Temperature-controlled anti-inflammatory and antibacterial skin wound dressing capable of promoting revascularization as well as preparation method and application thereof

Abstract

The invention discloses a temperature-controlled anti-inflammatory and antibacterial skin wound dressing capable of promoting revascularization and a preparation method and application thereof, and relates to the technical field of medical materials, the preparation method comprises the following steps: S1, preparing calcium peroxide microspheres; s2, the poloxamer 407, the chitosan and the forsythiaside A are jointly dissolved in an acetic acid solution, and mixed gel is obtained; s3, dispersing calcium peroxide microspheres into the mixed gel to obtain the skin wound dressing capable of controlling temperature, resisting inflammation and bacteria and promoting revascularization. The traditional Chinese medicine active ingredient forsythiaside A is loaded, so that the healing efficiency of diabetic wounds is improved; due to the temperature-sensitive property of the gel, the gel can be better attached to the wound, the antibacterial property can inhibit microorganism breeding at the wound surface, the oxygen release capacity can improve the anoxic environment at the wound, and the loaded traditional Chinese medicine active ingredient forsythiaside A has the anti-inflammatory capacity, promotes cell regeneration at the wound and accelerates the repair process.

Description

Technical Field

[0001] The present invention relates to the technical field of medical materials, and in particular to a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, as well as a preparation method and application thereof. Background Art

[0002] Accelerating the healing of skin wounds is a challenging issue in the field of diabetic complications research. Under the pathological environment of diabetic hyperglycemia, skin wound healing faces multiple challenges, including elevated oxidative stress, imbalanced energy metabolism, persistent chronic inflammatory responses, localized wound hypoxia, abnormal angiogenesis, epidermal nerve dysfunction, and decreased growth factor secretion. These factors collectively hinder the normal wound healing process. This invention aims to develop a temperature-sensitive antibacterial hydrogel with a slow oxygen release function, loaded with forsythiaside A, an active ingredient in Traditional Chinese Medicine, to accelerate the healing efficiency of diabetic wounds. Summary of the Invention

[0003] In response to the defects in the existing technology, the purpose of the present invention is to provide a temperature-controlled, anti-inflammatory, antibacterial and angiogenesis-promoting skin wound dressing, as well as its preparation method and application, which is loaded with the traditional Chinese medicine active ingredient Forsythiaside A to increase the healing efficiency of diabetic wounds; the temperature-sensitive properties of the gel enable it to fit the wound better, the antibacterial properties can inhibit the growth of microorganisms at the wound, and the oxygen-releasing capacity can improve the hypoxic environment at the wound; the loaded traditional Chinese medicine active ingredient Forsythiaside A has anti-inflammatory ability, promotes cell regeneration at the wound, and accelerates the repair process.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is: a method for preparing a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, comprising the following steps:

[0005] Step S1. preparing calcium peroxide microspheres;

[0006] Step S2. dissolving poloxamer 407, chitosan, and forsythiaside A in an acetic acid solution to obtain a mixed gel;

[0007] Step S3. Dispersing calcium peroxide microspheres into the mixed gel to obtain a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing.

[0008] Based on the above technical solution, step S1 specifically includes:

[0009] Step S101. Weigh 4.0 to 5.0 parts by weight of anhydrous calcium chloride and dissolve it in 100 parts by weight of distilled water;

[0010] Step S102. Add 75 to 90 parts by weight of a 1 mol / L sodium hydroxide solution to generate a precipitate;

[0011] Step S103. Add 90 to 120 parts by weight of a 3% hydrogen peroxide solution dropwise under stirring to generate nano calcium peroxide particles;

[0012] Step S104. Washing the precipitate with 30 to 50 parts by weight of a NaoH solution and distilled water, respectively;

[0013] Step S105: Drying and grinding to obtain calcium peroxide microspheres.

[0014] Based on the above technical solution, step S2 specifically includes: weighing 1.2-1.8 parts by weight of poloxamer 407, 0.04-0.08 parts by weight of chitosan, and 0.025-0.04 parts by weight of forsythiaside A, and dissolving them in 0.5% acetic acid solution with stirring at low temperature overnight to obtain a mixed gel.

[0015] On the basis of the above technical solution, the preparation process of the 0.5% acetic acid solution is as follows: 5.5 parts by weight of double distilled water are measured, and 0.0325 parts by weight of glacial acetic acid are added to obtain a 0.5% acetic acid solution.

[0016] Based on the above technical solution, step S3 specifically includes: dispersing 0.01 parts by weight of calcium peroxide microspheres into the above mixed gel, stirring at low temperature overnight to uniformly disperse them, and obtaining a temperature-controlled anti-inflammatory, antibacterial and angiogenesis-promoting skin wound dressing.

[0017] The present invention also provides a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, characterized in that the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing is prepared using the above-mentioned preparation method.

[0018] The present invention also provides a use of the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing prepared by the above-mentioned preparation method in the preparation of a medicine for repairing diabetic wounds.

[0019] The beneficial effects of the present invention are:

[0020] The present invention can realize a safe and effective skin wound repair strategy and can effectively prevent the inflammatory response that occurs during the body's damage repair process. This provides a new method for the rapid repair of surface injuries in diabetic patients and provides a new idea for the application of the traditional Chinese medicine ingredient forsythiaside A and the improvement of dressings.

[0021] It is loaded with the active ingredient of traditional Chinese medicine, Forsythiaside A, to increase the healing efficiency of diabetic wounds; the thermosensitive properties of the gel allow it to fit the wound better, its antibacterial properties can inhibit the growth of microorganisms at the wound surface, and its oxygen-releasing capacity can improve the hypoxic environment at the wound. The loaded active ingredient of traditional Chinese medicine, Forsythiaside A, has anti-inflammatory ability, promotes cell regeneration at the wound, and accelerates the repair process. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is an oxygen release curve of the calcium peroxide microspheres of the present invention;

[0023] Figure 2 This is a comparison chart of the antibacterial effect of the composite gel dressing of the present invention;

[0024] Figure 3 This is a comparison chart of the recovery of the skin wound on the back of mice in the present invention;

[0025] Figure 4 This is a graph showing the expression of CD31 in mouse skin wounds detected by immunofluorescence in the present invention;

[0026] Figure 5 This is a graph showing the expression of CD86 in mouse skin wounds detected by immunofluorescence in the present invention;

[0027] Figure 6 This is a comparison chart of the histomorphological detection of the mouse back skin wound in the present invention. DETAILED DESCRIPTION

[0028] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements with the same or similar functions.

[0029] The following description of the embodiments of the present invention is provided in conjunction with the accompanying drawings to further describe the specific embodiments of the present invention so that the technical solutions and beneficial effects of the present invention will be more clearly understood. The following description of the embodiments with reference to the accompanying drawings is illustrative and intended to explain the present invention, but is not to be construed as limiting the present invention.

[0030] The present invention provides a method for preparing a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, comprising the following steps:

[0031] Step S1. Preparing calcium peroxide microspheres; specifically, step S1 specifically includes: step S101. Weighing 4.3 g of anhydrous calcium chloride and dissolving it in 100 mL of distilled water; step S102. Adding 80 mL of 1 mol / L sodium hydroxide solution to generate a precipitate; step S103. Adding 100 mL of 3% hydrogen peroxide solution dropwise while stirring to generate nano calcium peroxide particles; step S104. Washing the precipitate with 30 mL of NaOH solution and distilled water, respectively; step S105. Drying and grinding to obtain calcium peroxide microspheres.

[0032] Step S2. Dissolving poloxamer 407, chitosan, and forsythiaside A in an acetic acid solution to obtain a mixed gel. Specifically, step S2 comprises weighing 1.2 g of poloxamer 407, 0.04 g of chitosan, and 0.025 g of forsythiaside A, and dissolving the mixture in a 0.5% acetic acid solution under stirring at low temperature overnight to obtain the mixed gel. Specifically, the 0.5% acetic acid solution is prepared by measuring 5.5 mL of double-distilled water and adding 32.5 μL of glacial acetic acid to obtain the 0.5% acetic acid solution.

[0033] Step S3. Dispersing calcium peroxide microspheres into the mixed gel to obtain a temperature-controlled, anti-inflammatory, antibacterial, and angiogenic skin wound dressing. Specifically, step S3 comprises dispersing 0.01 g of calcium peroxide microspheres into the mixed gel, stirring at low temperature overnight to uniformly disperse the microspheres, and obtaining a temperature-controlled, anti-inflammatory, antibacterial, and angiogenic skin wound dressing.

[0034] The present invention also provides a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, which is prepared using the above-mentioned preparation method.

[0035] The present invention also provides a use of the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing prepared by the above-mentioned preparation method in the preparation of a medicine for repairing diabetic wounds.

[0036] Forsythiaside A has excellent anti-inflammatory and antioxidant pharmacological properties. The inventors have found that Forsythiaside A can play a role in the improvement and treatment of diabetic wounds. However, after oral administration, the bioavailability of Forsythiaside A after gastrointestinal absorption into the blood circulation is low, and its targeting is also relatively limited, making it difficult to accurately enrich the wound site. To address this problem, the present invention uses a temperature-sensitive gel material as a drug carrier. The gel material has unique phase change properties. Under low temperature conditions, the interaction between its molecules is weak, and it exhibits good liquid fluidity. This liquid property gives the material a significant advantage in the drug application process, allowing the drug to be easily and evenly applied to the wound surface. At the same time, the liquid fluidity helps the drug to better penetrate into the deep layers of the wound tissue, increase the contact area between the drug and the wound cells, thereby increasing the initial concentration of the drug in the local wound, laying a solid foundation for subsequent treatment. When the ambient temperature rises to room temperature or close to human body temperature, the temperature-sensitive gel material undergoes a rapid phase change, from liquid to solid. This solidification process enables the material to form a stable covering layer at the wound site, fixing the loaded drug, Forsythiaside A, at the wound site. In order to further improve the repair effect on diabetic wounds, the inventors have improved and optimized the temperature-sensitive gel. By cleverly introducing chitosan, a natural polymer component, the gel is endowed with excellent antibacterial properties. Furthermore, the inventors dispersed calcium peroxide particles into the gel material. This move enables the gel to have the ability to slowly and continuously release oxygen, which plays a vital role in promoting tissue repair and accelerating wound healing. By adding multiple components, a slow-release oxygen, temperature-sensitive, antibacterial gel material is constructed, which is loaded with Forsythiaside A, an active ingredient in traditional Chinese medicine, and provides a new solution for the efficient and safe repair of diabetic wounds.

[0037] The present application will be further described below through several embodiments.

[0038] Example 1 Preparation of calcium peroxide microspheres and detection of their oxygen release capacity

[0039] A. Prepare calcium peroxide particles using a precipitation method. Weigh 4.3 g of anhydrous calcium chloride and dissolve it in 100 mL of distilled water. Add 80 mL of 1 mol / L sodium hydroxide solution to form a precipitate. Then, add 100 mL of 3% hydrogen peroxide solution dropwise while stirring to form nano-calcium peroxide particles. Wash the precipitate with 30 mL of NaOH solution and then with distilled water. Dry and grind to obtain calcium peroxide microspheres.

[0040] B. Weigh 0.1g of calcium peroxide microspheres and disperse them in 25mL of distilled water. Monitor the oxygen content in the water every 24 hours for 15 days.

[0041] See also Figure 1As shown, the oxygen release curve of calcium peroxide microspheres can be obtained through the above steps. The blue circle marks the dissolved oxygen content of calcium peroxide microspheres dispersed into water, and the red square marks the dissolved oxygen content in the dispersant water.

[0042] Example 2 Evaluation of the antibacterial effect of gel

[0043] A. Select a monoclonal colony of Staphylococcus aureus or Escherichia coli, inoculate it into 10 ml of LB liquid culture medium, and culture it in a shaking incubator at 37°C for 12 hours to obtain a bacterial liquid.

[0044] B. Divide the culture dishes with a diameter of 10 cm into three groups: control group (add 100uL of Staphylococcus aureus or Escherichia coli to the culture dish and spread it evenly); gel group (add 100uL of hydrogel without FTA loading and 100uL of Staphylococcus aureus or Escherichia coli to the culture dish and spread it evenly); composite gel dressing group (add 100uL of composite gel dressing loaded with FTA and 100uL of Staphylococcus aureus or Escherichia coli to the culture dish and spread it evenly).

[0045] C. After relevant treatment, place it in the bacterial incubator.

[0046] D. After 12 hours, the number of colonies in the control group, gel group, and composite gel dressing group was counted, and the antibacterial rate was calculated.

[0047] See also Figure 2 As shown, the antibacterial effect of the gel can be detected through the above steps. A. Colony distribution of different bacterial groups after culture using the spread plate method. B. Statistical analysis of the number of colonies formed in different groups. N = 4; One-way ANOVA test; **, p < 0.01; ***, p < 0.001.

[0048] Example 3 Evaluation of the effect of composite gel dressing on promoting diabetic wound healing

[0049] A. Establishment of diabetic wound injury model in mice;

[0050] B. Diabetic mice were further divided into three groups: control group (wounds were not treated); gel group (wounds were smeared with hydrogel not loaded with forsythiaside A); and treatment group (wounds were smeared with hydrogel loaded with forsythiaside A).

[0051] C. Take photos daily to observe and record the wound healing status.

[0052] See also Figure 3 As shown, the above operations can visually evaluate the healing effect of the composite gel dressing on the wound of diabetic mice, WT, wild-type mouse control group; DM, diabetic model group; OTH, thermosensitive oxygen-releasing hydrogel group; OTH-FTA, hydrogel dressing loaded with forsythiaside A treatment group.

[0053] Example 4 Evaluation of the ability of composite gel dressing to promote angiogenesis

[0054] A. Establishment of diabetic wound injury model in mice;

[0055] B. Diabetic mice were divided into two groups: a control group (wounds were smeared with normal saline) and a treatment group (wounds were smeared with a composite dressing loaded with forsythiaside A).

[0056] C. After 4 days, some mice in each group were killed, and approximately 1 cm2 of skin tissue around the wound was collected, paraffin-sectioned, and immunofluorescence staining was performed to detect the expression of CD31 at the wound site.

[0057] See also Figure 4 As shown in the figure, after the above manipulations, angiogenesis in the wound surface of mice was observed. Compared with the control group, more new blood vessels were detected in the treatment group on day 4, especially in the epidermis. Compared with the control group, the wound surface of the treatment group showed greater expression of CD31. While CD31 was undetectable in the epidermis of the control group, CD31 was detected in the epidermis of the treatment group, indicating a stronger angiogenesis capacity in the treatment group. Epi, epidermis; Derm, dermis. The lower row of images shows magnified images of the field of view within the dotted box. The dotted line in the lower row of images indicates the boundary between the epidermis and dermis.

[0058] Example 5 Evaluation of the anti-inflammatory ability of composite gel dressing

[0059] A. A diabetic wound injury model was constructed in mice as described in Example 4, and the mice were divided into a control group and a treatment group.

[0060] B. On day 4, the mice were killed and a sample of approximately 1 cm around the wound was taken. 2 The skin tissue was paraffin-embedded and immunofluorescence stained to detect the expression of CD86 in the wound.

[0061] See also Figure 5 As shown in the figure, after the above operation, the distribution of M1 macrophages in the wound surface of mice can be observed. Compared with the control group, the number of CD86-positive cells in the treatment group was less on day 4. Compared with the control group, the wound surface of the treatment group showed lower CD86 expression. CD86 expression in the treatment group was concentrated in the blood scab outside the skin, and was less in the dermis and epidermis. This indicates that the treatment group has fewer M1 macrophages and less inflammation.

[0062] Example 6 Evaluation of the ability of composite gel dressings to promote recovery

[0063] A. A diabetic wound injury model was constructed in mice as described in Example 4, and the mice were divided into a control group and a treatment group.

[0064] B. On day 10, the mice were killed and a sample of about 1 cm around the wound was taken. 2Skin tissues were sectioned into paraffin and stained with HE and Masson's staining.

[0065] See also Figure 6 As shown, after the above operations, the tissue repair at the wound surface of the mice can be observed. It can be seen that although the wound of the control group is closed, its internal structure is still not completely repaired; while the treatment group shows a stronger repair ability.

[0066] In the description of the specification, reference to the terms "one embodiment," "preferably," "example," "specific example," or "some examples" means that the specific features, structures, materials, or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. The schematic expressions of the above terms in this specification do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described may be combined in any appropriate manner in any one or more embodiments or examples.

[0067] The present invention is not limited to the above-described embodiments. Persons skilled in the art may make various improvements and modifications without departing from the principles of the present invention, and such improvements and modifications are deemed to be within the scope of protection of the present invention. Any matters not described in detail in this specification constitute prior art known to those skilled in the art.

Claims

1. A method for preparing a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, characterized in that: The following steps are involved: Step S1. preparing calcium peroxide microspheres; Step S2. dissolving poloxamer 407, chitosan, and forsythiaside A in an acetic acid solution to obtain a mixed gel; Step S3. Dispersing calcium peroxide microspheres into the mixed gel to obtain a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing.

2. The method for preparing the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing according to claim 1, characterized in that: Step S1 specifically includes: Step S101. Weigh 4.0 to 5.0 parts by weight of anhydrous calcium chloride and dissolve it in 100 parts by weight of distilled water; Step S102. Add 75 to 90 parts by weight of a 1 mol / L sodium hydroxide solution to generate a precipitate; Step S103. Add 90 to 120 parts by weight of a 3% hydrogen peroxide solution dropwise under stirring to generate nano calcium peroxide particles; Step S104. Washing the precipitate with 30 to 50 parts by weight of a NaoH solution and distilled water, respectively; Step S105: Drying and grinding to obtain calcium peroxide microspheres.

3. The method for preparing the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing according to claim 1, wherein: Step S2 specifically includes: weighing 1.2-1.8 parts by weight of poloxamer 407, 0.04-0.08 parts by weight of chitosan, and 0.025-0.04 parts by weight of forsythiaside A, and dissolving them in 0.5% acetic acid solution under stirring at low temperature overnight to obtain a mixed gel.

4. The method for preparing the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing according to claim 3, characterized in that: The preparation process of the 0.5% acetic acid solution is as follows: 5.5 parts by weight of double distilled water are measured, and 0.0325 parts by weight of glacial acetic acid are added to obtain the 0.5% acetic acid solution.

5. The method for preparing the temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing according to claim 1, wherein: Step S3 specifically includes: dispersing 0.01 parts by weight of calcium peroxide microspheres into the mixed gel, stirring at low temperature overnight to uniformly disperse the microspheres, and obtaining a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing.

6. A temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing, characterized in that: The temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing is prepared by the preparation method according to any one of claims 1 to 5.

7. Use of a temperature-controlled, anti-inflammatory, antibacterial, and angiogenesis-promoting skin wound dressing prepared by the preparation method according to any one of claims 1 to 5 in the preparation of a drug for repairing diabetic wounds.

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