Ganoderma lucidum exosome-loaded skin care gel and preparation method thereof

Abstract

The invention relates to the field of skincare products, and particularly discloses skincare gel loaded with ganoderma lucidum exosome and a preparation method of the skincare gel. The skin care gel loaded with the lucid ganoderma exosome is prepared from the following raw materials in parts by weight: 20 to 40 parts of hydrogel matrix, 2 to 4 parts of lucid ganoderma exosome nanoparticles, 2 to 6 parts of niobium carbide nanosheets, 1 to 3 parts of nicotinamide, 0.1 to 2 parts of phenethyl resorcinol and 0.1 to 3 parts of green tea extract, the Mailing exosome nanoparticles are of a core-shell structure, a core layer comprises Mailing exosome and papain in a mass ratio of 1: (0.1-0.2), and a shell layer comprises a polylactic acid-glycolic acid copolymer and a penetration enhancer in a mass ratio of 1: (0.1-0.3). The skin-care gel provided by the invention has remarkable whitening, moisturizing and skin brightening effects.

Description

Technical Field

[0001] This application relates to the field of beauty and skincare technology, and more specifically, it relates to a skincare gel loaded with Ganoderma lucidum exosomes and its preparation method. Background Technology

[0002] Beauty and skincare have always been a hot topic. With advancements in technology and increasing consumer demand for skincare products, various new skincare products have emerged on the market. The main active ingredients responsible for inhibiting melanin production in whitening skincare products are kojic acid, arbutin, retinoic acid, hydroquinone, and ascorbic acid. They reduce melanin expression by inhibiting tyrosinase activity, thus achieving a whitening effect. However, these compounds have some side effects during application, including cytotoxicity, poor stability, low bioavailability, and skin irritation that can easily trigger allergies or dermatitis, and even cause DNA damage and cancer. These drawbacks limit their application in the field of beauty and skincare.

[0003] Hydrogels, due to their unique advantages—high transparency and excellent viscoelasticity, a uniform gel system, a glossy and crystal-clear appearance—and their three-dimensional network structure effectively encapsulating active ingredients, prolonging their effects, are increasingly widely used in skincare products. In the beauty and skincare field, gels can be used as basic daily skincare products, such as moisturizing gels and cleansing gels, as well as as special care products, such as acne-fighting gels, whitening gels, and medical fillers.

[0004] However, since melanin production occurs in the basal layer of the skin, the ingredients that inhibit melanin production are degraded by factors such as sunlight after whitening products are applied, resulting in a reduced amount penetrating into the basal layer. This reduces their ability to effectively inhibit melanin production. Furthermore, the stratum corneum acts as a barrier to the transdermal absorption of drugs, hindering their absorption. The poor permeability of the active ingredients leaves some of them on the skin surface, preventing the drugs from fully exerting their effects and thus failing to achieve effective whitening and skincare benefits. Summary of the Invention

[0005] To achieve a more significant whitening effect, this application provides a skin care gel loaded with Ganoderma lucidum exosomes and a method for preparing the same.

[0006] In a first aspect, this application provides a skin care gel carrying Ganoderma lucidum exosomes, employing the following technical solution:

[0007] A skin care gel carrying Ganoderma lucidum exosomes comprises the following raw materials in parts by weight: 20-40 parts hydrogel matrix, 2-4 parts Ganoderma lucidum exosome nanoparticles, 2-6 parts niobium carbide nanosheets, 1-3 parts niacinamide, 0.1-2 parts phenylethyl resorcinol and 0.1-3 parts green tea extract;

[0008] The spirosome nanoparticles have a core-shell structure. The core layer consists of spirosomes and papain in a mass ratio of 1:0.1-0.2, and the shell layer consists of polylactic acid-glycolic acid copolymer and a penetration enhancer in a mass ratio of 1:0.1-0.3.

[0009] By adopting the above technical solutions, the thin texture of the hydrogel facilitates the penetration of active ingredients. Green tea extract has antioxidant properties, which can slow down photoaging, reduce inflammation and soothe, inhibit bacteria and moisturize. Niacinamide can promote ceramide synthesis, enhance the skin's water-locking ability, inhibit melanin transfer to the epidermis, even out skin tone, and promote collagen synthesis. Phenethyl resorcinol, commonly known as 377, can effectively whiten and brighten skin tone. Niobium carbide nanosheets have antioxidant and antibacterial effects, which can effectively scavenge hydroxyl radicals and superoxide anions. At the same time, in in vitro oxidation, they can avoid oxidative damage and inhibit the growth of Escherichia coli and Staphylococcus aureus.

[0010] The coating material, polylactic acid-glycolic acid copolymer, is insoluble in water but degrades within the body. By encapsulating Ganoderma lucidum exosomes and papain, the nanoparticles can penetrate through the gaps between cells in the intercellular layer or through appendages such as sweat glands and hair follicles, making it easier to reach the deep epidermis and even the dermis. This prolongs the residence time of the nanoparticles on the skin surface, reduces loss, and indirectly improves penetration efficiency. After the Ganoderma lucidum exosome nanoparticles enter the skin, the coating material degrades in the in vivo environment, releasing the internal active ingredients, Ganoderma lucidum exosomes and papain. This gives the Ganoderma lucidum exosome nanoparticles a pre-penetration and pre-release effect, preventing the ingredients from being oxidized or decomposed on the skin surface and improving bioavailability.

[0011] After Ganoderma lucidum exosomes and papain penetrate into the skin, the exosomes, rich in Ganoderma lucidum polysaccharides and triterpenoids, can remove free radicals on the skin surface, reduce dullness and yellowing of skin caused by oxidative stress, promote cell repair, enhance the skin barrier, inhibit tyrosinase activity, and reduce melanin synthesis, thus achieving a whitening effect.

[0012] Papain is a cysteine ​​protease that can hydrolyze keratin and desmosomes in the stratum corneum, disrupting the connections between keratinocytes, loosening the stratum corneum structure, increasing skin permeability, promoting the shedding of the stratum corneum, and assisting the penetration of Ganoderma lucidum exosomes. This enhances the permeability of Ganoderma lucidum exosomes, delivering more active ingredients to the basal layer of the epidermis (where melanocytes are located), directly acting on the target site, and enhancing the whitening effect. In addition, papain can also accelerate the metabolism of old and dead keratinocytes containing melanin, making the skin tone more even.

[0013] Therefore, papain removes existing melanin by metabolizing dead skin cells, while Ganoderma lucidum exosomes inhibit the formation of new melanin by delivering active ingredients. Both work synergistically to improve dull skin tone and pigmentation issues from different perspectives. Furthermore, the anti-inflammatory effects of papain (such as inhibiting the release of inflammatory factors) may reduce post-inflammatory hyperpigmentation caused by UV radiation or stimulation, indirectly aiding in skin whitening.

[0014] Encapsulating Ganoderma lucidum exosomes with encapsulating materials can prevent the vesicle structure from rupturing due to environmental factors and the loss of internal active ingredients. This allows for a process of penetration followed by release: after encapsulation, the exosomes remain stable on the skin surface. Once inside the epidermis or dermis, the encapsulating material dissolves under the influence of body temperature, enzymes, or pH, releasing the internal exosomes and their active ingredients. This prevents premature oxidation of the ingredients on the skin surface and improves bioavailability.

[0015] Optionally, the mass ratio of the shell solution to the core solution of the Ganoderma lucidum exosome nanoparticles is 1.5-2.5:1.

[0016] By employing the above technical solution and preparing Ganoderma lucidum exosome nanoparticles at the above mass ratio, the shell layer can form a complete encapsulation, isolating papain from external moisture, oxygen, and other substances that could damage it. A higher shell layer ratio results in a thicker shell layer, providing stronger protection for the core layer and improving the storage stability of Ganoderma lucidum exosomes and papain. Increasing the amount of penetration enhancer accelerates skin penetration, but an excessively thick shell layer can affect penetration, delaying shell dissolution and causing slow release of core components, thus reducing immediate efficacy. Conversely, a lower shell layer ratio and a higher core layer ratio, while resulting in higher active ingredient content and stronger efficacy, can lead to the shell layer failing to encapsulate the core layer, causing papain and Ganoderma lucidum exosomes to be exposed and prematurely inactivated, reducing bioavailability. Furthermore, insufficient penetration enhancer dosage reduces penetration efficiency.

[0017] Optionally, the preparation method of the Ganoderma lucidum exosome nanoparticles is as follows:

[0018] Papain from Ganoderma lucidum exosomes was dispersed in ultrapure water containing 0.1% Tween-80 and stirred evenly to obtain a core layer solution.

[0019] PLGA and a penetration enhancer were added to an acetone-ethanol mixed solvent and stirred until homogeneous to obtain a shell solution.

[0020] While stirring, the shell solution was slowly dripped into the core solution. After the addition was completed, stirring was continued for 30 minutes. Acetone / ethanol diffused into the aqueous phase, and PLGA gradually precipitated out and coated the core layer to form a nanoparticle suspension.

[0021] The nanoparticles were collected by centrifugation using an ultrafiltration membrane with a molecular weight cutoff of 10 kDa to remove free solvent and small molecules.

[0022] By adopting the above technical solution, the penetration enhancer in the shell can be combined with the penetration ability of nanoparticles to improve skin penetration efficiency through a dual mechanism of carrier penetration and ingredient-promoted penetration, without relying on piercing the skin, making it safer and gentler.

[0023] Optionally, the niobium carbide nanosheets are loaded with a tyrosinase / ZIF-8 complex.

[0024] By adopting the above technical solution, tyrosinase, a key enzyme in melanin synthesis, is loaded with ZIF-8. Its microporous structure provides excellent protection for the enzyme molecules, improving their storage performance and environmental tolerance. It can also inhibit melanin synthesis and reduce skin pigmentation by regulating tyrosinase activity, thus achieving a skin whitening effect. Furthermore, ZIF-8 has good anti-inflammatory effects, and niobium carbide nanosheets also help regulate skin inflammatory responses. The combination of the two can help reduce skin inflammation and alleviate problems such as redness and sensitivity. At the same time, ZIF-8 can also promote skin healing and help heal damaged skin, while niobium carbide nanosheets can help reduce skin moisture loss, resist the invasion of harmful external substances, improve the overall health of the skin, reduce wrinkles, and improve skin elasticity.

[0025] Optionally, the niobium carbide nanosheets loaded with the tyrosinase / ZIF-8 complex are prepared as follows:

[0026] Carboxylated niobium carbide nanosheets were mixed with zinc nitrate hexahydrate, tyrosinase, and deionized water, shaken for 10-20 min, 2-methylimidazole was added, and the mixture was shaken at a constant temperature for 10-20 min. The mixture was then washed with deionized water, filtered, and freeze-dried.

[0027] By adopting the above technical solution, zinc ions can fully chelate with the carboxyl groups in carbon nanosheets. Zinc ions preferentially adsorb on the surface of niobium carbide nanosheets, providing nucleation sites for ZIF-8 and guiding the directional growth of ZIF-8 on the nanosheet surface. Tyrosinase is bound to the porous structure of ZIF-8 through intermolecular forces and is encapsulated in the framework of ZIF-8. Moreover, the carboxyl groups on the surface of carboxylated niobium carbide nanosheets interact with the amino and carboxyl groups in the tyrosinase molecule, which can also indirectly fix the tyrosinase at the interface between ZIF-8 and the nanosheet.

[0028] Optionally, the penetration enhancer is selected from one or more of laurocapram, hyaluronidase, cyclohexanehexyl alcohol, polyethylene glycol 400 monostearate, isopropyl myristate, N-methylpyrrolidone, and sodium glycocholate.

[0029] By adopting the above technical solutions, the above penetrants work together to enhance penetration efficiency, relieve skin dryness, and improve the overall skin care experience.

[0030] Optionally, the penetration enhancer includes laurocapram and hyaluronidase in a mass ratio of 3-5:1.

[0031] By employing the above technical solution, laurocapram, being lipid-soluble, can be stably dispersed in the shell and act uniformly on the stratum corneum, opening channels for nanoparticle penetration. Hyaluronidase, being a water-soluble protein, is prone to uneven dispersion and aggregation and inactivation in the shell if the amount added is too large, while if the amount added is too small, it cannot effectively degrade the dermal matrix. During nanoparticle penetration, laurocapram first breaks through the stratum corneum, efficiently opening the stratum corneum pathway. Subsequently, hyaluronidase accumulates locally, making it easier to exert its degradation effect and promoting the diffusion of components into the dermis. Therefore, using laurocapram to disrupt stratum corneum lipids and hyaluronidase to degrade the intercellular matrix achieves better penetration.

[0032] Optionally, the hydrogel matrix is ​​selected from at least one of carbomer, sodium hyaluronate, sodium alginate, polyethylene glycol, gum arabic, polyvinylpyrrolidone, polyvinyl alcohol, sodium polyacrylate, hydroxypropyl methylcellulose, and carboxyethyl cellulose.

[0033] By adopting the above technical solutions, the above materials are skin-friendly and gentle, with high transparency, and can achieve moisturizing and soothing effects.

[0034] Optionally, the hydrogel matrix comprises sodium hyaluronate and hydroxypropyl methylcellulose in a mass ratio of 1:1-1.5.

[0035] By adopting the above technical solution, sodium hyaluronate has good moisturizing and biocompatibility, can relieve skin irritation, and provide a refreshing skin feel. The stable gel structure can be mixed. The hydroxyl groups of hydroxypropyl methylcellulose can synergistically enhance the dispersion stability of niobium carbide nanosheets with the carboxyl groups of niobium carbide nanosheets. It also has high compatibility with zinc ions and enzymes, ensuring that the ZIF-8 structure and tyrosinase activity are not damaged.

[0036] Secondly, this application provides a method for preparing a skin care gel loaded with Ganoderma lucidum exosomes, using the following technical solution:

[0037] A method for preparing a skin care gel loaded with Ganoderma lucidum exosomes includes the following steps:

[0038] Mix the hydrogel matrix and deionized water, heat to 65-80℃, and homogenize for 15-30 minutes to obtain a gel mixture.

[0039] The skin care gel was prepared by uniformly mixing the gel mixture with Ganoderma lucidum exosome nanoparticles, niobium carbide nanosheets, niacinamide, phenylethyl resorcinol and green tea extract.

[0040] By adopting the above technical solution, the hydrogel matrix is ​​dissolved in deionized water to prepare a gel mixture, which is then mixed with the remaining components. The preparation method is simple and can be used in skin care lotions, as a face mask liquid, or as an injectable medical cosmetic filler after homogenization, extrusion sieving, and high-temperature and high-pressure steam sterilization.

[0041] Optionally, the viscosity of the gel mixture is 10,000-30,000 mPa·s.

[0042] By adopting the above technical solution, the gel mixture of this viscosity can make the Ganoderma lucidum exosome nanoparticles disperse evenly and easy to apply evenly.

[0043] In summary, this application has the following beneficial effects:

[0044] 1. Because this application uses polylactic acid-glycolic acid copolymer as the coating material and adds a penetration enhancer to encapsulate Ganoderma lucidum exosomes and papain, it not only reduces the activity loss of Ganoderma lucidum exosomes and increases their stability, but also improves the penetration effect of nanoparticles by using a penetration enhancer. Furthermore, the hydrogel matrix balances the stability of PLGA and skin permeability. Through the dissolution of PLGA in the skin, the retention time of Ganoderma lucidum exosomes in the skin is prolonged, reducing loss, improving penetration efficiency and bioavailability, and improving the whitening, skin rejuvenation and brightening effects.

[0045] 2. In this application, a tyrosinase / ZIF-8 complex is preferably loaded on niobium carbide nanosheets. The microporous structure of ZIF-8 has a good protective effect on tyrosinase, improves the activity and stability of the enzyme, and ZIF-8 can also reduce skin inflammation. Tyrosinase can inhibit the synthesis of melanin, further enhancing the whitening effect of the skin care gel.

[0046] 3. In this application, laurocapram and hyaluronidase are preferred as penetration enhancers. Laurocapram is lipid-soluble and can increase the compatibility of PLGA with stratum corneum lipids and reduce rejection. Hyaluronidase can hydrolyze hyaluronic acid, which is one of the main components of the skin extracellular matrix, especially abundant in the dermis, thus playing a role in deep delivery. Therefore, laurocapram enhances the fluidity of stratum corneum lipids and helps nanoparticles enter, while hyaluronidase promotes diffusion in the dermis and reduces the hyaluronic acid barrier. Detailed Implementation

[0047] The following embodiments provide a further detailed description of this application.

[0048] Preparation Examples of Ganoderma lucidum exosome nanoparticles 1-7

[0049] The sources of each raw material in the preparation example are as follows: Papain was selected from Xi'an Xihai Biotechnology, catalog number XH504, with an activity of 100,000 / g; the ratio of lactic acid to glycolic acid in PLGA was 50:50, selected from MCE (MedChemExpress), model in-stock; hyaluronidase was selected from Beijing Huamaike Biotechnology, catalog number H3506; Ganoderma lucidum exosomes were prepared using the following method:

[0050] The Ganoderma lucidum spawn (selected from Shandong Luzhiguan Pharmaceutical Co., Ltd., product number 002) was transferred and activated: Sterilized PDA medium (potato 200g / l, KH2PO4 3g / l, Mg2SO4·7H2O 1.5g / l, glucose 20g / l, vitamin B1 10mg / l, agar powder 20g / l) was poured into a Petri dish. After the medium cooled, a piece of Ganoderma lucidum spawn with a diameter of 5mm was inoculated into the center of the plate and cultured at 25℃ for 7 days. A piece of activated spawn with a diameter of 5mm was picked from the plate and inoculated into liquid medium (potato 200g / l, KH2PO4 3g / l, Mg2SO4·7H2O 1.5g / l, vitamin B1 10mg / l, agar powder 20g / l). The mixture was then cultured in a shaker at 140r / min and 25℃ for 7 days.

[0051] Ganoderma lucidum strain fermentation culture: The activated Ganoderma lucidum was inoculated into PDA liquid culture medium for fermentation culture. The shaking speed was 140 r / min, the culture temperature was 25℃, and the culture was carried out for 2-3 days to obtain Ganoderma lucidum fermentation broth.

[0052] Ultracentrifugation: Collect the Ganoderma lucidum fermentation broth. After filtering the fermentation broth through a sterile eight-layer gauze, centrifuge at high speed: 4℃, 300g for 10min, take the supernatant, centrifuge at 4℃, 2000g for 15min, take the supernatant, centrifuge at 4℃, 10000g for 30min; take the supernatant and ultracentrifuge: 4℃, 110000g for 90min, resuspend the precipitate with PBS, centrifuge at 4℃, 11000g for 90min, resuspend the precipitate with an appropriate amount of PBS, and store at -80℃.

[0053] Preparation Example 1: (1) Ganoderma lucidum exosomes and papain were dispersed in ultrapure water containing 0.1 wt% Tween-80 and stirred evenly to obtain a core layer solution. The concentration of Ganoderma lucidum exosomes was 5 mg / ml, the concentration of papain was 1 mg / l, and the mass ratio of Ganoderma lucidum exosomes to papain was 1:0.2.

[0054] (2) Add 8 mg of PLGA (lactic acid / glycolic acid ratio 50:50) and the penetration enhancer to an acetone-ethanol mixed solvent (volume ratio 3:1), stir evenly to obtain a shell solution. The concentration of PLGA is 8 wt%, the mass ratio of PLGA to penetration enhancer is 1:0.3, and the penetration enhancer includes laurocapram and hyaluronidase in a mass ratio of 5:1.

[0055] (3) Under magnetic stirring (600 rpm), the shell solution was slowly added dropwise to the core solution. After the addition was completed, stirring was continued for 30 min. Acetone / ethanol diffused into the aqueous phase, and PLGA gradually precipitated and coated the core layer to form a nanoparticle suspension. The mass ratio of the shell solution to the core solution was 2.5:1.

[0056] (4) Centrifuge with an ultrafiltration membrane with a molecular weight cutoff of 10 kDa (3000 g, 10 min) to remove free solvent and small molecules, and collect Ganoderma exosome nanoparticles.

[0057] Preparation Example 2: (1) Ganoderma lucidum exosomes and papain were dispersed in ultrapure water containing 0.1 wt% Tween-80 and stirred evenly to obtain a core layer solution. The concentration of Ganoderma lucidum exosomes was 5 mg / ml, the concentration of papain was 0.5 mg / l, and the mass ratio of Ganoderma lucidum exosomes to papain was 1:0.1.

[0058] (2) Add 8 mg of PLGA (lactic acid / glycolic acid ratio 50:50) and the permeation enhancer to an acetone-ethanol mixed solvent (volume ratio 3:1), stir evenly to obtain a shell solution. The concentration of PLGA is 4 wt%, and the mass ratio of PLGA to permeation enhancer is 1:0.1. The permeation enhancer includes laurocapram and hyaluronidase in a mass ratio of 3:1.

[0059] (3) Under magnetic stirring (600 rpm), the shell solution was slowly added dropwise to the core solution. After the addition was completed, stirring was continued for 30 min. Acetone / ethanol diffused into the aqueous phase, and PLGA gradually precipitated and coated the core layer to form a nanoparticle suspension. The mass ratio of the shell solution to the core solution was 1.5:1.

[0060] (4) Centrifuge with an ultrafiltration membrane with a molecular weight cutoff of 10 kDa (3000 g, 10 min) to remove free solvent and small molecules, and collect Ganoderma exosome nanoparticles.

[0061] Preparation Example 3: The difference from Preparation Example 1 is that the penetration enhancer is laurocapram.

[0062] Preparation Example 4: The difference from Preparation Example 1 is that the permeation enhancer includes cyclohexanehexol and hyaluronidase in a mass ratio of 5:1.

[0063] Preparation Example 5: The penetration enhancer comprises laurocapram and cyclohexanehexol in a mass ratio of 5:1.

[0064] Preparation Example 6: The difference from Preparation Example 1 is that no papain was added to the core layer. The Ganoderma lucidum exosomes were dispersed in ultrapure water containing 0.1 wt% Tween-80 and stirred evenly to obtain a core layer solution with a concentration of 5 mg / ml.

[0065] Preparation Example 7: The difference from Preparation Example 1 is that no permeation enhancer was added to the shell layer. 8 mg of PLGA (lactic acid / glycolic acid ratio 50:50) was added to an acetone-ethanol mixed solvent (volume ratio 3:1), stirred evenly, and a shell layer solution was obtained with a PLGA concentration of 8 wt%.

[0066] Example

[0067] Example 1: A skin care gel loaded with Ganoderma lucidum exosomes. The raw material amounts are shown in Table 1. The hydrogel matrix includes sodium hyaluronate and hydroxypropyl methylcellulose in a mass ratio of 1:1.5. The sodium hyaluronate is selected from Guangzhou Fengyi Biotechnology, with a molecular weight of 120 kDa. The hydroxypropyl methylcellulose is selected from Guangzhou Diweisai New Materials, model HPMC. The Ganoderma lucidum exosome nanoparticles are prepared from Preparation Example 1. The green tea extract is selected from Hansuyuan (Shaanxi) Biotechnology, product number D98954545.

[0068] The preparation method of the above-mentioned skin care gel loaded with Ganoderma lucidum exosomes includes the following steps:

[0069] Sodium hyaluronate and hydroxypropyl methylcellulose were mixed and added to deionized water. After mixing, the mixture was heated to 80°C and homogenized for 15 minutes to prepare a gel mixture with a viscosity of 30,000 mPa·s.

[0070] The skin care gel was prepared by uniformly mixing the gel mixture with Ganoderma lucidum exosome nanoparticles, niobium carbide nanosheets, niacinamide, phenylethyl resorcinol and green tea extract.

[0071] Table 1. Amounts of raw materials used in skin care gels in Examples 1-3

[0072]

[0073] Example 2: A skin care gel loaded with Ganoderma lucidum exosomes. The raw material amounts are shown in Table 1. The hydrogel matrix includes sodium hyaluronate and hydroxypropyl methylcellulose in a mass ratio of 1:1. The sodium hyaluronate is selected from Guangzhou Fengyi Biotechnology, with a molecular weight of 120 kDa. The hydroxypropyl methylcellulose is selected from Guangzhou Diweisai New Materials, model HPMC. The Ganoderma lucidum exosome nanoparticles are prepared from Preparation Example 2. The green tea extract is selected from Hansuyuan (Shaanxi) Biotechnology, product number D98954545.

[0074] The preparation method of the above-mentioned skin care gel loaded with Ganoderma lucidum exosomes includes the following steps:

[0075] Sodium hyaluronate and hydroxypropyl methylcellulose were mixed and added to deionized water. After mixing, the mixture was heated to 65°C and homogenized for 30 minutes to prepare a gel mixture with a viscosity of 10000 mPa·s.

[0076] The skin care gel was prepared by uniformly mixing the gel mixture with Ganoderma lucidum exosome nanoparticles, niobium carbide nanosheets, niacinamide, phenylethyl resorcinol and green tea extract.

[0077] Example 3: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that the amount of raw materials used is shown in Table 1.

[0078] Example 4: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that the Ganoderma lucidum exosome nanoparticles are prepared by Example 3.

[0079] Example 5: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that the Ganoderma lucidum exosome nanoparticles are prepared by Example 4.

[0080] Example 6: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that the Ganoderma lucidum exosome nanoparticles are prepared by Example 5.

[0081] Example 7: A skin care gel loaded with Ganoderma lucidum exosomes, differing from Example 1 in that the surface of the niobium carbide nanosheets is loaded with a tyrosinase / ZIF-8 complex. The method for preparing the niobium carbide nanosheets with the tyrosinase / ZIF-8 complex loaded on the surface is as follows:

[0082] Niobium carbide nanosheets were added to deionized water and sonicated for 30 min to obtain a dispersion with a concentration of 2 mg / ml. A 0.5 M citric acid solution was added, and the mixture was stirred in a water bath at 60 °C for 12 h. After centrifugation at 10,000 rpm for 15 min, the mixture was washed three times with deionized water and dried under vacuum at 60 °C to obtain carboxylated niobium carbide nanosheets. The niobium carbide nanosheets were selected from Beijing Deco Island Gold Technology Co., Ltd., model number ZKK09, and the tyrosinase was selected from Hunan Wokai Biotechnology Co., Ltd., catalog number D10095-50ku.

[0083] 2g of carboxylated niobium carbide nanosheets were mixed with 0.46035g of zinc nitrate hexahydrate, 20mg of tyrosinase, and 25mL of deionized water. The mixture was shaken for 10min, then 5.125g of 2-methylimidazole was added. The mixture was shaken at a constant temperature for 10min, washed with deionized water, filtered, and freeze-dried at -56℃.

[0084] Example 8: A skin care gel loaded with Ganoderma lucidum exosomes, differing from Example 7 in that the surface of the niobium carbide nanosheets is loaded with tyrosinase. The method for preparing the niobium carbide nanosheets with tyrosinase loaded on the surface is as follows:

[0085] Niobium carbide nanosheets were added to deionized water and sonicated for 30 min to obtain a dispersion with a concentration of 2 mg / ml. A 0.5 M citric acid solution was added, and the mixture was stirred in a water bath at 60 °C for 12 h. After centrifugation at 10,000 rpm for 15 min, the mixture was washed three times with deionized water and dried under vacuum at 60 °C to obtain carboxylated niobium carbide nanosheets. The niobium carbide nanosheets were selected from Beijing Deco Island Gold Technology Co., Ltd., model number ZKK09.

[0086] 2g of carboxylated niobium carbide nanosheets were mixed with 20mg of tyrosinase and 25mL of deionized water, shaken for 10min, washed with deionized water, filtered, and freeze-dried at -56℃. The tyrosinase was selected from Hunan Wokai Biotechnology Co., Ltd., catalog number D10095-50ku.

[0087] Example 9: A skin care gel loaded with Ganoderma lucidum exosomes, differing from Example 7 in that the surface of the niobium carbide nanosheets is loaded with a tyrosinase / ZIF-8 complex. The method for preparing the niobium carbide nanosheets with the tyrosinase / ZIF-8 complex loaded on the surface is as follows:

[0088] 2g of niobium carbide nanosheets were mixed with 0.46035g of zinc nitrate hexahydrate, 20mg of tyrosinase, and 25mL of deionized water. The mixture was shaken for 10min, then 5.125g of 2-methylimidazole was added. The mixture was shaken at a constant temperature for 10min, washed with deionized water, filtered, and freeze-dried at -56℃. The niobium carbide nanosheets were selected from Beijing Deco Island Gold Technology Co., Ltd., model number ZKK09, and the tyrosinase was selected from Hunan Wokai Biotechnology Co., Ltd., catalog number D10095-50ku.

[0089] Comparative Example

[0090] Comparative Example 1: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that the Ganoderma lucidum exosome nanoparticles are prepared by Preparation Example 6.

[0091] Comparative Example 2: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that the Ganoderma lucidum exosome nanoparticles are prepared by Example 7.

[0092] Comparative Example 3: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that an equal amount of Ganoderma lucidum exosomes is used instead of Ganoderma lucidum exosome nanoparticles.

[0093] Comparative Example 4: A skin care gel loaded with Ganoderma lucidum exosomes, which differs from Example 1 in that it does not contain niobium carbide nanosheets.

[0094] Performance testing

[0095] Skin care gels were prepared according to the methods in the examples and comparative examples, and their performance was then tested according to the following methods.

[0096] I. Security Testing

[0097] (1) Number of test subjects: a total of 30 people, 4 males and 26 females; the youngest age was 24 years old and the oldest age was 45 years old. They were divided into 2 groups, with 2 males and 13 females in each group. The two groups of subjects were respectively tested with the skin care gels prepared in Example 1 and Example 6.

[0098] (2) Test method: Patch test was performed on Examples 1 and 6. 0.025 ml of the skin care gel prepared in Examples 1 and 6 was placed in the patch tester and applied to the back of the subject with medical tape. The test substance was removed after 24 hours. Skin reaction was observed at 0.5, 24 and 48 hours after patch removal. The results were recorded according to the skin reaction grading standard in the "Cosmetic Safety Technical Specifications" (2015 edition).

[0099] (3) The evaluation criteria are shown in Table 2.

[0100] Table 2

[0101]

[0102] The results of the human skin occlusive patch test showed that, according to the contents of Table 7, 0 out of 30 people showed a positive reaction. According to the "Cosmetic Safety Technical Specifications" (2015 edition), the skin care gels tested in Examples 1 and 6 will not cause adverse skin reactions in this batch of subjects.

[0103] II. Skin Whitening and Brightening Effect Test

[0104] (1) Test area: 130 healthy women (25-45 years old) were divided into 13 groups of 10 people each;

[0105] (2) Test area: face (left cheek blank sample, right cheek test sample);

[0106] (3) How to use: Use twice a day, morning and evening, for 4 weeks;

[0107] (4) Testing instrument: Delfin Skin Color Catch (Finland);

[0108] (5) Performance indicators: Test the change in color brightness (ΔL*) and ITA°. L* value (brightness) and ITA° (color individual type angle) are mainly used as indicators;

[0109] Calculate (ΔL*) using the following formula: (L*(test sample after 1 month) - L*(test sample at the beginning)) - [L*(blank sample after 1 month) - L*(blank sample at the beginning)], and then use the average value of each group for statistical analysis.

[0110] ITA° (represents the overall change in chromaticity; the greater the change in ITA°, the lighter the skin tone) was tested. The ITA° was measured before the start of each test and one month later. The average value was taken for each test. The color classification of ITA° is shown in Table 3. The test results were recorded in Table 4.

[0111] (III) Moisturizing effect test: The skin care gels obtained in each of the above embodiments and comparative examples were used individually, and their moisturizing and enhancing abilities were tested using a CK test instrument. The test method was as follows: First, test areas were marked on the inside of both arms, and the moisture content of the marked areas was measured using a CK test instrument. Then, equal amounts of the above samples were applied to the marked areas on the inside of the arms, and the moisture content value was measured after 60 minutes. Finally, the moisturizing effect was expressed by the calculation method of (value after sample application - value before sample application) / value before sample application × 100. The higher the value, the better the moisturizing effect.

[0112] Table 3

[0113] ITA° range Skin color classification 55-90 very shallow 41-54 shallow 28-40 medium 10-27 brown -30～9 brown -90～29 black

[0114] Table 4

[0115]

[0116]

[0117] As can be seen from the data in Tables 3 and 4, the skin care gel prepared using the raw materials in Examples 1-3 has a higher ITA° value after 4 weeks of use, a more uniform skin tone, and a larger color difference change value, which proves that the skin care gel loaded with Ganoderma lucidum exosomes prepared in this application has good whitening, moisturizing and brightening effects.

[0118] In Example 4, Ganoderma lucidum exosome nanoparticles prepared in Example 3 were used, with laurocapram alone as a penetration enhancer. It can be seen that compared with Example 1, the whitening and brightening effects of the skin care gel prepared in Example 4 were all reduced, indicating that laurocapram and hyaluronidase work together to improve the penetration of Ganoderma lucidum exosome nanoparticles and improve the whitening, moisturizing and skin-rejuvenating effects.

[0119] In Example 5, Ganoderma lucidum exosome nanoparticles prepared in Preparation Example 4 were used. Compared with Example 1, cyclohexanehexyl alcohol and hyaluronidase were used as penetration enhancers. In Example 6, compared with Example 1, Ganoderma lucidum exosome nanoparticles prepared in Preparation Example 5 were used. Lauryl diazoline and cyclohexanehexyl alcohol were used as penetration enhancers. As can be seen from the data in Table 4, after 4 weeks of use, the color brightness and ITA value of the skin care gels prepared in Examples 5 and 6 were not as good as those in Example 1. This indicates that using lauryl diazoline and hyaluronidase as penetration enhancers can effectively improve the penetration of Ganoderma lucidum exosome nanoparticles and enhance the skin care and whitening effect.

[0120] In Example 7, niobium carbide nanosheets were pretreated using a tyrosinase / ZIF-8 complex. As shown in Table 4, the skin care gel prepared in Example 7, after continuous use for 4 weeks, increased the color brightness of the facial skin, increased the ITA° value, and further improved the whitening, moisturizing, and brightening effects.

[0121] In Example 8, only tyrosinase was used to pretreat niobium carbide nanosheets. In Example 9, no carboxylation treatment was performed on the niobium carbide nanosheets. The data comparison shows that the skin care gels prepared in Examples 8 and 9 have a lower skin brightening and whitening effect compared with Example 7.

[0122] Comparative Example 1 used Ganoderma lucidum exosome nanoparticles prepared in Preparation Example 6, and Comparative Example 2 used Ganoderma lucidum exosome nanoparticles prepared in Preparation Example 7. Compared with Preparation Example 1 in Example 1, Preparation Examples 6 and 7 did not add papain and penetration enhancer, respectively. It can be seen that the skin care gels prepared in Comparative Examples 1 and 2 have reduced whitening and brightening effects compared with Example 1.

[0123] Compared with Example 1, Comparative Example 3 added Ganoderma lucidum exosomes but did not encapsulate them, which shows that the whitening and brightening effects of the skin care gel prepared in Comparative Example 3 were significantly weakened.

[0124] In Comparative Example 4, no niobium carbide nanosheets were added, and compared with Example 1, the skin care gel produced had a weaker skin-improving effect.

[0125] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A skin care gel carrying Ganoderma lucidum exosomes, characterized in that, The ingredients include the following parts by weight: 20-40 parts hydrogel matrix, 2-4 parts Ganoderma lucidum exosome nanoparticles, 2-6 parts niobium carbide nanosheets, 1-3 parts nicotinamide, 0.1-2 parts phenylethyl resorcinol and 0.1-3 parts green tea extract; The spirosome nanoparticles have a core-shell structure. The core layer consists of spirosomes and papain in a mass ratio of 1:0.1-0.2, and the shell layer consists of polylactic acid-glycolic acid copolymer and a penetration enhancer in a mass ratio of 1:0.1-0.

3.

2. The skin care gel carrying Ganoderma lucidum exosomes according to claim 1, characterized in that: The mass ratio of the shell solution to the core solution of the Ganoderma lucidum exosome nanoparticles is 1.5-2.5:

1.

3. The skin care gel carrying Ganoderma lucidum exosomes according to claim 1, characterized in that: The surface of the niobium carbide nanosheets is loaded with a tyrosinase / ZIF-8 complex.

4. The skin care gel carrying Ganoderma lucidum exosomes according to claim 3, characterized in that: The method for preparing the niobium carbide nanosheets loaded with the tyrosinase / ZIF-8 complex is as follows: Carboxylated niobium carbide nanosheets were mixed with zinc nitrate hexahydrate, tyrosinase, and deionized water, shaken for 10-20 min, 2-methylimidazole was added, and the mixture was shaken at a constant temperature for 10-20 min. The mixture was then washed with deionized water, filtered, and freeze-dried.

5. The skin care gel carrying Ganoderma lucidum exosomes according to claim 1, characterized in that: The penetration enhancer is selected from one or more of laurocapram, hyaluronidase, cyclohexanehexyl alcohol, polyethylene glycol 400 monostearate, isopropyl myristate, N-methylpyrrolidone, and sodium glycocholate.

6. The skin care gel carrying Ganoderma lucidum exosomes according to claim 5, characterized in that: The penetration enhancer comprises laurocapram and hyaluronidase in a mass ratio of 3-5:

1.

7. The skin care gel carrying Ganoderma lucidum exosomes according to claim 1, characterized in that: The hydrogel matrix is ​​selected from at least one of carbomer, sodium hyaluronate, sodium alginate, polyethylene glycol, gum arabic, polyvinylpyrrolidone, polyvinyl alcohol, sodium polyacrylate, hydroxypropyl methylcellulose, and carboxyethyl cellulose.

8. The skin care gel carrying Ganoderma lucidum exosomes according to claim 7, characterized in that: The hydrogel matrix comprises sodium hyaluronate and hydroxypropyl methylcellulose in a mass ratio of 1:1-1.

5.

9. A method for preparing the skin care gel loaded with Ganoderma lucidum exosomes according to any one of claims 1-8, characterized in that: Includes the following steps: Mix the hydrogel matrix and deionized water, heat to 65-80℃, and homogenize for 15-30 minutes to obtain a gel mixture. The skin care gel was prepared by uniformly mixing the gel mixture with Ganoderma lucidum exosome nanoparticles, niobium carbide nanosheets, niacinamide, phenylethyl resorcinol and green tea extract.

10. The method for preparing the skin care gel loaded with Ganoderma lucidum exosomes according to claim 9, characterized in that: The viscosity of the gel mixture is 10000-30000 mPa·s.

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