A long-lasting moisturizing and barrier-repairing exosome active gel and its preparation method

Abstract

This application relates to the fields of skin care and medical cosmetology, disclosing a long-lasting moisturizing and barrier-repairing exosome-based active gel and its preparation method. The gel comprises the following components by weight: 2-5 parts exosomes; 3-7 parts trehalose; 2-6 parts glycerol; 1-2 parts polyvinyl acrylate; 0.5-1.5 parts octyl glucoside; 0.8-1.5 parts sodium hyaluronate; 0.2-0.5 parts low-molecular-weight peptides; 1-3 parts polyacrylamide; and 0.02-0.05 parts tea polyphenol antioxidants. This invention utilizes the synergistic repair effects of exosomes and low-molecular-weight peptides, dual moisturizing properties of trehalose and glycerol, an innovative thickening system to enhance stability, and the addition of tea polyphenol antioxidants to effectively achieve deep repair, long-lasting moisturization, improved skin feel, and anti-aging properties, comprehensively addressing multiple bottlenecks in traditional skin care technologies.

Description

Technical Field

[0001] The present invention relates to the fields of skin care and medical cosmetology, and specifically relates to a long-lasting moisturizing and barrier-repairing exosome active gel and a preparation method thereof. Background Art

[0002] With people's increasing concern for skin health and beauty, a large number of skincare products have appeared on the market, including facial masks, lotions, serums, and other types. The main functions of these products include moisturizing, anti-aging, and repairing skin damage. However, many existing skincare products still have some significant drawbacks. First, many products experience film instability, easily cracking or falling off during use, which greatly reduces their effectiveness. In addition, some products have poor skin feel after application, and may be sticky, thick, or difficult to absorb, affecting the user experience.

[0003] On the other hand, while many skincare products have attempted to enhance the skin's protective abilities by adding antioxidants, existing antioxidant ingredients haven't achieved the desired results. While traditional antioxidants can inhibit free radical damage to the skin to a certain extent, their activity is weak or their stability is poor, preventing them from exerting their effects over a long period of time. This reduces the product's antioxidant efficacy and skin repair capacity. Furthermore, single antioxidants often enhance antioxidant capacity without fully addressing skin repair effects, leaving the product's performance in areas such as anti-aging and barrier repair still to be improved.

[0004] Currently, many skincare products lack systematic and innovative approaches in their ingredient combinations and technical processes. Traditional products often rely on a single active ingredient, and their formulations often overlook the interactions and synergistic effects between different ingredients. This results in suboptimal results in enhancing the skin barrier, improving touch, and delaying aging. Consequently, existing technologies commonly suffer from issues such as insufficient skin repair, short-lasting antioxidant effects, and poor skin feel. Therefore, the present invention proposes a long-lasting moisturizing and barrier-repairing exosome-activated gel and its preparation method to address these shortcomings. Summary of the Invention

[0005] In response to the shortcomings of the existing technology, the present invention provides a long-lasting moisturizing and barrier-repairing exosome active gel and a preparation method thereof, which solves the problems of traditional skin care products in terms of moisturizing, skin barrier repair and antioxidant effects, as well as poor touch.

[0006] To achieve the above objectives, the present invention is implemented through the following technical solutions: a long-lasting moisturizing and barrier-repairing exosome active gel, the gel comprising the following components in parts by mass:

[0007] Exosomes: 2-5;

[0008] Trehalose: 3-7 parts;

[0009] Glycerin: 2-6 parts;

[0010] Polyvinyl acrylate: 1-2 parts;

[0011] Octyl glucoside: 0.5-1.5 parts;

[0012] Sodium hyaluronate: 0.8-1.5 parts;

[0013] Low molecular weight peptide: 0.2-0.5 parts;

[0014] Polyacrylamide: 1-3 parts;

[0015] Tea polyphenol antioxidants: 0.02-0.05 parts;

[0016] Cross-linking agent: 0.1-0.3 parts;

[0017] Water: 70-80 parts.

[0018] Exosomes are derived from placenta-derived mesenchymal stem cells or umbilical cord-derived mesenchymal stem cells; exosomes are also derived from amnion-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells.

[0019] As an important medium for intercellular communication, exosomes can regulate the physiological functions of target cells by carrying bioactive molecules such as cell-specific proteins, nucleic acids, and lipids. Exosomes derived from placenta- and umbilical cord-derived mesenchymal stem cells can promote tissue repair and regeneration by releasing growth factors and cytokines. In addition, exosomes from amniotic membrane-derived mesenchymal stem cells are rich in immunomodulatory factors and anti-fibrotic proteins. By inhibiting pro-inflammatory signaling pathways and regulating macrophage polarization, they reduce inflammatory responses and promote scar-free tissue repair. Exosomes from adipose-derived mesenchymal stem cells activate the PI3K / AKT pathway by delivering pro-angiogenic factors and lipid metabolism-related miRNAs, improving local microcirculation, accelerating injury repair, and inhibiting oxidative stress. At the same time, they enhance skin elasticity and barrier function by regulating signals related to adipocyte differentiation.

[0020] Trehalose is a natural polysaccharide derived from α,α-trehalose in natural brown algae. It has excellent moisturizing properties. In this formula, it forms a protective film on the skin's surface, preventing moisture loss.

[0021] Trehalose can bind to the surface of skin cells through its polysaccharide structure, forming a moisturizing layer that effectively prevents water evaporation. Its unique structure can enhance the skin's barrier function, improve the skin's resistance to the external environment, and prevent water loss.

[0022] Glycerin is a common moisturizer with strong hydrophilicity, which can help the skin absorb moisture from the air and lock in moisture.

[0023] As a polyol, glycerin can form hydrogen bonds with water molecules through its hydroxyl groups, thereby forming a moisturizing film on the skin surface. This moisturizing film can effectively lock in moisture, increase the skin's hydration content, and enhance the skin's softness and elasticity.

[0024] Polyvinyl acrylate is a thickener and stabilizer that can increase the viscosity of the gel, giving it good spreadability and stability.

[0025] Through its cross-linked molecular chain structure, polyvinyl acrylate absorbs water molecules and increases the gel's viscosity, thereby improving the gel's stability and moisturizing ability. Its polymeric structure also forms a thin film during application, preventing rapid evaporation of the ingredients.

[0026] Octyl Glucoside is a mild nonionic surfactant that improves skin cleansing and is low in irritation.

[0027] Octyl Glucoside acts as a surfactant, reducing interfacial tension between water and oil, thereby helping other ingredients disperse and penetrate the skin, resulting in a gentle cleansing effect. Its non-ionic structure also reduces skin irritation and improves the skin compatibility of the formula.

[0028] Sodium Hyaluronate is a powerful moisturizing ingredient that provides excellent hydration.

[0029] Sodium hyaluronate, through its macromolecular structure, attracts large amounts of water molecules on the skin's surface, thereby maintaining skin hydration. Sodium hyaluronate binds to water in the skin, boosting hydration and helping maintain skin's moisture and elasticity while strengthening the skin's barrier function.

[0030] Low molecular weight peptides refer to small molecule proteins composed of shorter amino acid chains, with a molecular weight usually below 3000 Da.

[0031] Low-molecular-weight peptides can more easily penetrate the surface of the skin, permeating into the dermis and promoting the repair and regeneration of skin cells. By activating specific receptors in skin cells, low-molecular-weight peptides can stimulate collagen synthesis, improve skin elasticity and firmness, and have anti-aging effects.

[0032] Polyacrylamide is a water-soluble polymer that thickens and stabilizes gel formulations.

[0033] The molecular chain of polyacrylamide is highly hydrophilic, capable of absorbing water and increasing the viscosity of the gel. This helps the gel maintain its proper physical state, making it easy to spread, while also improving the stability of the formula and preventing ingredients from separating or settling.

[0034] Tea polyphenols antioxidants are mainly derived from green tea, have strong antioxidant effects, and can fight against damage caused by free radicals.

[0035] Tea polyphenols can effectively capture free radicals, reduce oxidative damage, and delay skin aging. Tea polyphenols react with free radicals through their phenolic hydroxyl structure, inhibiting the cellular oxidation process, protecting the skin from oxidative damage caused by environmental factors such as ultraviolet rays, and enhancing the skin's anti-aging ability.

[0036] Cross-linkers are used in this formulation to enhance the gel network structure and stability.

[0037] Crosslinkers react with polymer chains to form a more stable three-dimensional network structure. This crosslinking makes the gel more flexible when applied, reduces the fluidity of the ingredients, improves the gel's physical stability, and helps control the gel's release properties.

[0038] Water, as a solvent, is the basic solution for all ingredients. Its main function is to dissolve other ingredients and maintain the fluidity and stability of the formula.

[0039] Water can dissolve most hydrophilic ingredients and help other active ingredients to be evenly dispersed in the gel. As a moisturizing factor, water can form a protective film on the skin surface to prevent water loss and enhance the skin's hydration.

[0040] Preferably, the exosomes are derived from placenta-derived mesenchymal stem cells or umbilical cord-derived mesenchymal stem cells, the trehalose is α,α-trehalose derived from natural brown algae, the glycerol is plant-derived glycerol, the polyvinyl acrylate is a high molecular weight polymer, the octyl glucoside is a nonionic surfactant, and the sodium hyaluronate is purified sodium hyaluronate.

[0041] Preferably, the low-molecular-weight peptide is a collagen peptide with a molecular weight of less than 3000 Da, and its amino acid sequence contains Gly-Pro-Hyp repeating units. The polyacrylamide is a modified water-soluble polymer compound. The tea polyphenol antioxidant is a combination of epigallocatechin gallate and rosmarinic acid in a mass ratio of 2:1-3:2. The cross-linking agent is a biodegradable water-soluble cross-linking agent. The water is ultrapure water that has been deionized and has a resistivity of ≥18 MΩ·cm.

[0042] The present invention also provides a method for preparing a long-lasting moisturizing and barrier-repairing exosome active gel, comprising the following steps:

[0043] S1. Add trehalose, glycerol, sodium hyaluronate and polyacrylamide into water and stir until completely dissolved to obtain a uniform solution;

[0044] S2. Cooling the homogeneous solution, then adding exosomes to the solution and stirring until the exosomes are completely dispersed to obtain a preliminary solution;

[0045] S3, adding octyl glucoside and polyvinyl acrylate to the preliminary solution, stirring until completely mixed, to obtain a mixed solution;

[0046] S4. Add sodium hyaluronate, low molecular weight peptides and tea polyphenol antioxidants to the mixed solution under light-shielding conditions and continue stirring to obtain an active solution;

[0047] S5. Heat the active solution, add a cross-linking agent, and continue stirring to complete the reaction;

[0048] S6. After the reaction is completed, the reaction mixture is placed in a vacuum environment for degassing, and finally the obtained gel is cooled to room temperature.

[0049] Preferably, in step S1, trehalose, glycerol, 0.3 to 1.5 parts of sodium hyaluronate and polyacrylamide are added to purified water in parts by mass, and stirred at 60 to 70° C. until the components are dissolved. The stirring speed during the process is 200 to 300 rpm, and stirring is continued for 15 to 20 minutes to obtain a uniform solution.

[0050] Preferably, in step S2, the obtained uniform solution is cooled to 4-10°C, a cooling device is used or the container is placed in a low-temperature environment during the process, and then the exosomes are added to the uniform solution. At this time, the stirring speed is controlled at 150-250 rpm and the stirring time is 10-15 minutes, so that the exosomes are evenly dispersed in the uniform solution to obtain a preliminary solution.

[0051] Preferably, in step S3, octyl glucoside and polyvinyl acrylate are added to the preliminary solution, and stirring is maintained. The stirring temperature is controlled at 20-25° C. and the stirring time is 10-15 minutes, until octyl glucoside and polyvinyl acrylate are dissolved and evenly distributed in the solution to obtain a mixed solution.

[0052] Preferably, in step S4, 0.2 to 0.8 parts of sodium hyaluronate, low molecular peptides and tea polyphenol antioxidants are added to the mixed solution under light-proof conditions, and stirring is continued for 10 to 20 minutes by using a brown glass container or wrapping the container in an opaque material, and the stirring speed is maintained at 150 to 250 rpm, so that the sodium hyaluronate, low molecular peptides and tea polyphenol antioxidants are dissolved and evenly distributed in the solution to obtain an active solution.

[0053] Preferably, in step S5, the active solution is heated to 40-45° C., the heating temperature is controlled by a temperature control device, a crosslinking agent is added, and stirring is continued for 20-30 minutes, with the stirring speed controlled at 200-300 rpm to allow the crosslinking agent to fully react with the solution.

[0054] Preferably, in step S6, after the reaction is completed, the reaction mixture is placed in a vacuum environment for degassing, the pressure of the vacuum environment is controlled at -0.08 MPa to -0.095 MPa, the degassing time is 10 to 20 minutes, and the vacuum pump extracts the gas in the solution to reduce the generation of bubbles. After completion, the obtained gel is cooled to room temperature.

[0055] The present invention provides a long-lasting moisturizing and barrier-repairing exosome active gel and its preparation method. It has the following beneficial effects:

[0056] 1. This invention utilizes a combination of exosomes and low-molecular-weight peptides to significantly promote skin repair and regeneration. Compared to existing single moisturizing ingredients, exosomes provide a greater variety of bioactive factors, helping skin cells rejuvenate more quickly. Compared to traditional formulas, this approach addresses the drawback of relying solely on surface moisturization without deep repair.

[0057] 2. This invention utilizes a dual moisturizing technology of trehalose and glycerin, effectively improving long-term skin hydration. Compared to single moisturizing ingredients in existing technologies, the synergistic effect of trehalose and glycerin can better lock in moisture and enhance the skin's barrier function, avoiding the short-term moisturizing effect limitations of existing technologies.

[0058] 3. By using polyvinyl acrylate and polyacrylamide as thickeners, this invention achieves the technical effect of improving gel stability and application experience. This innovative formula avoids the excessive stickiness associated with traditional thickeners and addresses the issue of gel products on the market that are prone to flow and difficult to maintain a stable form, resulting in a smoother and more comfortable application.

[0059] 4. The present invention incorporates tea polyphenol antioxidants and a crosslinking agent into its formula, effectively slowing the progression of skin aging. Compared to existing products that lack effective antioxidant ingredients, this invention effectively captures free radicals, mitigating damage to the skin from environmental factors like ultraviolet rays, addressing the lack of anti-aging effects found in traditional formulas. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 It is a flow chart of the preparation method of the present invention. DETAILED DESCRIPTION

[0061] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the drawings in the present specification. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0062] See also Figure 1 :

[0063] Example 1: Basic repair exosome active gel

[0064] Formula composition (by mass):

[0065] Exosomes: 2 parts, trehalose: 3 parts, glycerol: 2 parts, polyvinyl acrylate: 1 part, octyl glucoside: 0.5 parts, sodium hyaluronate (S1 stage): 0.5 parts, sodium hyaluronate (S4 stage): 0.3 parts, low molecular weight peptide: 0.2 parts, polyacrylamide: 1 part, tea polyphenol antioxidant: 0.02 parts, cross-linking agent: 0.1 parts, deionized water: 70 parts;

[0066] Preparation steps:

[0067] Stage S1: Polyvinyl acrylate, 0.5 parts of high molecular weight sodium hyaluronate, trehalose, glycerol, and octyl glucoside were added to deionized water in sequence (25°C, stirring speed 800 rpm, 20 minutes) until completely dissolved;

[0068] S2 stage: The solution was cooled to 10-15°C, and the exosome solution was slowly added dropwise at a low speed (300 rpm) to avoid foaming;

[0069] S3 stage: add polyacrylamide and continue stirring for 10 minutes;

[0070] Stage S4: Add 0.3 parts of medium molecular weight sodium hyaluronate, low molecular weight peptides, and tea polyphenol antioxidants, and stir for 5 minutes in the dark;

[0071] Stage S5: heating to 40°C, adding the cross-linking agent, reacting for 15 minutes, and then transferring to a vacuum environment (−0.09 MPa) for degassing for 15 minutes;

[0072] S6 stage: Cool to room temperature and naturally form a transparent gel.

[0073] Example 2: Enhanced Moisturizing Exosome Active Gel

[0074] Formula composition (by mass):

[0075] Exosomes: 3 parts, trehalose: 4.5 parts, glycerol: 4 parts, polyvinyl acrylate: 1.5 parts, octyl glucoside: 1 part, sodium hyaluronate (S1 stage): 0.8 parts, sodium hyaluronate (S4 stage): 0.5 parts, low molecular weight peptide: 0.4 parts, polyacrylamide: 2 parts, tea polyphenol antioxidant: 0.03 parts, cross-linking agent: 0.2 parts, deionized water: 76 parts;

[0076] Preparation steps:

[0077] Stage S1: Polyvinyl acrylate was dispersed in water (40°C, 1000 rpm, 15 min), and 0.8 parts of high molecular weight sodium hyaluronate, trehalose, glycerol, and octyl glucoside were added;

[0078] S2 stage: cooling to 20°C, adding exosome solution dropwise at low speed (200 rpm);

[0079] S3 stage: add polyacrylamide and stir until uniform;

[0080] Stage S4: Add 0.5 parts of medium molecular weight sodium hyaluronate, low molecular weight peptides, and tea polyphenol antioxidants, and stir in the dark for 10 minutes;

[0081] S5 stage: heating to 45°C, adding cross-linking agent and reacting for 20 minutes, followed by degassing under a vacuum environment of −0.095 MPa for 20 minutes;

[0082] S6 stage: Let it stand at room temperature for 3 hours to form a highly transparent gel.

[0083] Example 3: Highly active barrier-repairing exosome gel

[0084] Formula composition (by mass):

[0085] Exosomes: 5 parts, trehalose: 7 parts, glycerol: 6 parts, polyvinyl acrylate: 2 parts, octyl glucoside: 1.5 parts, sodium hyaluronate (S1 stage): 1 part, sodium hyaluronate (S4 stage): 0.5 parts, low molecular weight peptide: 0.5 parts, polyacrylamide: 3 parts, tea polyphenol antioxidant: 0.05 parts, cross-linking agent: 0.3 parts, deionized water: 80 parts;

[0086] Preparation steps:

[0087] Stage S1: Polyvinyl acrylate, 1.0 part of high molecular weight sodium hyaluronate, trehalose, glycerol, and octyl glucoside were dispersed in deionized water at low temperature (25°C, stirring speed 700 rpm);

[0088] S2 stage: Add the mixed exosome solution and stir at low speed (150 rpm) to avoid shear damage;

[0089] S3 stage: add polyacrylamide and stir until completely dissolved;

[0090] Stage S4: Add 0.5 parts of low-molecular-weight sodium hyaluronate, low-molecular-weight peptides, and tea polyphenol antioxidants, and stir in the dark for 15 minutes;

[0091] Stage S5: heating to 50°C, adding the natural cross-linking agent, reacting for 30 minutes, and then degassing at −0.085 MPa for 12 minutes;

[0092] S6 stage: Cool to room temperature and form gel naturally.

[0093] Comparative Example 1

[0094] Compared with Example 1, the difference is that the exosomes are removed, and the rest are the same.

[0095] Comparative Example 2

[0096] Compared with Example 1, the difference is that the low molecular weight peptide is removed, and the rest are the same.

[0097] Comparative Example 3

[0098] Compared with Example 1, the difference is that the exosomes are replaced by liposomes to encapsulate EGF, and the rest are the same.

[0099] Comparative Example 4

[0100] Compared with Example 1, the difference is that the low molecular weight peptide is replaced by ordinary hydrolyzed collagen (molecular weight>5000Da), and the rest are the same.

[0101] Comparative Example 5

[0102] Compared with Example 1, the difference is that exosomes and trehalose are removed at the same time, and the rest are the same.

[0103] Comparative Example 6

[0104] Compared with Example 2, the difference is that trehalose is removed, and the rest are the same.

[0105] Comparative Example 7

[0106] Compared with Example 2, the difference is that glycerol is removed, and the rest are the same.

[0107] Comparative Example 8

[0108] Compared with Example 2, the difference is that trehalose is replaced by sorbitol, and the rest are the same.

[0109] Comparative Example 9

[0110] Compared with Example 2, the difference is that glycerol is replaced by butylene glycol, and the rest are the same.

[0111] Comparative Example 10

[0112] Compared with Example 3, the difference is that polyvinyl acrylate is removed, and the rest are the same.

[0113] Comparative Example 11

[0114] Compared with Example 3, the difference is that polyacrylamide is removed, and the rest are the same.

[0115] Comparative Example 12

[0116] Compared with Example 3, the difference is that polyvinyl acrylate is replaced by carbomer 940, and the rest are the same.

[0117] Comparative Example 13

[0118] Compared with Example 3, the difference is that polyacrylamide is replaced by xanthan gum, and the rest are the same.

[0119] Comparative Example 14

[0120] Compared with Example 1, the difference is that the tea polyphenol antioxidant is removed, and the rest are the same.

[0121] Comparative Example 15

[0122] Compared with Example 1, the difference is that the cross-linking agent is removed, and the rest are the same.

[0123] Comparative Example 16

[0124] Compared with Example 1, the difference is that the tea polyphenol antioxidant is replaced by vitamin C, and the rest are the same.

[0125] Comparative Example 17

[0126] Compared with Example 1, the difference is that the cross-linking agent is replaced by glutaraldehyde, and the rest are the same.

[0127] Test case 1: Verification of the synergistic repair effect of exosomes and low molecular weight peptides

[0128] Purpose of the experiment

[0129] Verify the synergistic repair effect of exosomes and low-molecular peptides, as well as the auxiliary enhancement function of trehalose on the repair effect.

[0130] This experiment was evaluated with reference to the following industry standards:

[0131] ISO 10993-5: Biocompatibility testing: In vitro cytotoxicity testing to ensure that the material has no toxic effects on cells.

[0132] ASTM E1440: Cell proliferation and migration ability test of biomaterials, providing standards for evaluating cell dynamic behavior.

[0133] OECD 439: Alternative skin irritation test to ensure that all exosome and low molecular weight peptide formulas are non-irritating to the skin.

[0134] Experimental groups

[0135] Example 1 (complete formulation)

[0136] Comparative Example 1 (Exosome Removal)

[0137] Comparative Example 2 (Removal of Low Molecular Weight Peptides)

[0138] Comparative Example 3 (exosomes replaced with liposomal EGF)

[0139] Comparative Example 4 (Low-molecular peptide replaced with ordinary collagen)

[0140] Comparative Example 5 (simultaneous removal of exosomes and trehalose)

[0141] Experimental materials and equipment

[0142] Animal model: SPF BALB / c mice (6 weeks old, half male and half female, n=8 per group);

[0143] Reagents: exosomes, low molecular weight peptides, trehalose;

[0144] Instruments: wound surface measurement instrument, ELISA detector, pathology slide scanner;

[0145] Detection kits: collagen quantification kit, TNF-α / IL-6 ELISA kit;

[0146] Experimental procedures

[0147] Wound model establishment:

[0148] The backs of mice were shaved, and after anesthesia, full-thickness skin defects were created using a 6 mm diameter biopsy needle.

[0149] Group processing:

[0150] The corresponding gel was applied once in the morning and evening every day (50 μL / time), while the blank control group was only applied with normal saline.

[0151] Data collection:

[0152] Wound healing time: record the time (days) when the wound area is reduced to 10%.

[0153] Collagen content: Wound tissue was collected on the 7th day, homogenized and tested by ELISA (μg / mg tissue).

[0154] Levels of inflammatory factors: Wound exudate was collected on the third day to detect TNF-α and IL-6 concentrations (pg / mL).

[0155] Experimental data

[0156] Table 1: Comparison of synergistic repair effects of exosomes and low molecular weight peptides

[0157]

[0158] Note:

[0159] The data are expressed as mean ± standard deviation.

[0160] Significance annotation: <0.001: extremely significant difference; <0.01: highly significant; <0.05: significant.

[0161] Experimental Summary

[0162] This study, through systematic removal and replacement of core components and cross-technical intervention, revealed the synergistic repair mechanism of exosomes and low-molecular-weight peptides. Exosomes, through their miRNAs (such as miR-21 and miR-29), directly activate fibroblast migration and collagen synthesis signaling pathways (TGF-β / Smad). Low-molecular-weight peptides (molecular weight <1000 Da), leveraging their small molecule penetration, rapidly inhibit the release of inflammatory factors such as TNF-α and IL-6 in the wound microenvironment, creating a two-way synergistic "repair-anti-inflammatory" effect. Data from Comparative Examples 1 (exosome removal) and 2 (peptide removal) showed that the absence of either component resulted in a 40%-50% decrease in collagen synthesis and a 2-3-fold increase in inflammatory factor levels, confirming the essential synergy of both components.

[0163] Further analysis revealed that the exosome's nanostructure (hollow lipid bilayer) is key to its functional specificity. While Comparative Example 3 (using liposomal EGF) exhibited similar initial repair rates, due to the lack of sustained regulation by endogenous miRNA within the exosomes, collagen deposition after 7 days was significantly lower than in Example 1 (27.9 vs. 35.6 μg / mg, P < 0.05), and the barrier repair index (TEWL) deteriorated by 30%. This result is highly consistent with the mechanism by which exosomes deliver active ingredients through vesicle-cell membrane fusion, whereas liposomes only provide short-term sustained drug release and cannot mimic the biological communication function of exosomes.

[0164] Comparative Example 5 (simultaneously removing exosomes and trehalose) showed a dramatic decrease in repair efficacy (collagen content 14.6 μg / mg, P < 0.001), revealing the hidden synergistic value of trehalose in the formula. Trehalose stabilizes the exosome membrane structure through the glass transition effect, preventing loss of activity during storage and transdermal penetration, indirectly ensuring repair efficacy.

[0165] Test Example 2: Verification of the synergistic moisturizing effect of trehalose and glycerin

[0166] Purpose of the experiment

[0167] Verify the synergistic moisturizing mechanism of trehalose and glycerol, and evaluate the impact of component replacement on moisturizing durability, skin feel and stability.

[0168] This experiment was evaluated with reference to the following industry standards:

[0169] ISO 11664-4: Standard for measuring hydration in cosmetic and personal care products, using a moisture meter to determine the hydration state of the skin and moisturizing effectiveness.

[0170] ISO 11930: Microbiological testing of cosmetics to verify the microbiological cleanliness of samples in terms of moisturizing effect and their stability.

[0171] ASTM D1002: This standard for evaluating surface tension is important for assessing a product's permeability and adhesion to the skin.

[0172] Experimental groups

[0173] Example 2 (complete formulation);

[0174] Comparative Example 6 (removal of trehalose);

[0175] Comparative Example 7 (removal of glycerol);

[0176] Comparative Example 8 (trehalose replaced with sorbitol);

[0177] Comparative Example 9 (glycerol replaced with butylene glycol);

[0178] blank control group (basic gel matrix);

[0179] Experimental materials and equipment

[0180] In vitro skin model: human abdominal skin (0.5 mm thick, stored in 4°C saline);

[0181] Reagents: trehalose, glycerol, sorbitol, butanediol;

[0182] Instruments: Corneometer® CM825 (hydration test), Tewameter® TM300 (TEWL value test), texture analyzer (TA.XTPlus);

[0183] Environmental conditions: constant temperature and humidity chamber with a temperature of 25°C and a humidity of 45%;

[0184] Experimental procedures

[0185] Sample preparation:

[0186] The ex vivo skin was cut into 3×3 cm segments, n=6 per group, and pretreated (delipidated and soaked in PBS for 30 minutes).

[0187] Processing and detection:

[0188] Evenly apply 0.1g of the corresponding group of gel and test at 0h, 4h, 12h, and 24h:

[0189] Hydration level (Corneometer®, units AU);

[0190] TEWL value (Tewameter®, unit: g / h·m²);

[0191] Skin feel rating (volunteer blind test: 1 point - extremely sticky, 5 points - refreshing).

[0192] The following is a detailed scoring rule for stickiness, which is used to quantify the skin comfort after application of the sample:

[0193]

[0194] Implementation Specifications

[0195] Test conditions:

[0196] Environment: constant temperature and humidity (25℃±1℃, humidity 50%±5%);

[0197] Application site: inner forearm (5×5cm area), clean with water and dry before each test.

[0198] Operation process:

[0199] Volunteers tested only one sample at a time, with a uniform dosage of 0.1g. They applied the sample 10 times in circles with their fingertips and then let it sit for 1 minute before scoring.

[0200] Assessment of stickiness requires simulating daily activities (eg, patting, rubbing clothing).

[0201] Data Records:

[0202] The rating must be based on immediate feelings and comparison with other samples is prohibited.

[0203] If the same volunteer scores the same sample multiple times with a difference of more than 1 point, retesting is required.

[0204] Example

[0205] Comparative Example 12 (Carbomer 940):

[0206] Rating 2 points (relatively sticky): After application, there is a noticeable sticky feeling, and there is a brief sticky sound when tapping the skin. The tester reported that it "feels like a layer of glue."

[0207] Example 3 (complete recipe):

[0208] Rating 4.5 points (close to refreshing): It feels slightly sticky at first, but completely dissipates after 30 seconds. The tester described it as "as comfortable as ordinary lotion."

[0209] Data Analysis:

[0210] The 24-hour hydration decrease rate and TEWL increase rate were calculated, and the differences in skin feel scores were statistically analyzed.

[0211] Experimental data

[0212] Table 2: Comparison of synergistic moisturizing properties between trehalose and glycerol (n=6)

[0213]

[0214] Note:

[0215] The data are expressed as mean ± standard deviation.

[0216] Significance annotation: <0.001: extremely significant difference; <0.01: highly significant; <0.05: significant.

[0217] Experimental Summary

[0218] The synergistic effect of trehalose and glycerol stems from their complementary moisturizing mechanisms: trehalose reduces water evaporation from the skin surface by forming a glassy film (TEWL value reduced to 8.5 g / h·m²), while glycerol's strong hygroscopicity rapidly increases stratum corneum hydration (hydration level reached 82.3 AU at 0 h). The TEWL value of Comparative Example 6 (trehalose removed) increased to 12.8 g / h·m² (P<0.001), demonstrating the irreplaceable role of trehalose in maintaining barrier integrity. In contrast, the initial hydration level of Comparative Example 7 (glycerol removed) decreased to 58.6 AU (P<0.01), confirming the key role of glycerol in immediate moisturization.

[0219] Component substitution experiments further revealed the need for structural specificity. Sorbitol (Comparative Example 8) failed to form a stable water-locking film due to differences in molecular polarity, resulting in a 24-hour hydration loss rate 20% higher than that of Example 2 (63.4 vs. 82.3 AU, P < 0.05). Butanediol (Comparative Example 9), while similar in hygroscopicity to glycerin, has a lower molecular weight (90.1 Da vs. 92.1 Da), making it more likely to migrate deeper into the skin, thereby reducing surface moisturization (skin feel score 2.6, P < 0.001). This result is consistent with glycerin's mechanism of anchoring water through a hydrogen bond network, highlighting the necessity of precise molecular structure design.

[0220] It is worth noting that the TEWL value of Comparative Example 9 (9.9 g / h·m²) is not significantly different from that of Example 2 (P=0.03), but the skin feel score is significantly deteriorated, indicating that the formula needs to take into account both efficacy and user experience.

[0221] Test Example 3: Verification of the synergistic thickening effect of polyvinyl acrylate and polyacrylamide

[0222] Purpose of the experiment

[0223] Verify the synergistic thickening effect of polyvinyl acrylate and polyacrylamide, and evaluate its impact on rheological properties, stability and user experience.

[0224] This experiment was evaluated with reference to the following industry standards:

[0225] ASTM D4693: Rheology test standard, using the "Rheometer Test Procedure" (Brookfield or similar instrument) to determine viscosity, flow and thixotropy.

[0226] ISO 3219: Determination of rheological properties of polymer solutions and dispersions provides methodological support for evaluating the flow properties of different polymers.

[0227] ISO 11058: Standard for the flowability of liquids and their properties, ensuring the scientific and reproducible test methods.

[0228] Experimental groups

[0229] Example 3 (complete recipe)

[0230] Comparative Example 10 (Removal of polyvinyl acrylate)

[0231] Comparative Example 11 (Removal of Polyacrylamide)

[0232] Comparative Example 12 (polyvinyl acrylate replaced by carbomer 940)

[0233] Comparative Example 13 (polyacrylamide replaced by xanthan gum)

[0234] Blank control group (basic solution without thickener added)

[0235] Experimental materials and equipment

[0236] Materials: polyvinyl acrylate, polyacrylamide, carbomer 940, xanthan gum;

[0237] Instruments: Brookfield RV rheometer (spindle 7, 20 rpm), texture analyzer (TA.XT Plus), constant temperature and humidity chamber (40 ° C / 75% RH);

[0238] Testing criteria: thixotropic index (TI = recovery time / initial viscosity), sensory score (blind test with 10 people: 1 point - very poor, 5 points - excellent);

[0239] Experimental procedures

[0240] Sample preparation:

[0241] Weigh the thickener according to the group, disperse it in deionized water, stir magnetically (700 rpm, 25°C) until completely dissolved, and let it stand to defoam.

[0242] Rheological properties test:

[0243] Initial viscosity: measured by Brookfield RV rheometer (20 rpm, 25°C, unit: cP);

[0244] Thixotropic index: After high-speed shear (10,000 s⁻¹, 1 minute), the viscosity is allowed to recover and the time (in seconds) required to recover to 50% is recorded.

[0245] Stability test:

[0246] Place the sample in a constant temperature and humidity chamber (40°C / 75%RH) and observe whether it delaminates or liquefies within 7 days.

[0247] User experience evaluation:

[0248] Volunteers blindly tested the smoothness of application, residual white marks and stickiness, and gave a comprehensive score (1-5 points).

[0249] The following are the detailed scoring rules for application smoothness, residual white marks, and stickiness in Experiment 3:

[0250] Smoothness of application:

[0251]

[0252] Residual white marks:

[0253]

[0254] Sticky feeling:

[0255]

[0256] Comprehensive score calculation:

[0257] Total score = (smoothness score + residual white mark score + stickiness score) / 3

[0258] Example: If the smoothness is 4 points, the residual white marks are 5 points, and the stickiness is 4 points, then the total score is (4+5+4) / 3, which is 4.3 points.

[0259] Implementation Specifications:

[0260] Blind test conditions:

[0261] Volunteers need to be tested in an environment of 25℃ and 50% humidity, and use a unified measuring tool (0.1g sample) after cleaning their hands.

[0262] Operation process:

[0263] Only one sample was tested at a time, and the application area was the inner forearm (5×5 cm). The next sample was tested 30 minutes after the test.

[0264] Data Records:

[0265] The score sheet must be submitted anonymously to avoid subjective interference. Abnormal data (such as a single item score difference > 2 points) must be retested and confirmed.

[0266] Experimental data

[0267] Table 3: Comparison of rheological properties and stability of thickening systems (n=3)

[0268]

[0269] Note:

[0270] The data are expressed as mean ± standard deviation.

[0271] Significance annotation: <0.001: extremely significant difference; <0.01: highly significant; <0.05: significant.

[0272] Experimental Summary

[0273] The synergistic thickening effect of polyvinyl acrylate and polyacrylamide stems from the complementary nature of their molecular structures. Polyvinyl acrylate forms a three-dimensional network through hydrophobic association, providing a high elastic modulus (initial viscosity of 20,300 cP), while the long linear chains of polyacrylamide enhance thixotropy (TI value of 5.3) through hydrogen bonding. The combination of these two achieves a perfect balance of "high viscosity and easy spreadability." The system in Comparative Example 10 (without polyvinyl acrylate) completely failed to gel, demonstrating its irreplaceable role as a structural backbone. Meanwhile, the thixotropy index of Comparative Example 11 (without polyacrylamide) dropped to 1.8 (P<0.001), resulting in excessive fluidity after application (smoothness score of 2.6), confirming the critical influence of thixotropy on the user experience.

[0274] Component substitution experiments further revealed material-specific requirements. Carbomer 940 (Comparative Example 12) exhibited excessively high viscosity (52,800 cP) and poor thixotropic recovery (TI=3.1) due to excessive cross-linking, resulting in a user experience rating of only 2.1 (P<0.001). While xanthan gum (Comparative Example 13) maintained a similar viscosity (18,700 cP, P=0.08), its shear-thinning properties were insufficient (TI=2.2), leaving a noticeable white mark after application (score 3.4 vs. 4.5). This result is consistent with the mechanism by which polyacrylamide dynamically responds to shear forces through entanglement and disentanglement, highlighting the necessity of designing for specific rheological properties.

[0275] The implicit synergistic effects of process parameters are also not negligible. The stirring rate (700 rpm) in Example 3 ensured uniform dispersion of the polymer, while in Comparative Example 13, under the same process, the xanthan gum was unevenly dispersed (the smoothness score decreased) due to its molecular rigidity.

[0276] Test Example 4: Verification of the synergistic antioxidant effect of tea polyphenols and cross-linking agents

[0277] Purpose of the experiment

[0278] Verify the synergistic antioxidant effect of tea polyphenols and cross-linkers, and evaluate their effects on free radical scavenging, UV repair and safety.

[0279] This experiment was evaluated with reference to the following industry standards:

[0280] ISO 10993-5: Biocompatibility testing: In vitro cytotoxicity testing to detect the toxic effects of materials on cells and ensure the safety of the ingredients.

[0281] OECD 451: In vitro antioxidant efficacy test standard for cosmetics and personal care products, evaluating the antioxidant activity of different combinations.

[0282] ASTM D1609: This test method is used to determine the relative stability and resistance of samples to damage under ultraviolet radiation.

[0283] ISO 16128: Naturalness and origin standards for cosmetic ingredients, testing the acceptability and certification specifications of tea polyphenols and cross-linking agents in skin care products.

[0284] Experimental groups

[0285] Example 1 (complete formulation)

[0286] Comparative Example 14 (Removal of Tea Polyphenols)

[0287] Comparative Example 15 (Removal of Cross-linking Agent)

[0288] Comparative Example 16 (Tea Polyphenols Replaced with Vitamin C)

[0289] Comparative Example 17 (cross-linking agent replaced by glutaraldehyde)

[0290] Blank control group (no antioxidant added)

[0291] Experimental materials and equipment

[0292] Materials: tea polyphenols, biodegradable cross-linking agent, vitamin C, glutaraldehyde;

[0293] Instruments: UV spectrophotometer (UV-2600), fluorescence microscope (Olympus IX83), cell culture incubator (37°C / 5% CO2);

[0294] Detection methods: DPPH free radical scavenging rate (517nm absorbance), SOD activity (xanthine oxidase method), MTT cytotoxicity assay (570nm absorbance);

[0295] Experimental procedures

[0296] Free radical scavenging experiment:

[0297] Each group of samples was mixed with DPPH solution (0.1 mM), reacted in the dark for 30 minutes, and the absorbance at 517 nm was measured to calculate the clearance rate.

[0298] UV damage repair experiment:

[0299] Human keratinocytes (HaCaT) were irradiated with UVB (30 mJ / cm²) and then treated with each sample group for 24 hours, and the SOD activity (U / mg protein) was detected.

[0300] Cytotoxicity test:

[0301] NIH / 3T3 cells were co-cultured with the samples for 48 h, and the cell viability (LD50 value) was determined by MTT assay.

[0302] Experimental data

[0303] Table 4: Comparison of antioxidant and safety of tea polyphenols and cross-linking agents (n=5)

[0304]

[0305] Note:

[0306] The data are expressed as mean ± standard deviation.

[0307] Significance annotation: <0.001: extremely significant difference; <0.01: highly significant; <0.05: significant.

[0308] Experimental Summary

[0309] The synergistic effect of tea polyphenols and cross-linkers is reflected in the dual improvement of antioxidant activity and controlled release ability. Tea polyphenols (EGCG) directly scavenges free radicals through phenolic hydroxyl groups (DPPH scavenging rate 84.6%) and activates cellular SOD enzyme activity (52.3U / mg), while the cross-linker forms a gel network through dynamic disulfide bonds, delaying the release of tea polyphenols (SOD activity of comparative example 15 dropped to 34.6U / mg, P<0.01), ensuring long-term repair. Removing tea polyphenols (comparative example 14) resulted in a collapse of free radical scavenging ability (29.8% vs. 84.6%), and the ultraviolet repair rate was close to that of the blank control group (8.5 vs. 5.1U / mg), confirming its irreplaceable core role.

[0310] The structural design of cross-linkers is crucial for safety. While glutaraldehyde (Comparative Example 17) maintains similar antioxidant efficacy (DPPH 82.4% vs. 84.6%), its irreversible cross-linking properties lead to a surge in cytotoxicity (LD50 = 185 μg / mL, P < 0.001). Biodegradable cross-linkers, on the other hand, achieve controlled release through enzyme-responsive cleavage (LD50 > 1000 μg / mL). This difference is directly related to the cross-linker's chemical bond type (dynamic vs. static), highlighting the critical importance of biocompatibility in molecular design.

[0311] The systemic synergy of the formula is further reflected in the balance between stability and efficacy. Vitamin C (Comparative Example 16) is easily oxidized and inactivated, with its clearance rate dropping below 50% after 6 hours (51.3% vs. 84.6%, P < 0.001), and is unable to activate the SOD pathway (22.7 U / mg). However, tea polyphenols remain stable under the protection of a crosslinker (LD50 > 1000 μg / mL in Example 1).

[0312] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A long-lasting moisturizing and barrier-repairing exosome active gel, characterized in that: The gel comprises the following components in parts by mass: Exosomes: 2-5; Trehalose: 3-7 parts; Glycerin: 2-6 parts; Polyvinyl acrylate: 1-2 parts; Octyl glucoside: 0.5-1.5 parts; Sodium hyaluronate: 0.8-1.5 parts; Low molecular weight peptide: 0.2-0.5 parts; Polyacrylamide: 1-3 parts; Tea polyphenol antioxidants: 0.02-0.05 parts; Cross-linking agent: 0.1-0.3 parts; Water: 70-80 parts; The exosomes are derived from placenta-derived mesenchymal stem cells or umbilical cord-derived mesenchymal stem cells, and the glycerol is plant-derived glycerol; The low-molecular peptide is a collagen peptide with a molecular weight of less than 3000Da, and its amino acid sequence contains Gly-Pro-Hyp repeating units. The tea polyphenol antioxidant is a combination of epigallocatechin gallate and rosmarinic acid in a mass ratio of 2:1-3:

2. The cross-linking agent is a biodegradable water-soluble cross-linking agent.

2. The long-lasting moisturizing and barrier-repairing exosome active gel according to claim 1, characterized in that: The polyvinyl acrylate is a high molecular polymer, the octyl glucoside is a nonionic surfactant, and the sodium hyaluronate is purified sodium hyaluronate.

3. The long-lasting moisturizing and barrier-repairing exosome active gel according to claim 1, characterized in that: The water is ultrapure water that has been deionized and has a resistivity of ≥18 MΩ·cm.

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