A quaternary ammonium silver agarose polysaccharide-based hydrogel for wound dressing and a preparation method and application thereof

Abstract

The application provides a quaternary aminized silver fungus polysaccharide-based hydrogel for wound dressings and a preparation method thereof. The prepared hydrogel has obvious porous structure and viscoelasticity characteristics, and has good gel strength, antibacterial property and degradability. As a wound dressing, the hydrogel can provide a moist healing environment for a wound, can promote cell proliferation and tissue repair, and can promote wound healing. The quaternary aminized silver fungus polysaccharide-based hydrogel prepared by the application will promote the application research of quaternary aminization modification in the field of silver fungus polysaccharide, and promote the application development of organic degradable hydrogel in wound dressings.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials technology, specifically to a quaternized tremella polysaccharide-based hydrogel for wound dressing, its preparation method, and its application. Background Technology

[0002] Skin trauma is a common condition, and wounds can form due to a variety of factors, including external stimuli or internal pathological conditions, such as scratches, abrasions, burns, and diabetes. During wound healing, bacterial infection can occur, leading to inflammation and significantly prolonging healing time. In severe cases, it can cause necrosis or other serious complications. Applying dressings to the wound can protect it, prevent microbial infection, and promote healing.

[0003] Wound dressings are essential medical supplies used to protect and promote wound healing. They serve multiple functions, such as absorbing exudate, keeping wounds moist, relieving pain, and preventing infection. Common gauze, cotton pads, and adhesive tape are all classified as traditional dressings. Traditional dressings have problems such as poor absorbency, easy saturation leading to infection, and irritation to the wound during dressing changes, potentially causing scar hyperplasia.

[0004] Novel wound dressings offer better options for wound care. Based on their different forms, novel wound dressings can be categorized into films, foams, crystals, and hydrogels, which are made from various materials, including natural or synthetic polymers, and combinations thereof.

[0005] Tremella fuciformis polysaccharide can significantly enhance the function of the mononuclear macrophage system in mice, promote lymphocyte proliferation, and strengthen the body's immune function. In terms of anti-tumor effects, Tremella fuciformis polysaccharide can inhibit tumor growth by inducing apoptosis in cancer cells. It can also be used to treat leukopenia caused by radiotherapy or chemotherapy for tumors. Furthermore, it can lower blood sugar and blood lipids by affecting glucose metabolism-related catalytic proteins and promoting insulin secretion. The inhibitory effect and mechanism of Tremella fuciformis polysaccharide on amylase digestive enzymes have also been extensively studied, revealing significant inhibitory effects on pancreatic α-amylase and α-glucosidase.

[0006] Quaternization modification is a chemical method of grafting quaternary ammonium groups onto organic molecules. When chitosan undergoes quaternization modification to introduce quaternary ammonium groups, its antibacterial properties are significantly enhanced, particularly its significant inhibitory effect on the growth of Pseudomonas aeruginosa biofilms. Hydrogels prepared using quaternized-ammoniated chitosan exhibit improved antibacterial properties and biodegradability, providing a moist healing environment for wounds and promoting cell proliferation and tissue repair.

[0007] Tremella fuciformis polysaccharides exert their anti-inflammatory effects by reducing the expression of pro-inflammatory factors. Quaternized Tremella fuciformis polysaccharides, after quaternization modification, exhibit significantly enhanced antibacterial properties. Hydrogels formed by cross-linking quaternized Tremella fuciformis polysaccharides with chitosan possess a more stable network structure and stronger antibacterial activity compared to simple chitosan hydrogels and Tremella fuciformis polysaccharide hydrogels, making them suitable for wound dressings. Research on quaternized Tremella fuciformis polysaccharides is still in its early stages, and there are no reports of using hydrogels formed by cross-linking quaternized Tremella fuciformis polysaccharides with chitosan for wound dressings. Summary of the Invention

[0008] Purpose of the invention The purpose of this invention is to provide a quaternized tremella polysaccharide-based hydrogel for wound dressings and its preparation method. The prepared hydrogel exhibits a distinct porous structure and viscoelastic characteristics, as well as good gel strength and antibacterial properties. When applied to wound dressings, it demonstrates significant hemostatic effects and promotes wound healing. The quaternized tremella polysaccharide-based hydrogel prepared by this invention and its applications will promote research on the application of quaternization modification in the field of tremella polysaccharides and enrich the product range of hydrogel wound dressings.

[0009] Solution Quaternized tremella polysaccharide (QTP) was prepared using 3-chloro-2-hydroxypropyltrimethylammonium chloride (CHPTAC). Quaternized tremella polysaccharide hydrogel (QTP-gel) was then prepared using QTP, chitosan (CS), and curcumin (Cur) as basic raw materials, and glutaraldehyde (GA) as a cross-linking agent.

[0010] Using the antibacterial effect of QTP-gel as a key indicator, the experiment verified four key factors: QTP concentration, QTP:CS ratio, Curl concentration, and GA concentration. The preparation conditions of QTP-gel were optimized, and the optimal process conditions for preparing QTP-gel were determined to be: QTP concentration of 1%, QTP:CS ratio of 1:3, Curl concentration of 1%, and GA concentration of 4%. The QTP-gel prepared with the optimal formula showed an inhibition zone diameter of 37.7 ± 0.2 mm for *Escherichia coli* and 28.5 ± 0.2 mm for *Staphylococcus aureus*.

[0011] Specific preparation method of quaternized Tremella fuciformis polysaccharide hydrogel: (1) Dissolve 1.62 g of Tremella fuciformis polysaccharide in 65 ml of pure water and stir thoroughly. When the Tremella fuciformis polysaccharide is completely dissolved, add 7.424 g of NaOH and stir continuously for 30 min for alkalization treatment. Then, under 57°C water bath conditions, slowly add 30.08 g of 65% CHPTAC aqueous solution dropwise to the Tremella fuciformis polysaccharide solution and stir continuously for 12.5 h. After the reaction is complete, adjust the pH to 5.5, and then dialyze the product using a dialysis bag for 3 days. The dialyzed product is then freeze-dried to obtain QTP.

[0012] (2) Dissolve a certain amount of quaternized Tremella polysaccharide in pure water to prepare a 1% quaternized Tremella polysaccharide solution (solution A).

[0013] (3) Dissolve chitosan in 1.5% acetic acid solution to prepare 1.2% chitosan solution (B solution).

[0014] (4) Dissolve curcumin in 80% ethanol solution to prepare 1% curcumin solution (C solution).

[0015] (5) Add 50% glutaraldehyde to pure water to prepare a 4% glutaraldehyde solution (D solution).

[0016] (6) Place 10 ml of solution A in a beaker, and add solution B with the aid of a magnetic stirrer to make the ratio of quaternized tremella polysaccharide to chitosan 1:3. After the solution is completely mixed, stir for 5 min. Add 2.5 ml of solution C and stir for another 5 min. After stirring, remove the solution and place it in an ultrasonic bath to defoam. After the bubbles are eliminated, add 2 ml of solution D, pour it into a mold and let it stand to form. After the hydrogel has stood and formed, wash the hydrogel repeatedly with pure water 3 times to obtain quaternized tremella polysaccharide hydrogel (QTP-gel).

[0017] Technical effect The quaternized tremella polysaccharide hydrogel prepared by this invention has the following outstanding beneficial effects: (1) The diameter of the inhibition zone of E. coli on the QTP-gel hydrogel was 37.7 ± 0.2 mm, and the diameter of the inhibition zone of Staphylococcus aureus was 28.5 ± 0.2 mm, indicating that it has good antibacterial properties.

[0018] (2) The experimentally measured strength of QTP-gel hydrogel was 268.13±4.72g, which shows that it has good gel strength and is suitable for use in wound dressing.

[0019] Structural characterization and physicochemical properties of the quaternized tremella polysaccharide hydrogel (QTP-gel) prepared in this invention: (1) Scanning electron microscopy (SEM) observation showed that the QTP-gel hydrogel exhibited a distinct porous structure, which significantly increased the specific surface area and porosity of the hydrogel, enhancing its adsorption and exchange capacity. In addition, while maintaining flexibility, it exhibited good mechanical properties and provided favorable conditions for cell migration and angiogenesis, which is beneficial to tissue regeneration.

[0020] (2) In the rheological property test, the storage modulus (G') of both QTP-gel and CS-gel without QTP was higher than its loss modulus (G'') under all test conditions, exhibiting typical viscoelastic characteristics, indicating that a stable elastic cross-linked network was successfully constructed. During the experiment, the G' of the hydrogel remained stable throughout the test, further confirming the stability of its internal structure. Furthermore, with the addition of QTP, the G' and G'' of the hydrogel increased sequentially, indicating an increase in cross-linking degree and a more compact network structure.

[0021] The application effects of the quaternized tremella polysaccharide hydrogel (QTP-gel) prepared in this invention in wound dressings: Pour the ungelled QTP-gel solution into a mold to form a hydrogel sheet with a diameter of about 1 cm. Wash the prepared QTP-gel sheet three times with pure water to remove excess monomer, and then sterilize it under a UV lamp for later use.

[0022] (1) Hemostatic performance test In the liver hemorrhage model, the average bleeding amount of QTP-gel was 0.16 g, significantly lower than that of the blank group (0.26 g) and the gauze group (0.24 g), demonstrating excellent hemostatic effect. In the tail hemorrhage model, QTP-gel also showed significant hemostatic effect, with an average bleeding amount of 0.08 g, far lower than that of the blank group (0.25 g) and the gauze group (0.24 g).

[0023] (2) Mouse wound healing experiment The performance of QTP-gel in wound healing was evaluated using a mouse model of full-thickness skin defect. On day 3, the healing rate of the QTP-gel group was 63.62%, while the healing rates of the control group (12.71%) and gauze group (25.48%) were lower. At this time, the wounds in the control group and gauze group showed yellow exudate and significant inflammatory response; while the wounds in the QTP-gel group showed milder inflammation and had formed scabs. By day 7, the healing rate of the QTP-gel group further increased to 73.62%, while the healing rates of the control group and gauze group were 45.22% and 46.75%, respectively, indicating a slower healing process. By day 14, the wounds in the QTP-gel group were almost completely healed, with a healing rate of 95.68%, while the control group and gauze group still had large unclosed areas, with healing rates of 76.06% and 72.53%, respectively. In summary, QTP-gel can provide a moist and occlusive environment for the wound, promote cell migration and tissue regeneration, and significantly accelerate the wound healing process. Attached Figure Description Figure 1 Scanning electron microscopy image of quaternized tremella polysaccharide hydrogel (QTP-gel) prepared according to the optimal formulation of this invention. Figure 2 Schematic diagram of the quaternization modification process of Tremella fuciformis polysaccharide Figure 3 The storage modulus (a) and loss modulus (b) of the hydrogel are shown. Figure 4 The amount of bleeding in the liver hemorrhage model (a) and the tail hemorrhage model (b) is shown. Figure 5 Image of a full-thickness dermal wound on the back of a mouse. Figure 6 This study used ImageJ software to measure and statistically analyze images of full-thickness dermal wounds on the back of mice and their image overlap variations. Figure 7 The healing rate of full-thickness skin wounds on the back of mice.

Claims

1. A quaternized tremella polysaccharide-based hydrogel for wound dressing, its preparation method, and its application. Its characteristics are: (1) Quaternized Tremella polysaccharide was prepared using 3-chloro-2-hydroxypropyltrimethylammonium chloride. (2) Quaternized Tremella polysaccharide, chitosan and curcumin were used as basic raw materials, and glutaraldehyde was used as a crosslinking agent to prepare quaternized Tremella polysaccharide hydrogel. (3) The optimal composition for preparing quaternized Tremella fuciformis polysaccharide hydrogel is as follows: quaternized Tremella fuciformis polysaccharide concentration of 1%, material ratio of quaternized Tremella fuciformis polysaccharide to chitosan of 1:3, curcumin concentration of 1%, and glutaraldehyde concentration of 4%. The quaternized Tremella fuciformis polysaccharide hydrogel prepared by the optimal formula.

2. The preparation steps of the quaternized Tremella fuciformis polysaccharide-based hydrogel according to claim (1) are as follows: (1) Dissolve 1.62 g of Tremella fuciformis polysaccharide in 65 ml of pure water and stir thoroughly. When the Tremella fuciformis polysaccharide is completely dissolved, add 7.424 g of NaOH and stir continuously for 30 min for alkalization treatment. Then, under 57°C water bath conditions, slowly add 30.08 g of 65% CHPTAC aqueous solution dropwise to the Tremella fuciformis polysaccharide solution and stir continuously for 12.5 h. After the reaction is complete, adjust the pH to 5.5, and then dialyze the product using a dialysis bag for 3 days. The dialyzed product is freeze-dried to obtain quaternized Tremella fuciformis polysaccharide (QTP). (2) Dissolve a certain amount of quaternized tremella polysaccharide in pure water to prepare a 1% quaternized tremella polysaccharide solution (Solution A). (3) Dissolve chitosan in 1.5% acetic acid solution to prepare 1.2% chitosan solution (B solution). (4) Dissolve curcumin in 80% ethanol solution to prepare 1% curcumin solution (C solution). (5) Add 50% glutaraldehyde to pure water to prepare a 4% glutaraldehyde solution (D solution). (6) Place 10 ml of solution A in a beaker, and add solution B with the aid of a magnetic stirrer to make the ratio of quaternized tremella polysaccharide to chitosan 1:

3. After the solution is completely mixed, stir for 5 min. Add 2.5 ml of solution C and stir for another 5 min. After stirring, remove the solution and place it in an ultrasonic bath to defoam. After the bubbles are eliminated, add 2 ml of solution D, pour it into a mold and let it stand to form. After the hydrogel has stood and formed, wash the hydrogel repeatedly with pure water 3 times to obtain quaternized tremella polysaccharide hydrogel (QTP-gel).

3. The quaternized tremella polysaccharide hydrogel prepared according to the above-mentioned synthetic components and methods has good antibacterial properties, shows good gel strength potential, is suitable for use in wound dressings, and has good wound hemostasis and wound healing promotion properties.

4. The scope of protection of this invention shall not be limited to the embodiments or comparative examples shown herein, but shall conform to the widest scope consistent with claim 1.

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