A temperature-sensitive hydrogel for promoting wound healing and a preparation method and application thereof

Abstract

The application discloses a temperature-sensitive hydrogel for promoting wound healing and a preparation method and application thereof, and relates to the technical field of hydrogels. The temperature-sensitive hydrogel for promoting wound healing is composed of FDA-approved dressings, and active ingredients are at least one of FDA-approved carotenoids and ginsenosides which can be used as food additives, and the content is 0.1‰-1‰. The gel can provide a moist environment for wound repair, and the active ingredients have various biological activities, free radical scavenging and anti-inflammatory abilities, and can promote cell repair. The two active ingredients are assembled with poloxamer and dopamine, and the solubility, bioavailability, biological activity and stability of the two active ingredients are increased, so that the wound healing is accelerated, and the gel is formed under the condition of body temperature, the skin surface viscosity effect is enhanced, and the gel is a liquid at room temperature, so that storage and transportation are facilitated. The hydrogel can be widely applied to the preparation of dressings for promoting wound healing.

Description

Technical Field

[0001] This invention relates to the field of excipient technology, and more specifically, to a thermosensitive hydrogel that promotes wound healing, its preparation method, and its application. Background Technology

[0002] The skin is a vital organ covering the human body surface and in direct contact with the external environment. It functions to sense external stimuli, regulate body temperature, and protect the body from external harm. Due to its direct contact with the outside world, the skin is also one of the most vulnerable tissues. While most common skin injuries can heal to a basic degree within a period of time, adult skin injuries rarely achieve 100% recovery of skin function like those in infants. This is often accompanied by the formation of scar tissue and significant loss of skin appendages such as hair, sweat glands, and sebaceous glands. Skin wound repair involves four continuous and coordinated processes: hemostasis, inflammation, proliferation, and remodeling. However, the wound healing process is usually not perfectly orderly. Various factors at any stage can lead to abnormal wound repair, such as excessive inflammation, burns or infections caused by extensive skin tissue loss, and diabetic ulcers.

[0003] Ion channels are large molecular pores on the cell membrane that can be excited by external signals. They universally influence important physiological processes such as glandular secretion, muscle movement, cell division, and cell proliferation. Ion channels are recognized as the second largest class of drug targets in the global drug development field. Calcium-activated chloride channels (CaCCs) are widely distributed in various epithelial and non-epithelial tissues and participate in numerous physiological processes, controlling and regulating ion homeostasis, cell volume, cell excitability, sensory signal transduction, and transepithelial electrolyte transport. In late 2008, three independent research groups independently identified transmembrane 16A (TMEM16A) as the encoding gene for CaCCs channels using different methods. Currently, nearly 500 articles on TMEM16A can be found in the National Center for Bioinformatics (NCBI) database. These studies focus on the tissue distribution, channel gating and permeability of this ion channel, its association with signal transduction pathways, and its relationship with various major diseases, including cancer, hypertension, cystic fibrosis, gastrointestinal dysfunction, and, most recently, multiple types of cancer. Notably, recent research indicates that ion channels, particularly TMEM16A, participate in regulating the proliferation of tail muscles and mesenchymal cells during tail regeneration in salamanders by mediating the p44 / 42MAPK-ERK1 / 2 signaling pathway. However, the molecular mechanism by which TMEM16A regulates wound healing remains unclear.

[0004] In recent years, research on novel wound dressings has become increasingly extensive, resulting in the development of numerous wound dressings to promote wound healing, such as semipermeable membranes, semipermeable foams, hydrocolloids, and hydrogels. Among these, hydrogels have emerged as the most competitive candidate material for wound dressings due to their excellent hydrophilicity and biocompatibility, attracting considerable interest from researchers. Research on hydrogels as wound dressings has also shown an increasing trend over the past decade. Many hydrophilic polymers, including natural polymers such as chitosan, gelatin, hyaluronic acid, alginate, and dextran, as well as synthetic hydrophilic polymers such as polyethylene glycol (PEG) and poloxamer, polyvinyl alcohol, olefin-containing monomers such as polyacrylamide (PAM), poly(acrylic acid), and peptides, have been used to construct hydrogels through various chemical or physical crosslinking methods. Dynamic chemical bonding, photocrosslinking in-situ polymerization, dual-network, semi-interpenetrating networks, and 3D printing have all demonstrated significant advantages. Furthermore, the function of hydrogels has evolved from simple physical coverage or a single function to a combination of multiple functions, exhibiting a trend towards greater intelligence.

[0005] Existing hydrogels that promote wound healing cannot simultaneously achieve the functions of temperature sensitivity, free radical scavenging, and promoting cell regeneration at the wound site.

[0006] In view of this, the present invention is proposed. Summary of the Invention

[0007] The purpose of this invention is to provide a thermosensitive hydrogel that promotes wound healing, its preparation method, and its application.

[0008] This invention is implemented as follows:

[0009] In a first aspect, the present invention provides a thermosensitive hydrogel that promotes wound healing, wherein the raw materials, by mass percentage, include 0.1‰-1‰ of active ingredients, 1%-40% of poloxamer 407, 1%-10% of dopamine, 1%-20% of poloxamer 188, and the balance being an alkaline solvent; the active ingredients include at least one of carotenoids and ginsenosides.

[0010] In an optional embodiment, the carotenoid includes at least one of astaxanthin, canthaxanthin, or lutein.

[0011] And / or, the ginsenosides include at least one of ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rg2, ginsenoside NGR1, ginsenoside Rf, and ginsenoside Rh2;

[0012] And / or, the pH of the alkaline solvent is 8-9.

[0013] Secondly, the present invention provides a method for preparing a temperature-sensitive hydrogel for promoting wound healing as described in the foregoing embodiments, comprising: mixing the raw materials evenly.

[0014] In an optional embodiment, mixing the raw materials uniformly includes:

[0015] Add the poloxamer 407 to the pre-cooled alkaline solvent and stir until clear;

[0016] Next, add the active ingredient and continue stirring;

[0017] The dopamine was then added and stirred to allow it to self-polymerize under alkaline conditions, forming an interpenetrating network with the poloxamer.

[0018] Finally, poloxamer 188 is added so that the wound-healing thermosensitive hydrogel is liquid at room temperature and gel at body surface temperature.

[0019] In an optional embodiment, the viscosity of the wound-healing thermosensitive hydrogel when it is a liquid is 1 Pa·s-100 Pa·s.

[0020] In an optional embodiment, the temperature-sensitive hydrogel that promotes wound healing has a viscosity of 1500 Pa·s-2000 Pa·s when it is in gel form.

[0021] In an optional embodiment, the mixing process of the raw materials is carried out at 3-5°C.

[0022] Thirdly, the present invention provides the use of a wound-healing thermosensitive hydrogel prepared by any of the foregoing embodiments or by any of the foregoing embodiments in the preparation of a wound-healing medicament.

[0023] In an optional embodiment, promoting wound healing includes promoting wound area reduction, promoting skin function recovery at the wound site, and promoting the recovery of sebaceous glands and hair follicles in the wound.

[0024] In an optional implementation, the wound used to promote wound healing includes wounds from non-diabetic patients or wounds from diabetic patients.

[0025] The present invention has the following beneficial effects:

[0026] The thermosensitive hydrogel for promoting wound healing provided in this application uses at least one of carotenoids and ginsenosides as active ingredients, which are combined with poloxamer 407, dopamine, and poloxamer 188 under alkaline conditions to form an assembly. Both carotenoids and ginsenosides possess various biological activities and free radical scavenging abilities, and can promote cell repair and stimulate TMEM16A channels to promote cell proliferation. Assembling these two active ingredients with poloxamer and dopamine increases their solubility, bioavailability, biological activity, and stability, thereby accelerating wound healing. Furthermore, the thermosensitive hydrogel for promoting wound healing provided in this application forms a gel at body temperature (37°C), enhancing the skin surface viscosity effect, and remains liquid at room temperature, facilitating storage and transportation. The thermosensitive hydrogel for promoting wound healing provided in this application can be widely used in the preparation of drugs that promote wound healing, and is beneficial for reducing wound area, promoting the recovery of skin function at the wound site, and promoting the recovery of sebaceous glands and hair follicles in the wound. The thermosensitive hydrogel for promoting wound healing provided in this application not only has a good healing effect on normal wounds, but also on wounds of diabetic patients. Attached Figure Description

[0027] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0028] Figure 1 This is a SEM image of the wound-healing thermosensitive hydrogel provided in Embodiment 1 of this application.

[0029] Figure 2 A schematic diagram illustrating the transition between liquid and gel states of the temperature-sensitive hydrogel for promoting wound healing provided in Embodiment 1 of this application;

[0030] Figure 3 The graph shows the composite viscosity of the wound-healing thermosensitive hydrogels provided in Examples 1-3 of this application as a function of temperature.

[0031] Figure 4 This is a schematic diagram illustrating the effects of different treatment groups on wound healing in mice, as provided in Experiment Example 1 of this application.

[0032] Figure 5 This is a schematic diagram illustrating the difference in wound area between different treatment groups in the first experimental example of this application, showing their effect on wound healing in mice.

[0033] Figure 6HE staining images of the effects of different treatment groups on wound healing in mice, provided in Experiment Example 1 of this application;

[0034] Figure 7 This is a schematic diagram illustrating the effects of different treatment groups on wound healing on the back of rats, as provided in Experimental Example 2 of this application.

[0035] Figure 8 This is a schematic diagram illustrating the difference in wound area between different treatment groups in Experiment Example 2 of this application, which is used to analyze the healing effect of wounds on the back of rats.

[0036] Figure 9 HE staining images of the effects of different treatment groups on wound healing on the back of rats provided in Experimental Example 2 of this application;

[0037] Figure 10 This is a schematic diagram illustrating the effects of different treatment groups on the healing of rat paw wounds, provided in Experimental Example 2 of this application.

[0038] Figure 11 This is a schematic diagram illustrating the difference in wound area between different treatment groups in Experiment Example 2 of this application, showing their effect on wound healing in rat paws.

[0039] Figure 12 HE staining images of the effects of different treatment groups on the healing of rat paw wounds, provided in Experimental Example 2 of this application;

[0040] Figure 13 This is a schematic diagram illustrating the effects of different treatment groups on wound healing in mice, as provided in Experimental Example 3 of this application.

[0041] Figure 14 This is a schematic diagram illustrating the difference in wound area between different treatment groups in the treatment of infected wounds in mice, as provided in Experimental Example 3 of this application.

[0042] Figure 15 HE staining images of the effects of different treatment groups on wound healing in mice, provided in Experimental Example 3 of this application. Detailed Implementation

[0043] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0044] This invention provides a thermosensitive hydrogel that promotes wound healing. The raw materials, by mass percentage, include 0.1‰-1‰ of active ingredient, 1%-40% of poloxamer 407, 1%-10% of dopamine, 1%-20% of poloxamer 188, and the balance being an alkaline solvent.

[0045] The active ingredients include at least one of carotenoids and ginsenosides.

[0046] Carotenoids possess a variety of biological activities and free radical scavenging abilities, exhibiting high baseline antioxidant capacity and the ability to activate the TMEM16A channel to promote cell proliferation. They have been approved by the FDA as bioactive additives. However, carotenoids are poorly soluble in water and have a short residence time in vivo, limiting their use. This application increases the solubility of carotenoids by assembling them with poloxamer and dopamine, and constructs a temperature-responsive hydrogel system that allows it to form a gel at body temperature (37°C), enhancing the skin surface viscosity effect, while remaining liquid at room temperature for easy storage and transportation. Specifically, the carotenoids include at least one of astaxanthin, canthaxanthin, or lutein.

[0047] Ginsenosides possess the ability to promote cell repair and stimulate TMEM16A channels to promote cell proliferation. This application, by assembling ginsenosides with poloxamer and dopamine, helps improve the bioavailability, bioactivity, and stability of ginsenosides, and can regulate their release and delivery properties. Furthermore, the high content of phenolic hydroxyl groups in ginsenosides enhances the tissue adhesion of the wound-healing thermosensitive hydrogel, significantly accelerating the wound healing rate, reducing immune inflammatory responses, and increasing the formation of new proteins and pores. Specifically, the ginsenosides include at least one of ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rg2, ginsenoside NGR1, ginsenoside Rf, and ginsenoside Rh2. The pH of the alkaline solvent is 8-9. This application utilizes an alkaline solvent to provide alkaline reaction conditions, facilitating the self-polymerization of dopamine under alkaline conditions to form an interpenetrating network with poloxamer.

[0048] The preparation method of the above-mentioned thermosensitive hydrogel that promotes wound healing includes: mixing the raw materials evenly.

[0049] Specifically, mixing the raw materials evenly includes the following steps:

[0050] S1. Add poloxamer 407 to a pre-cooled alkaline solvent and stir until clear.

[0051] In this process, the alkaline solvent is pre-prepared into an aqueous solution with a pH of 8-9, and the alkaline solvent is pre-cooled to 3-5°C. Then, poloxamer 407 is added, and the mixture is stirred thoroughly overnight at 3-5°C until it becomes a clear solution.

[0052] S2. Next, add the active ingredients and continue stirring.

[0053] The active ingredient can be added directly or prepared into a suspension before addition. When preparing into a suspension, it can be prepared directly using a partially alkaline solvent. The prepared suspension of the active ingredient is added to the solution in step S1 and stirred thoroughly overnight at 3-5°C.

[0054] S3. Then dopamine is added and stirred to allow dopamine to self-polymerize under alkaline conditions, forming an interpenetrating network with poloxamer.

[0055] S4. Finally, add poloxamer 188 to make the wound-healing thermosensitive hydrogel liquid at room temperature and gel at body surface temperature. Specifically, at room temperature (e.g., 20-30℃), the viscosity of the wound-healing thermosensitive hydrogel in liquid form is 1 Pa·s-100 Pa·s, and the viscosity of the wound-healing thermosensitive hydrogel in gel form is 1500 Pa·s-2000 Pa·s.

[0056] The aforementioned thermosensitive hydrogel for promoting wound healing can be widely used in the preparation of drugs that promote wound healing. Promoting wound healing includes reducing wound area, promoting the recovery of skin function at the wound site, and promoting the recovery of sebaceous glands and hair follicles within the wound. The wounds targeted for promoting healing include wounds from non-diabetic patients and wounds from diabetic patients. That is, the thermosensitive hydrogel for promoting wound healing provided in this application not only has a good healing-promoting effect on normal wounds but also on wounds from diabetic patients.

[0057] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0058] Example 1

[0059] This embodiment provides a method for preparing a thermosensitive hydrogel that promotes wound healing, which includes the following steps:

[0060] S1. Prepare an aqueous solution of NaOH with a pH of 8-9 as an alkaline solvent, and pre-cool the alkaline solvent to 4°C. Take 10 mL of the alkaline solvent, add 2 g of poloxamer 407, and stir thoroughly overnight at 4°C until it becomes a clear solution.

[0061] S2. Prepare 0.1 mg of carotenoid (lutein) with a partially alkaline solvent, add the prepared suspension of active ingredients to the solution in step S1, and stir thoroughly overnight at 4°C.

[0062] S3. Then add 50 mg of dopamine and stir to allow dopamine to self-polymerize under alkaline conditions, forming an interpenetrating network with poloxamer.

[0063] S4. Finally, add 100 μL of poloxamer 188 to make the wound-healing thermosensitive hydrogel liquid at room temperature and gel at body surface temperature.

[0064] Please see Figure 1 , Figure 2 , Figure 3 ,from Figure 1 It can be seen that the porous network structure of the composite hydrogel, from Figures 2-3 It can be seen that the temperature-sensitive hydrogel that promotes wound healing is a liquid at room temperature. As the temperature increases, its viscosity increases significantly, and it becomes a gel at body surface temperature, which is beneficial to improving the adhesion of the temperature-sensitive hydrogel that promotes wound healing to the wound.

[0065] Example 2

[0066] This embodiment provides a method for preparing a thermosensitive hydrogel that promotes wound healing, which is basically the same as that in Example 1, except that the amount of carotenoid used in this embodiment is 1‰.

[0067] Specifically, the steps include the following:

[0068] S1. Prepare an aqueous solution of NaOH with a pH of 8-9 as an alkaline solvent, and pre-cool the alkaline solvent to 4°C. Take 10 mL of the alkaline solvent, add 2 g of poloxamer 407, and stir thoroughly overnight at 4°C until it becomes a clear solution.

[0069] S2. Prepare 1 mg of carotenoid (lutein) with a partially alkaline solvent, add the prepared suspension of active ingredients to the solution in step S1, and stir thoroughly overnight at 4°C.

[0070] S3. Then add 50 mg of dopamine and stir to allow dopamine to self-polymerize under alkaline conditions, forming an interpenetrating network with poloxamer.

[0071] S4. Finally, add 100 μL of poloxamer 188 to make the wound-healing thermosensitive hydrogel liquid at room temperature and gel at body surface temperature.

[0072] Example 3

[0073] This embodiment provides a method for preparing a thermosensitive hydrogel that promotes wound healing, which is basically the same as that in Example 2, except that in this embodiment, the carotenoids in Example 2 are replaced with ginsenoside Rb1.

[0074] Specifically, the steps include the following:

[0075] S1. Prepare an aqueous solution of NaOH with a pH of 8-9 as an alkaline solvent, and pre-cool the alkaline solvent to 4°C. Take 10 mL of the alkaline solvent, add 2 g of poloxamer 407, and stir thoroughly overnight at 4°C until it becomes a clear solution.

[0076] S2. Prepare 1 mg of ginsenoside Rb1 with a partially alkaline solvent, add the prepared suspension of active ingredients to the solution in step S1, and stir thoroughly overnight at 4°C.

[0077] S3. Then add 50 mg of dopamine and stir to allow dopamine to self-polymerize under alkaline conditions, forming an interpenetrating network with poloxamer.

[0078] S4. Finally, add 100 μL of poloxamer 188 to make the wound-healing thermosensitive hydrogel liquid at room temperature and gel at body surface temperature.

[0079] Comparative Example 1

[0080] This comparative example provides a hydrogel that is essentially the same as that in Example 1, except that it does not contain carotenoids; instead, an alkaline solvent is used to replace the amount of carotenoids. The specific preparation method is as follows:

[0081] S1. Prepare an aqueous solution of NaOH with a pH of 8-9 as an alkaline solvent, and pre-cool the alkaline solvent to 4°C. Take 10 mL of the alkaline solvent, then add poloxamer 407, and stir thoroughly overnight at 4°C until it becomes a clear solution.

[0082] S2. Add 50 mg of dopamine and stir to allow dopamine to self-polymerize under alkaline conditions, forming an interpenetrating network with poloxamer.

[0083] S3. Finally, add 100 μL of poloxamer 188 to make the hydrogel liquid at room temperature and gel at body surface temperature.

[0084] Experiment Example 1: The effect of a thermosensitive hydrogel on wound healing in mice

[0085] KM mice were selected to establish a wound model. Under anesthesia, a 6 mm diameter full-thickness skin lesion was created on the back of the mice using a pore-forming device. After rinsing the wound with physiological saline, different treatments were administered, resulting in four groups: a control group (Control), a positive control group (hEGF gel, human epidermal growth factor gel, Guilin Huanowei Gene Pharmaceutical Co., Ltd.), a hydrogel group (Gel), Example 1 group (Gel C+low), Example 2 group (Gel C+high), and Example 3 group (Gel B).

[0086] In this study, the control group received saline treatment on the wound; the positive control group received human epidermal growth factor gel applied to the wound once a day; the hydrogel group received the carotenoid-free hydrogel provided in Comparative Example 1; and the Example 1, Example 2, and Example 3 groups received the temperature-sensitive hydrogels provided in Examples 1, 2, and 3, respectively, which promote wound healing.

[0087] Each wound was wrapped with sterile gauze and secured with elastic tape. Wound healing was recorded using a digital camera at 0, 3, 6, 9, and 12 days post-surgery, and ImageJ software was used to calculate the healing progress. Please refer to [link to relevant documentation]. Figure 4 and Figure 5 It was found that from day 3 onwards, the positive control group and the high-concentration hydrogel Example 2 group showed good therapeutic effects, followed by the low-concentration hydrogel Example 1 and Example 3 groups. After 12 days of treatment, both Example 1 (low concentration) and Example 3 groups achieved therapeutic effects comparable to the positive control, while the wound area of ​​Example 2 (high concentration) and Example 3 groups was smaller than that of the positive control.

[0088] Please see Figure 6 HE staining revealed that the wounds treated in Example 1, Example 2, and Example 3 healed faster, the skin repair was more complete, and the sebaceous glands and pores achieved structural and functional restoration on the wound surface.

[0089] Experimental Example 2: Effect of a temperature-sensitive hydrogel promoting wound healing on wounds on the back and paws of diabetic rats.

[0090] Eight-week-old healthy male SD rats were routinely housed at a constant temperature of 22℃ and humidity of 55%. Type I diabetic rats were induced under aseptic conditions by a single intraperitoneal injection of 65 mg / kg streptozotocin (STZ). Rats with a blood glucose level exceeding 16.7 mmol / L one week after administration were considered diabetic. A wound model was established. Under anesthesia, a 15 mm diameter full-thickness skin lesion was created on the dorsal skin of the rats, and an 8 mm diameter full-thickness skin lesion was created on the dorsum of the right hind paw.

[0091] After rinsing the wound with saline solution, different treatments were administered, resulting in four groups: a control group (Control), a positive control group (hEGFgel), a hydrogel group (Gel), the Example 2 group (Gel+C), and a normal group (Normal, non-diabetic group). Wound healing was recorded using a digital camera at 0, 3, 6, 9, 12, and 15 days post-surgery, and the wound healing status was calculated using ImageJ software.

[0092] (1) Effect on healing back wounds

[0093] Please see Figure 7 and Figure 8 The experiment showed that from the sixth day onwards, the skin wound area of ​​rats in the Example 2 group and the positive control group was significantly smaller than that in the control group. After 15 days of treatment, the wounds on the backs of rats in the positive control group, Example 2 group, and normal group had basically healed, and the wound area was significantly different from that in the control group.

[0094] Please see Figure 9 HE staining revealed that the wound area in the positive control group and the Example 2 group was smaller, and the wounds healed more smoothly. The recovery of sebaceous glands and hair follicles in the wounds was also faster than in the control group. Notably, after treatment, the number of sebaceous glands and hair follicles in the Example 2 group was higher than that in the positive control group, indicating that the Example 2 group was more effective in promoting the healing of diabetic wounds.

[0095] (2) Effect on foot wound healing

[0096] Please see Figure 10 and Figure 11 Different drug treatments revealed that on day 3, all diabetic rats showed varying degrees of redness and swelling in their paw wounds, while normal rats did not (the wound area was smaller than the control group). From day 6, the redness and swelling in the positive control group and the Example 2 group subsided, and the wound area was significantly different from the control group. At day 15, compared with the control group, the wound area in the positive control group was significantly reduced, and the wound area in the Example 2 group was significantly reduced. Notably, the Example 2 group was better than the positive control drug, human epidermal growth factor gel, in promoting the healing of diabetic foot skin; on days 9, 12, and 15, the wound area in the Example 2 group was smaller than that in the positive control group.

[0097] Please see Figure 12 HE staining revealed that in the skin of diabetic rats, only the sebaceous glands and hair follicles in the group of Example 2 showed significant recovery, while the recovery was slower in the other groups. Furthermore, the recovery of sebaceous glands and hair follicles in the group of Example 2 was only slightly worse than that in normal rats. This indicates that the treatment in the group of Example 2 not only promotes faster wound healing in diabetic foot ulcers but, more importantly, restores skin function at the wound site.

[0098] Experiment Example 3: The effect of a thermosensitive hydrogel on wound healing in mice with skin infections.

[0099] Six-week-old healthy male KM mice were routinely housed in a constant temperature environment of 22°C and 55% humidity. An infected wound model was established by creating full-thickness skin lesions (8 mm in diameter) on the backs of mice under anesthesia using a pore-forming device. 100 μL of activated Staphylococcus aureus bacterial solution with an OD value of 1 was instilled into each wound. If the surrounding area of ​​the wound began to redden and swell within 48 hours, and the mouse cried out in pain upon pressure, along with pus discharge from the wound, indicating infection, the inflammatory wound model was considered complete.

[0100] The patients were divided into a control group, a positive control group, and an experimental group. The control group received no treatment, the positive control group was coated with hEGF gel (human epidermal growth factor gel) from Guilin Huanowei Gene Pharmaceutical Co., Ltd., and the experimental group was coated with the drug provided in Example 2. Wound healing was recorded using a digital camera at 0, 3, 6, 9, and 12 days post-surgery, and the wound healing status was calculated using ImageJ software.

[0101] Please see Figure 13 and Figure 14 The experiment found that within 6 days after the successful preparation of the inflammatory wound model, there was no significant difference in the healing rate among the three groups of mice, and the changes in wound area were basically the same. However, after 6 days, the wound area of ​​the experimental group mice decreased significantly, and the wound healing rate was significantly different from the other two groups, while there was no significant difference between the control group and the positive drug group from beginning to end.

[0102] Please see Figure 15 HE staining revealed that, due to the inflammatory response at the wound site, subcutaneous tissue did not form in any of the three groups of mice after 6 days of treatment. After 12 days of treatment, epithelial tissue had formed in all three groups of wounds and separated from the scab. The epithelial healing in the positive control group and the experimental group was smoother than that in the control group. Compared with the positive control group, the experimental group showed a significant increase in the number of fibroblasts around the wound and a more significant recovery of hair follicle function. Therefore, the experimental group has a significant effect on promoting wound healing in inflammatory wounds and can significantly promote epithelial cell repair and hair follicle regeneration, with better effects than the positive control group using human epidermal growth factor gel.

[0103] In summary, the thermosensitive hydrogel for promoting wound healing provided in this application uses at least one of carotenoids and ginsenosides as active ingredients, which are combined with poloxamer 407, dopamine, and poloxamer 188 under alkaline conditions to form an assembly. Both carotenoids and ginsenosides possess various biological activities and free radical scavenging abilities, and can promote cell repair and stimulate TMEM16A channels to promote cell proliferation. Assembling these two active ingredients with poloxamer and dopamine increases their solubility, bioavailability, biological activity, and stability, thereby accelerating wound healing. Furthermore, the thermosensitive hydrogel for promoting wound healing provided in this application forms a gel at body temperature (37°C), enhancing the skin surface viscosity effect, and remains liquid at room temperature, facilitating storage and transportation. The thermosensitive hydrogel for promoting wound healing provided in this application can be widely used in the preparation of drugs that promote wound healing, which is beneficial for reducing wound area, promoting the recovery of skin function at the wound site, and promoting the recovery of sebaceous glands and hair follicles in the wound. The thermosensitive hydrogel for promoting wound healing provided in this application not only has a good healing effect on normal wounds, but also on wounds of diabetic patients and skin wounds infected with bacteria.

[0104] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A temperature-sensitive hydrogel that promotes wound healing, characterized in that, The raw materials, by mass percentage, include 0.1‰-1‰ of active ingredient, 1%-40% of poloxamer 407, 1%-10% of dopamine, 1%-20% of poloxamer 188, and the balance being an alkaline solvent; the active ingredient is lutein or ginsenoside Rb1, and the alkaline solution is an aqueous solution of NaOH with a pH of 8-9.

2. A method for preparing a temperature-sensitive hydrogel for promoting wound healing as described in claim 1, characterized in that, It includes: Mix the raw materials thoroughly; Mixing the raw materials evenly includes: Add the poloxamer 407 to the pre-cooled alkaline solvent and stir until clear; Next, add the active ingredient and continue stirring; The dopamine was then added and stirred to allow it to self-polymerize under alkaline conditions, forming an interpenetrating network with the poloxamer. Finally, poloxamer 188 is added so that the wound-healing thermosensitive hydrogel is liquid at room temperature and gel at body surface temperature. The mixing process of the raw materials was carried out at 3-5℃.

3. The method for preparing the wound-healing thermosensitive hydrogel according to claim 2, characterized in that, The viscosity of the thermosensitive hydrogel that promotes wound healing when it is in liquid form is 1 Pa·s-100 Pa·s.

4. The method for preparing the wound-healing thermosensitive hydrogel according to claim 2, characterized in that, The thermosensitive hydrogel that promotes wound healing has a viscosity of 1500 Pa·s-2000 Pa·s when it is in gel form.

5. The application of the wound-healing thermosensitive hydrogel prepared by the method of claim 1 or any one of claims 2-4 in the preparation of a wound-healing medicament.

6. The application according to claim 5, characterized in that, The promotion of wound healing includes promoting the reduction of wound area, promoting the recovery of skin function at the wound site, and promoting the recovery of sebaceous glands and hair follicles in the wound.

7. The application according to claim 5, characterized in that, The wounds mentioned in the context of promoting wound healing include wounds from non-diabetic patients or wounds from diabetic patients.

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