A traditional Chinese medicine nano composite gel dressing for surgical wound repair and a preparation method thereof

Abstract

The present application relates to the technical field of medical dressings, and particularly relates to a traditional Chinese medicine nano composite gel dressing for surgical wound repair and a preparation method thereof. The preparation method of the gel dressing comprises the following steps: adding soybean lecithin and cholesterol into a mixed solvent, and then adding resveratrol and isoferulactone to obtain a liposome membrane; resuspending the liposome membrane in a PBS solution, and oscillating to hydrate, so as to obtain resveratrol-isoferulactone nano-liposomes; adding methacrylated gelatin and a photoinitiator into deionized water to obtain a gel matrix; uniformly mixing the resveratrol-isoferulactone nano-liposomes, Cr-ACP1 polypeptides and the gel matrix, degassing, and then injecting into a mold, and then performing ultraviolet light curing, so as to obtain the gel dressing. The method is simple in operation, mild in conditions and suitable for industrial production. The obtained gel dressing has excellent healing promotion effect and anti-scar effect.

Description

Technical Field

[0001] This invention relates to the field of medical dressing technology, and in particular to a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair and its preparation method. Background Technology

[0002] As the body's largest protective barrier, the skin plays an irreplaceable role in maintaining homeostasis. However, it is highly susceptible to damage from factors such as trauma, surgical incisions, burns, and microbial infections. Surgical wound repair is a common clinical problem, especially for chronic, difficult-to-heal wounds such as postoperative wounds and diabetic foot ulcers. The healing process is often hindered by persistent inflammation, oxidative stress, insufficient local blood supply, and a lack of growth factors, severely impacting patients' quality of life and imposing a heavy medical burden. Wound healing is a complex, multi-step biological process involving hemostasis, inflammation, proliferation, and tissue remodeling. During this process, excessive accumulation of reactive oxygen species (ROS) can lead to persistent oxidative stress, thereby inhibiting fibroblast migration and proliferation, hindering granulation tissue formation and re-epithelialization, and ultimately causing delayed healing or even healing failure. Therefore, effectively clearing excessive ROS from the wound site and suppressing the inflammatory response is one of the key strategies for promoting functional recovery of surgical wounds, especially chronic, difficult-to-heal wounds.

[0003] Hydrogel dressings, with their three-dimensional network structure similar to the extracellular matrix, exhibit unique advantages in wound repair. Their three-dimensional cross-linked network not only isolates the wound and increases breathability but also actively maintains a moist wound microenvironment, providing suitable physiological conditions for cell proliferation, migration, and tissue regeneration. However, traditional hydrogel dressings have relatively limited functions, primarily providing physical barrier protection and lacking the ability to actively regulate pathological factors such as oxidative stress and excessive inflammation in the wound microenvironment, making it difficult to meet the dynamic needs of different stages in the healing process of complex wounds. In recent years, researchers have attempted to endow dressings with multiple bioactivities by loading functional ingredients with antioxidant, anti-inflammatory, or antibacterial activities into hydrogels. However, existing systems still suffer from low loading efficiency of active ingredients, uncontrollable release behavior, and insufficient bioavailability. Therefore, developing a multifunctional hydrogel dressing that combines efficient ROS scavenging ability, good anti-inflammatory activity, and healing-promoting functions has significant clinical significance and application value. Summary of the Invention

[0004] In order to overcome the shortcomings of the prior art, the primary objective of this invention is to provide a method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair. This preparation method is simple to operate, has mild conditions, and is suitable for industrial production.

[0005] Another objective of this invention is to provide a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair, which has excellent healing-promoting and anti-scarring effects.

[0006] The objective of this invention is achieved through the following technical solution: A method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair includes the following steps: (1) Soybean lecithin and cholesterol were added to a mixed solvent, and then resveratrol and isocauterin were added. After mixing evenly, the solvent was removed to obtain a liposome membrane. The liposome membrane was resuspended in PBS solution, hydrated by shaking, dispersed by ultrasound, centrifuged, and the supernatant was collected to obtain resveratrol-isocauterin nanoliposomes. (2) Methacrylamide gelatin and photoinitiator were added to deionized water to prepare a gel matrix; (3) Mix the resveratrol-isobrinol-lactone nanoliposomes, Cr-ACP1 polypeptide and the gel matrix evenly, degas and inject into the mold, and cure by ultraviolet light to obtain the product.

[0007] Preferably, the mass ratio of isobrominolactone, soybean lecithin, cholesterol and resveratrol in step (1) is 1:(15-20):(3-5):(1-2).

[0008] Preferably, the mass concentration of the Cr-ACP1 polypeptide in step (3) in the gel matrix is ​​0.1-0.3%; and the mass concentration of the resveratrol-isobrinol nanoliposomes in the gel matrix is ​​2-4%.

[0009] Preferably, the temperature of the oscillation hydration in step (1) is 37°C and the time is 20-30 min.

[0010] Preferably, the liposome membrane in step (1) has a mass concentration of 0.1-0.3 g / mL in PBS solution; the mixed solvent is composed of anhydrous ethanol and chloroform in a volume ratio of 1:(2-3).

[0011] Preferably, the concentration of the PBS solution is 0.01 mol / L and the pH is 7.2-7.4.

[0012] Preferably, the mass concentration of the methacrylamide gelatin in step (2) in deionized water is 5-10%; and the mass concentration of the photoinitiator in deionized water is 0.1-0.2%.

[0013] Preferably, the photoinitiator is lithium phenyl (2,4,6-trimethylbenzoyl) phosphate.

[0014] Preferably, the UV curing conditions in step (3) are: radiation intensity 5-10 mW / cm². 2 The wavelength is 365nm and the radiation time is 40-60s.

[0015] A traditional Chinese medicine nanocomposite gel dressing for surgical wound repair was prepared using the above-mentioned preparation method.

[0016] The present invention has the following advantages over the prior art: 1. This invention provides a method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair. The preparation method is simple to operate, the conditions are mild, and the prepared gel dressing has excellent healing-promoting and anti-scarring effects.

[0017] 2. This invention enhances the healing and anti-scarring effects of gel dressings by adding isobryolide. Both resveratrol and isobryolide are active ingredients in traditional Chinese medicine. This invention improves the bioavailability of resveratrol and isobryolide through liposome encapsulation modification. The combination of these two ingredients exerts antioxidant and anti-inflammatory effects: on the one hand, it effectively removes excess ROS free radicals generated by oxidative stress in the wound, reduces oxidative damage to the wound, and protects fibroblast activity; on the other hand, it promotes the expression of vascular endothelial growth factor, induces angiogenesis, improves local blood supply to the wound, and reduces postoperative inflammatory edema of surgical wounds, thereby comprehensively promoting postoperative wound repair.

[0018] 3. This invention also enhances the healing and anti-scarring effects of the gel dressing by adding Cr-ACP1 peptide. Cr-ACP1 peptide can promote orderly collagen deposition in the wound, accelerate epithelial tissue closure, reduce disordered collagen accumulation, effectively shorten wound healing time, and inhibit excessive scar proliferation.

[0019] 4. This invention uses methacrylamide gelatin as a gel matrix, which can rapidly cross-link and solidify under ultraviolet light irradiation in the presence of a photoinitiator, forming a hydrogel with a three-dimensional network structure. Resveratrol-isolinolactone nanoliposomes and Cr-ACP1 peptides are uniformly dispersed in the gel matrix and then photocured, allowing the liposomes and peptides to be in situ embedded in the three-dimensional network of the gel. This three-dimensional network structure can, on the one hand, further delay the release rate of the nanoliposomes, significantly prolong the action time of the active ingredients on the wound surface, reduce the frequency of dressing changes, and avoid secondary traction damage to the wound surface caused by frequent dressing changes; on the other hand, the gel dressing can closely adhere to the wound tissue, forming a physical barrier, isolating external contaminants, further reducing the risk of infection, and providing an ideal moist microenvironment for wound healing. Attached Figure Description

[0020] Figure 1 This is a graph showing the in vitro cytotoxicity results of the blank control group; Figure 2This is a graph showing the in vitro cytotoxicity results of the gel dressing obtained in Example 1; Figure 3 This is a graph showing the in vitro cytotoxicity results of the positive control group. Detailed Implementation

[0021] The present invention will be further described in detail below with reference to embodiments, but the implementation of the present invention is not limited thereto. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field. Test methods in the following embodiments that do not specify specific experimental conditions are generally performed according to conventional experimental conditions or experimental conditions recommended by the manufacturer. Unless otherwise specified, the reagents and raw materials used in the present invention are commercially available.

[0022] The amino acid sequence of the Cr-ACP1 polypeptide in this invention is: AWKLFDDGV; the concentration of the PBS solution is 0.01 mol / L and the pH is 7.2.

[0023] Example 1 A method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair includes the following steps: (1) Soybean lecithin, cholesterol and resveratrol were added to a mixed solvent (composed of anhydrous ethanol and chloroform in a volume ratio of 1:2.5) according to the mass ratio of isocauterin lactone, soybean lecithin, cholesterol and resveratrol 1:18:4:1.5. Then, resveratrol and isocauterin lactone were added and mixed evenly. The solvent was removed by rotary evaporation to obtain liposome membranes. The above liposome membranes were resuspended in PBS solution to make the mass concentration of liposome membranes in PBS solution 0.2 g / mL. The membranes were hydrated by shaking at 37℃ for 25 min. After ultrasonic dispersion and centrifugation, the supernatant was collected to obtain resveratrol-isocauterin lactone nanoliposomes. (2) The mass concentration of methacrylamide gelatin in deionized water is 8%, and the mass concentration of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate in deionized water is 0.15%. The two are added to deionized water and stirred to mix evenly to obtain a gel matrix. (3) With a resveratrol-isobrinol-lactone nanoliposomes concentration of 3% and a Cr-ACP1 peptide concentration of 0.2% in the gel matrix, the two were mixed evenly with the above gel matrix, degassed, and injected into a mold. The mixture was then heated under a nitrogen atmosphere at 365 nm and 7 mW / cm². 2 Under illumination, radiation for 50 seconds yields the result.

[0024] A traditional Chinese medicine nanocomposite gel dressing for surgical wound repair was prepared using the above-mentioned preparation method.

[0025] Example 2 A method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair includes the following steps: (1) Soybean lecithin, cholesterol and resveratrol were added to a mixed solvent (composed of anhydrous ethanol and chloroform in a volume ratio of 1:2) according to the mass ratio of isocauterin lactone, soybean lecithin, cholesterol and resveratrol 1:15:3:1. Then, resveratrol and isocauterin lactone were added and mixed evenly. The solvent was removed by rotary evaporation to obtain liposome membranes. The above liposome membranes were resuspended in PBS solution to make the mass concentration of liposome membranes in PBS solution 0.1 g / mL. The membranes were hydrated by shaking at 37℃ for 20 min. After ultrasonic dispersion and centrifugation, the supernatant was collected to obtain resveratrol-isocauterin lactone nanoliposomes. (2) With the mass concentration of methacrylamide gelatin in deionized water being 5% and the mass concentration of lithium phenyl (2,4,6-trimethylbenzoyl) phosphate in deionized water being 0.1%, the two were added to deionized water and stirred to mix evenly to obtain a gel matrix. (3) With a resveratrol-isobrinol-lactone nanoliposomes concentration of 2% and a Cr-ACP1 polypeptide concentration of 0.1% in the gel matrix, the two were mixed evenly with the above gel matrix, degassed, and injected into a mold. The mixture was then heated under a nitrogen atmosphere at 365 nm and 5 mW / cm². 2 Under illumination, radiation for 60 seconds yields the result.

[0026] A traditional Chinese medicine nanocomposite gel dressing for surgical wound repair was prepared using the above-mentioned preparation method.

[0027] Example 3 A method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair includes the following steps: (1) Soybean lecithin, cholesterol and resveratrol were added to a mixed solvent (composed of anhydrous ethanol and chloroform in a volume ratio of 1:3) according to the mass ratio of isocauterin lactone, soybean lecithin, cholesterol and resveratrol 1:20:5:2. Then, resveratrol and isocauterin lactone were added and mixed evenly. The solvent was removed by rotary evaporation to obtain liposome membranes. The above liposome membranes were resuspended in PBS solution to make the mass concentration of liposome membranes in PBS solution 0.3 g / mL. The membranes were hydrated by shaking at 37℃ for 30 min. After ultrasonic dispersion and centrifugation, the supernatant was collected to obtain resveratrol-isocauterin lactone nanoliposomes. (2) The gel matrix was prepared by adding the methacrylamide gelatin to deionized water at a mass concentration of 10% and the lithium phenyl (2,4,6-trimethylbenzoyl) phosphate at a mass concentration of 0.2% in deionized water and stirring until homogeneous. (3) The resveratrol-isobrinol-lactone nanoliposomes were mixed with the gel matrix at a mass concentration of 4% and the Cr-ACP1 peptide was mixed with the gel matrix at a mass concentration of 0.3%. After degassing, the mixture was injected into a mold and heated under a nitrogen atmosphere at 365 nm and 10 mW / cm². 2 Under illumination, radiation for 40 seconds yields the result.

[0028] A traditional Chinese medicine nanocomposite gel dressing for surgical wound repair was prepared using the above-mentioned preparation method.

[0029] Comparative Example 1 The difference between this comparative example and Example 1 is that isocauliflorin was not added in step (1).

[0030] Comparative Example 2 The difference between this comparative example and Example 1 is that no Cr-ACP1 polypeptide was added in step (3).

[0031] Experimental Example 1 L929 cells were fed at a rate of 1×10⁻⁶ 6 Cells were seeded at a density of 100 cells / well in 6-well plates and cultured for 24 h. Then, H2O2 at a final concentration of 100 μmol / L was added for 1 h of stimulation. The original culture medium was discarded, and the cells were treated with the soaking solution of the gel dressing from the examples or comparative examples (prepared with serum-free medium at a concentration of 10 mg / mL). A blank control group (treated with an equal volume of fresh culture medium only, without H2O2 stimulation) and a model control group (treated with H2O2 followed by an equal volume of fresh culture medium) were also set up. After 2 h of treatment, cells were collected and resuspended in serum-free medium containing 10 μmol / L DCFH-DA fluorescent probe, and incubated at 37°C in the dark for 20 min. After incubation, the cells were washed three times with PBS to thoroughly remove any probe that had not entered the cells. Finally, the intracellular reactive oxygen species (ROS) levels were detected by flow cytometry, and the results are shown in Table 1.

[0032] Table 1 As shown in Table 1, compared with the blank control group, the ROS level of the model control group increased sharply after being stimulated by H2O2, indicating that the oxidative damage model was successfully constructed.

[0033] Compared with the model control group, the ROS levels in each embodiment group were significantly reduced, indicating that the gel dressing of the present invention has excellent ROS scavenging ability and can effectively protect cells from oxidative stress damage.

[0034] Compared to Example 1, Comparative Example 1 lacked isolinacin, resulting in a significantly increased ROS level. These experimental results indicate that the combination of resveratrol and isolinacin can effectively eliminate excessive ROS generated by oxidative stress in wounds, reduce cellular oxidative damage, and create a favorable immune microenvironment for subsequent wound healing.

[0035] Experimental Example 2 Fifty SPF-grade male mice, weighing 20-22g, were randomly divided into five groups (Example 1, Example 2, Example 3, Comparative Example 1, and Comparative Example 2, n=10 per group) after one week of acclimatization. After anesthesia, a full-thickness skin defect model with a diameter of 10mm was created on the midline of the back of each mouse. Following modeling, the corresponding gel dressing was applied to the wound, and the dressing was changed every two days. The residual wound area was observed and recorded daily. After complete wound healing, the width of the widest point of the scar tissue in each group was measured. The results are shown in Table 2.

[0036] Table 2 As shown in Table 2, the wound healing speed of Examples 1-3 at each time point was significantly better than that of Comparative Example 1 and Comparative Example 2, and the scar width was controlled within 0.91-0.94 mm, indicating that the gel dressing obtained by the present invention has excellent healing-promoting and anti-scarring effects.

[0037] Compared to Example 1, Comparative Example 1 showed a significant decrease in the healing-promoting and anti-scarring effects. These results indicate that the combined use of resveratrol and isolinone can enhance the healing-promoting and anti-scarring effects of the dressing. Specifically, resveratrol and isolinone can effectively remove excess ROS generated by oxidative stress in the wound, reduce oxidative damage, and protect fibroblast activity; simultaneously, they promote vascular endothelial growth factor expression, induce angiogenesis, improve local blood supply to the wound, and reduce postoperative inflammatory edema, thereby comprehensively promoting wound repair.

[0038] Compared to Example 1, Comparative Example 2 showed reduced healing and anti-scarring effects. These results indicate that the addition of Cr-ACP1 peptide can shorten wound healing time and effectively inhibit excessive scar formation. This may be because the Cr-ACP1 peptide can promote orderly collagen deposition in the wound, accelerate epithelial tissue closure, and reduce disordered collagen accumulation.

[0039] Experimental Example 3 Log-phase L929 mouse fibroblasts were harvested, digested with trypsin, and then the cell density was adjusted to 1×10⁻⁶. 5Cells were seeded at a density of 100 μL / mL in 96-well cell culture plates. Each well was incubated at 37°C with 5% CO2 for 24 hours. After cell attachment, the original culture medium was discarded, and 100 μL of the gel dressing (50 μg / mL, as described in Example 1) was added to each well. The plates were then incubated for another 24 hours. Cell morphology was observed under an inverted microscope after the incubation period. A blank control group (DMEM serum-free medium) and a positive control group (DMEM complete medium containing 5% phenol) were also included. Results are shown below. Figure 1-3 .

[0040] Figure 1 This is a graph showing the in vitro cytotoxicity results of the blank control group. Figure 2 This is a graph showing the in vitro cytotoxicity results of the gel dressing obtained in Example 1. Figure 3 This is a graph showing the in vitro cytotoxicity results for the positive control group. (Observation) Figure 1-3 Know: Figure 1 The cells are typically elongated spindle-shaped, with uniform density and normal morphology; Figure 2 Cell morphology and Figure 1 Similar, with no obvious shrinkage; Figure 3 The cells shrank, became rounded, and decreased in density, exhibiting typical toxicity characteristics. The above experimental results demonstrate that the gel dressing obtained in this invention has high safety.

[0041] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.

Claims

1. A method for preparing a traditional Chinese medicine nanocomposite gel dressing for surgical wound repair, characterized in that, Includes the following steps: (1) Soybean lecithin and cholesterol were added to a mixed solvent, and then resveratrol and isocauterin were added. After mixing evenly, the solvent was removed to obtain a liposome membrane. The liposome membrane was resuspended in PBS solution, hydrated by shaking, dispersed by ultrasound, centrifuged, and the supernatant was collected to obtain resveratrol-isocauterin nanoliposomes. (2) Methacrylamide gelatin and photoinitiator were added to deionized water to prepare a gel matrix; (3) Mix the resveratrol-isobrinol-lactone nanoliposomes, Cr-ACP1 polypeptide and the gel matrix evenly, degas and inject into the mold, and cure by ultraviolet light to obtain the product.

2. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 1, characterized in that, The mass ratio of isobrominol, soybean lecithin, cholesterol and resveratrol in step (1) is 1:(15-20):(3-5):(1-2).

3. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 1, characterized in that, The mass concentration of the Cr-ACP1 polypeptide in step (3) in the gel matrix is ​​0.1-0.3%; the mass concentration of the resveratrol-isobrinol nanoliposomes in the gel matrix is ​​2-4%.

4. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 1, characterized in that, The temperature for oscillating hydration in step (1) is 37°C and the time is 20-30 min.

5. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 1, characterized in that, The liposome membrane in step (1) has a mass concentration of 0.1-0.3 g / mL in PBS solution; the mixed solvent consists of anhydrous ethanol and chloroform in a volume ratio of 1:(2-3).

6. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 5, characterized in that, The concentration of the PBS solution was 0.01 mol / L, and the pH was 7.2-7.

4.

7. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 1, characterized in that, The mass concentration of the methacrylamide gelatin in step (2) in deionized water is 5-10%; the mass concentration of the photoinitiator in deionized water is 0.1-0.2%.

8. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 7, characterized in that, The photoinitiator is lithium phenyl (2,4,6-trimethylbenzoyl) phosphate.

9. The method for preparing the traditional Chinese medicine nanocomposite gel dressing for surgical wound repair according to claim 1, characterized in that, The UV curing conditions described in step (3) are: radiation intensity 5-10 mW / cm². 2 The wavelength is 365nm and the radiation time is 40-60s.

10. A traditional Chinese medicine nanocomposite gel dressing for surgical wound repair, characterized in that, It is prepared by the preparation method according to any one of claims 1-9.

Images