Hydrogel dressing for wound healing and preparation method thereof

Abstract

The invention discloses a hydrogel dressing for wound healing and a preparation method thereof, and 100 g of finished product comprises the following components: 0.50-2.00 g of carbomer 940, 0.50-2.00 g of chitosan, 0.50-2.00 g of chitosan, 0.50-2.00 g of sodium alginate, 0.50-2.00 g of The content of xanthoxylin WGX-50 is 0.01 to 0.50 mg; the content of the exosome preparation is 50000 to 500000AU; the content of sodium hydroxide is 0.02 to 0.50 g; the balance is normal saline. The gel dressing has the advantages that stable wet coverage can be formed on the surface of a wound surface, and a gel dressing system which can mildly compound and deliver xanthoxylin WGX-50 and an exosome preparation can be formed, so that the comprehensive performance and preparation feasibility of wound nursing are improved.

Description

Technical Field

[0001] This invention relates to the field of medical dressings and topical preparations, and particularly to a hydrogel dressing for wound healing and its preparation method. Background Technology

[0002] Skin wounds (such as abrasions, lacerations, superficial burns, and some chronic, difficult-to-heal wounds) require a suitable moist microenvironment, effective physical barriers, and local microenvironment regulation that promotes epithelialization and tissue remodeling during the repair process. Traditional dry dressings tend to adhere to the wound and may cause pain during dressing changes and secondary damage; hydrogel dressings can improve moisturization and coverage, but there is still room for improvement in their overall support for the wound repair process.

[0003] Zanthoxylin (WGX-50) is a representative compound of zanthoxylin-related active molecules. Previous studies have suggested its potential application in improving skin hydration and barrier indicators. In keratinocyte models, it has also been shown to have a certain dosage window, that is, it can improve cell viability at lower doses but shows an inhibitory trend at higher doses, suggesting that the dosage / concentration needs to be reasonably controlled to obtain better compatibility and suitability.

[0004] Mesenchymal stem cell-derived exosomes serve as intercellular information carriers, carrying bioactive substances such as proteins and nucleic acids, and have potential applications in tissue repair and inflammation regulation. However, exosomes are sensitive to high temperatures and strong shearing, requiring a mild and controllable preparation process to achieve stable loading and delivery. Summary of the Invention

[0005] The purpose of this invention is to provide a hydrogel dressing for wound healing and its preparation method, thereby forming a stable moist coverage on the wound surface and a hydrogel dressing system that can be gently compounded and delivered with zanthoxylin WGX-50 and exosome preparations, so as to improve the overall performance and preparation feasibility of wound care.

[0006] The above-mentioned technical objective of the present invention is achieved through the following technical solution:

[0007] A hydrogel dressing for wound healing, characterized in that, per 100g of finished product, it comprises the following components: Carbomer 940 is 0.50–2.00 g; The concentration of xanthocyanin WGX-50 is 0.01–0.50 mg. The exosome formulation is 50,000–500,000 AU; Sodium hydroxide is 0.02–0.50 g; The remainder is physiological saline.

[0008] Preferably, the hydrogel dressing has a pH of 5.0 to 7.5.

[0009] Preferably, the exosome preparation is a mesenchymal stem cell-derived exosome.

[0010] Preferably, the purity of the xanthocyanin WGX-50 is ≥98%.

[0011] A method for preparing a hydrogel dressing for wound healing includes the following steps: S1, Dispersion and swelling: Carbomer 940 is added to the prescribed amount of physiological saline, dispersed and mixed, and allowed to stand to swell to obtain a carbomer dispersion system; S2, Introduction of xanthoside: Xanthoside WGX-50 was prepared into a uniform dispersion with physiological saline and then added to the carbomer dispersion system and mixed. S3, Neutralization and gel formation: Add sodium hydroxide solution dropwise to adjust the pH to 5.0-7.5 to form a gel matrix; S4, Low-temperature addition of exosomes: The gel matrix is ​​added to the exosome formulation and mixed uniformly under low shear. S5, Defoaming and Filling: Defoaming and filling the obtained system and sealing it to obtain a hydrogel dressing.

[0012] Preferably, the operating environment, equipment, and packaging containers used in the preparation method are all sterilized by ultraviolet irradiation.

[0013] Preferably, in step S4, the exosome preparation is added after the gel matrix is ​​cooled to 2-25°C.

[0014] Preferably, the system obtained in step S5 is degassed and then filled and sealed under aseptic conditions.

[0015] Preferably, the degassing of the system obtained in step S5 is carried out under a vacuum of -0.04 to -0.10 MPa.

[0016] In summary, the present invention has the following beneficial effects: 1. Carbomer hydrogel forms a continuous, adherent gel covering on the wound surface, reducing wound exposure and moisture loss, and maintaining the appropriate moisture required for wound exudate; at the same time, it reduces the adhesion between traditional dry dressings and granulation tissue / new epithelium, reducing the risk of secondary damage and rebleeding caused by mechanical peeling during dressing changes, thereby facilitating the continuous progress of key healing steps such as epithelial cell migration, keratinization reconstruction and granulation tissue formation.

[0017] 2. This invention utilizes hydrogel as a local delivery platform, enabling the WGX-50 cinnamon and exosome preparations to form a relatively stable retention and release environment on the wound surface, enhancing their supportive effect on the local wound microenvironment. WGX-50 cinnamon, as a small-molecule active ingredient, is added in controlled amounts through formulation to improve compatibility and suitability. Exosome preparations, as nanoscale bioactive carriers, carry signaling molecules such as proteins / nucleic acids, potentially exerting their effects by influencing inflammatory responses, cell migration and proliferation, and tissue remodeling. The hydrogel provides a moist retention and barrier mechanism, WGX-50 cinnamon provides microenvironmental regulation support at the small-molecule level, and the exosome preparations provide regulatory support at the biological signal level, thus forming a multi-layered synergistic effect at the wound site.

[0018] 3. Since xanthocyanin WGX-50 may have different effects on cell viability under different dosage conditions, this invention controls its local exposure level through formulation range design and pre-dissolution / dispersion process, making it more likely to be within a compatible dosage range; at the same time, considering the sensitivity of exosomes to high temperature and strong shear, this invention adopts the process of "adding exosomes at low temperature and low shear after gelation" to reduce the risk of damage to exosome structure and activity, and improve system homogeneity and batch-to-batch reproducibility.

[0019] 4. In this invention, an ultraviolet irradiation sterilization control point is set in the preparation process to sterilize the operating environment, equipment and packaging containers with ultraviolet irradiation, and the reagent addition and filling and sealing are completed under aseptic conditions, so as to improve the hygiene control level of the preparation process. Attached Figure Description

[0020] Figure 1 Schematic diagram of the live / dead staining of L929 cells by different concentrations of xanthocyanin WGX-50; Figure 2 This is a schematic diagram showing the change in water retention rate of each group of samples over time in this invention; Figure 3 This is a schematic diagram of in vitro hemolysis of various groups of samples in this invention; Figure 4 This is a schematic diagram showing the change of wound area over time in each group of samples of the present invention. Detailed Implementation

[0021] The specific embodiments of the present invention will be further described below with reference to the accompanying drawings. These embodiments do not constitute a limitation on the present invention.

[0022] Hydrogel dressings for wound healing were prepared according to the components of each embodiment in Table 1 below.

[0023] Table 1

[0024] This invention only describes the actual preparation of the formulation shown in Example 1, and the evaluation was conducted under non-clinical conditions such as a rat wound model. To facilitate understanding of the adjustability of the component content in this invention's technical solution, and to illustrate that different gel properties and application suitability can be obtained by adjusting the amount of xanthocyanin WGX-50, exosome potency, and carbomer 940 content, physiological saline was used as the dispersion medium in all formulations of this invention. A 10% (mass fraction) NaOH aqueous solution was added dropwise to adjust the pH of the system to 5.0–7.5 to form a gel. The "sodium hydroxide dosage" listed in the table is converted to the mass of NaOH soda ash. Actual preparation was based on adjusting to the target pH. Exosomes were measured in activity units (AU); AU is the unit of exosome potency, which can be calibrated by the supplier or an equivalent unit can be established through in vitro biological evaluation.

[0025] Preparation process: The specific steps of the preparation process in Example 1 are as follows: (1) Dispersion and swelling: Carbomer 940 is slowly sprinkled into the required volume of physiological saline, stirred at 300-800 rpm for 30-60 min, and then allowed to stand for swelling for 1-12 h to obtain a uniform carbomer dispersion system.

[0026] (2) Introduction of xanthoside: xanthoside WGX-50 was pre-dissolved in physiological saline system and then added to carbomer dispersion system and stirred evenly.

[0027] (3) Neutralization and gel formation: Add NaOH solution to adjust the pH to 6.0-7.0 to form a gel matrix.

[0028] (4) Adding exosomes at low temperature: Cool the gel to 2-25°C, add mesenchymal stem cell-derived exosomes according to AU, and mix at 100-300 rpm for 5-20 min with low shear.

[0029] (5) Degassing and filling: Degassing under vacuum conditions of -0.04 to -0.10 MPa, aseptic filling and sealing.

[0030] Ultraviolet irradiation sterilization control point: Before preparation, the operating environment, equipment and packaging containers are sterilized by ultraviolet irradiation, specifically 254nm is preferred, the distance is 10-50cm, the irradiation time is 15-60min, and step (4) and filling and sealing are completed under aseptic conditions.

[0031] To evaluate the material properties, compatibility, and non-clinical performance of the hydrogel dressing in Example 1 of this invention, this invention conducted water retention tests, hemolysis tests, and rat wound model evaluations, and provided the observation of live / dead staining of L929 cells with xanthocyanin WGX-50 as an in vitro reference.

[0032] Test Example 1: Observation of the live / dead staining effect of xanthocyanin WGX-50 on L929 cells Objective: To observe the effect of different concentrations of xanthoside WGX-50 on the survival of L929 cells, and to provide an in vitro reference for the selection of xanthoside WGX-50 dosage.

[0033] Grouping and Materials: Cells: L929 cells. Grouping: Piperine WGX-50 concentration gradient (0, 0.25, 0.5, 1, 2 μg / mL); Group 0 was a control containing the same volume fraction of solvent. Staining: Calcein-AM / PI.

[0034] Test steps: (1) Seed L929 cells into culture plates and culture until adherent. (2) Replace the culture medium with different concentrations of xanthocyanin WGX-50 and treat for 24 h. (3) Perform live / dead staining according to the reagent instructions, incubate for 20 min, and wash gently with PBS. (4) Acquire images under the same exposure conditions using a fluorescence microscope, and randomly select ≥3 fields of view for each group.

[0035] Live / dead staining results are shown in Figure 1 At concentrations of WGX-50 (0–2 μg / mL), cells showed predominantly green live cell signals and fewer red dead cell signals. These results suggest that WGX-50 exhibits good biocompatibility within this range.

[0036] Note: This test case is an in vitro reference for "piperazine WGX-50 on cells" and is not equivalent to "cell compatibility evaluation of the finished gel".

[0037] Test Example 2: Water Retention Performance Test of Three Groups of Hydrogel Dressings (Zanthoxylate Gel / Exosome Gel / Combined Gel) Test objective: To evaluate the ability of different combinations of active components in carbomer 940 hydrogels to maintain a moist microenvironment under certain environmental conditions, and to compare the effects of xanthocyanin, exosomes and their combined formulations on the water retention performance of the gel.

[0038] Grouping and sample definition: Each group was supplemented with physiological saline to a final volume of 100g, and the pH was adjusted to 5.0–7.5 (preferably 6.0–7.0) by adding 10% NaOH solution dropwise to form a gel. The amount of Carbomer 940 used was consistent (1.0g / 100g). WGX-50 (Zanthoxylate Gel Set): Carbomer 940 + Zanthoxylate WGX-50 (0.10mg / 100g) EXOSOME (Exosome Gel Set): Carbomer 940 + Mesenchymal Stem Cell-Derived Exosomes (100,000 AU / 100g) WGX-50 / EXOSOME (Combined Gel Set): Carbomer 940 + Zanthoxylin WGX-50 + Mesenchymal Stem Cell-Derived Exosomes (0.10mg + 100000AU / 100g) Test conditions: Ambient temperature: room temperature; Relative humidity: 45%; Container: 2 2 1.2cm circular container; repeat n=3 times per group.

[0039] Test steps: (1) Weigh the initial mass m0 (g) of each group of samples and record it to 0.01g. (2) Place the samples in the same container and place them under the set environment. (3) Weigh the sample mass m at 4, 8, 12, 24 and 30h respectively. t (g). (4) Calculate the water retention rate.

[0040] Calculation formula: Water retention rate (%) = m t / m0×100% The results of the water retention experiment are shown below Figure 2 There was no significant difference in water retention between the combined gel group and the single-component gel group within the 4–30 h range. These results suggest that the compound formulation does not weaken the water retention performance of the matrix and is beneficial for maintaining a moist microenvironment in the wound.

[0041] Test Example 3: Hemolysis Test (Gel Extract; Three Control Groups) Test objective: To evaluate the effects of different combinations of active components of carbomer 940 hydrogel on erythrocytes, to verify the blood compatibility of the material, and to compare the differences between xanthocyanin gel, exosome gel and their combined gel.

[0042] Grouping and Samples: Carbomer 940 hydrogels were prepared for each group, and tests were performed using the gel extraction solution. WGX-50 Zanthoxylin Gel Set: Carbomer 940 hydrogel containing Zanthoxylin WGX-50; EXOSOME exosome gel kit: Carbomer 940 hydrogel containing exosomes derived from mesenchymal stem cells; WGX-50 / EXOSOME combined gel group: Carbomer 940 hydrogel containing both xanthocyanin WGX-50 and mesenchymal stem cell-derived exosomes.

[0043] Control settings: Negative control: physiological saline (0% hemolysis); Positive control: double-distilled water (100% hemolysis).

[0044] Preparation of gel extract: Add 0.2 g / mL of each group of gels to the extraction medium physiological saline and extract at 37℃ for 24 h; after extraction, centrifuge and collect the supernatant to obtain the extract of each group.

[0045] Note: The extraction conditions were kept consistent across all three groups to ensure the interpretability of the comparison.

[0046] Test steps: (1) Collect anticoagulated whole blood, centrifuge to remove plasma; wash red blood cells repeatedly with physiological saline until the supernatant is clear, and prepare 4% (v / v) red blood cell suspension. (2) Mix each group of extracts with 4% red blood cell suspension at a volume ratio of 1:1 and incubate at 37℃ for 3h. (3) After incubation, centrifuge and collect the supernatant, measure the absorbance OD value at 540nm, and retain a photo of the appearance of the supernatant for evidence.

[0047] Calculation formula: Hemolysis rate (%) = (OD sample) OD negative) / (OD positive) OD negative) × 100% The results of the hemolysis test are shown below. Figure 3 The hemolysis rates of all three groups of gel extracts were below 2%, and the differences between the groups were not significant; no obvious hemolysis was observed in the supernatant photographs. These results indicate that the hydrogel system of this invention has good blood compatibility, and the combination of xanthocyanin and exosomes does not increase the risk of hemolysis. (Generally, a hemolysis rate below 5% is considered to indicate good blood compatibility; the results in this experiment were all below 2%, further supporting its compatibility.) Test Example 4: Non-clinical evaluation of a rat skin wound model Test objective: To evaluate the effect of the combined gel (piperazine + exosome composite carbomer gel) of this invention on the wound repair process in a rat skin wound model by conducting a non-clinical comparison of different treatment methods, and to compare it with single-component gel and positive control human epidermal growth factor gel (HEGF).

[0048] Grouping and Sample Definition: Five groups are set up in total. NS saline group: The wound was treated with normal saline. WGX-50 Zanthoxylin Gel Assembly: Zanthoxylin WGX-50 was added to a Carbomer 940 hydrogel matrix (the amount used was the same as in this invention). EXOSOME exosome gel assembly: Mesenchymal stem cell-derived exosomes (measured in AU, the amount is consistent with the present invention) are added to a carbomer 940 hydrogel matrix. WGX-50 / EXOSOME (a type of cinnamon) + exosome combined gel group: WGX-50 cinnamon and exosomes derived from mesenchymal stem cells were added to a carbomer 940 hydrogel matrix (corresponding to Example 1 of this invention). HEGF human epidermal growth factor gel group: human epidermal growth factor gel preparation, as a positive control group.

[0049] The WGX-50 group, EXOSOME group, and combined gel group all used the same Carbomer 940 gel matrix and the same pH adjustment method (10% NaOH solution was added dropwise to adjust the pH to 5.0-7.5, preferably 6.0-7.0), with only the active components being different, in order to ensure variable control.

[0050] Animals and Model: Six-week-old female SD rats; n=3 per group. After anesthesia, the back was shaved and disinfected to prepare a full-thickness skin defect with a diameter of 1.5 cm. The day of modeling was recorded as Day 0.

[0051] Administration and treatment methods: NS group: the wound was treated with normal saline once a day; WGX-50 group, EXOSOME group, WGX-50 / EXOSOME group: the corresponding gel was applied, with a single application amount of about 1.5g to cover the wound, once a day; HEGF group: about 1.5g to cover the wound, once a day.

[0052] Photos were taken on days 4, 8, 14, and 21.

[0053] Comparison of rat wound photos Figure 4 The wounds in each group gradually shrank over time; the combined gel group and the exosome gel group healed faster than the other groups.

[0054] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art can make various modifications or equivalent substitutions to the present invention within the scope of its essence and protection. Such modifications or equivalent substitutions should also be considered to fall within the protection scope of the present invention.

Claims

1. A hydrogel dressing for wound healing, characterized in that, Based on 100g of finished product, it contains the following components: Carbomer 940 is 0.50–2.00 g; The concentration of xanthocyanin WGX-50 is 0.01–0.50 mg. The exosome formulation is 50,000–500,000 AU; Sodium hydroxide is 0.02–0.50 g; The remainder is physiological saline.

2. The wound healing hydrogel dressing according to claim 1, characterized in that: The hydrogel dressing has a pH of 5.0 to 7.

5.

3. The wound healing hydrogel dressing according to claim 1, characterized in that: The exosome preparation is a mesenchymal stem cell-derived exosome.

4. The wound healing hydrogel dressing according to claim 1, characterized in that: The purity of the xanthocyanin WGX-50 is ≥98%.

5. A method for preparing a hydrogel dressing for wound healing according to any one of claims 1 to 4, characterized in that, Includes the following steps: S1, Dispersion and swelling: Carbomer 940 is added to the prescribed amount of physiological saline, dispersed and mixed, and allowed to stand to swell to obtain a carbomer dispersion system; S2, Introduction of xanthoside: After preparing a uniform dispersion of xanthoside WGX-50 with physiological saline, add it to the carbomer dispersion system and mix well. S3, Neutralization and gel formation: Add sodium hydroxide solution dropwise to adjust the pH to 5.0-7.5 to form a gel matrix; S4, Low-temperature addition of exosomes: The gel matrix is ​​added to the exosome formulation and mixed uniformly under low shear. S5, Defoaming and Filling: The obtained system is defoamed, filled, and sealed to obtain a hydrogel dressing.

6. The method for preparing a hydrogel dressing for wound healing according to claim 5, characterized in that: The operating environment, equipment, and packaging containers used in the preparation process are all sterilized by ultraviolet irradiation.

7. The method for preparing a hydrogel dressing for wound healing according to claim 5, characterized in that: In step S4, the exosome preparation is added after the gel matrix is ​​cooled to 2-25°C.

8. The method for preparing a hydrogel dressing for wound healing according to claim 5, characterized in that: The system obtained in step S5 is degassed and then filled and sealed under aseptic conditions.

9. The method for preparing a hydrogel dressing for wound healing according to claim 5, characterized in that: The degassing of the system obtained in step S5 was carried out under a vacuum of -0.04 to -0.10 MPa.

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