Hydrogel dressing for promoting healing of diabetic wound as well as preparation method and application of hydrogel dressing
The hydrogel constructed by the self-assembly of glycyrrhizic acid and S-allyl-L-cysteine solves the problem of insufficient adaptability of diabetic wound dressings to the dynamic wound microenvironment, achieving continuous protection and efficient healing of the wound. It has flexibility and bioactivity, and promotes the repair of diabetic wounds.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY (GUANGZHOU SEVERE MATERNAL TREATMENT CENTER GUANGZHOU ROUJI HOSPITAL)
- Filing Date
- 2026-03-30
- Publication Date
- 2026-05-12
AI Technical Summary
Existing diabetic wound dressings have limited adaptability to the dynamic wound microenvironment, making it difficult to effectively promote the healing of diabetic wounds. Traditional H2S delivery systems lack flexibility, which limits the therapeutic effect.
A novel drug-active molecule hydrogel was constructed by intermolecular interaction between glycyrrhizic acid and S-allyl-L-cysteine. Utilizing its three-dimensional network structure and synergistic effect, an intelligent bioactive platform capable of responding to wound needs was formed, generating endogenous H2S signaling molecules to regulate inflammation and oxidative stress.
It enables the hydrogel to adapt to dynamic physiological conditions, providing continuous protection, promoting the healing of diabetic wounds, and has the flexibility to adapt to different healing stages. It also reduces inflammatory response, activates angiogenesis, and improves wound repair efficiency.
Smart Images
Figure CN122005906A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of medical technology, specifically relating to a hydrogel dressing that promotes the healing of diabetic wounds, its preparation method, and its application. Background Technology
[0002] Chronic diabetic wounds (such as foot ulcers) are often in a stagnant inflammatory state due to the high-glucose microenvironment, persistent inflammation, and severe hypoxia, making them extremely difficult to heal. Although modern wound dressings (such as hydrogels, hydrocolloids, and polymer films) can provide physical protection and moisturizing effects—for example, Chinese patent application CN106267302A discloses a smart moisture-management hydrogel composite dressing comprising a substrate layer and a hydrogel layer covering the surface of the substrate layer. The hydrogel layer undergoes a sol-gel transition, containing 40-80% water throughout, and its surface becomes a hydrophobic interface, combined with an absorbent nonwoven substrate—their adaptability to the dynamic wound microenvironment is limited, often leading to poor healing, especially in full-thickness skin injuries. Studies have shown that a dual strategy of reducing ROS levels and guiding M2 macrophage polarization holds promise for inhibiting inflammation, stimulating angiogenesis, and restoring effective wound regeneration.
[0003] In recent years, self-assembled hydrogel systems based on small natural drug molecules have become a research frontier in the biomedical field. Compared with traditional hydrogels that rely on inert polymers, the core advantage of this system lies in realizing the integrated carrier-drug design concept: natural drug molecules not only serve as the structural framework for building the three-dimensional network of the gel, but also retain and exert their endogenous pharmacological activity, thereby avoiding the introduction of a large number of exogenous auxiliary materials and significantly improving biosafety and therapeutic efficiency.
[0004] Hydrogen sulfide (H2S), as a key gaseous signaling molecule, shows great potential in regulating inflammation, oxidative stress, and extracellular matrix remodeling—processes crucial for effective wound healing. Cystathione-γ-lyase (CSE) is the core enzyme in its synthesis. However, conventional H2S delivery systems lack the flexibility to adapt to the dynamic needs of different healing stages, thus limiting their therapeutic efficacy. Summary of the Invention
[0005] To address the shortcomings of existing technologies, the present invention aims to provide a hydrogel dressing for promoting the healing of diabetic wounds, its preparation method, and its applications. This invention constructs a novel pharmaceutically active molecular hydrogel through the intermolecular interaction between glycyrrhizic acid (GA) and S-allyl-L-cysteine (SAC). This hydrogel not only utilizes the three-dimensional network structure of GA to achieve physical encapsulation and chemical stability of SAC, but also, through the synergistic effect of the two, constructs an intelligent bioactive platform capable of responding to the needs of the wound. The hydrogel of this invention exhibits tissue-like viscoelastic characteristics, adapting to wound movement and thus providing continuous protection for chronic wounds under dynamic physiological conditions, possessing the flexibility to adapt to the dynamic needs of different healing stages.
[0006] The technical solution of this invention is: A hydrogel dressing that promotes the healing of diabetic wounds is prepared by using glycyrrhizic acid and S-allyl-L-cysteine as raw materials, buffer solution as dispersion medium, and controlling the mass ratio of raw materials, reaction temperature and reaction time, and pH to enable the self-assembly of glycyrrhizic acid and S-allyl-L-cysteine.
[0007] Furthermore, the buffer solution is a phosphate buffer solution.
[0008] Furthermore, the buffer solution is a 0.1× phosphate buffer solution with a pH of 7-8.
[0009] Furthermore, the buffer solution is a 0.1× phosphate buffer solution with pH=7.4, which not only provides a mild, stable, and physiologically close pH dissolution environment, but also neutralizes the acidic pathological environment of diabetic wounds.
[0010] Furthermore, the mass ratio of glycyrrhizic acid to S-allyl-L-cysteine is 10:2~8.
[0011] Furthermore, the mass ratio of glycyrrhizic acid to S-allyl-L-cysteine is 10:2.
[0012] The present invention also provides a method for preparing the aforementioned hydrogel dressing for promoting the healing of diabetic wounds, comprising the following steps: S1 Take glycyrrhizic acid, add buffer solution, and vortex sonicate until completely dissolved to obtain a clear and transparent glycyrrhizic acid solution; S2 Take S-allyl-L-cysteine, add buffer solution, vortex and sonicate until completely dissolved to obtain a clear and transparent S-allyl-L-cysteine solution; S3. Place the glycyrrhizic acid solution obtained in step S1 in a water bath and stir continuously. Add the S-allyl-L-cysteine solution obtained in step S2 and continue stirring until the pH of the system stabilizes at 7-8. Transfer the solution to a refrigerator to form a hydrogel dressing.
[0013] Furthermore, the concentration of the glycyrrhizic acid solution in step S1 is 10~20 mg / mL.
[0014] Further, the buffer solution in step S1 is a phosphate buffer solution.
[0015] Further, the buffer solution in step S1 is a 0.1× phosphate buffer solution with a pH of 7-8.
[0016] Further, the buffer solution in step S1 is a 0.1× phosphate buffer solution with pH = 7.4.
[0017] Furthermore, the concentration of the S-allyl-L-cysteine solution in step S2 is 2~8 mg / mL.
[0018] Furthermore, in step S1, the ultrasonic power is 200~300W and the ultrasonic time is 1~10 minutes.
[0019] Furthermore, in step S2, the ultrasonic power is 100~300W and the ultrasonic time is 1~10 minutes.
[0020] Furthermore, in step S3, the temperature of the water bath environment is 60~80℃; the continuous stirring time is 10~60 minutes; and the stirring time continues for 10~60 minutes.
[0021] Furthermore, in step S3, the temperature of the water bath environment is 70°C; the continuous stirring time is 30 minutes; and the stirring time continues for another 30 minutes.
[0022] Furthermore, the stirring speed in step S3 is 800~1500 rpm.
[0023] Further, step S3 involves transferring it to a 4°C refrigerator.
[0024] Compared with existing technologies that employ covalent cross-linking or simple physical doping, this invention achieves molecular self-assembly of glycyrrhizic acid and SAC by adjusting the charge balance and hydrogen bond network between components. The preparation process of this invention, without damaging the active structure of SAC, not only improves the drug encapsulation efficiency but also regulates and dynamically remodels the microenvironment of diabetic wounds.
[0025] Glycyrrhizic acid (GA), a natural pentacyclic triterpenoid compound, possesses good biocompatibility and inherent anti-inflammatory effects. S-allyl-L-cysteine (SAC) can serve as a substrate and, through the catalysis of cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE), generate endogenous H2S, making it not only an ideal biological endogenous H2S donor but also exhibiting excellent antioxidant and cytoprotective effects. However, there are currently no reports of constructing a hydrogel from these two compounds via self-assembly and applying it to the treatment of diabetic wounds.
[0026] Diabetic skin wounds require both moisture retention and adequate breathability and an anti-infection barrier. Single-glycoside (GA) gels are often brittle and lack sufficient mechanical strength. This invention, through extensive inventive experiments, constructs a novel pharmaceutically active hydrogel by utilizing the intermolecular interaction between glycyrrhizic acid (GA) and S-allyl-L-cysteine (S-Cysteine). This hydrogel not only utilizes the three-dimensional network structure of GA to achieve physical encapsulation and chemical stability of S-Cysteine, but also, through the synergistic effect of both, creates an intelligent bioactive platform that responds to the needs of the wound. The hydrogel of this invention exhibits tissue-like viscoelastic characteristics, adapting to wound movement and providing continuous protection for chronic wounds under dynamic physiological conditions, demonstrating flexibility to adapt to the dynamic needs of different healing stages. The introduction of S-Cysteine during assembly not only introduces the drug but also enhances the cross-linking density between GA fiber bundles due to the amino groups in the S-Cysteine's chemical structure. This carrier-free hydrogel significantly improves the hydrogel's viscoelasticity, enabling it to closely adhere to complex wounds, forming a stable physical barrier while maintaining an ideal moist healing environment.
[0027] Another object of the present invention is to provide the use of the hydrogel dressing for promoting diabetic wound healing, or the hydrogel dressing prepared by the method thereof, in the preparation of a medicament for promoting diabetic wound healing.
[0028] Compared with the prior art, the present invention has the following advantages: (1) The hydrogel prepared by the present invention has a three-dimensional porous structure that allows it to closely fit the contour of the wound, effectively manage the accumulation of exudate, and at the same time help maintain a suitable moist environment for the healing process.
[0029] (2) In this invention, S-allyl-L-cysteine can be catalyzed by cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) to generate endogenous H2S in vivo, which has functions such as cell protection and immune regulation. It can promote the polarization of M1 macrophages to M2 type, effectively reduce the inflammatory response of wounds, and thus accelerate the repair and healing of diabetic skin wounds.
[0030] (3) The endogenous hydrogen sulfide generated by H2S signal remodeling can dynamically neutralize excess reactive oxygen species (ROS) and reduce oxidative stress levels. Simultaneously, H2S can activate the vascular endothelial growth factor pathway, remodeling the angiogenesis environment damaged by high glucose levels and providing blood supply for tissue repair. The hydrogel dressing of this invention, due to its ability to dynamically remodel the pathological microenvironment, has become a promising new alternative in the field of diabetic wound treatment. Attached Figure Description
[0031] Figure 1 Photographs of the appearance of the hydrogels obtained in Examples 1, 2, 3 and Comparative Example 1, taken with a digital camera.
[0032] Figure 2 The images are scanning electron microscope images of the hydrogels obtained in Examples 1, 2, 3, and Comparative Example 1.
[0033] Figure 3 To draw an image of the letters "SAC" using a syringe on the hydrogel prepared in Example 1 of this invention.
[0034] Figure 4 Fourier transform infrared spectra of GA, SAC, and the hydrogel GA-SAC prepared in Example 1 of this invention.
[0035] Figure 5 The amplitude scanning results are for the hydrogel prepared in Example 1 of this invention.
[0036] Figure 6 The results show the shear thinning behavior of the hydrogel prepared in Example 1 of this invention.
[0037] Figure 7 The hemolytic safety evaluation results for Triton X-100, PBS, GA, and the hydrogel GA-SAC prepared in Example 1 of this invention.
[0038] Figure 8 The wound healing status of mice in different groups at different time points.
[0039] Figure 9 A bar chart showing the relative wound area of mice in different groups at different time points.
[0040] Figure 10 HE staining images of skin wound tissue from different groups of mice.
[0041] Figure 11 The results show the hematological parameters of mice in different groups.
[0042] Figure 12 Results of key biochemical indicators of liver and kidney function in mice from different groups.
[0043] Figure 13 HE staining images of major organ tissues (heart, liver, spleen, lung, and kidney) from different groups of mice.
[0044] Figure 14 This image shows the changes in wound area in each group during the wound healing process of Bama pigs, as well as representative wound images at different stages.
[0045] Figure 15 The wound closure rate of each group during the wound healing process of Bama pigs.
[0046] Figure 16 HE staining image of wound tissue from Bama pigs on day 18. Detailed Implementation
[0047] The present invention will be further described below through specific embodiments, but this is not a limitation of the present invention. Those skilled in the art can make various modifications or improvements based on the basic idea of the present invention, but as long as they do not depart from the basic idea of the present invention, they are all within the scope of the present invention.
[0048] Example 1: A method for preparing a hydrogel dressing (GA-SAC) to promote the healing of diabetic wounds. The method for preparing the hydrogel dressing that promotes the healing of diabetic wounds includes the following steps: S1 Weigh 10 mg of glycyrrhizic acid, add 1 mL of 0.1× phosphate buffer solution with pH = 7.4, and vortex sonicate at 70℃ until completely dissolved. The sonication power is 250 W and the sonication time is 5 minutes to obtain a clear and transparent glycyrrhizic acid solution. S2 Weigh 2 mg of S-allyl-L-cysteine, add 1 mL of 0.1× phosphate buffer solution with pH=7.4, and vortex sonicate until completely dissolved. The sonication power is 250 W and the sonication time is 5 minutes to obtain a clear and transparent S-allyl-L-cysteine solution. S3. Slowly add the glycyrrhizic acid solution obtained in step S1 into a vial and stir continuously in a 70°C water bath for 30 minutes at a stirring speed of 1500 rpm. Add the S-allyl-L-cysteine solution obtained in step S2. After the two are thoroughly mixed, continue stirring under magnetic stirring for 30 minutes until the pH of the system stabilizes at 7.4. Transfer it to a 4°C refrigerator. After 30 minutes, it is observed that glycyrrhizic acid and S-allyl-L-cysteine gradually form a gel and do not flow when inverted.
[0049] Example 2: A method for preparing a hydrogel dressing to promote the healing of diabetic wounds. The preparation method of the hydrogel dressing for promoting diabetic wound healing is similar to that of Example 1. The difference from Example 1 is that the mass ratio of glycyrrhizic acid to S-allyl-L-cysteine is 10:4, that is, 10 mg of glycyrrhizic acid and 4 mg of S-allyl-L-cysteine.
[0050] Example 3: A method for preparing a hydrogel dressing to promote the healing of diabetic wounds. The preparation method of the hydrogel dressing for promoting diabetic wound healing is similar to that of Example 1. The difference from Example 1 is that the mass ratio of glycyrrhizic acid to S-allyl-L-cysteine is 10:8, that is, 10 mg of glycyrrhizic acid and 8 mg of S-allyl-L-cysteine.
[0051] Comparative Example 1: A method for preparing a hydrogel dressing The method for preparing the hydrogel dressing includes the following steps: S1 Weigh 10 mg of glycyrrhizic acid, add 1 mL of 0.1× phosphate buffer solution with pH = 7.4, and vortex sonicate until completely dissolved. The sonication power is 250 W and the sonication time is 5 minutes to obtain a clear and transparent glycyrrhizic acid solution. S2. Slowly add the glycyrrhizic acid solution obtained in step S1 into a vial, stir continuously in a 70°C water bath for 30 minutes, transfer it to a 4°C refrigerator, and gradually form a gel that does not flow when inverted.
[0052] Experimental Example 1: Hydrogel Characterization The hydrogels obtained in Examples 1, 2, 3, and Comparative Example 1 were photographed using a digital camera. The appearance photographs of the hydrogels obtained in Examples 1, 2, 3, and Comparative Example 1 taken with a digital camera are shown below. Figure 1 As shown. By Figure 1 As can be seen, the hydrogel is uniform, stable, and transparent.
[0053] The morphology of the hydrogels obtained in Examples 1, 2, 3, and Comparative Example 1 was characterized using scanning electron microscopy. Scanning electron micrographs of the hydrogels obtained in Examples 1, 2, 3, and Comparative Example 1 are shown below. Figure 2 As shown. By Figure 2 As can be seen, the hydrogel obtained by crosslinking glycyrrhizic acid and S-allyl-L-cysteine at a mass ratio of 10:2 in Example 1 exhibits a honeycomb-like three-dimensional network structure. In Example 2, the gel has a sheet-like internal structure without a porous three-dimensional network. In Example 3, the gel has a fragmented internal structure with a discontinuous network. In Comparative Example 1, the gel may collapse internally.
[0054] The hydrogel prepared in Example 1 of this invention was used to draw the letter "SAC" using a syringe. An image of the "SAC" lettering drawn using the hydrogel prepared in Example 1 of this invention using a syringe is shown below. Figure 3 As shown. By Figure 3 As can be seen, the hydrogel prepared in Example 1 of this invention can be used to draw the letter "SAC" with a syringe without significant collapse or disintegration, indicating that it can quickly maintain its shape in the in vitro / in vivo environment. After contacting the wound, it can conform to the contour of the wound and is not easily lost, ensuring the effective retention and action time of the dressing on the wound. This directly proves that the hydrogel has good injectability, plasticity and rheological properties, which is suitable for the actual application needs of diabetic wound dressings, and provides a structural basis for its subsequent anti-inflammatory, healing and other biological functions on the wound.
[0055] Fourier transform infrared (FTIR) spectra of GA (glycyrrhizic acid raw material powder), SAC, and the hydrogel GA-SAC prepared in Example 1 of this invention were analyzed using a Fourier transform infrared spectroscopy (FTIR) detector. The FTIR spectra of GA, SAC, and the hydrogel GA-SAC prepared in Example 1 of this invention are shown below. Figure 4 As shown. By Figure 4 It can be seen that GA-SAC is at 3430.75cm - The broad peak shift at ¹ confirms the formation of a dense intermolecular hydrogen bond network between the components. GA at 1708.89 cm⁻¹ - The characteristic peak of the carboxyl group at ¹ was significantly weakened after gelation, while that of SAC at 1487.73 cm⁻¹ was significantly weakened. - ¹ -NH 3+ Displacement occurred. This was confirmed by comparing the SAC value to 1487.73 cm. - ¹With GA-SAC 1494.5cm - The characteristic peak shift of ¹ confirms the -NH of SAC 3+ There is a strong electrostatic attraction between it and the COO- of GA.
[0056] The hydrogel prepared in Example 1 of this invention was subjected to amplitude scanning and shear thinning behavior tests. GA-SAC hydrogels with GA fixed at 10.0 mg / mL and SAC varying between 2 mg / mL were tested using a rotational rheometer. The modulus test included amplitude scanning at a fixed frequency of 1 Hz and a specified strain (stress) range of 0.1%-100%. The logarithmic relationship of the shear rate was determined in the kinetic viscosity test. The measured shear rate range was 0.1~100 s⁻¹. -1 The amplitude scanning results of the hydrogel prepared in Example 1 of this invention are as follows: Figure 5 As shown. The shear-thinning behavior of the hydrogel prepared in Example 1 of this invention is as follows. Figure 6 As shown. Figure 5The amplitude scan (strain scan) results showed that when the strain exceeded 14%, the elastic modulus (G') dropped below the loss modulus (G''), indicating that the GA-SAC hydrogel transitioned from a gel state (G'>G'') to a sol state (G''>G'). Figure 6 It can be seen that the shear viscosity of GA-SAC hydrogel decreases with increasing shear rate, indicating that it has good shear thinning behavior and good injectability.
[0057] Experimental Example 2: Evaluation of the hemolytic safety of hydrogels The hemolytic safety of the hydrogels prepared in Example 1 and Comparative Example 1 of this invention was evaluated. They were continuously extracted in PBS buffer at pH 7.4 for 48 hours. The extract was then co-incubated with fresh red blood cell suspension. After centrifugation, the absorbance of the supernatant was measured, and the hemolysis rate was calculated. Triton X-100 was used as a positive control, and PBS buffer at pH 7.4 was used as a negative control. The hemolytic safety evaluation results for Triton X-100, PBS, GA, and the hydrogel GA-SAC prepared in Example 1 of this invention are as follows: Figure 7 As shown. By Figure 7 It can be seen that the hemolysis rate of the positive group, which consists of lysed red blood cells from Triton X-100, is 100%. The hemolysis rates of the samples treated with PBS and the experimental groups (GA and GA-SAC groups) are all below 5%. Theoretically, a hemolysis rate below 5% is considered normal, indicating that the drug is non-hemolytic and has good blood compatibility.
[0058] Experimental Example 3: The effect of the hydrogel prepared in this invention on the healing of mouse skin wounds. 1. Test materials 1.1 Experimental animals: 15 male Balb / c mice (20-25g, 6-7 weeks old) were selected.
[0059] 1.2 Experimental drugs: hydrogels prepared according to the raw materials and methods of Example 1 of the present invention, and hydrogels prepared according to Comparative Example 1.
[0060] 2. Test methods 2.1 Grouping and Administration: (1) Model control group (Model); (2) GA group (10 mg / mL); (3) GA-SAC group (10mg / mL / 2mg / mL).
[0061] Five mice in each group.
[0062] 2.2 Test Procedure (1) Establishment of full-thickness skin defect model: All mice were anesthetized by intraperitoneal injection of 4% chloral hydrate, the back hair was shaved, the back skin of the mice was lifted with tweezers, and a circular wound with a diameter of about 8 mm was cut on the back, and then wiped with sterile physiological saline and iodine.
[0063] (2) On the day of modeling, the experimental group was given 50 µL of hydrogel to evenly cover the wound to prevent overflow. The control group mice were given 50 µL of PBS buffer evenly on the wound site.
[0064] (3) During the treatment process, the wound was photographed on days 0, 3, 7, 10 and 14 after surgery to observe the wound healing of the mice.
[0065] 2.3 Test Results Wound healing status of mice in different groups at different time points, as follows Figure 8 As shown in the figure. The bar chart shows the relative wound area of skin wound healing in mice of different groups at different time points. Figure 9 As shown. By Figure 8 and Figure 9 The experimental results show that the wound healing speed of the GA group and the GA-SAC group is significantly better than that of the model control group. At the same time, the wound area of the GA-SAC group shrinks more. The wound of the GA-SAC group is almost completely healed on the 14th day after surgery. The healing speed and integrity are better than those of the Model group and the single GA group. This shows that the hydrogel dressing provided by the present invention has excellent wound healing ability, and the hydrogel of the present invention has good application prospects in clinical practice as a new type of wound dressing.
[0066] Mouse skin wound tissue was fixed, embedded, sectioned, and stained with hematoxylin and eosin (HE). The tissue was then photographed and observed under a microscope. HE staining images of mouse skin wound tissue from different groups are shown below. Figure 10 As shown. By Figure 10 It can be seen that inflammatory cells still accumulated at the wound site in the Model group, while the number of inflammatory cells in GA was reduced. However, the GA-SAC epidermal structure was the most intact, and the number of inflammatory cells was significantly reduced, with the appearance of regenerated epithelium, epithelial layer, and dermal tissue. The results confirm that the hydrogel dressing prepared in this invention not only has excellent anti-inflammatory effects but also significantly accelerates the structural remodeling of damaged tissue. Therefore, the hydrogel prepared in this invention can be used as a highly effective healing-promoting dressing for difficult-to-heal wounds such as those in diabetic dermatitis.
[0067] Blood was collected from the eyeballs of mice before sacrifice. Serum was separated by centrifugation, and liver function indicators (such as ALT and AST), kidney function indicators (such as urea and Crea), and routine blood counts were measured. The results of different hematological indicators in mouse blood and serum were analyzed. The results of various hematological indicators in different groups of mice are shown below. Figure 11 As shown. By Figure 11As can be seen, the hematological parameters (red blood cell count, hemoglobin, hematocrit / hematocrit, mean corpuscular volume, mean corpuscular hemoglobin content, mean corpuscular hemoglobin concentration, mean corpuscular platelet count, and mean platelet volume, MPV) of the GA-SAC group mice remained at normal levels, with no significant difference from the model control group. The results of key biochemical indicators of liver and kidney function in different groups of mice are as follows: Figure 12 As shown. By Figure 12 As can be seen, the key biochemical indicators of liver and kidney function (alanine aminotransferase / alanine transaminase (ALT), aspartate aminotransferase / aspartate transaminase (AST), urea / blood urea nitrogen (Urea), and creatinine (Crea) in the GA-SAC group mice remained at normal levels, with no significant differences compared to the model control group. Therefore, it can be concluded that there were no statistically significant differences in liver and kidney function and blood routine indicators between the mice and the model control group, and no obvious liver and kidney toxicity or hematological toxicity was observed.
[0068] In summary, the hydrogel prepared by this invention not only promotes healing but also has no toxic side effects on the internal organs of mice, meeting the safety standards for biomedical materials.
[0069] Fourteen days after treatment, the mice were euthanized, and their major organs (heart, liver, spleen, lung, and kidney) were collected for hematoxylin and eosin (H&E) staining analysis. HE staining images of major organs (heart, liver, spleen, lung, and kidney) from different groups of mice are shown below. Figure 13 As shown. By Figure 13 As can be seen, after 14 days of treatment with the GA / SAC hydrogel prepared in this invention, the major organs of the mice in the GA / SAC group showed no histopathological changes compared with the model control group. These data indicate that the GA-SAC hydrogel prepared in this invention has good in vivo biocompatibility.
[0070] To further verify the efficacy and translational potential of GA / SAC hydrogel in wound healing, this invention uses the Bama pig model, whose skin structure and wound healing process are very similar to those of humans.
[0071] Experimental Example 4: The effect of the hydrogel prepared in this invention on wound healing in Bama pigs 1. Test materials 1.1 Experimental animal: A small Bama pig, with a total of 9 wounds created.
[0072] 1.2 Experimental drug: Hydrogel prepared according to the raw materials and method of Example 1 of this invention.
[0073] 2. Test methods 2.1 Grouping and Administration: (1) Negative control group (Control); (2) GA-SAC group (10mg / mL / 2mg / mL); (3) Clinical control group: alginate dressing group.
[0074] 2.2 Test Procedure (1) Establishment of full-thickness skin defect model: Pigs were rapidly sedated with ketamine (10~15mg / kg) and diazepam, and continuously anesthetized with inhaled isoflurane. A series of square full-thickness wounds (2cm*2cm) were formed on both sides of the back of the pigs. The wound depth was about 1cm and the distance between each wound was 3cm. The wounds were disinfected. The medication was administered on the day of modeling, twice, with a volume of about 1mL. After the wounds were administered, they were covered with PU film and then sutured.
[0075] (2) Nine wounds were created on the backs of pigs and randomly divided into three groups, with three wounds in each group. Drug administration: On the day the full-thickness skin injury model was established, 1 mL of the hydrogel dressing prepared in Example 1 was uniformly applied to the wounds of the GA-SAC group, 1 mL of alginate dressing was uniformly applied to the wounds of the clinical control group, and 1 mL of PBS buffer was uniformly applied to the wounds of the negative control group. The medication was administered twice. After administration, the dressings were covered and fixed with a PU film. Wounds were photographed on postoperative days 0, 3, 6, 12, and 18 to observe wound healing.
[0076] 2.3 Test Results The changes in wound area in different groups during the wound healing process of Bama pigs and representative wound images at different stages are shown below. Figure 14 As shown in the figure. The wound closure rate of each group during the wound healing process in Bama pigs is as follows. Figure 15 As shown. By Figure 14 , Figure 15 As can be seen, compared with the clinical control group, the GA-SAC group treated with the hydrogel dressing prepared by the method of the present invention had a faster wound shrinkage rate and a smoother wound repair process, showing a good wound repair effect.
[0077] HE staining image of wound tissue from Bama pigs on day 18. Figure 16 As shown. By Figure 16 It can be seen that the hydrogel treatment group prepared by the method of the present invention completed the regeneration of dermal tissue, and a large number of collagen fibers began to form in the newly formed wound tissue.
Claims
1. A hydrogel dressing for promoting the healing of diabetic wounds, characterized in that, The hydrogel dressing is prepared by self-assembly of glycyrrhizic acid and S-allyl-L-cysteine using glycyrrhizic acid and S-allyl-L-cysteine as raw materials and buffer solution as dispersion medium, by controlling the mass ratio of raw materials, reaction temperature and reaction time and pH.
2. The hydrogel dressing for promoting diabetic wound healing according to claim 1, characterized in that, The buffer solution is a phosphate buffer solution.
3. The hydrogel dressing for promoting diabetic wound healing according to claim 2, characterized in that, The buffer solution is a 0.1× phosphate buffer solution with a pH of 7-8.
4. The hydrogel dressing for promoting diabetic wound healing according to claim 1, characterized in that, The mass ratio of glycyrrhizic acid to S-allyl-L-cysteine is 10:2~8.
5. The method for preparing the hydrogel dressing for promoting diabetic wound healing according to any one of claims 1-4, characterized in that, Includes the following steps: S1 Take glycyrrhizic acid, add buffer solution, and vortex sonicate until completely dissolved to obtain a clear and transparent glycyrrhizic acid solution; S2 Take S-allyl-L-cysteine, add buffer solution, vortex and sonicate until completely dissolved to obtain a clear and transparent S-allyl-L-cysteine solution; S3. Place the glycyrrhizic acid solution obtained in step S1 in a water bath and stir continuously. Add the S-allyl-L-cysteine solution obtained in step S2 and continue stirring until the pH of the system stabilizes at 7-8. Transfer the solution to a refrigerator to form a hydrogel dressing.
6. The method for preparing the hydrogel dressing for promoting diabetic wound healing according to claim 5, characterized in that, The concentration of glycyrrhizic acid solution in step S1 is 10~20 mg / mL.
7. The method for preparing the hydrogel dressing for promoting diabetic wound healing according to claim 5, characterized in that, In step S2, the concentration of the S-allyl-L-cysteine solution is 2~8 mg / mL.
8. The method for preparing the hydrogel dressing for promoting diabetic wound healing according to claim 5, characterized in that, In step S1, the ultrasonic power is 200-300W and the ultrasonic time is 1-10 minutes; in step S2, the ultrasonic power is 100-300W and the ultrasonic time is 1-10 minutes.
9. The method for preparing the hydrogel dressing for promoting diabetic wound healing according to claim 5, characterized in that, The temperature of the water bath environment in step S3 is 60~80℃; the continuous stirring time is 10~60 minutes; and the stirring time continues for 10~60 minutes.
10. The use of the hydrogel dressing for promoting diabetic wound healing according to any one of claims 1-4 or the hydrogel dressing for promoting diabetic wound healing prepared by any one of claims 5-9 in the preparation of a medicament for promoting diabetic wound healing.