix solution, utilizing the metal coordination sites in the matrix to selectively adsorb active TCM groups. This achieves uniform dispersion of components and regulates the release sequence of different components through dynamic coordination equilibrium. A 72-hour sustained drug release is achieved through aluminum ion-phenolic hydroxyl coordination and hydrogen bond interpenetration networks, ensuring stable drug concentration in the wound microenvironment. This three-step process, through physical cross-linking and solvent synergy, solves the phase separation problem of traditional blending methods while avoiding high-temperature damage to active components, and balances cross-linking density with drug loading space requirements.

[0032] Furthermore, a mixed solvent system with gradient polarity of glycerol-water-ethanol was constructed. Glycerol, as a polyol solvent, has high viscosity, which reduces the volatility of the solvent system and decreases the risk of thermal degradation of active ingredients during high-temperature processing. Water, as a polar solvent, can dissolve hydrophilic components, while ethanol, as a moderately polar solvent, can solubilize lipophilic components. The three solvents are mixed in a volume ratio to create a solvent environment with continuously changing polarity. By adjusting the solubility balance of different polar components, phase separation caused by differences in solubility in multi-component traditional Chinese medicine extracts is avoided.

[0033] Furthermore, through a temperature control strategy involving pre-crosslinking and dynamic mixing, the porosity of the gel network can be precisely controlled, with batch-to-batch performance differences of less than 8%. During production, only the stirring rate and the B / C mixing ratio (2:1) need to be adjusted to adapt to different wound types, and a single batch can prepare 10-15 m³ of gel network. 2 Dressing, compared to traditional ultrasonic methods (<2 m) 2 Efficiency is increased by more than 5 times per time.

[0034] Furthermore, after being kept warm and left to stand, an interpenetrating network structure is formed, giving the dressing excellent resistance to shear deformation and allowing it to conform to irregular wound surfaces.

[0035] Furthermore, the low-temperature loading process (20-30 ℃) combined with low-speed stirring (200-400 rpm) ensures that the activity retention rate of heat-sensitive ginsenosides is >95% (compared to <70% by the traditional high-temperature method), and the uniformity of component distribution (RSD=3.2%) meets the ISO 13485 medical device manufacturing standard. Attached Figure Description

[0036] Figure 1 for Figure 1The image shows the 24-hour antibacterial ability of the dressing in Example 1 of this invention. The Wound group in the image is the blank control group. Product 1 and Product 2 are two hydrogel dressing products currently on the market. The Hydrogel group is a hydrogel without added Chinese medicine. Groups 1, 2 and 3 are hydrogel dressings loaded with different Chinese medicines. Figure 2 The image shows the hemostatic effect of the dressing in Example 1 of this invention on mouse wounds. The Control group in the image is a blank control group. Product 1 and Product 2 are two hydrogel dressing products currently on the market. The Hydrogel group is a hydrogel without added Chinese medicine. Groups 1, 2 and 3 are hydrogel dressings loaded with different Chinese medicines. Figure 3 The image shows the effect of the dressing in Example 1 of this invention on promoting wound healing in mice. The Wound group in the image is a blank control group. Product 1 and Product 2 are two hydrogel dressing products currently on the market. The Hydrogel group is a hydrogel without added Chinese medicine. Groups 1, 2 and 3 are hydrogel dressings loaded with different Chinese medicines. Figure 4 The image shows the healing of scratches on mouse fibroblast L-929 cells in the dressing in Example 1 of this invention. DMEM is the negative control, FBS is the positive control, Product 1 and Product 2 are two hydrogel dressing products currently on the market, Hydrogel group is hydrogel without added Chinese medicine, and groups 1, 2 and 3 are hydrogel dressings loaded with different Chinese medicines. Figure 5 The survival of fibroblasts in Example 1 of this invention is shown in DMEM (serum-free culture medium, as a negative control group), FBS (fetal bovine serum, as a positive control group) and various dressings. Product 1 and Product 2 are two hydrogel dressing products currently on the market. The Hydrogel group is a hydrogel without added Chinese medicine, while groups 1, 2 and 3 are hydrogel dressings loaded with different Chinese medicines. Detailed Implementation

[0037] To enable those skilled in the art to understand the features and effects of the present invention, the following descriptions and definitions are only general descriptions of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in the event of any conflict, the definitions in this specification shall prevail.

[0038] Unless otherwise defined, the technical terms used in the following embodiments have the same meanings as commonly understood by those skilled in the art to which this invention pertains. Unless otherwise specified, the experimental reagents used in the following embodiments are conventional biochemical reagents; and the experimental methods described are conventional methods.

[0039] The raw materials used in this invention are: glycerol (Maclean, 99%), ethanol (Sinopharm Chemical Reagent, 99.5%), (polyvinyl alcohol 1799 type (PVA), Aladdin, molecular weight 44.05260), tannic acid (TA) (Aladdin, USP purity grade), anhydrous aluminum trichloride (Aladdin, molecular weight 133.34), matrine (Aladdin, 98%), honeysuckle extract (Ron, 10:1), scutellaria baicalensis extract (Ron, 10:1), centella asiatica extract (Ron, 10:1), salvia miltiorrhiza extract (Ron, 10%), onion extract (Ron, 10:1), ginsenoside Rb1 (Yuanye Biotechnology, ≥98%), and ginsenoside Rg1 (Yuanye Biotechnology, ≥98%).

[0040] Example 1 This embodiment provides an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing. The specific preparation method of this traditional Chinese medicine hydrogel dressing is as follows: Add 30 mL of glycerol, 40 mL of water and 50 mL of ethanol to a 200 mL beaker. Place the beaker on a magnetic stirrer and set the stirring speed to 200 rpm. Stir for 10 minutes to obtain solution A.

[0041] Weigh out 6 g of polyvinyl alcohol (PVA), 6 g of tannic acid (TA), and 1.2 g of aluminum trichloride and add them to 60 mL of solution A. Place the beaker containing the mixed solution in a constant temperature water bath and heat it to 70°C. At the same time, use a magnetic stirrer to stir at 200 rpm for 0.5 hours to dissolve the solution. After standing at 70°C for 1 hour, allow it to cool naturally to room temperature to eliminate air bubbles in the solution, and obtain solution B.

[0042] Weigh 3 g of honeysuckle extract, 300 mg of matrine, 6 g of scutellaria baicalensis extract, and 4 g of centella asiatica extract and add them sequentially to 20 mL of solution A. Place the solution on a magnetic stirrer, set the stirring speed to 300 rpm, and the stirring temperature to 26℃. Stir for 10 minutes to obtain solution C1. Take 20 mL of solution A and add 14 mL of tanshinone extract, 4 g of onion extract, 60 mg of ginsenoside Rb1, and 40 mg of ginsenoside Rg1. Stir the above mixture at 300 rpm and 26℃ for 10 minutes using a magnetic stirrer to obtain solution C2. Take 20 mL of solution A and add 3 g of honeysuckle extract, 300 mg of matrine, 6 g of scutellaria baicalensis extract, 4 g of centella asiatica extract, 14 mL of tanshinone extract, 4 g of onion extract, 60 mg of ginsenoside Rb1, and 40 mg of ginsenoside Rg1. The above mixture was stirred with a magnetic stirrer at 300 rpm for 10 minutes at 26 °C to obtain solution C3. Mix 20 mL of solution B and 20 mL of solution C1; mix 20 mL of solution B and 20 mL of solution C2; mix 20 mL of solution B and 20 mL of solution C3. Stir each of the three solutions for 15 minutes using a magnetic stirrer at 200 rpm to obtain herbal hydrogel dressings BC1, BC2 and BC3. These three herbal hydrogel dressings are used as experimental group 1, experimental group 2 and experimental group 3, respectively.

[0043] (1) Antibacterial test This embodiment evaluates the antibacterial properties of traditional Chinese medicine hydrogel dressings using antibacterial experiments and colony counting methods. *Escherichia coli* and *Staphylococcus aureus* were used as test strains. The three types of traditional Chinese medicine hydrogel dressings prepared above were compared with a blank control group, commercially available wound dressings Product 1 (Yifu Hyaluronic Acid Gel Dressing, Nanjing Tianzong Yikang Biotechnology Co., Ltd.) and Product 1 (Haishi Hainuo Medical Silicone Gel Dressing, Zhengzhou Heji Biotechnology Co., Ltd.), and a hydrogel without added traditional Chinese medicine ingredients. *Escherichia coli* was cultured (…). E. coli ) and Staphylococcus aureus ( S. aureus The strain was prepared into a concentration of 10. 6 CFU / mL bacterial suspension. Antibacterial test procedure: Collect 50 μL of colony count. 6A CFU / mL suspension of *E. coli* or *S. aureus* was added to bacterial culture medium containing 500 μL of each group of hydrogel dressings: experimental groups (Sample A, B, C) and control groups (blank control without dressing, Product 1, Product 2, and blank hydrogel). The culture was incubated at 180 rpm and 37°C for 7 hours in a shaker. The culture was then diluted 100 times, and 100 μL of the diluted bacterial solution was evenly dropped into the center of a nutrient agar plate. Using a sterile spreader, the plate was evenly spread in a clockwise-counterclockwise-cross pattern. The plate was left uncovered in a clean bench for 15 minutes to allow the bacterial solution to be absorbed. Finally, all plates were inverted and placed in a 37°C incubator for static incubation for 18–24 hours.

[0044] The results are as follows Figure 1 As shown, the herbal hydrogel dressing prepared in this invention exhibits significant antibacterial effects against two strains, with bacterial density significantly lower than the blank control group and the hydrogel group without added herbal ingredients. The inhibition against Staphylococcus aureus is particularly pronounced, with sparse Staphylococcus aureus distribution in the herbal hydrogel group. Comparing the antibacterial ability of the hydrogel dressing prepared in this invention with commercially available hydrogel dressings, it was found that the antibacterial ability of the hydrogel dressing with only the first four herbal ingredients was not significantly different from that of commercially available products. However, the antibacterial effect of the hydrogel dressing with the last four herbal ingredients was better than that of commercially available products. Further addition of eight herbal ingredients further enhanced the antibacterial ability of the hydrogel dressing, significantly outperforming commercially available products. This also demonstrates the synergistic effect of multiple herbal ingredients, which significantly improved the antibacterial ability of the hydrogel dressing. Figure 1 The results showed that the third group of herbal hydrogel dressings exhibited the best antibacterial performance. Compared with the blank control group, the inhibition rates of the herbal hydrogel dressings in experimental group 3 against Escherichia coli and Staphylococcus aureus were 71.8% and 85.3%, respectively. The experimental results indicate that the synergistic effect of traditional Chinese medicine enhanced the antibacterial ability of the herbal hydrogel dressings. Combined with the membrane disruption mechanisms of matrine, honeysuckle, and onion extracts, the ability of Scutellaria baicalensis extract to interfere with bacterial DNA replication, the ability of Centella asiatica extract to promote fibroblast proliferation, and the angiogenic activity of Salvia miltiorrhiza extract, a broad-spectrum antibacterial spectrum covering Gram-positive bacteria, Gram-negative bacteria, and fungi was formed, resulting in stronger antibacterial capabilities.

[0045] (2) Hemostasis test The hemostatic function of the herbal hydrogel dressing was further evaluated. Using a mouse wound model, the herbal hydrogel dressing prepared above was compared with a blank control group, commercially available wound dressing products Product 1 and Product 2, and hydrogel without added herbal ingredients. To further evaluate the hemostatic function of the herbal hydrogel dressing, a mouse liver hemorrhage model was used in the experiment. Healthy SPF-grade ICR mice (weighing 20-22g, half male and half female) were selected. Before the experiment, the mice were fasted for 12 hours but allowed free access to water. They were then euthanized by an intraperitoneal injection of an excessive amount of sodium pentobarbital, and their weight was recorded. The mice were dissected, and their livers were exposed. A circular filter paper with a diameter of 4cm was placed under the liver. The liver was punctured (6-8 punctures) using a sterile syringe needle, and 300μL of each group of hydrogel dressings (experimental groups (Sample A, B, C) and control groups (blank control without dressing, Product 1, Product 2, and blank hydrogel)) were immediately applied to the damaged liver. After standing for 1 hour, the filter paper and dressings were removed, and the mice were weighed and their weight was recorded. The difference in weight before and after liver hemorrhage was calculated to represent the amount of blood loss and the hemostatic efficacy of each dressing group. The average hemostasis time of each group was statistically analyzed using GraphPad Prism. The software was used to perform one-way ANOVA to compare the differences in hemostatic effects among different groups (P<0.05 was considered statistically significant), thereby evaluating the hemostatic advantages of the traditional Chinese medicine hydrogel dressing.

[0046] The results are as follows Figure 2 As shown, compared to the blank control group, the third group of hydrogel dressings prepared in this invention exhibited a significant hemostatic effect, with blood flow reduced by 82.3%. This indicates that the dressing possesses a significant hemostatic effect. Through the analysis of... Figure 2 In groups 1, 2, and 3, it was observed that the hemostatic effect of the hydrogel dressing became more significant with the increase of medicinal components in the herbal hydrogel. This demonstrates that the synergistic effect of multiple herbal medicines improved the hemostatic effect of the herbal hydrogel dressing. By comparing the hemostatic effect of the hydrogel dressing prepared in this invention with commercially available hydrogel dressings, it was found that the antibacterial effect of the hydrogel dressing prepared in this invention was significantly better than that of commercially available hydrogel products. Furthermore, the hydrogel dressing with more herbal medicines exhibited stronger hemostatic ability, indicating that the herbal components in the hydrogel played a significant hemostatic role, and that multiple herbal medicines exerted a synergistic effect, thereby improving the hemostatic ability of the hydrogel dressing.

[0047] (3) Experiment on wound healing effect To further evaluate the wound healing function of the herbal hydrogel dressing, a mouse wound model was used. The three groups of herbal hydrogel dressings prepared above were compared with a blank control group, commercially available wound dressing products Product 1 and Product 2, and hydrogel without added herbal ingredients. Healthy mice with similar weights were randomly divided into the Wound group (blank control group), Product 1 group (commercially available hydrogel dressing product 1), Product 2 group (commercially available hydrogel dressing product 2), Hydrogel group (hydrogel without added herbal ingredients), Group 1 (hydrogel dressing loaded with herbal ingredient 1), Group 2 (hydrogel dressing loaded with herbal ingredient 2), and Group 3 (hydrogel dressing loaded with herbal ingredient 3), with 15 mice in each group. The backs of all mice were shaved to expose the skin. Circular wounds of the same diameter were made at the same location on the back of the mice using a uniform punch, and the initial wound area was recorded. The Wound group received no dressing; Product 1 and Product 2 groups received their respective commercially available hydrogel dressings; the Hydrogel group received a hydrogel dressing without added traditional Chinese medicine; Groups 1, 2, and 3 received hydrogel dressings loaded with their respective traditional Chinese medicines, ensuring complete coverage of the wound. Dressings were applied on day 0 (D0), day 3 (D3), day 7 (D7), and day 14 (D8) after wound creation. 14 ) and the 18th day (D 18 The wounds of mice in each group were photographed and recorded, and the wound area was measured and recorded using image analysis software.

[0048] The results are as follows Figure 3 As shown, with the increase of traditional Chinese medicine (TCM) components in the TCM hydrogel dressing, the wound showed a faster healing speed. After 18 days of observation, the wound treated with the third type of hydrogel dressing had completely closed, and the regenerated skin showed no significant difference from the surrounding normal skin tissue. The TCM hydrogel dressing of this invention can significantly accelerate wound closure and epithelialization. Through comparisons of experimental groups 1, 2, and 3, it was found that the TCM hydrogel dressing with more complex TCM components had a more significant effect on promoting wound repair and reconstruction, thus demonstrating that the synergistic effect of multiple TCM components can significantly improve the wound healing ability of the TCM hydrogel dressing. By comparing the healing results of the hydrogel dressing prepared in this invention with commercially available hydrogel dressings, it was found that the wound covered with the TCM hydrogel dressing prepared in this invention healed faster, and the reconstructed skin showed almost no difference in appearance from normal skin tissue. This indicates that the TCM hydrogel dressing prepared in this invention has a superior wound healing ability.

[0049] (4) Effect of scratch healing on mouse fibroblast L-929 cells Further cell scratch healing experiments were conducted using mouse fibroblast L929 cells (YC-C091, Yuanjing Biotechnology). L929 mouse fibroblasts were seeded in culture flasks containing appropriate culture medium and cultured at 37°C with 5% CO2 until the cells reached 80%–90% confluence. Scratches were created on the monolayer of cells using a sterile pipette tip, ensuring uniform scratch width. The cells were then gently washed with PBS to remove cell debris generated during the scratching process. Cells after scratching were randomly divided into the following groups: DMEM group (negative control, using serum-free DMEM medium), FBS group (positive control, using medium containing a certain concentration of fetal bovine serum), Product 1 group (applied with extract of commercially available hydrogel dressing product 1), Product 2 group (applied with extract of commercially available hydrogel dressing product 2), Hydrogel group (applied with hydrogel extract without added Chinese medicine), Group 1 (applied with hydrogel dressing extract loaded with Chinese medicine 1), Group 2 (applied with hydrogel dressing extract loaded with Chinese medicine 2), and Group 3 (applied with hydrogel dressing extract loaded with Chinese medicine 3). The corresponding treatment solution was added to each well according to the group, and then the cell plates were returned to the incubator for further culture. Cell plates were removed at 0 h, 12 h, 24 h, 36 h, 48 h, and 72 h after scratch creation, and cell migration and healing in the scratched area were observed and photographed under a microscope. The relative area of ​​the scratches at different time points was measured using image analysis software.

[0050] The results are as follows Figure 4 As shown, DMEM (serum-free culture medium) was the negative control, and FBS (fetal bovine serum) was the positive control. It can be seen that compared to the negative control group, the traditional Chinese medicine components in the herbal hydrogel played a positive role in promoting the proliferation and migration of mouse fibroblasts, and this effect became more significant with the increase of herbal components. This indicates that the synergistic effect of multiple herbal components can promote cell proliferation and migration, thereby accelerating wound healing and skin regeneration. The results of experimental group 3 show that the cell migration ability was significantly enhanced from 48 to 72 hours, indicating the sustained-release effect of the herbal hydrogel. By releasing components that promote cell migration during this period, it can accelerate cell migration and even significantly promote wound healing in the wound environment. Comparing the experimental results of commercially available products with the hydrogel dressing prepared in this invention, it was found that the commercially available product 1 had a better effect on promoting cell migration than the hydrogel dressing without added traditional Chinese medicine, while the commercially available product 2 had a significantly worse effect on promoting cell migration than the hydrogel prepared in this invention. Overall, the hydrogel dressing with added eight kinds of traditional Chinese medicine showed the most significant effect in promoting cell migration, indicating that the synergistic effect of traditional Chinese medicine promoted cell migration, and the traditional Chinese medicine hydrogel dressing prepared by this invention has excellent ability to promote cell migration.

[0051] (5) Experiments on the effect on fibroblast survival Cell viability was further evaluated by culturing fibroblasts (L-929 cells or other suitable fibroblast lines) to the logarithmic growth phase, digesting them with trypsin to prepare single-cell suspensions, and seeding them at appropriate densities in multi-well plates. The plates were then incubated at 37°C with 5% CO2 to allow cell adhesion. After cell adhesion, the cultured cells were randomly divided into the following groups: DMEM group (using serum-free DMEM medium as a negative control), FBS group (using medium containing a certain concentration of fetal bovine serum as a positive control), Product 1 group, Product 2 group, Hydrogel group, Group 1 (applied with hydrogel dressing extract loaded with traditional Chinese medicine 1), Group 2 (applied with hydrogel dressing extract loaded with traditional Chinese medicine 2), and Group 3 (applied with hydrogel dressing extract loaded with traditional Chinese medicine 3). The corresponding treatment solutions were added to the cell culture systems of each group, and the cells were incubated for another 24 hours.

[0052] Cell viability was assessed using a double staining method with calcein-AM and propidium iodide (PI). Calcein-AM can penetrate the membrane of living cells and is converted to calcein by esterases within the cells, emitting green fluorescence. PI can only penetrate the membrane of dead cells and emits red fluorescence after binding to nucleic acids. Appropriate concentrations of Calcein-AM and PI staining solutions were added to the cell culture systems of each group, and the cells were incubated at 37°C in the dark for 20 min. After staining, the cells were gently washed 2–3 times with PBS to remove unbound staining solution. Multiwell plates containing stained cells were observed and photographed under a fluorescence microscope. Images were acquired in both the green fluorescence channel (detecting Calcein-AM-labeled living cells) and the red fluorescence channel (detecting PI-labeled dead cells). Image analysis software was used to quantitatively analyze the green fluorescence intensity (representing the number or activity of living cells) and the red fluorescence intensity (representing the number of dead cells) in the images of each group, and the average fluorescence intensity was calculated.

[0053] The results are as follows Figure 5As shown, DMEM (serum-free culture medium) served as a negative control, and FBS (fetal bovine serum) served as a positive control. The results indicated that the cell survival rate in the first type of herbal hydrogel was significantly higher than that in the negative control group and the hydrogel group without added herbal medicine, and slightly higher than the positive control group. Comparison of experimental groups 1, 2, and 3 revealed that the cell survival rate significantly increased with the increase of herbal components in the herbal hydrogel dressing, indicating that multiple herbal medicines play a positive role in cell survival. Through the combined effect of multiple herbal medicines, the cell survival-promoting effect of the herbal hydrogel dressing was significantly enhanced. Furthermore, a comparison of cell survival between commercially available hydrogel dressings and the hydrogel dressing prepared in this invention showed that the cell survival rate in the commercially available product was higher than that in the hydrogel without added herbal medicine. The addition of herbal components significantly improved the cell survival rate in the hydrogel dressing, and the cell survival rate in the herbal hydrogel dressing was higher than that in the commercially available product, further demonstrating that herbal components have the function of promoting cell survival.

[0054] The above evaluation and testing demonstrated that the herbal hydrogel dressing, rich in various traditional Chinese medicines, possesses excellent antibacterial and hemostatic functions. Simultaneously, the dressing promotes wound cell survival and proliferation, thus achieving significant antibacterial and wound-healing effects, surpassing commonly used commercially available hydrogel dressings in these aspects. Furthermore, the experimental results also proved that the various herbal components in the herbal hydrogel dressing exert a synergistic effect, significantly enhancing its antibacterial and wound-healing properties.

[0055] Example 2 This embodiment provides an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing. The specific preparation method of this traditional Chinese medicine hydrogel dressing is as follows: Add 20 mL of glycerol, 30 mL of water, and 40 mL of ethanol to a 200 mL beaker. Place the beaker on a magnetic stirrer and set the stirring speed to 400 rpm. Stir for 10 minutes to obtain solution A.

[0056] Weigh out 6g of polyvinyl alcohol (PVA), 6g of tannic acid (TA), and 1.2g of aluminum trichloride, and add them to solution A at concentrations of 100 mg / mL, 100 mg / mL, and 20 mg / mL, respectively. Place the beaker containing the mixed solution in a constant temperature water bath and heat it to 70°C. At the same time, stir with a magnetic stirrer at 400 rpm for 0.5 hours to dissolve the solution. After standing at 70°C for 1 hour, eliminate the air bubbles in the solution to obtain solution B.

[0057] Weigh out 9g of honeysuckle extract, 750 mg of matrine, 12 g of scutellaria baicalensis extract, 12 g of centella asiatica extract, 6 g of onion extract, 30 mL of salvia miltiorrhiza extract, 120 mg of ginsenoside Rb1 and 60 mg of ginsenoside Rg1 and add them sequentially to 30 mL of solution A. Place the solution on a magnetic stirrer, set the stirring speed to 400 rpm, control the stirring temperature at 24℃, and stir for 10 minutes to obtain solution C.

[0058] Measure 30 mL of solution B and 30 mL of solution C, add them to a beaker, place the beaker on a magnetic stirrer, set the stirring speed to 400 rpm, and stir for 30 minutes to ensure that the two solutions are fully mixed and homogeneous, and finally obtain solution D, which is the antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing.

[0059] Example 3 This embodiment provides an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing. The specific preparation method of this traditional Chinese medicine hydrogel dressing is as follows: Add 30 mL of glycerol, 40 mL of water and 40 mL of ethanol to a beaker, place the beaker on a magnetic stirrer, set the stirring speed to 400 rpm, and stir for 10 minutes to obtain solution A.

[0060] Weigh out 6g of polyvinyl alcohol (PVA), 6g of tannic acid (TA), and 1.2g of aluminum trichloride and add them to 60mL of solution A. Place the beaker containing the mixed solution in a constant temperature water bath and heat it to 70℃. At the same time, use a magnetic stirrer to stir at 400 rpm for 0.5 hours to dissolve the solution. After standing at 70℃ for 1 hour, eliminate the air bubbles in the solution to obtain solution B.

[0061] Weigh 18 g of honeysuckle extract, 1.2 g of matrine, 25 g of scutellaria baicalensis extract, 35 g of centella asiatica extract, add 45 mL of salvia miltiorrhiza extract, 25 g of onion extract, 250 mg of ginsenoside Rb1, and 150 mg of ginsenoside Rg1 to 50 mL of solution A. Place the solution on a magnetic stirrer, set the stirring speed to 400 rpm, and the stirring temperature to 22 ℃. Stir for 10 minutes to obtain solution C.

[0062] Measure 50 mL of solution B and 50 mL of solution C, add them to a beaker, place the beaker on a magnetic stirrer, set the stirring speed to 400 rpm, and stir for 30 minutes to ensure that the two solutions are fully mixed and homogeneous. The final product is solution D, which is an antibacterial and healing-promoting herbal hydrogel dressing.

[0063] The antibacterial and healing-promoting herbal hydrogel dressings of Examples 2 and 3 were tested and verified. The antibacterial and healing-promoting effects of the herbal hydrogel dressings of Examples 2 and 3 were significantly better than those of commercially available products and the control group, better than experimental group 1 and experimental group 2, and close to those of experimental group 3. The above results further illustrate that multiple herbal medicines played a synergistic role, which improved the antibacterial and healing-promoting effects of the hydrogel dressing.

[0064] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A traditional Chinese medicine hydrogel dressing with antibacterial and healing-promoting properties, characterized in that, The herbal hydrogel dressing is made by loading herbal extracts onto polyvinyl alcohol-tannic acid-Al. 3+ The scaffold matrix is ​​obtained, and the herbal extract is composed of two or more of the following: honeysuckle extract, matrine, scutellaria baicalensis extract, centella asiatica extract, salvia miltiorrhiza extract, onion extract, ginsenoside Rb, and ginsenoside Rg; the polyvinyl alcohol-tannic acid-Al... 3+ The scaffold matrix was obtained by crosslinking polyvinyl alcohol, tannic acid and aluminum trichloride.

2. The antibacterial and healing-promoting herbal hydrogel dressing according to claim 1, characterized in that, The concentrations of the following extracts are as follows: honeysuckle extract (100-400 mg / mL), matrine (10-30 mg / mL), scutellaria baicalensis extract (200-700 mg / mL), centella asiatica extract (200-700 g / mL), salvia miltiorrhiza extract (0.7-1 mL / mL), onion extract (200-500 mg / mL), ginsenoside Rb1 (3-5 mg / mL), and ginsenoside Rg (2-3 mg / mL).

3. The method for preparing an antibacterial and healing-promoting herbal hydrogel dressing according to claim 1 or 2, characterized in that, include: Step 1: Polyvinyl alcohol, tannic acid, and aluminum trichloride are added to a ternary solvent, mixed, heated and stirred to react, and then kept at the temperature and allowed to stand to obtain polyvinyl alcohol-tannic acid-Al. 3+ Matrix solution; Step 2: Add the Chinese herbal extract to the ternary solvent and stir to obtain a mixed solution of Chinese herbal medicine; Step 3: Add the mixed Chinese medicine solution to polyvinyl alcohol-tannic acid-Al 3+ The matrix solution was mixed to obtain an antibacterial and healing-promoting herbal hydrogel dressing.

4. The method for preparing an antibacterial and healing-promoting herbal hydrogel dressing according to claim 3, characterized in that, In step 1, the ternary solvent is prepared by mixing glycerol, water and ethanol in a volume ratio of (2~3):(3~4):(4~5).

5. The method for preparing an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing according to claim 3, characterized in that, In step 1, the mass ratio of polyvinyl alcohol, tannic acid and aluminum trichloride is (5~6):(5~6):

1.

6. The method for preparing an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing according to claim 3, characterized in that, In step 1, the heating and stirring reaction temperature is 60~80℃, the reaction time is 0.5~1h, the stirring speed is 200~400rpm, and the standing time is 1~2h.

7. The method for preparing an antibacterial and healing-promoting herbal hydrogel dressing according to claim 3, characterized in that, In step 2, the stirring temperature is 20-30℃, the speed is 200-400 rpm, and the time is 10-30 min.

8. The method for preparing an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing according to claim 3, characterized in that, In step 3, the traditional Chinese medicine mixture solution is reacted with polyvinyl alcohol-tannic acid-Al 3+ The volume ratio of the skeleton matrix solution is 1:1~2.

9. The method for preparing an antibacterial and healing-promoting traditional Chinese medicine hydrogel dressing according to claim 3, characterized in that, The mixing speed is 200-400 rpm, and the time is 10-30 min.

10. The application of the antibacterial and healing-promoting herbal hydrogel dressing according to claim 1 or 2 in the preparation of a treatment for diabetic foot ulcers or burn infected wounds.

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