Composition for skin repair as well as preparation method and application thereof

Abstract

The invention relates to a composition for skin repair and a preparation method and application thereof, the composition comprises gallic acid or a derivative thereof and trehalose, and can be used for skin superficial repair; the composition further comprises Ectoine or a salt thereof and sodium hyaluronate, so that the promoting effect of the composition on skin wounds can be further improved, uncomfortable symptoms are effectively relieved, the skin absorptivity is improved, and the sensitization risk is reduced; the composition can be prepared into a preparation which is used for recovery of superficial wounds, surgical suture wounds, small wounds, scratches and incision wounds or used as a product after micro-plastic surgery, and has important clinical value and practical significance.

Description

Technical Field

[0001] This invention belongs to the field of skin repair technology, and in particular relates to a composition for skin repair, its preparation method and application. Background Technology

[0002] In the current field of cosmetic medicine, laser therapy and chemical peels have become mainstream methods for improving skin appearance and treating various skin problems, with widespread clinical applications and clear effects. Among them, medical laser therapy, utilizing the precise targeting of high-energy lasers, can act on specific layers of the skin, effectively improving skin imperfections such as pigmentation, acne scars, and wrinkles by breaking down pigment particles and stimulating collagen regeneration. Chemical peels, on the other hand, involve applying professionally concentrated glycolic acid, using chemical exfoliation to reduce the adhesion of keratinocytes, promoting rapid renewal of stratum corneum and superficial epidermal cells, and simultaneously stimulating collagen synthesis in the dermis, showing significant improvement effects on acne, melasma, and photoaging.

[0003] However, while these cosmetic procedures offer therapeutic benefits, they can also damage the skin barrier to varying degrees, leading to a series of adverse post-operative reactions that severely impact patients' treatment experience and quality of life. Specifically, after cosmetic laser treatments, the skin often experiences acute inflammatory reactions, manifesting as redness, swelling, heat, and pain; some patients may also experience peeling due to damage to the epidermis and disordered stratum corneum metabolism, accompanied by significant itching.

[0004] After a glycolic acid peel, the integrity of the skin's stratum corneum is damaged, and the barrier function is significantly weakened, resulting in abnormally fragile skin. Not only is it prone to inflammation such as redness, swelling, heat, and pain, but it can also cause dryness and peeling due to increased moisture loss. Furthermore, its sensitivity to external stimuli is greatly increased, and it may easily trigger adverse reactions such as allergies if not handled carefully.

[0005] While some repair products exist on the market to address the aforementioned post-operative skin problems, these products generally have limitations and fail to meet the actual needs of clinicians and consumers. Most repair products are unsatisfactory in their effects, failing to effectively treat or alleviate symptoms such as redness, itching, pigmentation, and inflammation. This results in a prolonged skin repair cycle and slow symptom relief, causing significant physical and psychological stress for patients. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides a composition for skin repair, its preparation method, and its application.

[0007] The technical solution adopted in this invention is: a composition for skin repair, comprising gallic acid or its derivatives, trehalose and ectoine.

[0008] Preferably, ectoine is replaced with ectoine salt, which is ectoine hydrochloride, ectoine sulfate, or ectoine phosphate.

[0009] Preferably, the gallic acid derivative is propyl gallate or bismuth subgallate.

[0010] Preferably, the mass fractions of gallic acid or its derivatives are 0.01-6, the mass fractions of trehalose are 0.01-6, and the mass fractions of ectoine or its salts are 0.01-5.

[0011] Preferably, it further includes sodium hyaluronate; wherein the mass fraction of gallic acid or its derivative is 0.01-6, the mass fraction of trehalose is 0.01-6, the mass fraction of ectoine or its salt is 0.01-5, and the mass fraction of sodium hyaluronate is 0.005-2.

[0012] An formulation for skin repair, comprising a composition for skin repair.

[0013] Preferably, the preparation is a paste dressing, a liquid dressing, or a patch dressing.

[0014] A method for preparing a skin repair formulation involves mixing the active ingredients of the composition with an emulsifier, emollient, thickener, preservative, and water to prepare a paste dressing.

[0015] Alternatively, a liquid dressing can be prepared by mixing the active ingredients of the composition with a humectant, a pH adjuster, a thickener, a preservative, and water.

[0016] Alternatively, the active ingredient in the composition can be mixed with absorbent and adhesive materials to prepare a patch dressing.

[0017] The application of compositions or preparations for skin repair in the repair of superficial wounds, including surgical suture wounds, small incisions, abrasions, or cuts.

[0018] The use of compositions or preparations for skin repair in products following minimally invasive cosmetic procedures.

[0019] Preferably, it is used as a post-treatment repair product for one or more of the following: laser, photon, chemical peel, mesotherapy, and microneedling.

[0020] The advantages and positive effects of this invention are as follows: It provides a composition that can be used for superficial skin repair. By combining the effects of gallic acid or its derivatives with trehalose, it can create a moist healing environment for superficial skin and promote skin recovery. The addition of ectoine (or its salt) and sodium hyaluronate to the composition can further enhance its promoting effect on skin wounds, effectively relieve discomfort symptoms, improve skin absorption, and reduce the risk of sensitization. This composition can be formulated into a preparation for the recovery of superficial wounds, surgical suture wounds, small wounds, abrasions, and cuts, or used as a postoperative product for minimally invasive cosmetic procedures, which has important clinical value and practical significance. Attached Figure Description

[0021] Figure 1 Effects of different compositions on morphological changes of wounds in mice;

[0022] Figure 2 Effects of different compositions on morphological changes of wounds in mice;

[0023] Figure 3 Effects of different compositions on wound healing rate in mice;

[0024] Figure 4 Expression of various mRNAs in mouse wound skin; where A: IL-1α, B: IL-1β, C: IL-6, D: CXCL1, E: CXCL2; compared with the blank control group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; compared with the positive control group, # P<0.05, ## P<0.01, ### P<0.001;

[0025] Figure 5 mRNA expression in mouse wounds after treatment with experimental groups 12, 15, 16, and 17;

[0026] Figure 6 mRNA expression in mouse wounds after treatment with experimental groups 13, 18, 19 and 20;

[0027] Figure 7 mRNA expression in mouse wounds after treatment with experimental groups 14, 21, 22 and 23;

[0028] Figure 8Changes in microvessel density of different compositions and their formulations in mouse ear-damaged skin; compared with the blank control group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; compared with the positive control group, #P<0.05, ##P<0.01, ###P<0.001;

[0029] Figure 9 Relative mRNA levels of VEGF-A in mouse ear lesions of different compositions and their formulations; compared with the blank control group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; compared with the positive control group, #P<0.05, ##P<0.01, ###P<0.001;

[0030] Figure 10 Diagrams showing tube formation in in vitro induced inflammatory cells by different compositions and their formulations;

[0031] Figure 11 Bar graph showing the changes in tube formation length in in vitro induced inflammatory cells by different compositions and their formulations; compared with the blank control group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; compared with the positive control group, #P<0.05, ##P<0.01, ###P<0.001. Detailed Implementation

[0032] The embodiments of the present invention will now be described with reference to the accompanying drawings.

[0033] This invention relates to a composition for skin repair, its preparation method, and its application. The composition comprises gallic acid or its derivatives, and trehalose. The composition can be used for superficial skin repair, and is particularly suitable for repairing superficial skin wounds, non-chronic wounds after laser, photon, and minimally invasive cosmetic procedures.

[0034] Lasers and photons, through photothermal effects; fruit acids, through chemical peels; and mesotherapy and microneedling, through physical punctures, all create micro-wounds or irritations on the skin's surface or superficial layers. These irritations disrupt the skin's natural homeostasis, releasing "trauma" signals to skin cells. These signals initially trigger a transient inflammatory response, which mobilizes immune cells to clear damaged tissue and releases various growth factors. These growth factors further stimulate keratinocytes to accelerate division and migration, repairing superficial wounds and achieving epidermal renewal. Simultaneously, they activate fibroblasts in the dermis, prompting them to produce more collagen and elastin fibers, completing the structural repair and remodeling of the dermis, ultimately improving skin texture, elasticity, and overall appearance.

[0035] Gallic acid or its derivatives possess signal inhibition, cell regulation, and anti-inflammatory and antioxidant effects. Trehalose, one of the most stable natural disaccharides, effectively maintains the stability and integrity of intracellular biomembranes, proteins, and bioactive peptides under adverse conditions, earning it the title of "sugar of life." It has wide applications in various industries, including biopharmaceuticals, pharmaceuticals, food, and cosmetics. Trehalose has strong moisturizing properties and can be added to face masks and liquid dressings as a humectant and protectant. Combining the effects of gallic acid or its derivatives with trehalose, a protective film is formed on the wound surface, protecting it and providing a moist healing environment, thereby promoting rapid superficial skin repair and effectively alleviating discomfort caused by minimally invasive procedures. Compositions containing gallic acid or its derivatives and trehalose can create a moist healing environment for superficial skin, promoting skin recovery, accelerating the inflammatory response rate, effectively relieving discomfort, and improving skin absorption.

[0036] In this composition, gallic acid can be used as the active ingredient, or a gallic acid derivative can be used instead. The gallic acid derivative can be propyl gallate or bismuth subgallate. In the composition, the mass fraction of gallic acid or its derivative is 0.01-6, and the mass fraction of trehalose is 0.01-6.

[0037] In some embodiments of the present invention, the composition further includes ectoine or its salts and / or sodium hyaluronate, which can further enhance the repair properties of the composition and effectively reduce or alleviate the risks of redness, swelling, and infection. Ectoine, as a small organic molecule, is widely found in aerobic, chemoheterotrophic, and halophilic microorganisms. As an osmotic pressure compensating solute, ectoine can link with biomolecules such as proteins through hydrogen bonds at dehydrated sites, forming a new protective film to replace the original water film. This protects the hydrogen bond sites from direct exposure to the surrounding environment, allowing biomolecules to maintain their natural structure and functional properties even under dehydrated conditions. Ectoine salts are ectoine hydrochloride, ectoine sulfate, or ectoine phosphate. Unlike other moisturizers that are short-lasting and dependent, the relatively small molecular weight of ectoine allows it to penetrate deep into the epidermis, enhancing the skin's water-locking capacity and mitigating rapid water loss caused by skin barrier damage. It has excellent long-lasting moisturizing and anti-drying effects. Sodium hyaluronate is a high molecular weight polymer with excellent moisturizing properties. It has an extremely strong water absorption capacity, with each molecule capable of absorbing hundreds of times its own weight in water. This results in significant effects on maintaining skin hydration and improving skin elasticity. At the same time, it has a strong lubricating and film-forming property, and is widely used in the fields of beauty, medicine, and health care.

[0038] When the composition includes gallic acid or its derivatives, trehalose, and ectoine (or its salt), the mass fractions of gallic acid or its derivatives are 0.01-6, the mass fractions of trehalose are 0.01-6, and the mass fractions of ectoine (or its salt) are 0.01-5; when the composition includes gallic acid or its derivatives, trehalose, ectoine (or its salt), and sodium hyaluronate, the mass fractions of gallic acid or its derivatives are 0.01-6, the mass fractions of trehalose are 0.01-6, the mass fractions of ectoine (or its salt) are 0.01-5, and the mass fractions of sodium hyaluronate are 0.005-2. In some embodiments of the present invention, the composition may include, by weight parts, 0.01, 0.05, 0.1, 0.3, 0.5, 0.7, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, or 6 parts of gallic acid or its derivatives, and 0.01, 0.05, 0.1, 0.3, 0.5, 0.7, 0.9, 1, 1 0.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5 or 6 parts; 0.01, 0.05, 0.1, 0.3, 0.5, 0.7, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5 or 5 parts; 0.005, 0.01, 0.05, 0.1, 0.5, 1.5 or 2 parts of sodium hyaluronate.

[0039] To better utilize the effects of each active ingredient, the above composition can be formulated into a preparation, such as a cream dressing, liquid dressing, or patch dressing. When the composition is formulated into a cream dressing, it also includes one or more of emulsifiers, emulsifier auxiliaries, emollients, thickeners, preservatives, and water; when the composition is formulated into a liquid dressing, it also includes one or more of humectants, pH adjusters, thickeners, preservatives, and water; when the composition is formulated into a patch dressing, it also includes absorbent materials and / or adhesive substances. The excipients in the formulation can be selected from different raw materials according to existing processes. Different types of excipients can be mixed with the active ingredients in the composition to prepare the formulation according to the following selection. The emollient and moisturizer can be glycerin and / or propylene glycol; the preservative can be hydroxypropyl methyl ester and / or phenoxyethanol; the emulsifier is one or more of lactic acid fatty acid glycerides, ES-5300, glyceryl stearate, and PEG-100 stearate; the emulsifier auxiliaries are one or more of cocoa butter, caprylic / capric triglycerides, cetearyl alcohol, and beeswax; the thickener is one or more of sodium carboxymethyl cellulose, carbomer, and hydroxyethyl cellulose; the pH adjuster is sodium hydroxide and / or triethanolamine; the water-absorbing material can be sodium carboxymethyl cellulose, hydroxyethyl cellulose, or sodium alginate; and the viscous substance includes one or two of thermoplastic rubber and hot melt adhesive.

[0040] The above-described compositions or preparations can be used for the repair of superficial wounds, including surgical sutures, small abrasions, and cuts. The compositions or preparations are particularly suitable for post-operative care following minimally invasive cosmetic procedures, such as laser treatments, phototherapy, chemical peels, mesotherapy, and microneedling. The compositions or preparations precisely target superficial skin problems caused by laser treatments, chemical peels, and other cosmetic procedures, including redness, swelling, heat, pain, peeling, itching, dryness, scaling, and increased sensitivity. They can be used to treat skin barrier damage or superficial wounds, including surgical sutures, small abrasions, and cuts, or as post-operative care following laser / phototherapy / chemical peels / mesotherapy / microneedling / minimally invasive cosmetic procedures. They effectively relieve discomfort, improve skin absorption, and reduce the risk of sensitization, demonstrating significant clinical value and practical importance.

[0041] The quaternary active system composed of gallic acid derivatives, trehalose, ectoine (or its salts), and sodium hyaluronate is not a simple superposition of the effects of individual components. Instead, it forms a closed-loop network of action—"protection-repair-anti-inflammatory-moisturizing"—through a multi-target, multi-pathway synergistic mechanism. This network mutually empowers the core aspects of skin repair, significantly improving the healing efficiency of superficial skin wounds and the quality of barrier reconstruction. The composition promotes wound healing while simultaneously building the skin barrier, providing a relatively moist and stable environment for the healing process.

[0042] Sodium hyaluronate's "hydration gel barrier" and trehalose's "molecular protective film" work synergistically to lock in moisture and protect wounds. Sodium hyaluronate, as a high-molecular polymer, can absorb hundreds of times its own weight in water, forming a transparent, moist gel-like film on the wound surface. This reduces wound moisture evaporation and maintains the moist microenvironment necessary for healing (a moist environment can increase wound healing speed by more than 50%). It also physically isolates external dust and bacteria, thus reducing the risk of infection. Trehalose, as a natural disaccharide, can bind to proteins and lipids on the surface of wound epidermal cells through hydrogen bonds, forming a dense molecular protective film on the extracellular layer. This compensates for the shortcomings of sodium hyaluronate's gel film in "small molecule permeability protection." Especially when the skin barrier is damaged, leading to widening of intercellular spaces, trehalose can embed itself in the gaps and stabilize the cell membrane structure, preventing cell rupture due to osmotic pressure imbalance. This complements sodium hyaluronate's "macro-micro" protective barrier.

[0043] Gallic acid derivatives can reduce the production of inflammatory mediators such as prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) by inhibiting the activity of cyclooxygenase (COX-2) and lipoxygenase (LOX), thereby alleviating wound inflammation at its source and preventing tissue damage caused by excessive inflammatory activation. Simultaneously, its phenolic hydroxyl structure can scavenge reactive oxygen species (ROS) generated during wound healing, reducing the damage of oxidative stress to newly formed cells. Ectoin, as an "osmolarity-compensating solute," can bind to intracellular proteins and nucleic acids through a "hydration shell" mechanism when wound cells are in a state of dehydration and inflammatory stress, maintaining the natural conformation of biomolecules. While gallic acid derivatives inhibit inflammation, ectoin can protect fibroblasts, epidermal stem cells, and other repair-related cells in the wound from stress damage, ensuring the activity and function of repair cells. The two form a synergistic "anti-inflammatory-cell-protective" chemical protection, laying the cellular foundation for subsequent repair. Furthermore, after skin injury, the wound site releases large amounts of pro-inflammatory factors such as IL-1α, IL-1β, and IL-6, recruiting neutrophils and macrophages to aggregate and triggering acute inflammation. Gallic acid derivatives' "inhibition of inflammatory factors" and ectoine's "regulation of immune cell chemotaxis" synergistically reduce inflammation. Gallic acid derivatives can directly inhibit the transcription and release of pro-inflammatory factors by downregulating the NF-κB signaling pathway (the core regulatory pathway for pro-inflammatory factor expression); while ectoine regulates the expression of chemokines CXCL1 and CXCL2 (experimental data show that the expression levels of CXCL1 / CXCL2 in the quaternary system group were reduced by more than 60% compared to the control group, and lower than the single-component group), avoiding the "cytokine storm" caused by excessive aggregation of immune cells. Under the synergistic effect of gallic acid or its derivatives and ectoine, acute inflammatory responses can be rapidly controlled while maintaining appropriate immune cell infiltration (e.g., macrophages can secrete transforming growth factor β (TGF-β), promoting fibroblast proliferation), achieving a balance of "controllable inflammation and orderly repair."

[0044] Ectoin's "deep hydration" and sodium hyaluronate's "surface moisturization" work together to create a gradient moisturizing network: Sodium hyaluronate mainly acts on the skin's surface (stratum corneum), achieving "instant moisturization" through a gel film, but it is easily lost due to factors such as sweat and friction; while ectoin, as a small molecule (molecular weight of approximately 221 Da), can penetrate the stratum corneum and enter the epidermis, binding with keratin in keratinocytes and enhancing intercellular hydration; its unique "osmotic pressure compensation" mechanism can form a "moisture reservoir" inside the skin, slowly releasing moisture to replenish the moisturizing effect of sodium hyaluronate on the surface.

[0045] Gallic acid derivatives, through their "lipid synthesis promotion," and trehalose, through its "cell junction repair," synergistically rebuild the skin barrier. The core of the skin barrier is the "brick-and-mortar" structure of the stratum corneum (keratinocytes are the "bricks," and intercellular lipids are the "mortar"). Gallic acid derivatives can activate peroxisome proliferator-activated receptor (PPAR-γ), promoting the synthesis of intercellular lipids such as ceramides and cholesterol by keratinocytes, thus replenishing the "mortar." Meanwhile, trehalose promotes the expression and assembly of tight junction proteins (such as occludin and claudin-1) between keratinocytes, enhancing the stability of the connections between the "bricks." The synergistic effect of gallic acid and trehalose not only rapidly replenishes barrier components but also optimizes the integrity of the barrier structure, reducing the skin's sensitivity to external stimuli after repair (e.g., shortening the duration of erythema after laser treatment by 30%). Meanwhile, the gel membrane of sodium hyaluronate can provide a "quiet microenvironment" for lipid synthesis and tight junction repair, while ectoine maintains the activity of keratinocytes, further enhancing this synergistic effect and forming a closed loop of barrier reconstruction of "component replenishment - structural repair - environmental protection".

[0046] Analysis of the characteristics of each active component revealed that gallic acid or its derivatives, trehalose, ectoine (or its salt), and sodium hyaluronate can combine and complement each other in promoting repair and protecting the skin, thus providing a suitable repair environment and promoting the repair of superficial skin. Experiments using formulations with different components yielded similar conclusions: formulations containing the four active ingredients showed higher repair and healing rates, inhibited the accumulation of inflammatory factors, and suppressed the formation of new microvessels.

[0047] The present invention will now be described with reference to the accompanying drawings. Experimental methods not specifically described in the accompanying drawings are performed according to the corresponding product instructions. Unless otherwise specified, the instruments, reagents, and consumables used in the examples can be purchased from commercial companies. Specifically, ectoine hydrochloride is (S)-2-methyl-3,4,5,6-tetrahydropyrimidine-4-carboxylate hydrochloride, ectoine sulfate is (S)-2-methyl-3,4,5,6-tetrahydropyrimidine-4-carboxylate sulfate, and ectoine phosphate is (S)-2-methyl-3,4,5,6-tetrahydropyrimidine-4-carboxylate phosphate.

[0048] Example 1: Ointment containing bismuth subgallate

[0049] Bismuth subgallate and petrolatum were mixed at a mass ratio of 1:20 and heated and stirred at a temperature of 40℃-45℃ and a stirring speed of 30-45r / min for 15-20min. Then, the mixture was gradually cooled to 30℃ while stirring to obtain the preparation of experimental group 1.

[0050] Example 2: Ointment containing gallic acid

[0051] Gallic acid and petrolatum were mixed at a mass ratio of 1:20 and heated and stirred at a temperature of 40℃-45℃ and a stirring speed of 30-45 r / min for 15-20 min. Then, the mixture was gradually cooled to 30℃ while stirring to obtain preparation 2 of experimental group.

[0052] Example 3: Ointment containing propyl gallate

[0053] Propyl gallate and petrolatum were mixed at a mass ratio of 1:20 and heated and stirred at a temperature of 40℃-45℃ and a stirring speed of 30-45r / min for 15-20min. Then, the mixture was gradually cooled to 30℃ while stirring to obtain preparation 3 of experimental group.

[0054] Example 4: Paste formulation containing trehalose

[0055] Trehalose and petrolatum were mixed at a mass ratio of 1:20 and heated and stirred at a temperature of 40℃-45℃ and a stirring speed of 30-45r / min for 15-20min. Then, the mixture was gradually cooled to 30℃ while stirring to obtain preparation 4 of the experimental group.

[0056] Example 5: Ointment dressing containing bismuth subgallate and trehalose

[0057] Components A and B were prepared separately. Component A: Trehalose, glycerol, methylparaben, and purified water were mixed in a mass ratio of 1:5:0.2:43.8, heated and stirred until dissolved and homogeneous at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A, B, and bismuth subgallate were mixed and homogenized in a mass ratio of 50:42:8 for 5 minutes, then cooled to obtain preparation 5 of experimental group.

[0058] Components A and B were prepared separately. Component A: Trehalose, glycerol, methylparaben, and purified water were mixed in a mass ratio of 0.5:5:0.2:44.3, heated and stirred until dissolved and homogeneous at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:17:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A, B, and bismuth hypogallate were mixed and homogenized in a mass ratio of 50:49:1 for 5 minutes, then cooled to obtain the experimental group 5-1 formulation.

[0059] Example 6: Ointment dressing containing gallic acid and trehalose

[0060] Components A and B were prepared separately. Component A: Gallic acid, trehalose, glycerol, methylparaben, and purified water were mixed in a mass ratio of 0.1:1:5:0.2:45.7, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and then cooled to obtain preparation 6 of experimental group.

[0061] Components A and B were prepared separately. Component A: Gallic acid, trehalose, glycerol, methylparaben, and purified water were mixed in a mass ratio of 6:1:5:0.2:45.8, heated and stirred until dissolved and homogeneous at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 58:42 for 5 minutes, and then cooled to obtain the experimental group 6-1 formulation.

[0062] Example 7: Ointment dressing containing propyl gallate and trehalose

[0063] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, glycerol, methylparaben, and purified water were mixed in a mass ratio of 0.1:1:5:0.2:45.7, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and then cooled to obtain preparation 7 of experimental group.

[0064] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, glycerol, methylparaben, and purified water were mixed in a mass ratio of 6:1:5:0.2:45.8, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 58:42 for 5 minutes, and then cooled to obtain the experimental group 7-1 formulation.

[0065] Example 8: Ointment containing ectoine

[0066] Ectoin and petrolatum were mixed at a mass ratio of 1:20 and heated and stirred at a temperature of 40℃-45℃ and a stirring speed of 30-45r / min for 15-20min. Then, the mixture was gradually cooled to 30℃ while stirring to obtain preparation 8 of the experimental group.

[0067] Example 9: Ointment dressing containing gallic acid, trehalose and ectoine

[0068] Components A and B were prepared separately. Component A: Gallic acid, trehalose, ectoine, glycerin, methylparaben, and purified water were mixed in a mass ratio of 0.1:1:0.1:5:0.2:45.6, heated and stirred until dissolved uniformly at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and then cooled to obtain preparation 9 of experimental group.

[0069] Components A and B were prepared separately. Component A: Gallic acid, trehalose, ectoine, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:2:5:0.2:43.8, heated and stirred until dissolved and homogeneous at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 58:42 for 5 minutes, and then cooled to obtain the experimental group 9-1 formulation.

[0070] Example 10: Ointment dressing containing propyl gallate, trehalose and ectoine

[0071] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, ectoine, glycerin, methylparaben, and purified water were mixed in a mass ratio of 0.1:1:0.1:5:0.2:45.6, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and cooled to room temperature to obtain formulation 10 of experimental group.

[0072] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, ectoine, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:2:5:0.2:43.8, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 58:42 for 5 minutes, and cooled to room temperature to obtain the experimental group 10-1 formulation.

[0073] Example 11: Ointment dressing containing gallic acid, trehalose, sodium hyaluronate and ectoine

[0074] Components A and B were prepared separately. Component A: Gallic acid, trehalose, ectoine, sodium hyaluronate, glycerin, methylparaben, and purified water were mixed in a mass ratio of 0.1:1:0.1:0.01:5:0.2:45.59, heated and stirred until dissolved uniformly at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and cooled to room temperature to obtain preparation 11 of experimental group.

[0075] Components A and B were prepared separately. Component A: Gallic acid, trehalose, ectoine, sodium hyaluronate, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:2:0.5:5:0.2:43.3, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 58:42 for 5 minutes, and cooled to room temperature to obtain formulation 11-1.

[0076] Example 12: Ointment dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine

[0077] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, ectoine, sodium hyaluronate, glycerin, methylparaben, and purified water were mixed in a mass ratio of 0.1:1:0.1:0.01:5:0.2:45.59, heated and stirred until dissolved uniformly at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and cooled to room temperature to obtain preparation 12 of the experimental group.

[0078] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, ectoine, sodium hyaluronate, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:2:0.5:5:0.2:43.3, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:10:20, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 58:42 for 5 minutes, and cooled to room temperature to obtain the experimental group 12-1 formulation.

[0079] Example 13: A patch dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine

[0080] Propyl gallate, trehalose, ectoine, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, carbomer, triethanolamine, and purified water were mixed in a mass ratio of 6:1:1.5:0.5:2:10:0.2:0.2:0.01:78.59. The mixture was heated and stirred until it was evenly dissolved. The heating temperature was 80℃-90℃, and the mixture was heated for 30-40 minutes. After cooling to room temperature, the inner material of the dressing was obtained. The inner material was combined with non-woven fabric to form the experimental group 13 formulation.

[0081] Example 14: Liquid dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine

[0082] Propyl gallate, trehalose, ectoine, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:1.5:0.5:0.2:10:0.2:80.6. The mixture was heated and stirred until it was evenly dissolved. The heating temperature was 80℃-90℃, and the mixture was heated for 30-40 minutes. After cooling to room temperature, the liquid dressing was obtained, which was the experimental group 14 preparation.

[0083] Example 15: Ointment dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine hydrochloride Components A and B were prepared separately. Component A: Propyl gallate, trehalose, sodium hyaluronate, ectoine hydrochloride, glycerin, methylparaben, and purified water were mixed in a mass ratio of 0.2:1:0.2:0.5:5:0.2:45.6, heated and stirred until uniformly dissolved at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and cooled to room temperature to obtain the experimental group 15 formulation.

[0084] Example 16: Ointment dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine sulfate

[0085] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, sodium hyaluronate, ectoine sulfate, glycerin, methylparaben, and purified water were mixed in a mass ratio of 0.2:1:0.2:0.4:5:0.2:45.6, heated and stirred until dissolved uniformly at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and cooled to room temperature to obtain the experimental group 16 formulation.

[0086] Example 17: Ointment dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine phosphate

[0087] Components A and B were prepared separately. Component A: Propyl gallate, trehalose, sodium hyaluronate, ectoine phosphate, glycerol, methylparaben, and purified water were mixed in a mass ratio of 0.2:1:0.2:1:5:0.2:45.6, heated and stirred until dissolved uniformly at 80℃-90℃ for 30-40 minutes. Component B: Emulsifier PEG-100 stearate, propylene glycol, cocoa butter, and caprylic / capric triglycerides were mixed in a mass ratio of 8:4:12:24, heated and stirred until melted at 80℃-90℃ for 30-40 minutes. Components A and B were then mixed and homogenized in a mass ratio of 52:44 for 5 minutes, and cooled to room temperature to obtain the experimental group 17 formulation.

[0088] Example 18: A patch dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine hydrochloride

[0089] Propyl gallate, trehalose, ectoine hydrochloride, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, carbomer, triethanolamine, and purified water were mixed in a mass ratio of 6:1:0.5:0.5:2:10:0.2:0.2:0.015:78.59. The mixture was heated and stirred until it dissolved evenly at 80℃-90℃ for 30-40 minutes. After cooling to room temperature, the inner material of the dressing was obtained. The inner material was then combined with nonwoven fabric to obtain the experimental group 18 formulation.

[0090] Example 19: A patch dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine sulfate

[0091] Propyl gallate, trehalose, ectoine sulfate, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, carbomer, triethanolamine, and purified water were mixed in a mass ratio of 6:1:0.4:0.5:2:10:0.2:0.2:0.015:78.59. The mixture was heated and stirred until it dissolved evenly at 80℃-90℃ for 30-40 minutes. After cooling to room temperature, the inner material of the dressing was obtained. The inner material was then combined with nonwoven fabric to obtain the experimental group 19 formulation.

[0092] Example 20: A patch dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine phosphate

[0093] Propyl gallate, trehalose, ectoine phosphate, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, carbomer, triethanolamine, and purified water were mixed in a mass ratio of 6:1:1:0.5:2:10:0.2:0.2:0.015:78.59. The mixture was heated and stirred until it dissolved evenly at 80℃-90℃ for 30-40 minutes. After cooling to room temperature, the inner material of the dressing was obtained. The inner material was then combined with nonwoven fabric to obtain the experimental group 20 formulation.

[0094] Example 21: Liquid dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine hydrochloride

[0095] Propyl gallate, trehalose, ectoine hydrochloride, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:0.3:0.5:0.2:10:0.2:80.6. The mixture was heated and stirred until it was evenly dissolved. The heating temperature was 80℃-90℃ for 30-40 minutes. After cooling to room temperature, the product was obtained, which is the preparation of experimental group 21.

[0096] Example 22: Liquid dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine sulfate

[0097] Propyl gallate, trehalose, ectoine sulfate, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:0.3:0.5:0.2:10:0.2:80.6. The mixture was heated and stirred until it was evenly dissolved. The heating temperature was 80℃-90℃, and the mixture was heated for 30-40 minutes. After cooling to room temperature, the product was obtained, which is the experimental group 22 preparation.

[0098] Example 23: Liquid dressing containing propyl gallate, trehalose, sodium hyaluronate and ectoine phosphate

[0099] Propyl gallate, trehalose, ectoine phosphate, sodium hyaluronate, sodium carboxymethyl cellulose, glycerin, methylparaben, and purified water were mixed in a mass ratio of 6:1:0.5:0.5:0.2:10:0.2:80.6. The mixture was heated and stirred until it was evenly dissolved. The heating temperature was 80℃-90℃, and the mixture was heated for 30-40 minutes. After cooling to room temperature, the product was obtained, which is the preparation of experimental group 23.

[0100] The effects of the preparations prepared in Examples 1-23 were verified by grouping. When comparing multiple preparations, the mass fraction of gallic acid or its derivatives in each preparation was the same. Among the preparations prepared in Examples 5, 6, 7, 9, 10, 11, and 12, preparations in experimental groups 5, 6, 7, 9, 10, 11, and 12 were selected for verification and comparison.

[0101] Example 24: Application of the composition in post-laser skin repair

[0102] 24.1 Selection of patients for facial laser treatment and treatment methods using combination therapy

[0103] The study included 360 patients who had just undergone laser cosmetic surgery, with no gender restriction. There were 90 males and 270 females, aged 18 to 60 years. They were randomly divided into 12 groups of 30 patients each. There were no significant differences in the basic characteristics of the patients among the groups (P>0.05), which allowed for comparison.

[0104] After cleaning the affected areas daily, patients in each group used the preparations obtained in Examples 1-12, applying them twice daily, morning and evening, for a total of 2 weeks. The single application amount was 0.5-1g, evenly covering the entire face with a thickness of approximately 0.1mm.

[0105] 24.2 The skin condition of each group of patients was scored after treatment.

[0106] The scoring criteria are shown in Table 1.

[0107] Table 1. Skin Symptom Scoring Criteria

[0108]

[0109] The efficacy evaluation criteria are divided into four levels: cured, significantly effective, effective, and ineffective. The efficacy index is calculated as follows: [(pre-treatment score - post-treatment score) ÷ pre-treatment score] × 100%. Among them, an efficacy index ≥ 95% indicates a cured condition; an efficacy index of 70%-94% indicates a significantly effective condition; an efficacy index of 30%-69% indicates an effective condition; and an efficacy index < 30% indicates an ineffective condition.

[0110] Patients in each group using experimental preparations 1-12 were scored and their efficacy index was calculated. The results are shown in Table 2.

[0111] Table 2

[0112]

[0113] 24.3 Transepidermal water loss and stratum corneum moisture content were measured in patients in each group after two weeks of the above treatment.

[0114] Transepidermal water loss and stratum corneum moisture content were measured before and after treatment. The instruments used for testing were a transepidermal water loss tester and a stratum corneum moisture content (SCWC) measuring instrument. The results are shown in Figure 3.

[0115] Table 3

[0116]

[0117] The above data reflects the skin condition of patients treated with laser therapy on the face after using different types of formulations. Table 2 shows that experimental groups 12, 11, and 10 showed the best results. When the formulation contains 3-4 active ingredients, due to the synergistic effect of each active component, even with a reduced amount of active ingredient, the formulation with multiple types of active substances is significantly more effective than the formulation with a single active ingredient. Table 3 shows that, in terms of transdermal water loss (gm³),... -2 .h -1 In the detection of ) and stratum corneum moisture content (au), experimental groups 11 and 12 also showed the best results.

[0118] Example 25: Application of the composition in wound skin repair

[0119] 25.1 Construction of a mouse model of wound healing and use of the combined drug

[0120] Healthy male SPF clean-grade Balb / c mice, 6-8 weeks old, were selected. The Balb / c mice were randomly divided into 25 groups, including 23 experimental groups, one blank control group, and one positive control group. After acclimatization, the back hair of the mice was removed using an electric shaver and depilatory cream. The mice were observed after shaving. One day after hair removal, the following procedure was performed: After the mice were stretched and fasted, they were anesthetized via intraperitoneal injection. A silicone ring with an inner diameter of 10 mm and an outer diameter of 16 mm was fixed to the skin in the middle of the mouse's back using sutures, symmetrically distributed along the midline, with a 1-2 cm gap between the two rings to ensure no overlap. After fixing the silicone ring, the skin inside the ring was disinfected with iodine. A 6 mm diameter punch was used to make a hole in the skin inside the ring on the mouse's back. The circular skin was then cut off with scissors down to the myofascia. The wound was then hemostatically stopped and disinfected.

[0121] In this study, mice in the experimental group were administered the formulations prepared in Examples 1-23, respectively. The blank control group received no treatment, and the positive control group received 5% panthenol. The dosage for each group was 30 μl per administration. The wounds of mice in the experimental and positive control groups were first cleaned with physiological saline, and then the different formulations were applied and fixed with polyurethane film. The dressings were changed every 2 days, and the wound healing was observed.

[0122] After a wound forms, fibroblasts differentiate from granulation tissue at the wound site to pull the wound edges together, while epidermal stem cells differentiate into epidermal cells for epidermal repair and reconstruction. The rate of wound closure slows down as the wound surface is filled with new tissue, epidermal repair is nearing completion, and the barrier separating the internal and external environments has been re-established. At this point, the wound can be considered closed. The red appearance of the wound, visible to the naked eye, is due to the incomplete maturation of the newly formed stratum corneum. The most direct indicator of wound healing is the healing rate, which clearly demonstrates the healing status and is often used as an important indicator for evaluating the efficacy of related drugs. On days 0, 4, 8, 10, and 15 of the model, photos were taken using a mobile phone and ruler to record and observe the dynamic changes of the skin wound on the rat's back. The results are as follows: Figure 1 and Figure 2 As shown. According to Figure 1 Figure 2 The wound area was analyzed using ImageJ software to calculate the wound healing rate. Wound healing rate % = (Original wound area - Post-treatment wound area) / Original wound area. Results are as follows... Figure 3 As shown.

[0123] according to Figure 1 and Figure 2As shown, on day 8 after modeling, the wound area of ​​mice in each experimental group and the positive control group gradually decreased, and the healing rate also showed a gradual upward trend. There was no infection or suppuration. Among them, the blank control group began to show scab formation, which was dark red, and the healing rate was relatively low. On day 10 after modeling, the scabs on the wounds of the blank control group continued to harden, the surface of the skin tissue around the wound was uneven, and the skin at the wound edge contracted. At the same time, no scab formation was observed in each experimental group and the positive control group, and the wound area further decreased. Among them, the wounds in experimental groups 5, 6, 7, 9, 10, 11, 12, 13, and 14 recovered faster than those in the positive control group, and the healing rate was faster. By day 15, the wound healing speed of the blank control group was slower, while the epithelialization of each experimental group and the control group was basically completed. Among them, the healing speed of experimental group 12 was faster, showing flesh-colored new skin with complete new flesh-colored tissue, and the healing effect was more obvious.

[0124] 25.2 Effects of different compositions and their formulations on the mRNA expression of IL-1α, IL-1β, IL-6, CXCL1 and CXCL2 in mouse wounds

[0125] Wound healing is a highly coordinated and dynamic biological process, typically divided into four overlapping phases: hemostasis, inflammation, proliferation, and remodeling. The inflammatory response is a crucial part of the healing process. The production and aggregation of inflammatory cells are controlled to some extent by inflammatory factors and chemokines, while inflammatory cells, in turn, can produce or influence the body's production of inflammatory factors and chemokines. These cytokines, like inflammatory cells, play important roles in the inflammatory response phase of wound healing. Key inflammatory factors include IL-1, IL-1β, and IL-6, while chemokines (CXCL1 and CXCL2), as chemotactic cytokines, play a recruitment role and can also participate in regulating the inflammatory response. The expression of IL-1α, IL-1β, IL-6, CXCL1, and CXCL2 in the skin of a mouse model was detected using quantitative real-time PCR.

[0126] Fifteen days after modeling, mice in each group were euthanized by cervical dislocation. Skin tissue from the dorsal lesions was cut using sterile scissors, quickly collected, and labeled. The tissue was then rapidly frozen in liquid nitrogen for 1 hour and transferred to a -80°C freezer for later use in experiments. A suitable amount of skin was placed in a 2ml RNase-free EP tube, with 500μl of Trizol reagent and a suitable amount of grinding beads added to each tube. The mixture was rapidly homogenized using a tissue homogenizer. After the foam disappeared, the mixture was centrifuged at 12000 rpm for 15 minutes at 4°C. The supernatant was transferred to a new RNase-free EP tube, and 100μl of chloroform was added. The mixture was rapidly vortexed and allowed to stand for 10 minutes, followed by centrifugation at 12000 rpm for 15 minutes at 4°C. The upper aqueous phase from 200μl of the EP tube was slowly transferred to a new RNase-free EP tube, and 200μl of isopropanol was added. The mixture was inverted and mixed thoroughly, allowed to stand for 10 minutes, and then centrifuged at 12000 rpm for 10 minutes at 4°C. Pour out the liquid, invert and air dry, then add 200 μl of 75% anhydrous ethanol to the tube, shake well, centrifuge at 12000 r for 5 min at 4 °C, discard the supernatant, allow the precipitate to dry at room temperature in a clean bench, and add 20 μl of DEPC water to dissolve the precipitate.

[0127] RNA was reverse transcribed into cDNA using a reverse transcription kit. GAPDH was used as an internal control, SYBR Green was used as the dye, and the cDNA was used as a template for quantitative real-time PCR amplification. The reaction volume was 20 μL, and the amplification conditions were: 95℃ pre-denaturation for 10 min; 95℃ denaturation for 15 s, 60℃ annealing for 1 min, 40 amplification cycles; melting curve amplification: 95℃ for 5 s, 60℃ for 5 s; extension at 95℃ for 5 s. The results were calculated using 2... －ΔΔCt Relative expression levels were analyzed to compare the differences in IL-1α, IL-1β, IL-6, CXCL1, and CXCL2 expression among the groups. Results are as follows: Figure 4 As shown.

[0128] like Figure 4 As shown, the mRNA levels of IL-1α, IL-1β, IL-6, CXCL1, and CXCL2 in the blank control group were abnormally high. Compared with the blank control group, the levels of the above mRNAs in each experimental group and the positive control group were significantly reduced to varying degrees (P<0.05, P<0.01, P<0.001, P<0.0001). Compared with the positive control group, experimental groups 5, 6, 7, 9, 10, 11, 12, 13, and 14 significantly inhibited the expression of IL-1α, IL-1β, IL-6, CXCL1, and CXCL2 (P<0.05, P<0.01, P<0.001), with experimental group 12 showing the best effect. This indicates that it can significantly inhibit the accumulation of inflammatory factors, thereby promoting rapid wound healing.

[0129] The above experimental results revealed significant differences in the efficacy of different compositions and formulations in promoting wound healing. Quantitative analysis and statistical comparison were performed on key indicators such as wound healing rate and related inflammatory factors. Experimental groups 11-14, containing gallic acid or its derivatives, trehalose, ectoine, and sodium hyaluronate, showed better results. Among these groups, experimental group 12, containing propyl gallate ointment dressings, demonstrated the best effect, indicating that ointment formulations are more advantageous in promoting wound healing. Based on the comprehensive test indicators, the efficacy ranking of the experimental groups was as follows: Experimental group 12 >> Experimental group 13 > Experimental group 14 > Experimental group 11 > Experimental group 10 > Experimental group 9 > Experimental group 7 > Experimental group 6 > Experimental group 5 > Positive control group > Experimental group 3 > Experimental group 2 > Experimental group 4 ≈ Experimental group 8 > Experimental group 1 > Blank control group.

[0130] The composition in experimental group 12 was significantly superior to that in experimental groups 7 and 10 in promoting wound healing and inhibiting inflammatory factors, indicating that the composition containing all four active substances produced a significant synergistic effect. While gallic acid derivatives, trehalose, and ectoine alone have some effect on wound healing, they are far less effective than any combination of two or three substances, and even less effective than the composition containing all four active substances.

[0131] like Figure 5 As shown, experimental groups 12, 15, 16, and 17 were all ointment dressings. Compared with experimental group 12, experimental group 17 showed a significant decrease in the mRNA levels of IL-1α and IL-1β (P<0.05), while there was no significant difference between experimental groups 15 and 16. Figure 6 As shown, experimental groups 13, 18, 19, and 20 were all patch dressings. Compared with experimental group 13, experimental group 20 showed a significant decrease in mRNA levels of IL-1α and IL-1β (P<0.05), while there was no significant difference between experimental groups 18 and 19. Figure 7 As shown, experimental groups 14, 21, 22, and 23 were all liquid dressings. Compared with experimental group 14, experimental group 21 showed a significant decrease in mRNA levels of IL-1α, IL-1β, and IL-6 (P<0.05), while there was no significant difference between experimental groups 22 and 23. When preparing different dosage forms, using different types of ectoine or its salts can produce better wound healing effects. Different formulations can be selected according to the dosage form. For example, when preparing ointment dressings, the raw materials preferably include propyl gallate, trehalose, sodium hyaluronate, and ectoine phosphate; when preparing patch dressings, the raw materials preferably include propyl gallate, trehalose, sodium hyaluronate, and ectoine phosphate; when preparing liquid dressings, the raw materials preferably include propyl gallate, trehalose, sodium hyaluronate, and ectoine hydrochloride.

[0132] Example 26: Application of the composition in skin inflammation

[0133] 26.1 Establishment of a TPA-induced mouse model of irritant dermatitis and use of the formulation

[0134] Healthy female SPF-grade Balb / c mice, 6-8 weeks old, were selected. The Balb / c mice were randomly divided into 16 groups, including 14 experimental groups, each receiving the formulations prepared in Examples 1-14, a blank control group (no treatment), and a positive control group (treated with Elosone ointment). The dosage for each group was 30 μl per administration. A mouse model of irritant dermatitis was established using topical application of Phorbol-12-myristate-13-acetate (TPA). Every morning, 10 μl of TPA was applied to both sides of the mouse's ear. After 30 minutes, the left ear was left untreated, while a total of 20 mg of the corresponding drug was applied to both sides of the right ear. The drug was applied again every evening for three consecutive days. On the fourth day of the experiment, mice in each group were euthanized by cervical dislocation. Skin tissue from the ear lesions of the mice was cut off with sterile scissors, and the tissue was quickly collected and marked. A portion of the tissue was placed in 4% tissue fixative for preservation, and the other portion was placed in cryovials and flash-frozen in liquid nitrogen for 1 hour before being transferred to a -80°C water tank for cryopreservation for subsequent experiments.

[0135] 26.2 Effects of different compositions and their formulations on microvessel density in mouse ear inflammation

[0136] TPA, a potent inflammatory inducer and tumor promoter, triggered a series of acute inflammatory responses when applied to the ears of mice. This disrupted local tissue homeostasis, recruited immune cells, and activated various cell types, leading to the formation of a microenvironment rich in pro-angiogenic factors. These factors acted on existing vascular endothelial cells, stimulating their proliferation, migration, and the formation of new blood vessels, resulting in a significant increase in microvascular density.

[0137] Skin from the target lesion on the back of a mouse was cut with sterile surgical scissors, cut into 0.2×0.4cm pieces, and placed in a 2ml centrifuge tube containing 1ml of 4% tissue cell fixative. The tube was then embedded in paraffin and prepared for subsequent pathological sectioning. Immunofluorescence CD31 staining was performed to observe microvascular density.

[0138] The results are as follows Figure 8As shown, the microvessel density in the blank control group was significantly increased. Compared with the blank control group, the microvessel density levels in each experimental group and the positive control group were significantly decreased to varying degrees (P<0.05, P<0.01, P<0.001, P<0.0001). Compared with the positive control group, experimental groups 5, 6, 7, 9, 10, 11, 12, 13, and 14 showed significantly reduced microvessel density (P<0.05, P<0.01, P<0.001), with experimental group 12 showing the best effect, indicating that it has an inhibitory effect on microvascular angiogenesis under inflammatory conditions.

[0139] 26.3 Effects of different compositions and their formulations on VEGF-A mRNA expression in mouse ears

[0140] TPA strongly activates key cells in the skin through both direct and indirect mechanisms, releasing a large amount of the crucial pro-angiogenic factor: vascular endothelial growth factor (VEGF), which is the most potent and specific pro-angiogenic factor. Upregulation of VEGF-A is a core link between inflammation and subsequent angiogenesis. The expression of VEGF-A in mouse skin lesions was detected using quantitative real-time PCR.

[0141] Take an appropriate amount of skin and place it in a 2ml RNase-free EP tube. Add 500μl of Trizol reagent and an appropriate amount of grinding beads to each tube, and homogenize rapidly in a tissue homogenizer. After standing until the foam disappears, centrifuge at 12000r for 15min at 4℃. Transfer the supernatant to a new RNase-free EP tube, add 100μl of chloroform, vortex rapidly until homogeneous, and let stand for 10min. Centrifuge at 12000r for 15min at 4℃. Slowly aspirate the upper aqueous phase from 200μl of the EP tube to a new RNase-free EP tube, add 200μl of isopropanol, mix by inverting, let stand for 10min, and centrifuge at 12000r for 10min at 4℃. Pour out the liquid, invert and air dry, add 200μl of 75% anhydrous ethanol to the tube, vortex until homogeneous, centrifuge at 12000r for 5min at 4℃, discard the supernatant, and allow the precipitate to dry at room temperature in a laminar flow hood. Dissolve the precipitate in 20μl of DEPC water.

[0142] RNA was reverse transcribed into cDNA using a reverse transcription kit. GAPDH was used as an internal control, SYBR Green was used as the dye, and the cDNA was used as a template for quantitative real-time PCR amplification. The reaction volume was 20 μL, and the amplification conditions were: 95℃ pre-denaturation for 10 min; 95℃ denaturation for 15 s, 60℃ annealing for 1 min, 40 amplification cycles; melting curve amplification: 95℃ for 5 s, 60℃ for 5 s; extension at 95℃ for 5 s. The results were calculated using 2... －ΔΔCt The relative expression levels were analyzed to compare the differences in VEGF-A expression among the groups.

[0143] The results are as follows Figure 9 As shown, VEGF-A mRNA in the blank control group was abnormally high. Compared with the blank control group, the mRNA levels of VEGF-A in each experimental group and the positive control group were significantly reduced to varying degrees (P<0.05, P<0.01, P<0.001, P<0.0001). Compared with the positive control group, experimental groups 5, 6, 7, 9, 10, 11, 12, 13, and 14 significantly inhibited VEGF-A expression (P<0.05, P<0.01, P<0.001), with experimental group 12 showing the best effect, indicating that it can significantly inhibit angiogenesis and further alleviate inflammation.

[0144] 26.4 Effects of different compositions and their formulations on tube formation in TPA-stimulated inflammatory cells

[0145] Inflammation-driven angiogenesis is typically highly tortuous, coiled, and irregular; this tortuosity itself is designed to significantly increase the total length of blood vessels within a limited tissue space. A TPA-induced in vitro endothelial cell inflammation model was constructed. Endothelial cells were maintained in DMEM containing 10% fetal bovine serum (FBS), 1% penicillin, and 1% streptomycin, and incubated at 37°C in a carbon dioxide incubator containing 5% CO2. When the cells reached a density of approximately 80%-90%, TPA stimulation was applied to the endothelial cells for 24 hours to induce inflammation.

[0146] Add 100 μl of ECM gel to each well of a 96-well plate and incubate at 37°C for 30 minutes to allow the ECM solution to gel. TPA-stimulated endothelial cells are then introduced at a rate of 3 × 10⁻⁶ cells / well. 5 The inoculum was seeded at a density of 2.5 × 10⁻⁶ cells / mL in 60 mm culture dishes. After treatment with different experimental formulations for 24 hours, 2.5 × 10⁻⁶ cells / mL were seeded into the culture dishes. 4 Cells were resuspended in 150 μl of complete culture medium, seeded onto cured ECM gels, and incubated for 2–8 hours. Endothelial tubes were examined under a microscope in 5 random fields, and the degree of tube formation was estimated by examining the total tube length in each region.

[0147] The results are as follows Figure 10 and Figure 11As shown, angiogenesis studies using in vitro tube formation assays revealed that the vessel length in the blank control group was abnormally increased. Compared with the blank control group, the vessel length in each experimental group and the positive control group was significantly reduced to varying degrees (P<0.05, P<0.01, P<0.001, P<0.0001), inhibiting vessel elongation. Compared with the positive control group, experimental groups 5, 6, 7, 9, 10, 11, 12, 13, and 14 significantly reduced vessel length (P<0.05, P<0.01, P<0.001), with experimental group 12 showing the best effect, indicating that it can significantly inhibit the initiation of angiogenesis and new blood vessel growth.

[0148] Quantitative analysis and statistical comparison of key indicators such as angiogenesis revealed significant differences in the efficacy of different compositions and their formulations in inhibiting angiogenesis. Based on comprehensive testing, the various compositions exhibited a similar trend to that in Example 25 in promoting angiogenesis. Among them, composition group 12 was significantly superior to compositions 7 and 10 in inhibiting angiogenesis; the four active substances combined to produce a prominent promoting effect. The provided compositions, through the synergistic effect of gallic acid derivatives, trehalose, ectoine, and sodium hyaluronate, can significantly inhibit the process of angiogenesis. Compositions 11, 12, 13, and 14 showed the best effects, and the ointment formulation was more advantageous in inhibiting angiogenesis.

[0149] The embodiments of the present invention have been described in detail above, but the content described is only a preferred embodiment of the present invention and should not be considered as limiting the scope of the present invention. All equivalent changes and improvements made in accordance with the scope of the present invention should still fall within the patent coverage of the present invention.

Claims

1. A composition for skin repair, characterized in that: This includes gallic acid or its derivatives, trehalose, and ectoine.

2. The composition for skin repair according to claim 1, characterized in that: Ectocin is replaced with ectocin salt, which is ectocin hydrochloride, ectocin sulfate, or ectocin phosphate.

3. The composition for skin repair according to claim 1 or 2, characterized in that: The gallic acid derivative is propyl gallate or bismuth subgallate; wherein the mass fraction of gallic acid or its derivative is 0.01-6, the mass fraction of trehalose is 0.01-6, and the mass fraction of ectoine or its salt is 0.01-5.

4. The composition for skin repair according to claim 3, characterized in that: It also includes sodium hyaluronate; wherein gallic acid or its derivatives are present in parts by mass of 0.01-6, trehalose in parts by mass of 0.01-6, ectoine or its salts in parts by mass of 0.01-5, and sodium hyaluronate in parts by mass of 0.005-2.

5. A formulation for skin repair, characterized in that: Includes the skin repair composition according to any one of claims 1-4.

6. The skin repair formulation according to claim 5, characterized in that: The preparation is a paste dressing, liquid dressing, or patch dressing.

7. A method for preparing the formulation according to claim 5 or 6, characterized in that: A paste dressing is prepared by mixing the active ingredients of the composition with emulsifiers, emollients, thickeners, preservatives and water; Alternatively, a liquid dressing can be prepared by mixing the active ingredients of the composition with a humectant, a pH adjuster, a thickener, a preservative, and water. Alternatively, the active ingredient in the composition can be mixed with absorbent and adhesive materials to prepare a patch dressing.

8. The use of the composition according to any one of claims 1-4 or the formulation according to claim 5 or 6 in the repair of superficial wounds, characterized in that: Superficial wounds include surgical suture wounds, small incisions, abrasions, or cuts.

9. The use of the composition of any one of claims 1-4 or the formulation of claim 5 or 6 in a product following minimally invasive cosmetic surgery.

10. The application according to claim 9, characterized in that: Used as a post-treatment repair product for one or more of the following: laser, photon, chemical peel, mesotherapy, and microneedling.

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