Ergothioneine-containing special skin repair hydrogel for pets and preparation method of ergothioneine-containing special skin repair hydrogel

Abstract

The invention discloses a special ergothioneine-containing skin repair hydrogel for pets and a preparation method of the special ergothioneine-containing skin repair hydrogel, and belongs to the technical field of pet skin care hydrogels. The invention relates to a skin care product which comprises the following components in parts by weight: 1.6-3.8 parts of a functional active component consisting of ergothioneine, quaternized chitosan and oat beta-glucan, 0.15-0.35 part of a cross-linking agent, 5-10 parts of 1, 3-propylene glycol, 5-10 parts of glycerol, 0.2-0.5 part of a preservative and the balance of water, totaling 100 parts. Through precise compatibility of functional active components and synergistic construction of the ionic cross-linked gel matrix, systematic integration of three effects of oxidation resistance, bacteria resistance and healing promotion is achieved, and the technical problems that an existing pet skin care product is single in function, insufficient in synergy and limited in repairing effect are solved.

Description

Technical Field

[0001] This invention relates to the field of pet skin care hydrogel technology, and in particular to a pet-specific skin repair hydrogel containing ergothioneine and its preparation method. Background Technology

[0002] Pet skin health management has become a hot research topic at the intersection of veterinary medicine, pet care product development, and materials science. Dogs and cats have thin stratum corneum, dense fur, and unique sebaceous gland distribution, making them susceptible to external stimuli that can trigger multiple problems such as dysbiosis, oxidative stress, inflammatory responses, and slow wound healing, creating a vicious cycle of "dysbiosis → inflammatory response → oxidative stress → slow repair." Existing technologies, such as Chinese patent CN202310256789.1, disclose a pet skin care gel loaded with aloe vera extract, which focuses only on anti-inflammatory effects and lacks antioxidant and long-lasting antibacterial functions; Chinese patent CN202211034567.2 discloses a pet wound repair hydrogel containing bioactive peptides, which mainly focuses on tissue regeneration and does not address the synergistic problem of oxidative stress and microbial infection. Furthermore, many existing products neglect the need for surface adhesion, making them prone to detachment due to pet activity, leading to interrupted efficacy. There is currently no technological solution that can systematically integrate the four core needs of highly effective anti-oxidation, broad-spectrum and long-lasting antibacterial properties, antipruritic and soothing effects, and long-lasting adhesion to the skin surface, making it difficult to comprehensively address the complex skin problems of pets.

[0003] Existing topical formulations often focus on a single dimension such as anti-infection or basic moisturizing, making it difficult to break through this cycle. Therefore, developing an innovative formulation that can simultaneously achieve the synergistic effects of highly effective anti-oxidation, broad-spectrum antibacterial properties, and active healing promotion has become a key path to overcome the current bottleneck in pet skin care effects, and is of great significance for improving the comprehensive ability to cope with common skin problems in dogs and cats such as trauma, eczema, and dermatitis.

[0004] The hydrophilic three-dimensional network structure of hydrogels can provide an ideal moist healing environment for wounds, thus many studies have been dedicated to applying hydrogel technology in this field. For example, some studies have attempted to achieve anti-inflammatory and microecological regulation by loading aloe vera extract with specific probiotics; others have focused on preparing water-based gels using bioactive peptides to promote tissue regeneration in canine open wounds. In addition, to improve the user experience of products on the irregular skin surfaces of pets, some studies have focused on constructing dual-network structures to enhance the mechanical strength and adhesion properties of hydrogel coatings.

[0005] However, existing technologies still have significant limitations: First, there is insufficient synergistic effect; most products can only achieve single or dual functions, lacking the ability to simultaneously address the four core issues of oxidative stress, microbial infection, itching and discomfort, and tissue repair. Second, the delivery system has poor compatibility; traditional gel matrices (such as carbomer) are not compatible with active ingredients, resulting in excessively rapid release of active ingredients and difficulty in maintaining long-lasting effects. Third, the surface adhesion is poor, making it unsuitable for use on pets' irregular skin surfaces and in frequently active environments, affecting the repair effect. This lack of a synergistic mechanism makes it difficult for existing products to simultaneously address the multiple challenges of oxidative stress, microbial infection, and delayed tissue repair coexisting in pet skin.

[0006] Therefore, there is an urgent need to develop a new type of pet-specific skin repair hydrogel. It not only needs to have excellent biocompatibility and skin adhesion, but more importantly, it needs to achieve a true unity of antioxidant, antibacterial and healing-promoting effects through the precise formulation and synergy of active ingredients, so as to provide a more comprehensive and efficient solution for pet skin health management. Summary of the Invention

[0007] The purpose of this invention is to overcome the technical shortcomings of existing pet skin care products, such as limited functionality, insufficient synergy, poor adhesion, and limited repair effects, and to provide a pet-specific skin repair hydrogel containing ergothioneine and its preparation method. Through a precise ratio of ergothioneine, quaternized chitosan, and oat β-glucan, combined with the synergistic construction of an ionic cross-linked gel matrix, a synergistic unity of four effects—antioxidant, broad-spectrum antibacterial, antipruritic and soothing, and healing-promoting—is achieved. Simultaneously, the product's surface adhesion and the sustained-release performance of active ingredients are improved, providing a more comprehensive, long-lasting, and efficient solution for common pet skin problems such as trauma, eczema, and dermatitis.

[0008] To achieve the above objectives, the present invention provides the following solution:

[0009] A pet-specific skin repair hydrogel containing ergothioneine, characterized in that the hydrogel comprises the following components in parts by weight: 2.0 to 4.5 parts of gel matrix, 1.6 to 3.8 parts of functional active components composed of ergothioneine, quaternized chitosan and oat β-glucan, 0.15 to 0.35 parts of crosslinking agent, 5 to 10 parts of 1,3-propanediol, 5 to 10 parts of glycerin, 0.2 to 0.5 parts of preservative, and water to 100 parts.

[0010] Preferably, the mass ratio of the above-mentioned ergothioneine, oat β-glucan and quaternized chitosan is (0.15-0.25):(1.2-1.8):1.

[0011] Preferably, the gel matrix is ​​composed of carboxymethyl chitosan, sodium hyaluronate, gellan gum, and carrageenan in a mass ratio of (2.0–3.0):(0.5–1.0):(0.3–0.6):1.

[0012] Specifically, the number average molecular weight of the aforementioned sodium hyaluronate is 80kDa to 150kDa.

[0013] Specifically, the aforementioned quaternized chitosan is any one of hydroxypropyltrimethylammonium chloride chitosan, carboxymethyl chitosan quaternary ammonium salt, or N-trimethyl chitosan quaternary ammonium salt; the aforementioned crosslinking agent is calcium chloride; and the aforementioned preservative is any one or a combination of ethylhexylglycerin, octyl glycol, or phenoxyethanol.

[0014] The key to the preparation method of the above-mentioned pet-specific skin repair hydrogel containing ergothioneine lies in including the following steps:

[0015] S1. Polymer phase preparation: The gel matrix is ​​dispersed in water under stirring and swollen to obtain a viscous liquid;

[0016] S2. Preparation of active phase: Ergothioneine was dissolved in 1,3-propanediol to obtain an ergothioneine solution; oat β-glucan was dissolved in warm water to obtain a glucan solution; quaternized chitosan was dissolved in water to obtain a chitosan solution.

[0017] S3. Mixing: Under stirring, add ergothioneine solution, dextran solution, chitosan solution, glycerin and preservative to the viscous liquid, stir and mix to obtain a premixed gel solution;

[0018] S4. Ionic crosslinking: Dissolve calcium chloride in water to prepare a calcium chloride solution. Add the calcium chloride solution to the premixed gel solution obtained in step S3 while stirring to form a uniform gel.

[0019] S5. Post-processing: The gel obtained in step S4 is degassed and filled to obtain the above-mentioned hydrogel.

[0020] Furthermore, in step S1, the swelling is carried out at a temperature of 22°C to 30°C for 4 to 8 hours.

[0021] Furthermore, in step S2, the temperature of the warm water is 42℃~48℃.

[0022] Furthermore, in step S3, the stirring speed is 50 r / min to 150 r / min, and the mixing time is 15 min to 30 min.

[0023] Furthermore, in step S4, the mass concentration of the calcium chloride solution is 5% to 10%.

[0024] The present invention discloses the following technical effects:

[0025] This invention provides a pet-specific skin repair hydrogel containing ergothioneine and its preparation method. Through the precise formulation of functional active components and the synergistic construction of an ion-crosslinked gel matrix, it achieves a systematic integration of three effects: anti-oxidation, antibacterial and healing promotion. This solves the technical problems of existing pet skin care products having single functions, insufficient synergy, and limited repair effects.

[0026] Specific technical effects include:

[0027] First, this invention establishes a synergistic mechanism of antioxidant defense and cell protection. By introducing ergothioneine as a core antioxidant, it effectively neutralizes free radicals and reduces oxidative stress damage, building a robust protective barrier for skin cells. This component not only possesses excellent antioxidant capabilities on its own but also works synergistically with other active ingredients to enhance the overall stability and effectiveness of the formulation.

[0028] Secondly, this invention constructs a broad-spectrum and long-lasting multi-target antibacterial system. Utilizing the cationic properties of quaternized chitosan and the auxiliary antibacterial activity of ergothioneine, a synergistic inhibitory network is formed against common pet pathogens such as Staphylococcus aureus and Malassezia. This system can disrupt the cell structure of microorganisms and interfere with their normal metabolism, thereby achieving rapid onset of action and long-lasting protection.

[0029] Third, this invention achieves a dual improvement in promoting healing and relieving itching. By combining the immunomodulatory function of oat β-glucan with the anti-inflammatory properties of ergothioneine, it effectively inhibits the release of pruritus-inducing factors, relieves skin discomfort, and at the same time promotes fibroblast migration and proliferation, accelerating the wound healing process and forming a complete repair loop.

[0030] Fourth, this invention innovatively designs an ion-crosslinked gel delivery system. A three-dimensional network structure is constructed by crosslinking carboxymethyl chitosan with calcium chloride, providing a stable environment for the active ingredients while ensuring good adhesion and biocompatibility of the product on the skin surface. This matrix can continuously release active ingredients, maintain a long-lasting effect, and create an ideal microenvironment for skin repair.

[0031] In summary, the hydrogel prepared by this invention, through the organic combination of precise formulation of active ingredients and optimized design of delivery system, exhibits significant advantages in multiple aspects such as anti-oxidation, antibacterial, and healing promotion. It effectively breaks through the technical bottlenecks of single efficacy and insufficient synergy in existing technologies, and provides an innovative solution for the comprehensive management of complex skin problems in pets. Detailed Implementation

[0032] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0033] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0035] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0036] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0037] The specific specifications of the raw materials used in this invention are as follows: sodium hyaluronate with a number average molecular weight of 80 kDa (purity ≥ 98%) was purchased from Shandong Zhongshan Biotechnology Co., Ltd.; sodium hyaluronate with a number average molecular weight of 115 kDa (purity ≥ 98%) and 150 kDa (purity ≥ 98%) was purchased from Jiangsu Xianfeng Nanomaterials Technology Co., Ltd.; gellan gum (purity ≥ 99%) was purchased from Zhejiang Tianwei Biotechnology Co., Ltd.; carrageenan (κ-carrageenan, purity ≥ 98%) was purchased from Fujian Huanhai Biotechnology Co., Ltd.; oat β-glucan (purity ≥ 95%) was purchased from Shaanxi Pufengyuan Biotechnology Co., Ltd.; ergothioneine (purity ≥ 99.9%) was donated by Shanghai Ergothioneine Biotechnology Group Co., Ltd.; quaternized chitosan, 1,3-propanediol, glycerin, and preservatives were all food-grade or cosmetic-grade pure (≥ 99%).

[0038] Example 1

[0039] This embodiment provides a pet-specific skin repair hydrogel containing ergothioneine, comprising the following components in parts by weight: 3.2 parts gel matrix, 2.7 parts functional active ingredient, 0.25 parts crosslinking agent calcium chloride, 7.5 parts 1,3-propanediol, 7.5 parts glycerin, 0.35 parts preservative (ethylhexylglycerin: caprylyl glycol = 1:1), and water to 100 parts.

[0040] The functional active component consists of ergothionein, oat β-glucan, and quaternized chitosan in a mass ratio of 0.2:1.5:1; the gel matrix consists of carboxymethyl chitosan, sodium hyaluronate, gellan gum, and carrageenan in a mass ratio of 2.5:0.75:0.45:1; the number average molecular weight of the sodium hyaluronate used in this embodiment is 115 kDa.

[0041] The quaternized chitosan is hydroxypropyltrimethylammonium chloride chitosan; the preservative is a 1:1 mass ratio mixture of ethylhexylglycerin and octyl glycol.

[0042] The preparation process of the hydrogel in this embodiment includes the following steps:

[0043] S1. Preparation of polymer phase: 1.70 parts of carboxymethyl chitosan, 0.51 parts of sodium hyaluronate, 0.31 parts of gellan gum and 0.68 parts of carrageenan were sequentially dispersed in 65 parts of water at a speed of 800 r / min. After swelling at 26℃ for 6 h, a viscous liquid was obtained.

[0044] S2. Preparation of active phase: 0.20 parts of ergothioneine were dissolved in 7.5 parts of 1,3-propanediol to obtain an ergothioneine solution; 1.50 parts of oat β-glucan were dissolved in 5 parts of warm water at 45℃ to obtain a glucan solution; 1.00 parts of hydroxypropyltrimethylammonium chloride chitosan were dissolved in 10 parts of water to obtain a chitosan solution;

[0045] S3. Mixing: Add ergothioneine solution, dextran solution, chitosan solution, 7.5 parts glycerol and 0.35 parts preservative to the viscous liquid at a speed of 120 r / min, and mix for 22 min at a stirring speed of 100 r / min to obtain a premixed gel solution.

[0046] S4. Ionic crosslinking: Dissolve 0.25 parts of calcium chloride in 2.5 parts of water to prepare a calcium chloride solution with a mass concentration of 10%. Add the calcium chloride solution to the premixed gel solution obtained in step S3 at a speed of 60 r / min to form a uniform gel.

[0047] S5. Post-processing: The gel obtained in step S4 is degassed and filled to obtain a hydrogel, which is denoted as sample 1.

[0048] Example 2

[0049] This embodiment provides a pet-specific skin repair hydrogel containing ergothioneine, comprising the following components in parts by weight: 4.5 parts gel matrix, 3.8 parts functional active ingredients, 0.15 parts crosslinking agent calcium chloride, 5 parts 1,3-propanediol, 10 parts glycerin, 0.2 parts preservative (ethylhexylglycerin: caprylyl glycol = 1:1.2 and water to 100 parts).

[0050] The functional active component consists of ergothionein, oat β-glucan, and quaternized chitosan in a mass ratio of 0.15:1.8:1; the gel matrix consists of carboxymethyl chitosan, sodium hyaluronate, gellan gum, and carrageenan in a mass ratio of 3.0:0.5:0.6:1; the number average molecular weight of the sodium hyaluronate used in this embodiment is 150 kDa.

[0051] Among them, quaternized chitosan is carboxymethyl chitosan quaternary ammonium salt; the preservative is caprylyl glycol.

[0052] The preparation process of the hydrogel in this embodiment includes the following steps:

[0053] S1. Preparation of polymer phase: 2.64 parts of carboxymethyl chitosan, 0.44 parts of sodium hyaluronate, 0.53 parts of gellan gum and 0.88 parts of carrageenan were sequentially dispersed in 55 parts of water at a speed of 1000 r / min. After swelling at 22℃ for 8 h, a viscous liquid was obtained.

[0054] S2. Preparation of active phase: 0.19 parts of ergothioneine were dissolved in 5 parts of 1,3-propanediol to obtain an ergothioneine solution; 2.32 parts of oat β-glucan were dissolved in 8 parts of warm water at 42℃ to obtain a glucan solution; 1.29 parts of carboxymethyl chitosan quaternary ammonium salt were dissolved in 12 parts of water to obtain a chitosan solution;

[0055] S3. Mixing: Add ergothioneine solution, dextran solution, chitosan solution, 10 parts glycerol and 0.2 parts preservative to the viscous liquid at a speed of 150 r / min, and mix for 30 min while maintaining a stirring speed of 50 r / min to obtain a premixed gel solution;

[0056] S4. Ionic crosslinking: Dissolve 0.15 parts of calcium chloride in 3.0 parts of water to prepare a calcium chloride solution with a mass concentration of 5%. Add the calcium chloride solution to the premixed gel solution obtained in step S3 at a rotation speed of 80 r / min to form a uniform gel.

[0057] S5. Post-processing: The gel obtained in step S4 is degassed and filled to obtain a hydrogel, which is denoted as sample 2.

[0058] Example 3

[0059] This embodiment provides a pet-specific skin repair hydrogel containing ergothioneine, comprising the following components in parts by weight: 2.0 parts gel matrix, 1.6 parts functional active ingredient, 0.35 parts crosslinking agent calcium chloride, 10 parts 1,3-propanediol, 5 parts glycerin, 0.5 parts preservative (ethylhexylglycerin: caprylyl glycol = 1:0.8), and water to 100 parts.

[0060] The functional active component consists of ergothionein, oat β-glucan, and quaternized chitosan in a mass ratio of 0.25:1.2:1; the gel matrix consists of carboxymethyl chitosan, sodium hyaluronate, gellan gum, and carrageenan in a mass ratio of 2.0:1.0:0.3:1; the number average molecular weight of the sodium hyaluronate used in this embodiment is 80 kDa.

[0061] Among them, quaternized chitosan is N-trimethyl chitosan quaternary ammonium salt; the preservative is phenoxyethanol.

[0062] The preparation process of the hydrogel in this embodiment includes the following steps:

[0063] S1. Preparation of polymer phase: 0.93 parts of carboxymethyl chitosan, 0.47 parts of sodium hyaluronate, 0.14 parts of gellan gum and 0.47 parts of carrageenan were sequentially dispersed in 68 parts of water at a speed of 500 r / min. After swelling at 30℃ for 4 h, a viscous liquid was obtained.

[0064] S2. Preparation of active phase: 0.16 parts of ergothioneine were dissolved in 10 parts of 1,3-propanediol to obtain an ergothioneine solution; 0.78 parts of oat β-glucan were dissolved in 3 parts of warm water at 48℃ to obtain a glucan solution; 0.65 parts of N-trimethyl chitosan quaternary ammonium salt were dissolved in 8 parts of water to obtain a chitosan solution;

[0065] S3. Mixing: Add ergothioneine solution, dextran solution, chitosan solution, 5 parts glycerol and 0.5 parts preservative to the viscous liquid at a stirring speed of 50 r / min, and mix for 15 min at a stirring speed of 150 r / min to obtain a premixed gel solution.

[0066] S4. Ionic crosslinking: Dissolve 0.35 parts of calcium chloride in 3.5 parts of water to prepare a calcium chloride solution with a mass concentration of 10%. Add the calcium chloride solution to the premixed gel solution obtained in step S3 at a rotation speed of 30 r / min to form a uniform gel.

[0067] S5. Post-processing: The gel obtained in step S4 is degassed and filled to obtain a hydrogel, which is denoted as sample 3.

[0068] Comparative Example 1

[0069] This comparative example provides a hydrogel, which is implemented in the same way as in Example 1, except that ergothioneine is not added. It is referred to as control 1.

[0070] The control sample 1 of this comparative example contains the following components in parts by weight: 3.2 parts of gel matrix, 2.43 parts of functional active component, 0.25 parts of cross-linking agent calcium chloride, 7.5 parts of 1,3-propanediol, 7.5 parts of glycerol, 0.35 parts of preservative, and water to 100 parts.

[0071] The functional active component consists of oat β-glucan and quaternized chitosan in a mass ratio of 1.7:1; the gel matrix consists of carboxymethyl chitosan, sodium hyaluronate, gellan gum and carrageenan in a mass ratio of 2.5:0.75:0.45:1; the number average molecular weight of the sodium hyaluronate used is 115 kDa.

[0072] The quaternized chitosan is hydroxypropyltrimethylammonium chloride chitosan; the preservative is a 1:1 mass ratio mixture of ethylhexylglycerin and octyl glycol.

[0073] The preparation method for this comparative example is the same as that for Example 1, except that in step S2, ergothioneine solution is not prepared and added, but only 7.5 parts of 1,3-propanediol are added; the specific preparation method is as follows:

[0074] S1. Polymer phase preparation: Same as step S1 in Example 1;

[0075] S2. Preparation of active phase: 1.46 parts of oat β-glucan were dissolved in 3 parts of warm water at 48℃ to obtain a glucan solution; 0.97 parts of hydroxypropyltrimethylammonium chloride chitosan were dissolved in 8 parts of water to obtain a chitosan solution;

[0076] S3, Mixing: Same as step S3 in Example 1;

[0077] S4, Ionic crosslinking: Same as step S4 in Example 1;

[0078] S5. Post-processing: Same as step S5 in Example 1, denoted as control sample 1.

[0079] Comparative Example 2

[0080] This comparative example provides a hydrogel, and the specific implementation method is the same as that of Example 1, except that the mass ratio of the functional active components is changed, and it is referred to as control 2.

[0081] The control standard 2 of this comparative example contains the following components in parts by weight: 3.2 parts of gel matrix, 2.7 parts of functional active ingredient control standard, 0.25 parts of cross-linking agent calcium chloride, 7.5 parts of 1,3-propanediol, 7.5 parts of glycerol, 0.35 parts of preservative, and water to 100 parts.

[0082] The functional active ingredient reference standard was composed of ergothioneine, oat β-glucan and quaternized chitosan in a mass ratio of 0.1:2.5:1; the gel matrix was composed of carboxymethyl chitosan, sodium hyaluronate, gellan gum and carrageenan in a mass ratio of 2.5:0.75:0.45:1; the number average molecular weight of the sodium hyaluronate used was 115 kDa.

[0083] The quaternized chitosan is hydroxypropyltrimethylammonium chloride chitosan; the preservative is a 1:1 mass ratio mixture of ethylhexylglycerin and octyl glycol.

[0084] The preparation method for this comparative example is the same as that for Example 1, except that the functional active components are weighed and dissolved according to the adjusted ratio. The specific preparation method is as follows:

[0085] S1. Polymer phase preparation: Same as step S1 in Example 1;

[0086] S2. Preparation of active phase: 0.08 parts of ergothioneine were dissolved in 7.5 parts of 1,3-propanediol to obtain an ergothioneine solution; 1.88 parts of oat β-glucan were dissolved in 5 parts of warm water at 45℃ to obtain a glucan solution; 0.75 parts of hydroxypropyltrimethylammonium chloride chitosan were dissolved in 10 parts of water to obtain a chitosan solution.

[0087] S3, Mixing: Same as step S3 in Example 1;

[0088] S4, Ionic crosslinking: Same as step S4 in Example 1;

[0089] S5. Post-processing: Same as step S5 in Example 1, denoted as control sample 2.

[0090] Comparative Example 3

[0091] This comparative example provides a hydrogel, which is implemented in the same way as in Example 1, except that carbomer is used to replace the original gel matrix and the ionic crosslinking step is omitted. It is referred to as control 3.

[0092] The control sample 3 of this comparative example contains the following components in parts by weight: 3.2 parts of carbomer, 2.7 parts of the functional active ingredient, 7.5 parts of 1,3-propanediol, 7.5 parts of glycerol, 0.35 parts of preservative, and water to 100 parts.

[0093] The preparation method for this comparative example is as follows:

[0094] S1. Polymer phase preparation: 3.2 parts of carbomer were dispersed in 65 parts of water at 500 r / min and swollen for 12 h to obtain a carbomer dispersion.

[0095] S2. Preparation of active phase: 0.20 parts of ergothioneine were dissolved in 7.5 parts of 1,3-propanediol to obtain an ergothioneine solution; 1.50 parts of oat β-glucan were dissolved in 5 parts of warm water at 45℃ to obtain a glucan solution; 1.00 parts of hydroxypropyltrimethylammonium chloride chitosan were dissolved in 10 parts of water to obtain a chitosan solution;

[0096] S3. Mixing: Add ergothioneine solution, dextran solution, chitosan solution, 7.5 parts glycerol and 0.35 parts preservative to carbomer dispersion at 120 r / min. Mix at 100 r / min for 22 min. Then slowly add triethanolamine to adjust the pH to 6.5, allowing the system to gradually form a gel.

[0097] S4. Post-processing: The gel obtained in step S3 is degassed and filled to obtain a hydrogel, which is designated as reference standard 3.

[0098] Analysis and Testing

[0099] I. Antioxidant Performance Test

[0100] To verify the antioxidant capacity of the hydrogel of the present invention, the DPPH free radical scavenging method (refer to GB / T 31740.2-2015 "Determination of antioxidant properties in cosmetics - Part 2: DPPH method") and the intracellular reactive oxygen species (ROS) detection method were used for evaluation.

[0101] The DPPH free radical scavenging method directly reflects the antioxidant strength of a sample by measuring its scavenging rate of stable free radicals; intracellular ROS detection is achieved by establishing an H2O2-induced oxidative stress model in canine skin fibroblasts (cell concentration 1×10⁻⁶). 5 (samples / mL), simulating the antioxidant effect of the test sample in vivo, and detected using the DCFH-DA fluorescent probe method.

[0102] The test results are shown in Table 1.

[0103] Table 1: Results of Antioxidant Properties Test of Hydrogel

[0104]

[0105] As shown in Table 1, the samples prepared in the embodiments of the present invention all exhibited excellent antioxidant properties. Among them, sample 1 showed the best antioxidant effect, with a DPPH free radical scavenging rate of 95.2% and an intracellular ROS inhibition rate of 89.7%.

[0106] In contrast, control 1 lacked ergothioneine, resulting in a significant reduction in its antioxidant capacity, which confirms the crucial role of ergothioneine as a core antioxidant component. Control 2 altered the proportion of active ingredients, leading to a significant impairment in its antioxidant performance, indicating that a specific mass ratio is essential for maintaining optimal antioxidant effects. Although control 3 maintained a certain level of antioxidant activity, it was still inferior to the embodiments of the present invention, demonstrating that the gel matrix of the present invention facilitates the full utilization of the antioxidant function of the active ingredients, and that the two have a synergistic effect.

[0107] Test results show that the antioxidant performance of the samples in the embodiments of the present invention is significantly better than that of the comparative examples: the DPPH free radical scavenging rate of control 1 (ergothioneine deficiency) is only 45.6%, confirming that ergothioneine is the core antioxidant component; the scavenging rate of control 2 (with changed active component ratio) is reduced to 68.3%, indicating that the mass ratio of (0.15~0.25):(1.2~1.8):1 is the key to maintaining the optimal antioxidant effect; the scavenging rate of control 3 (carbomer matrix) is 85.7%, which is lower than that of the sample in the embodiments, confirming that the ion-crosslinked gel matrix of the present invention has a synergistic antioxidant effect with the active ingredients and can promote the efficacy of the active ingredients.

[0108] II. Antibacterial Performance Test

[0109] To accurately evaluate the antibacterial properties of the hydrogel of this invention, the agar diffusion method (referring to QB / T 2738-2012 "Microbiological Examination of Cosmetics - Determination of Inhibition Zone") and the plate count method (referring to Appendix C of GB 15979-2002 "Hygienic Standard for Disposable Sanitary Products") were used for evaluation. The specific methods are as follows:

[0110] Agar diffusion method: A bacterial suspension of Staphylococcus aureus (ATCC 6538) and Malassezia (ATCC 14521) (1×10⁻⁶) is diffused into the agar. 6 The hydrogel sample (CFU / mL) was evenly spread onto the corresponding agar plate. A sterile Oxford cup was placed in the center of the plate, and 0.5g of the sample to be tested was then added to the cup, ensuring full contact between the sample and the agar surface. For Staphylococcus aureus, the inhibition zone diameter was measured after incubation at 37℃ for 24 hours, and for Malassezia, the inhibition zone diameter was measured after incubation at 37℃ for 48 hours.

[0111] Plate coating and counting method (quantitative assessment of 24-hour antibacterial rate): 1.0 g of hydrogel sample is mixed with 1 mL of bacterial suspension (1×10⁻⁶). 6 The CFU / mL solution was thoroughly mixed in sterile EP tubes and incubated at 37°C for 24 h (Staphylococcus aureus) and 48 h (Malassezia), respectively. After incubation, the solutions were serially diluted with physiological saline and plated. After culturing, the number of colonies was counted, and the inhibition rate was calculated.

[0112] The test results are shown in Table 2.

[0113] Table 2: Test results of antibacterial properties of hydrogel

[0114]

[0115] As shown in Table 2, the samples prepared in the embodiments of the present invention exhibited significant, sustained, and highly effective inhibitory effects against both pathogenic bacteria. Among them, Sample 1 showed the best overall antibacterial effect, characterized by the largest inhibition zone diameter, with inhibition rates of 99.2% and 98.7% against Staphylococcus aureus and Malassezia at 24h and 48h, respectively.

[0116] Test results show that the antibacterial efficacy of the samples in this invention is significantly better than that of the comparative examples: Control 1 (ergothioneine deficiency) had inhibition zone diameters of only 12.3 mm and 10.5 mm against Staphylococcus aureus and Malassezia, respectively, and the inhibition rates at 24 h and 48 h decreased to 65.3% and 58.9%, respectively, a decrease of more than 34% compared to sample 1. This confirms that ergothioneine and quaternized chitosan have a synergistic antibacterial effect and are key components for enhancing the antibacterial effect; the inhibition zone diameter of Control 2 (with altered active component ratio) was reduced to 10 mm. Samples with diameters of 0.8mm and 9.3mm showed an antibacterial rate of less than 73%, indicating that the combination of ergothioneine, oat β-glucan, and quaternized chitosan at a mass ratio of (0.15–0.25):(1.2–1.8):1 is the core prerequisite for achieving broad-spectrum antibacterial activity. While control 3 (carbomer replacing the original matrix) showed better antibacterial effects than the other two, its antibacterial rate was still 14%–19% lower than sample 1, confirming that the ion-crosslinked gel matrix of this invention can more stably carry active antibacterial components, reducing their loss and thus maintaining long-lasting antibacterial ability. In summary, the samples of this invention can achieve rapid and long-lasting inhibitory effects against common pet pathogens, meeting the needs of complex skin infection scenarios.

[0117] III. In vitro evaluation of antipruritic efficacy

[0118] Itching is closely related to skin inflammation, and histamine and inflammatory factors (such as TNF-α and IL-6) are key signaling molecules that trigger and exacerbate itching. To evaluate the antipruritic potential of the hydrogel of this invention, enzyme-linked immunosorbent assay (ELISA) was used to analyze the inhibitory ability of the sample on pruritogenic factors. The test results are shown in Table 3.

[0119] Sample preparation: 1.0 g of the sample to be tested was incubated with 10 mL of cell culture medium at 37 °C for 24 h to prepare an extract containing gel release components.

[0120] Cell model: The mouse mononuclear macrophage cell line (J774A.1, purchased from Qingqi (Shanghai) Biotechnology Development Co., Ltd.) was used to establish an inflammation model by stimulating cells with LPS. The control group was treated with LPS and then the culture medium was replaced with one without the sample extract. The experimental group was treated with LPS and then the culture medium was replaced with one containing different sample extracts.

[0121] Detection and Evaluation: Different concentrations of sample extracts were added to the cell culture system and co-cultured for 24 hours. The supernatant was then collected, and the concentrations of histamine, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were measured. The inhibition rate of pruritogenic factors was used as an indirect indicator to evaluate the antipruritic effect. The calculation formula is as follows:

[0122] Inhibition rate (%) = (1 - concentration of experimental group / concentration of model control group) × 100%.

[0123] Table 3: Inhibitory effect of hydrogels on the release of pruritogenic factors

[0124]

[0125] As shown in Table 3, the inhibitory ability of the samples in this invention against pruritus factors is far superior to that of the comparative examples: Control 1 (lacking ergothioneine) showed inhibition rates of only 35.2%, 32.6%, and 38.9% against histamine, TNF-α, and IL-6, respectively, a decrease of 53%–62% compared to Sample 1, clearly demonstrating that the potent anti-inflammatory properties of ergothioneine are the core factor in blocking the release of pruritus factors; Control 2 (inappropriate ratio of active ingredients) showed an inhibition rate of less than 50%, indicating that a specific ratio of the three active ingredients can synergistically enhance immune regulation and anti-inflammatory effects, and arbitrary adjustments will disrupt the antipruritic mechanism; Control 3 (replaced with carbomer matrix) showed an inhibition rate 23%–29% lower than that of Sample 1, demonstrating that the original ion-crosslinked matrix can release active ingredients more efficiently, allowing them to act on inflammatory cells and improve antipruritic efficiency. Therefore, the samples of this invention can alleviate skin itching from the source and improve scratching behavior in pets caused by skin discomfort.

[0126] IV. In vitro evaluation of healing-promoting efficacy

[0127] The healing rate of skin wounds mainly depends on the migration and proliferation capacity of epidermal cells. Cell scratch assays and cell proliferation assays allow for direct observation and quantification of the promoting effect of the hydrogel sample of this invention on cell repair behavior.

[0128] Sample preparation: Same as in section "III. In vitro evaluation of antipruritic efficacy", prepare hydrogel extract.

[0129] Cell model: Rat skin fibroblast cell line (YB-73430RC, purchased from Shanghai Yubo Biotechnology Co., Ltd.) was used.

[0130] Scratch test: After a monolayer of cells is formed in the culture plate, a uniform scratch is created using a sterile pipette tip to simulate a skin wound. The culture medium is then replaced with one containing the sample extract. The scratches are photographed under a microscope at 0h, 24h, and 48h to measure the scratch width and calculate the scratch healing rate. No sample is added to the blank control group.

[0131] Cell proliferation assay: The effect of sample extract on fibroblast proliferation activity was detected using the CCK-8 assay.

[0132] The test results are shown in Table 4.

[0133] Table 4: The promoting effect of hydrogels on cell migration and proliferation

[0134]

[0135] As shown in Table 4, the healing-promoting ability of the samples in this invention is significantly better than that of the comparative examples and the blank control group: Control 1 (ergothioneine deficiency) had a scratch healing rate of only 45.6% and 70.8% at 24h and 48h, respectively, and a cell proliferation rate close to that of the blank control group (105.2%), a decrease of 33%–28% compared to sample 1. This demonstrates that ergothioneine can protect fibroblast activity and promote their migration and proliferation, which is key to accelerating wound healing. Control 2 (incorrect ratio of active components) had a scratch healing rate of less than 66% and a cell proliferation rate of less than 100%, indicating that precise ratio of active ingredients can synergistically activate cell repair function, while an imbalanced ratio leads to decreased repair efficiency. Control 3 (carbomer matrix) had a healing rate and proliferation rate 20%–14% lower than that of sample 1, reflecting that the three-dimensional network structure of the original gel matrix can provide a better growth microenvironment for cells, aiding wound repair. In summary, the samples of this invention can effectively shorten the healing cycle of pet skin injuries and reduce the risk of scar formation.

[0136] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A pet-specific skin-repairing hydrogel containing ergothioneine, characterized in that, The hydrogel comprises the following components in parts by weight: 2.0 to 4.5 parts of gel matrix, 1.6 to 3.8 parts of functional active components composed of ergothioneine, quaternized chitosan and oat β-glucan, 0.15 to 0.35 parts of crosslinking agent, 5 to 10 parts of 1,3-propanediol, 5 to 10 parts of glycerol, 0.2 to 0.5 parts of preservative, and water to 100 parts.

2. The hydrogel according to claim 1, characterized in that, The mass ratio of ergothionein, oat β-glucan, and quaternized chitosan is (0.15–0.25):(1.2–1.8):

1.

3. The hydrogel according to claim 1, characterized in that, The gel matrix is ​​composed of carboxymethyl chitosan, sodium hyaluronate, gellan gum, and carrageenan in a mass ratio of (2.0–3.0):(0.5–1.0):(0.3–0.6):

1.

4. The hydrogel according to claim 3, characterized in that, The number-average molecular weight of the sodium hyaluronate is 80kDa to 150kDa.

5. The hydrogel according to claim 1, characterized in that, The quaternized chitosan is any one of hydroxypropyltrimethylammonium chloride chitosan, carboxymethyl chitosan quaternary ammonium salt, or N-trimethyl chitosan quaternary ammonium salt; the crosslinking agent is calcium chloride; and the preservative is any one or a combination of ethylhexylglycerin, octyl glycol, or phenoxyethanol.

6. The method for preparing the pet-specific skin repair hydrogel containing ergothioneine as described in any one of claims 1 to 5, characterized in that, Includes the following steps: S1. Polymer phase preparation: The gel matrix is ​​dispersed in water under stirring and swollen to obtain a viscous liquid; S2. Preparation of active phase: Ergothioneine was dissolved in 1,3-propanediol to obtain an ergothioneine solution; oat β-glucan was dissolved in warm water to obtain a glucan solution; quaternized chitosan was dissolved in water to obtain a chitosan solution. S3. Mixing: Under stirring, add ergothioneine solution, dextran solution, chitosan solution, glycerin and preservative to the viscous liquid, stir and mix to obtain a premixed gel solution; S4. Ionic crosslinking: Dissolve calcium chloride in water to prepare a calcium chloride solution, and add the calcium chloride solution to the premixed gel solution obtained in step S3 under stirring to form a uniform gel. S5. Post-processing: The gel obtained in step S4 is degassed and filled to obtain the hydrogel.

7. The preparation method according to claim 6, characterized in that, The swelling described in step S1 is carried out at a temperature of 22℃~30℃ for 4h~8h.

8. The preparation method according to claim 6, characterized in that, The temperature of the warm water mentioned in step S2 is 42℃~48℃.

9. The preparation method according to claim 6, characterized in that, The stirring speed in step S3 is 50 r / min to 150 r / min, and the mixing time is 15 min to 30 min.

10. The preparation method according to claim 6, characterized in that, The mass concentration of the calcium chloride solution in step S4 is 5% to 10%.

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