Essence for repairing skin aging and preparation method thereof

Abstract

The invention discloses essence for repairing skin aging and a preparation method thereof. The essence is prepared from the following raw materials in percentage by mass: 10 to 30 percent of umbilical cord tissue extract, 1 to 5 percent of lactobacillus / ginseng root fermentation product filtrate, 0.2 to 0.8 percent of solubilizer, 5 to 15 percent of humectant, 0.01 to 0.1 percent of antioxidant, 0.5 to 1 percent of preservative, 0.05 to 0.1 percent of pH regulator, 0.1 to 0.3 percent of essence and the balance of deionized water. From the aspects of promoting collagen synthesis, inhibiting inflammatory factor expression, scavenging free radicals, repairing DNA damage and the like, the essence disclosed by the invention plays a remarkable effect of repairing skin aging, and can eliminate wrinkles, tighten skin, improve skin elasticity and luster and make the skin youthful.

Description

Technical Field

[0001] The present invention belongs to the field of cosmetics, and particularly relates to a serum for repairing skin aging and a preparation method thereof. Background Art

[0003] At present, the anti-aging skin care products on the market mainly rely on the following active ingredients: 1) Vitamins (such as vitamin C, vitamin E): They have antioxidant effects, but poor stability, and are easily inactivated by light and temperature; 2) Peptides (such as palmitoyl pentapeptide-4): They can promote collagen synthesis, but have high costs and limited permeability; 3) Plant extracts (such as green tea extract, grape seed extract): They have antioxidant and anti-inflammatory effects, but their efficacy is relatively single and it is difficult to comprehensively solve the problem of skin aging. In addition, due to cost and technical limitations, the content of active ingredients in many products is low, making it difficult to achieve the ideal repair effect. Therefore, there is an urgent need to develop a serum that is easy to absorb, has good stability, is safe in nature and can effectively repair skin aging to meet the market demand. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a serum for repairing skin aging and a preparation method thereof. The serum takes umbilical cord tissue extract and filtrate of Lactobacillus ginseng root fermentation product as core active ingredients, combines with moisturizers, antioxidants and other ingredients, and through scientific proportioning and preparation processes, can effectively promote collagen production, scavenge free radicals, improve skin elasticity, effectively repair skin aging, has significant anti-aging effects, and has good stability and is safe in nature.

[0005] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0006] A serum for repairing skin aging is composed of the following raw materials in mass percentages: umbilical cord tissue extract 10 - 30%, filtrate of Lactobacillus ginseng root fermentation product 1 - 5%, solubilizer 0.2 - 0.8%, moisturizer 5 - 15%, antioxidant 0.01 - 0.1, preservative 0.5 - 1%, pH regulator 0.05% - 0.1%, essence 0.1 - 0.3%, and the balance is deionized water.

[0007] Further, the preparation steps of the umbilical cord tissue extract are as follows: Homogenize fresh porcine umbilical cord tissue, add an equal volume of trypsin solution with a concentration of 0.2 - 0.4% thereto, stir and mix evenly, incubate in an incubator at 35 - 40 °C for 24 - 72 h, then centrifuge, discard the supernatant, disperse the centrifuged precipitate in serum-free medium, filter, collect the filtrate, add HIF-1 and astragalus polysaccharide to the filtrate, cultivate under hypoxic conditions for 48 - 72 h, then centrifuge, collect the supernatant, filter the supernatant, collect the filtrate, and lyophilize the filtrate to obtain the umbilical cord tissue extract.

[0008] Further, the HIF-1 is hypoxia-inducible factor-1 (HIF-1), and its final concentration is 10 - 20 ng / mL. The concentration of astragalus polysaccharide is 5 - 15 μg / mL.

[0009] The HIF-1 is purchased from Shanghai Bolson Biotechnology Co., Ltd.

[0010] The astragalus polysaccharide is purchased from Shanghai Macklin Biochemical Co., Ltd.

[0011] Further, the conditions for hypoxic cultivation are static cultivation in a 5% O2, 37 °C CO2 incubator.

[0012] Preferably, the solubilizer is octyl / decyl glucoside.

[0013] Preferably, the humectant is one or more of glycerin, 1,3-propanediol, butanediol, glycerol polyether-26, dipalmitoyl hydroxyproline, hyaluronic acid hydroxypropyltrimonium chloride, trehalose, and water-soluble jojoba oil.

[0014] Preferably, the antioxidant is one or more of tocopheryl acetate, vitamin E, ergothioneine, ascorbyl tetraisopalmitate, and magnesium ascorbyl phosphate.

[0015] Preferably, the preservative is a mixture of ethylhexylglycerin and phenoxyethanol. More preferably, the preservative is euxyl pe 9010 from Ashland. TM pe 9010.

[0016] Preferably, the pH regulator is one or several of lactic acid, citric acid, sodium citrate, triethanolamine, and tromethamine.

[0017] Preferably, the filtrate of Lactobacillus / Panax ginseng root fermentation product (INCI: Butanediol, filtrate of Lactobacillus / Panax ginseng root fermentation product, hydrolyzed ginsenosides) is purchased from Guangzhou Yunli Biotechnology Co., Ltd.

[0018] The present invention also provides a method for preparing the above essence, comprising the following steps: dissolving the umbilical cord tissue extract, Lactobacillus / Panax ginseng root ferment filtrate, and solubilizer in deionized water, stirring to dissolve, then adding a humectant, antioxidant, preservative, pH regulator, and fragrance, and stirring well to dissolve to obtain the essence.

[0019] The umbilical cord tissue extract is a common anti-aging active ingredient, which is rich in various active ingredients such as cytokines, enzymes, exosomes, etc. It exerts anti-aging effects by synergistically acting through multiple mechanisms such as promoting collagen synthesis, antioxidant, promoting cell regeneration, improving microcirculation, anti-inflammatory immune regulation, and moisturizing barrier repair, and repairing skin aging problems at the cellular level. However, the types and contents of active ingredients contained in umbilical cord tissue extracts prepared by different processes vary, resulting in uneven anti-aging effects. The present invention has found through research that compared with umbilical cord tissue extracts prepared by conventional processes, the umbilical cord tissue extract prepared by the process of the present invention has more excellent anti-aging effects. It is speculated that this is because the extract is rich in cytokines and antioxidant enzymes, especially the contents of vascular endothelial growth factor A (VEGF-A) and insulin-like growth factor-1 (IGF-1), and the activity of superoxide dismutase (SOD) is relatively high. Specifically, in the process of the present invention, trypsin is first used to digest umbilical cord tissue, and after high-speed centrifugation, it is dispersed in a culture medium, and then filtered and other steps are used to obtain a filtrate rich in microtissues. Then, hypoxia-inducible factor-1 and astragalus polysaccharide are added to the filtrate rich in microtissues, and cultured under hypoxic conditions with an oxygen volume fraction of 5% to stimulate the expression and secretion of VEGF-A and IGF-1. Then, pathogens are removed by centrifugation, filtration and other means to obtain an umbilical cord tissue extract rich in VEGF-A and IGF-1. The astragalus polysaccharide can indirectly promote the expression of VEGF-A and IGF-1 by upregulating endothelial nitric oxide synthase (eNOS) and downstream signaling pathways, and at the same time enhance the activity of SOD, ultimately enabling the obtained umbilical cord tissue extract to exert significant effects in repairing skin aging and anti-aging.

[0020] The Lactobacillus / Panax ginseng root ferment filtrate contains rare ginsenosides, is easy to absorb, and has high bioavailability. It has the following effects: promoting the migration of fibroblasts, accelerating wound healing; reducing scars, making the newly formed skin smooth and flat, and helping to make the skin texture delicate; reducing skin atrophy caused by ultraviolet rays and alleviating photoaging; significantly increasing the collagen content of the skin, promoting the synthesis of skin fibroblast fibronectin, improving the basal layer of the skin, and firming the skin.

[0021] The present invention takes cord tissue extract and Lactobacillus / Ginseng Root Ferment Filtrate as core active ingredients, combines ingredients such as humectants and antioxidants, and through scientific proportioning and preparation processes, successfully prepares a serum with significant repair and anti-aging effects. Starting from multiple perspectives such as promoting collagen synthesis, inhibiting the expression of inflammatory factors, scavenging free radicals, and repairing DNA damage, this serum can significantly repair skin aging, eliminate wrinkles, tighten the skin, improve skin elasticity and luster, and make the skin glow with youthful radiance. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Detection results of the contents of cytokines VEGF-A and IGF-1 and SOD activity in cord tissue extracts prepared by different processes. Compared with UCE, *p < 0.05. DETAILED DESCRIPTION OF THE INVENTION

[0023] The following examples are further illustrations of the present invention rather than limitations thereof. Unless otherwise specified, the components of the formulations in the following examples are all commercially available products.

[0024] Example 1

[0025] (1) Preparation of cord tissue extract:

[0026] In a laminar flow hood, take fresh porcine umbilical cord, place it in PBS containing penicillin (100 U / mL) and streptomycin (0.1 mg / mL) and wash it 3 times, 5 minutes each time. Cut the washed porcine umbilical cord tissue into small pieces with sterile surgical scissors, then add sterile pure water 3 times the weight of the cord tissue, perform homogenization treatment under sterile conditions, add an equal volume of 0.3% m / v trypsin solution to the homogenized tissue, stir and mix evenly, incubate in an incubator at 40 °C for 48 h, then centrifuge at 6000 r / min for 10 min, discard the supernatant, disperse the centrifuged precipitate in serum-free medium (brand: GIBCO), filter it using a 200 μm filter, collect the filtrate, add HIF-1 with a final concentration of 10 ng / mL and astragalus polysaccharide with a final concentration of 10 μg / mL to the filtrate, statically culture it in a 5% O2, 37 °C, CO2 incubator for 48 h, then centrifuge at 6000 r / min for 10 min, collect the supernatant, filter the supernatant with a 0.22 μm filter, collect the filtrate, and freeze-dry the filtrate to obtain cord tissue extract (UCE).

[0027] (2) Preparation of cord tissue extract:

[0028] Prepare umbilical cord tissue extract (UCE-NA) without adding astragalus polysaccharide according to the method in step (1) above. The specific preparation steps are generally the same as those in step (1) above, except that astragalus polysaccharide is not added for treatment.

[0029] (3) Preparation of umbilical cord tissue extract (conventional process):

[0030] In a laminar flow hood, take fresh porcine umbilical cord, place it in PBS containing penicillin (100 U / mL) and streptomycin (0.1 mg / mL) and wash it 3 times, 5 minutes each time. Cut the washed porcine umbilical cord tissue into pieces with sterile surgical scissors, then add sterile pure water 3 times the weight of the umbilical cord tissue, and perform homogenization treatment under sterile conditions. Add an equal volume of trypsin solution with a concentration of 0.3% m / v to the homogenized tissue, stir and mix evenly, incubate in an incubator at 40 °C for 48 h, then centrifuge at 6000 r / min for 10 min, collect the supernatant, and retain the precipitate at the same time; add trypsin solution with a concentration of 1% m / v 3 times the volume to the centrifuged precipitate, stir and react for 48 h, then add an equal volume of sterile pure water and mix evenly, centrifuge at 6000 r / min for 15 min, collect the supernatant, combine the supernatants obtained twice, filter through a 0.22 μm filter, collect the filtrate, and lyophilize the filtrate to obtain umbilical cord tissue extract (UCE-CP).

[0031] Example 2

[0032] Detect the contents of cytokines VEGF-A and IGF-1 and the activity of antioxidant enzyme SOD in the umbilical cord tissue extracts UCE, UCE-NA, and UCE-CP of Example 1 respectively. The results are shown in Figure 1 .

[0033] It can be seen from Figure 1 that compared with the umbilical cord tissue extract (UCE) of the present invention, the contents of VEGF-A and IGF-1 in the umbilical cord tissue extract (UCE-NA) without adding astragalus polysaccharide for treatment and the umbilical cord tissue extract (UCE-CP) prepared by the conventional process are significantly reduced, and the SOD activity is also reduced. This result indicates that the contents of anti-aging cytokines and the activity of antioxidant enzymes in the umbilical cord tissue extract prepared by the process of the present invention are relatively high, which helps to improve its effect of repairing skin aging.

[0034] Example 3

[0035] A serum for repairing skin aging, which is composed of the following raw materials in mass percentages: umbilical cord tissue extract (UCE) 10%, filtrate of Lactobacillus / Panax ginseng root fermentation product 5%, octyl / decyl glucoside 0.8%, glycerol 10%, hyaluronic acid hydroxypropyltrimonium chloride 5%, tocopheryl acetate 0.05%, euxylTM pe 9010 0.5%, sodium citrate 0.05%, essence 0.1%, and the balance is deionized water.

[0036] Preparation steps: Dissolve the umbilical cord tissue extract, Lactobacillus / ginseng root fermentation product filtrate, and octyl / decyl glucoside in deionized water, stir to dissolve, and then add glycerol, hyaluronic acid hydroxypropyltrimonium chloride, tocopheryl acetate, euxyl TM pe 9010, sodium citrate, and essence, and stir well to dissolve to obtain the essence.

[0037] Example 4

[0038] An essence for repairing skin aging is composed of the following raw materials in mass percentages: umbilical cord tissue extract (UCE) 20%, Lactobacillus / ginseng root fermentation product filtrate 1%, octyl / decyl glucoside 0.2%, glycerol 2%, hyaluronic acid hydroxypropyltrimonium chloride 3%, tocopheryl acetate 0.01%, euxyl TM pe 9010 0.5%, sodium citrate 0.05%, essence 0.1%, and the balance is deionized water.

[0039] The preparation steps refer to Example 3.

[0040] Example 5

[0041] An essence for repairing skin aging is composed of the following raw materials in mass percentages: umbilical cord tissue extract (UCE) 30%, Lactobacillus / ginseng root fermentation product filtrate 5%, octyl / decyl glucoside 0.6%, glycerol 5%, hyaluronic acid hydroxypropyltrimonium chloride 5%, tocopheryl acetate 0.05%, euxyl TM pe 9010 0.5%, sodium citrate 0.1%, essence 0.3%, and the balance is deionized water.

[0042] The preparation steps refer to Example 3.

[0043] Example 6

[0044] An essence for repairing skin aging is composed of the following raw materials in mass percentages: umbilical cord tissue extract (UCE) 15%, Lactobacillus / ginseng root fermentation product filtrate 5%, octyl / decyl glucoside 0.6%, glycerol polyether - 26 5%, dipalmitoyl hydroxyproline 5%, ergothioneine 0.1%, euxyl TM pe 9010 0.5%, sodium citrate 0.05%, essence 0.2%, and the balance is deionized water.

[0045] The preparation steps refer to Example 3.

[0046] Example 7

[0047] An essence for repairing skin aging, which is composed of raw materials in the following mass percentages: umbilical cord tissue extract (UCE-NA) 30%, filtrate of Lactobacillus / ginseng root fermentation product 5%, octyl / decyl glucoside 0.6%, glycerol 5%, hyaluronic acid hydroxypropyltrimonium chloride 5%, tocopheryl acetate 0.05%, euxyl TM pe 90100.5%, sodium citrate 0.1%, essence 0.3%, and the balance is deionized water.

[0048] The preparation steps refer to Example 3.

[0049] Example 8

[0050] An essence for repairing skin aging, which is composed of raw materials in the following mass percentages: umbilical cord tissue extract (UCE-CP) 30%, filtrate of Lactobacillus / ginseng root fermentation product 5%, octyl / decyl glucoside 0.6%, glycerol 5%, hyaluronic acid hydroxypropyltrimonium chloride 5%, tocopheryl acetate 0.05%, euxyl TM pe 90100.5%, sodium citrate 0.1%, essence 0.3%, and the balance is deionized water.

[0051] The preparation steps refer to Example 3.

[0052] Comparative Example 1

[0053] An essence for repairing skin aging, which is composed of raw materials in the following mass percentages: umbilical cord tissue extract (UCE-CP) 28%, astragalus polysaccharide 2%, filtrate of Lactobacillus / ginseng root fermentation product 5%, octyl / decyl glucoside 0.6%, glycerol 5%, hyaluronic acid hydroxypropyltrimonium chloride 5%, tocopheryl acetate 0.05%, euxyl TM pe90100.5%, sodium citrate 0.1%, essence 0.3%, and the balance is deionized water.

[0054] The preparation steps refer to Example 3.

[0055] Comparative Example 2

[0056] The difference between Comparative Example 2 and Example 5 is that it does not contain umbilical cord tissue extract (UCE).

[0057] Comparative Example 3

[0058] The difference between Comparative Example 3 and Example 5 is that it does not contain filtrate of Lactobacillus / ginseng root fermentation product.

[0059] Application Example 1. Human skin patch test

[0060] The essence obtained in Examples 3 - 8 of the present invention was tested according to the relevant requirements and determination methods of "Human Skin Occlusive Patch Test" in the "Technical Specifications for Cosmetics Safety" (2015 Edition): The product was placed in the small chamber of the patch tester, with a dosage of 0.020 - 0.025 mL. The patch tester was applied to the flexor side of the forearm of the subject. One side was the blank control area, and the other side was the test area, lasting for 24 h. The results were observed and recorded 30 min (waiting for the indentation to disappear), 24 h, and 48 h after removing the test patch, and the results were evaluated according to the grading standard (Table 1) of the "Technical Specifications for Cosmetics Safety" (2015 Edition). The patch test results showed that all the subjects had negative reactions, indicating that the essence of the present invention has high safety and will not cause adverse reactions to the human body.

[0061] Table 1 Skin Reaction Grading Standard

[0062]

[0063] Application Example 2. Detection of Anti - aging Effect

[0064] Healthy female volunteer subjects, aged 45 - 60 years old, with no history of skin allergies, meeting the subject inclusion criteria, were divided into 9 groups, with 8 people in each group, namely Examples 3 - 8 groups and Comparative Examples 1 - 3 groups. Among them, the subjects in Examples 3 - 8 groups used the essence obtained in Examples 3 - 8, and the subjects in Comparative Examples 1 - 3 groups used the essence obtained in Comparative Examples 1 - 3.

[0065] Usage method: After cleansing and toning the face in the morning and evening, take an appropriate amount of the essence and apply it to the face, evenly apply and massage until absorbed. Before use and 28 days after use, a skin elasticity Cotometer MPA580 test instrument manufactured by Courage + Khazaka GmbH (German CK company) was used to measure the skin elasticity R2 value of the stratum corneum in the cheek area of the subject (the larger the R2 value, the better the skin elasticity), and the skin firmness F4 value (the smaller the F4 value, the better the skin firmness). The Sa (roughness) and Sq (texture unevenness) values of the skin were measured using VC98. The smaller the Sa and Sq values, the fewer skin wrinkles and the smoother the skin. According to the formula: change rate = (measurement value 28 days after use - measurement value before use) / measurement value before use * 100%, the change rates of the above - mentioned various indicators were calculated, and the results are shown in Table 2 below:

[0066] Table 2 Anti - aging Effect Detection Results

[0067] Group Change rate of R2 value (%) Change rate of F4 value (%) Change rate of Sa value (%) Change rate of Sq value (%) Example 3 24.62 -38.44 -20.06 -14.75 Example 4 27.15 -42.26 -22.17 -17.48 Example 5 29.77 -44.80 -24.29 -19.62 Example 6 26.84 -41.75 -21.63 -16.91 Example 7 17.52 -28.32 -16.14 -10.06 Example 8 16.11 -26.97 -15.30 -9.25 Comparative Example 1 19.07 -30.15 -17.82 -11.57 Comparative Example 2 10.92 -19.48 -9.64 -5.22 Comparative Example 3 14.38 -22.60 -11.75 -7.84

[0068] The results showed that the essence prepared in Examples 3-6 could increase the R2 value of the skin elasticity of the subjects, reduce the F4 value of skin firmness and the Sa and Sq values of the skin, and had good effects in enhancing skin elasticity, improving firmness, and reducing skin wrinkles, with obvious anti-aging effects. It can be seen from Examples 7 and 8 that compared with the umbilical cord tissue extract (UCE-NA) without adding astragalus polysaccharide and the umbilical cord tissue extract (UCE-CP) prepared by conventional processes, the umbilical cord tissue extract (UCE) of the present invention had better anti-aging effects. It can be seen from Comparative Example 1 that when the combination of the umbilical cord tissue extract (UCE-CP) prepared by conventional processes and astragalus polysaccharide was used as the raw material to replace the umbilical cord tissue extract (UCE) of the present invention to prepare the essence, its anti-aging effect was weakened, indicating that compared with the simple mixture of astragalus polysaccharide and umbilical cord tissue extract, using astragalus polysaccharide in the preparation process of umbilical cord tissue extract was more helpful to improve the anti-aging effect of the essence. It can be seen from Comparative Examples 2 and 3 that when the umbilical cord tissue extract (UCE) or the filtrate of Lactobacillus / Ginseng Root Fermentation Product was not added to the formula respectively, the anti-aging effect of the prepared essence was significantly reduced, indicating that the above two components played a synergistic effect in anti-aging.

[0069] The above detailed description is a specific description of the embodiments that can be implemented in the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or change without departing from the present invention shall be included in the patent scope of this case.

Claims

1. An essence for repairing skin aging, characterized in that, It is composed of the following raw materials by mass percentage: umbilical cord tissue extract 10 - 30%, Lactobacillus / Ginseng Root Fermentation Product Filtrate 1 - 5%, solubilizer 0.2 - 0.8%, humectant 5 - 15%, antioxidant 0.01 - 0.1%, preservative 0.5 - 1%, pH regulator 0.05% - 0.1%, essence 0.1 - 0.3%, and the balance is deionized water.

2. The essence according to claim 1, characterized in that, The preparation steps of the umbilical cord tissue extract are as follows: Homogenize fresh porcine umbilical cord tissue, add an equal volume of trypsin solution with a concentration of 0.2 - 0.4% to the homogenized tissue, stir and mix evenly, incubate in an incubator at 35 - 40 °C for 24 - 72 h, then centrifuge, discard the supernatant, disperse the centrifuged precipitate in serum-free medium, filter, collect the filtrate, add HIF-1 and astragalus polysaccharide to the filtrate, cultivate under hypoxic conditions for 48 - 72 h, then centrifuge, collect the supernatant, filter the supernatant, collect the filtrate, and lyophilize the filtrate to obtain the umbilical cord tissue extract.

3. The essence according to claim 2, characterized in that, The final concentration of the HIF-1 is 10 - 20 ng / mL, and the concentration of astragalus polysaccharide is 5 - 15 μg / mL.

4. The essence according to claim 2, characterized in that, The conditions for hypoxic cultivation are static cultivation in a CO2 incubator at 5% O2 and 37 °C.

5. The essence according to claim 1, characterized in that, The solubilizer is octyl / decyl glucoside.

6. The essence according to claim 1, characterized in that, The humectant is one or more of glycerol, 1,3-propanediol, butanediol, glycerol polyether-26, dipalmitoyl hydroxyproline, hyaluronic acid hydroxypropyltrimonium chloride, trehalose, and water-soluble jojoba oil.

7. The essence according to claim 1, characterized in that, The antioxidant is one or more of tocopheryl acetate, vitamin E, ergothioneine, ascorbyl tetraisopalmitate, and magnesium ascorbyl phosphate.

8. The essence according to claim 1, characterized in that, The preservative is a mixture of ethylhexylglycerin and phenoxyethanol.

9. The essence according to claim 1, characterized in that, The pH regulator is one or several of lactic acid, citric acid, sodium citrate, triethanolamine, and tromethamine.

10. A method for preparing the essence according to any one of claims 1-9, characterized in that, It includes the following steps: Dissolve the umbilical cord tissue extract, Lactobacillus / Ginseng Root Fermentation Product Filtrate, and solubilizer in deionized water, stir to dissolve, then add the humectant, antioxidant, preservative, pH regulator, and essence, and stir well to dissolve to obtain the essence.

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