Preparation of anti-wrinkle and moisturizing freeze-dried powder composition

Abstract

The present invention relates to the preparation of an anti-wrinkle and moisturizing freeze-dried powder composition, wherein the freeze-dried powder comprises an extract of a fermentation product of Alteromonas, soluble collagen, a porcine umbilical cord extract and a callus cell extract of Nymphoides; the essence comprises a composition and a solvent; the two components are prepared and stored separately, and the freeze-dried powder and the solvent are mixed before use. In order to solve the interaction between the increase of free radicals and the loss of collagen and water during skin aging, the present invention uses plant callus cell extracts, animal umbilical cord extracts and fungus fermentation product extracts as the main substances, and is matched with soluble collagen to achieve moisturizing the skin, deep hydration, brightening the skin tone, and anti-wrinkle and face-lifting.

Description

Technical Field

[0001] The invention relates to the technical field of cosmetics, and in particular to the preparation of an anti-wrinkle and moisturizing freeze-dried powder composition. Background Art

[0002] As a bright new star in the cosmetics field, freeze-dried powder is showing a strong development trend with its unique technical advantages and remarkable application effects.

[0003] With the continuous advancement of freeze-drying technology and the reduction of costs, more and more cosmetics companies have begun to adopt this technology to store highly active skin care ingredients in the form of freeze-dried powder, thereby ensuring the purity and stability of the ingredients. At the same time, as consumers pursue mild and effective skin care products, freeze-dried powder has won the favor of more and more consumers with its mild, non-irritating and highly effective characteristics.

[0004] In cosmetics, freeze-dried powder is widely used in products with various functions such as whitening, moisturizing, anti-aging, and repair, becoming an important carrier for customized skin care. In the future, with the continuous growth of market demand and the continuous promotion of technological innovation, the application of freeze-dried powder in the cosmetics field will be more extensive, bringing consumers more safe and effective skin care options.

[0005] The present invention aims to provide an anti-wrinkle and moisturizing freeze-dried powder composition and a preparation method thereof, so as to solve the problem that existing skin care products need to provide anti-wrinkle properties on top of the most basic moisturizing properties. Summary of the invention

[0006] In order to achieve the above-mentioned purpose of the invention, the present invention provides an anti-wrinkle and moisturizing freeze-dried powder composition, which comprises an extract of a fermentation product of Alteromonas, soluble collagen, a porcine umbilical cord extract and a Nymphoides callus cell extract; in addition, it also comprises an antioxidant ascorbic acid.

[0007] Alteromonas macleodii fermentation product extract, Alteromonas macleodii fermentation product extract, the antimicrobial peptides, enzymes, polysaccharides, alkaloids, ketones, phenols and other compounds contained in it provide anti-wrinkle, moisturizing, antibacterial and other effects to the skin.

[0008] Soluble collagen (SOLUBLE COLLAGEN), a type of collagen obtained through genetic recombination and bioengineering technology, has excellent water solubility and is widely used in the field of skin care. Because its amino acid sequence is highly similar to human collagen, it has good tissue compatibility, making it easier to be absorbed by the human body.

[0009] Umbilical cord extract (pig) is a substance extracted from the umbilical cord of animals. It is rich in various nutrients and growth factors, has multiple biological activities, and is widely used in the field of beauty. It has many advantages, such as nourishing, enhancing skin's moisturizing ability, reducing free radical damage to the skin, and delaying skin aging.

[0010] Nymphoides peltatum callus cell extract is a bioactive ingredient obtained by extracting and freeze-drying callus cells in the division stage of Nymphoides peltatum leaves. It is rich in polysaccharides, proteins, flavonoids, polyphenols, carotene and other active substances, and has significant antioxidant, antibacterial, anti-inflammatory and other biological activities.

[0011] In order to achieve the above object, the present invention adopts the following technical solutions:

[0012] An anti-wrinkle and moisturizing composition, comprising the following components in parts by mass:

[0013] 0.1-2 parts of Alteromonas fermentation product extract;

[0014] 0.5-4 parts of soluble collagen;

[0015] 0.1-0.5 parts of pig umbilical cord extract;

[0016] 0.1-1 part of Nymphoides callus cell extract.

[0017] In particular, the preparation method of the Nymphoides callus cell extract is: using its leaves as explant materials, sterilizing, and implanting into a culture medium; the specific operations are as follows:

[0018] Select fresh tender buds and leaves of water lily plants with high purity, typical characteristics of water lily, healthy growth, no injuries, and no diseases or insects. After wiping the leaf surface with 75% ethanol for disinfection, cut the tender bud and leaf tissue into 2-6 mm square pieces.

[0019] In the disinfected inoculation room, the young bud and leaf tissues are inoculated into culture bottles using aseptic operation procedures.

[0020] The inoculated culture bottle is placed in a light-proof, constant-temperature incubator. When the callus cells are in the division stage, they are taken out, ultrasonically broken, the cell fluid is filtered and extracted, and freeze-dried to obtain a callus cell extract.

[0021] In particular, the culture medium configuration method is specifically performed as follows:

[0022] S1. Weigh MS medium, sucrose, and plant growth hormone;

[0023] S2. Heat water to boiling under normal pressure, add S1, stir, and adjust the pH to 5.5-6;

[0024] S3. Package S2, sterilize, and wait for explant inoculation.

[0025] Preferably, the present invention provides a leave-on skin external preparation, characterized in that it comprises 3% of the composition and 97% of the solvent.

[0026] Preferably, the vehicle is characterized by comprising water and sodium hyaluronate.

[0027] The present invention also provides a method for preparing an essence, characterized in that the preparation method is as follows:

[0028] Add the composition into the solvent and stir evenly to obtain an essence;

[0029] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention innovatively uses water lily callus cell extracts mixed with alternatormonas fermentation product extracts, soluble collagen and porcine umbilical cord extracts, and the carotene, ketones and polyphenols rich in water lily callus can effectively improve the rough state of the skin and make the skin more delicate and smooth. Secondly, carotene, as a Va precursor, can be converted into Va in the human body. Va is essential for skin health and can effectively stimulate mucosal cells to continuously secrete mucus, cover the cell surface, prevent bacterial erosion, maintain the integrity of the skin mucosa, and Va can also enhance skin toughness. The water lily callus extracts, soluble collagen and alternatormonas fermentation products can be used together to improve the skin's moisture content, moisture retention and inhibition rate of elastase in the skin while repairing the rough state of the skin, making the skin more energetic. At the same time, the chondroitin sulfate, glycogen, lipids and a large amount of protein and collagen contained in the umbilical cord extract nourish the skin tissue in time, and work synergistically with the water spinach callus cell extract, soluble collagen and Alteromonas fermentation product extract to maximize the efficacy of the composition.

[0030] It has been found through experiments that the composition can repair skin damage caused by various reasons, has the effects of nourishing, anti-wrinkle and moisturizing, and is suitable for various skin types. The present invention has found that under the premise of the same usage amount, the specific ratio of Nymphoides callus cell extract and Alteromonas fermentation product extract, soluble collagen and porcine umbilical cord extract defined in the present invention has better effects than using them alone or using any three or more of them together, producing a synergistic effect, making its nourishing, anti-wrinkle and moisturizing effects better. The present invention also provides a preferred formula and preparation method of the essence containing the anti-wrinkle and moisturizing composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1Example 4-6 HaCaT survival rate.

[0032] Figure 2 Hyaluronidase inhibition rate of Examples 1-3 and Comparative Examples 1-5.

[0033] Figure 3 TNF-α inflammatory factor inhibition rate of Examples 1-3 and Comparative Examples 1-5.

[0034] Figure 4 The inhibition rate of zebrafish tail area reduction in Examples 1-3 and Comparative Examples 1-5.

[0035] Figure 5 Hacat cell migration rate of Examples 1-3 and Comparative Examples 1-5.

[0036] Figure 6 Relative transepidermal water loss rates of Examples 4-6 and Comparative Examples 6-7. DETAILED DESCRIPTION

[0037] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention is further described by the following examples. It should be understood that the examples of the present invention are only used to illustrate the technical effects of the present invention, but not to limit the scope of protection of the present invention. In the examples, the methods used are conventional methods unless otherwise specified.

[0038] Some of the raw materials in the present invention come from the following sources:

[0039] Alteromonas fermentation product extract: purchased from Shaanxi Jinhan Biotechnology Co., Ltd.

[0040] Soluble collagen: purchased from Hubei Maidehao Chemical Co., Ltd.

[0041] Pig umbilical cord extract: purchased from Shaanxi Zelang Biotechnology Co., Ltd.

[0042] Nymphoides: Shandong Jiaxiang Aquatic Plant Planting Co., Ltd.

[0043] Nymphoides extract: purchased from Shaanxi Baichuan Biotechnology Co., Ltd.

[0044] MS medium: purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0045] The specific configuration steps of the culture medium configuration method are as follows:

[0046] Weighing medicines:

[0047] S1: MS medium 4.5 g, sucrose 30 g, NAA (0.3 mg / L) 1.5 mL, 6-BA (1.0 mg / L) 5 mL;

[0048] S2: MS medium 4.5 g, sucrose 30 g, NAA (0.3 mg / L) 2 mL, 6-BA (1.0 mg / L) 5 mL;

[0049] S3: MS medium 4.5 g, sucrose 30 g, NAA (0.3 mg / L) 2.5 mL, 6-BA (1.0 mg / L) 5 mL

[0050] Add the weighed medicine into 1L deionized water at 90-100°C; stir evenly until transparent; adjust the pH to 5.8-6.0; dispense into culture bottles and then sterilize; wait for explant inoculation.

[0051] The preparation of Nymphoides callus extract comprises the following steps:

[0052] Select fresh tender buds and leaves of water lily plants with high purity, typical characteristics of water lily, healthy growth, no injuries, and no diseases or insects. After wiping the leaf surface with 75% ethanol for disinfection, cut the tender bud and leaf tissue into 2-6 mm square pieces.

[0053] In the disinfected inoculation room, the tender shoot and leaf tissues were inoculated into culture bottles containing S1, S2, and S3 culture media respectively through aseptic operation procedures.

[0054] The inoculated culture bottle is placed in a light-proof, constant temperature incubator. When the callus cells are in the division stage, they are taken out, ultrasonically broken, filtered and cell fluid is extracted, and the cell fluids extracted from S1, S2 and S3 are combined and freeze-dried to obtain callus cell extracts.

[0055] The preparation of a solvent comprises the following steps:

[0056] The solvent can be obtained by adding sodium hyaluronate into water and stirring evenly.

[0057] A preparation method of an essence, comprising the steps of:

[0058] The composition is added into the solvent and stirred evenly to obtain the essence.

[0059] The specific mass distribution ratio of the composition is shown in Table 1.

[0060] The specific mass distribution ratio of the essence is shown in Table 2.

[0061] Table 1 Composition mass ratio

[0062]

[0063] Table 2 Essence mass percentage ratio

[0064]

[0065] Safety Testing

[0066] With reference to the 2015 Technical Specifications for Safety of Cosmetics, the compositions of Examples 4-6 were evaluated for cosmetic irritation. The test method was a skin patch test, and 30 randomly distributed people aged 16-45 years were tested.

[0067] Test method: Put the test substance into the patch tester, the dosage is 0.020-0.025g, cover the patch tester with the test substance on the back or forearm of the subject with non-irritating cloth-based tape, and gently press it with the palm to make it evenly adhere to the skin surface for 24 hours. After removing the test patch tester, observe the skin reaction after the indentation disappears 30 minutes later. If the result is negative, observe again 24 hours and 48 hours after the patch test.

[0068] Evaluation criteria:

[0069] Grade 0: negative reaction;

[0070] Grade 1: Suspicious reaction, only slight erythema;

[0071] Grade 2: weak positive reaction, erythema, infiltration, edema, and papules may be present;

[0072] Grade 3: Strong positive reaction, erythema, infiltration, edema, papules, and the reaction may extend beyond the test area;

[0073] Grade 4: Very strong positive reaction, obvious erythema, severe infiltration, edema, confluent herpes, and the reaction exceeds the test area.

[0074] Test results: Skin reactions were negative in all subjects.

[0075] The above test results show that the composition provided by the present invention is mild and non-irritating to the skin and is safe to use.

[0076] Cytotoxicity test

[0077] Take the human immortalized epidermal cell line (HaCaT) with stable growth state and adjust the cell concentration to 1×10 5 / mL, 100μL per well. Inoculate into 96-well cell culture plates (except the outermost wells and blank control wells). Add 200μL of PBS to the outermost wells of the cell culture plate to reduce the evaporation of the culture medium. Place in a carbon dioxide incubator and culture at 37℃±1℃, CO2 concentration 5.0%±1% until the cell fusion reaches more than 90%. Add samples, the sample concentration is 10mg / L, set up blank control and sample groups, 100μL per well. Start adding samples (sample group) and PBS (blank control) at the same time interval between each two columns to ensure that the intervention time of each test substance on the cells is 5min. After 5min, remove the test substances in turn, add PBS to each well to wash the test substances, and wash twice. When using a vacuum suction pump or a discharge gun to remove the liquid in the well, be careful to avoid sucking away the surviving cells and affecting the experimental results. Add 100μL of complete culture medium and 10μL of CCK-8 solution to each well, and place in an incubator to culture for about 2h. At a wavelength of 450 nm, the absorbance (OD) value was measured and the cell viability was calculated; the results are shown in Table 3.

[0078]

[0079] Table 3 Cytotoxicity test

[0080] Experimental Group HaCaT survival rate Example 4 (96.64±0.09)％ Example 5 (97.42±1.27)％ Example 6 (97.32±0.17)％ blank (100)％

[0081] According to the experimental results in Table 3, the cell survival rate was maintained above 95%. The combination of the present experiment had little effect on the viability of HaCaT cells.

[0082] Hyaluronidase activity inhibition assay

[0083] Hyaluronic acid, also known as hyaluronic acid, is a powerful moisturizer. It can attract and lock in moisture, maintain skin moisture, and improve dry, rough and dehydrated skin conditions. Hyaluronic acid can also enhance the biological activity of cells and has a strong synergistic effect on repairing and protecting the skin. Therefore, by inhibiting hyaluronidase, the decomposition rate of hyaluronic acid in the skin is reduced, thereby achieving long-term moisture retention and repair of the skin, which is of great significance.

[0084] (1) Take four groups of clean test tubes, label them A, B, C, and D, add 50 μL of the sample (10 mg / L) to test tubes A and B, add 50 μL of distilled water to test tubes C and D, add 50 μL of 500 U / mL hyaluronidase solution to test tubes A and C, add 50 μL of pH 5.6 acetate buffer to test tubes B and D, and place the test tubes in a 37°C incubator for 20 min.

[0085] (2) Add 10 μL of 2.5 mol / L CaCl2 solution to each of the four test tubes and place them in an incubator at 37°C for 20 min.

[0086] (3) Add 50 μL of 0.5 mg / mL sodium hyaluronate solution to test tubes A and C, and add 50 μL of pH 5.6 acetate buffer to test tubes B and D. Place the test tubes in a 37°C incubator for 40 min and then take them out and place them at room temperature for 10 min.

[0087] (4) Add 50 μL of distilled water, 10 μL of 5 mol / L NaOH solution, and 50 μL of acetylacetone solution to the four test tubes respectively. Place the test tubes in a boiling water bath for 30 min, then in an ice bath for 10 min, and finally at room temperature for 10 min.

[0088] (5) Add 100 μL of P-DAB reagent to each test tube, and then measure the absorbance at 530 nm using an ELISA reader.

[0089]

[0090] Wherein: A is the OD value of the sample solution; B is the OD value of the sample blank; C is the OD value of the control sample solution; D is the OD value of the control sample blank.

[0091] Table 4 Hyaluronidase activity inhibition rate

[0092] Experimental Group Hyaluronidase inhibition rate Example 1 84.32％ Example 2 91.75％ Example 3 85.41％ Comparative Example 1 41.22％ Comparative Example 2 74.23％ Comparative Example 3 53.29％ Comparative Example 4 37.35％ Comparative Example 5 78.19％

[0093] From the results in Table 4, it can be seen from the results of Examples 1-3 that the compositions provided by the embodiments of the present invention have good hyaluronidase inhibition efficacy, especially the effect of Example 2 is better.

[0094] From the results of Example 2 and Comparative Examples 1-5, it is shown that there is a certain synergistic effect when using the components defined in the present invention, and the effect achieved by the present invention cannot be achieved by omitting or replacing any one of them.

[0095] Inflammatory factor inhibition test

[0096] TNF-α is the most common inflammatory factor in inflammatory reactions. It can induce the phenotype of vascular endothelial cells to change to an inflammatory type, leading to changes in the body's metabolism and hemodynamics, and promoting the production of inflammatory mediators. Inflammatory mediators can increase vascular permeability, causing local congestion and edema, and may stimulate nerve endings to cause itching and pain. Therefore, the inhibition of TNF-α helps soothe and repair the skin.

[0097] RAW264.7 cells grown to the logarithmic phase were cultured at 1.5×10 5Cells were inoculated at a density of 100 μg / mL in a 24-well plate, with 800 μL of cell suspension per well, and cultured in a 37°C, 5% CO2 incubator for 24 hours. When the cells grew to near fusion, 50 μL of the sample concentration of 10 mg / L was administered, with 3 replicates per group. The positive control was 50 μL of 10 μg / mL dexamethasone, and 1 μg / mL lipopolysaccharide (LPS) was used for stimulation 1 hour later. After incubation in a cell culture incubator for 24 hours, the cell culture fluid was transferred to a centrifuge tube, centrifuged at 5000 r / min for 10 minutes, and the cell supernatant was collected and tested according to the instructions of the mouse TNF-α enzyme-linked immunosorbent assay kit.

[0098]

[0099] Table 5 Results of RAW secretion of TNF-α

[0100] Experimental Group Inhibition rate Positive group 100％ Blank Group 0.00％ Example 1 77.24％ Example 2 86.37％ Example 3 79.47％ Comparative Example 1 67.52％ Comparative Example 2 64.17％ Comparative Example 3 58.04％ Comparative Example 4 55.32％ Comparative Example 5 75.19％

[0101] From the results of Examples 1-3 in Table 5, it can be seen that the composition has a significant effect of inhibiting TNF-α inflammatory factor, and Example 2 has the best effect of inhibiting TNF-α inflammatory factor.

[0102] Comparing Example 2 with Comparative Examples 1-5, Example 2 has the best effect, indicating that the composition of different components can have a certain degree of influence on the efficacy of inhibiting TNF-α inflammatory factors, indicating that there is a certain synergistic effect in the composition. Any replacement or omission of the raw materials or components defined in the present invention cannot achieve the efficacy proposed by the present invention.

[0103] Effect of the composition on moisturizing of zebrafish

[0104] The zebrafish epidermis has a certain osmotic pressure tolerance range, and water loss will occur if the tolerance range is exceeded. Using a high osmotic pressure solution to treat zebrafish will cause its tail to shrink and become smaller due to water loss. Therefore, an induced model of zebrafish hydration and moisturizing can be established to evaluate the effect of moisturizing products.

[0105] (1) Breeding: Zebrafish were paired at a male-female ratio of 2:1, and the eggs were collected and incubated overnight (28°C incubator).

[0106] (2) Experimental groups: The experiment set up a blank control group and an experimental group. The experimental groups included a model group, a positive group, and a sample group, with 8 2 dpf fry in each group.

[0107] (3) Solution preparation: Accurately weigh 1 mg of sample and prepare the sample solution to be tested, use urea (0.05%) as the positive control for the experiment, and add 1 mg of sample to each well. After the administration is completed, place the 6-well plate in a 28°C incubator in the dark for 24 hours.

[0108]

[0109] Test results: as shown in Table 6 below.

[0110] Table 6 Zebrafish moisturizing effect test

[0111] Experimental Group Zebrafish tail area reduction inhibition rate Positive group 1 blank 0 Example 1 0.53 Example 2 0.62 Example 3 0.53 Comparative Example 1 0.17 Comparative Example 2 0.41 Comparative Example 3 0.37 Comparative Example 4 0.11 Comparative Example 5 0.49

[0112] From the results in Table 6, it can be seen from the results of Examples 1-3 that the compositions provided by the embodiments of the present invention have a good effect of inhibiting the reduction of the tail area of ​​zebrafish, especially Example 2 has a better effect.

[0113] From the results of Example 2 and Comparative Examples 1-5, it is shown that there is a certain synergistic effect when using the components defined in the present invention, and the effect achieved by the present invention cannot be achieved by omitting or replacing any one of them.

[0114] Effects of the composition on the migration of Hacat cells through scratch wound

[0115] The degree of cell migration has an important impact on wound healing and skin repair. In the process of wound healing, cell migration is an important step for epidermal cells to cover the wound, so the higher the cell migration rate, the faster the wound heals. At the same time, cell migration is also involved in the repair process after skin damage. If cell migration is impaired, it will lead to skin repair disorders, which in turn cause various skin diseases.

[0116] Inoculate cells into 12-well plates at the corresponding inoculation density and incubate overnight at 37°C in a CO2 incubator. When the cell plating rate in the 12-well plate reaches about 80%, scratch the sample group and the solvent control group. After the scratching, wash the cells with PBS to remove dead cells, add fresh EGF-free keratinocyte culture solution, add 50μL of 10mg / L according to Example 1-3 and Comparative Example 1-5, and take photos and record using an inverted microscope. Incubate in a 37°C, 5% CO2 incubator for 24h, take photos and record again, and use ImageJ software for statistical analysis.

[0117] Table 7 Hacat cell migration rate

[0118] Experimental Group Mobility Blank Group 0 Example 1 29.33％ Example 2 28.92％ Example 3 31.43％ Comparative Example 1 18.01％ Comparative Example 2 17.49％ Comparative Example 3 18.31％ Comparative Example 4 12.09％ Comparative Example 5 24.26％

[0119] From the results of Table 7, it can be seen from the results of Examples 1-3 that the compositions provided by the embodiments of the present invention have a good effect of improving cell migration rate, especially Example 2 has a better effect.

[0120] From the results of Example 2 and Comparative Examples 1-5, it is shown that there is a certain synergistic effect when using the components defined in the present invention, and the effect achieved by the present invention cannot be achieved by omitting or replacing any one of them.

[0121] Preparation and stability testing of skin care gel

[0122] According to the above formula design, Examples 4-6 and Comparative Examples 6-7 were prepared and tested.

[0123] 1. Heat resistance test: Adjust the constant temperature incubator to 40℃, put the sample into a transparent glass bottle with a volume of 10ml / bottle, seal it and place it in a constant temperature incubator. Take it out after three months, return it to room temperature, and observe the changes in appearance.

[0124] 2. Cold resistance test: Adjust the constant temperature incubator to -10℃, put the sample into a transparent glass bottle with a volume of 10ml / bottle, seal it and place it in a constant temperature incubator. Take it out after three months, return it to room temperature, and observe the changes in appearance.

[0125] 3. Room temperature test: Take a transparent glass bottle, fill it with 10 ml of sample per bottle, seal it and place it at room temperature for 6 months, then observe the changes in appearance.

[0126] 4. Light aging test: Put the sample into a transparent glass bottle with a volume of 10 ml / bottle. After sealing, place it under incandescent light for three months and observe the changes in appearance.

[0127] No precipitation or settling was observed in the heat resistance test, cold resistance test, room temperature test, and light aging test, and the original physical properties were maintained.

[0128] Test on the immediate repair effect of skin care gel on human body

[0129] Transepidermal water loss (TEWL) is an important parameter for evaluating skin barrier function. It indicates the loss of water from the deep dermis through evaporation of the epidermis. The higher the TEWL, the more water is lost through the skin and the weaker the skin's moisturizing ability.

[0130] The transepidermal water loss rate is closely related to the hydration of the stratum corneum and is a commonly used indicator reflecting the barrier function of the stratum corneum. In the development of cosmetics, the efficacy of moisturizing cosmetics can be evaluated by testing the TEWL value of skin water loss. In addition, the transepidermal water loss rate can also be used in allergic patch tests, contact dermatitis, physical therapy, burns and new tissue monitoring to promptly detect whether the protective function of the skin has been damaged.

[0131] Therefore, transepidermal water loss rate is an important parameter for assessing skin health and is of great significance to the research, development and evaluation of moisturizing products.

[0132] 50 healthy subjects aged 18-60 years old with no injuries on the inner arm were selected and divided into groups, with 10 people in each group, including 5 males and 5 females. A sample group and a blank group (i.e., negative control group: no test sample) were set up, and the test area was randomly distributed 1 cm inside the volunteer's arm. 2Area. The forearms (two areas) of the above-mentioned subjects were torn off with medical tape 20 times to induce damage to the skin barrier function, and then 5 mg / mL histamine was added to model for 10 minutes (obvious erythema appeared). One area was treated with 1g of the test product, and the other area was blank as a negative control area. The transepidermal water loss rate (TEWL) of the forearm skin was collected before tape tearing off (T0), before using the product (i.e., immediately after histamine) (T1), and 2 hours after using the product (T2). The difference in skin index changes between the product-treated area and the negative control area before and after using the product was used to evaluate the short-term repair effect of the product on skin barrier damage caused by tape tearing and histamine-induced skin irritation, and the average value was taken. The results are shown in Table 8 below.

[0133]

[0134] Table 8 Relative water loss rate of epidermis

[0135] Experimental Group Relative transepidermal water loss Example 4 -4.01％ Example 5 -11.29％ Example 6 -10.26％ Comparative Example 6 2.53％ Comparative Example 7 -11.32％

[0136] Result analysis: From the results of Examples 4-6, the essence prepared by the present invention has significant moisturizing and anti-wrinkle effects, and Example 5, i.e., when the active ingredient is 3 wt%, has the best moisturizing and repairing effects.

[0137] Comparing Example 5 with Comparative Example 6, Example 5 has a better effect, which indicates that the essence prepared by the present invention has better moisturizing and anti-wrinkle effects.

[0138] Comparing Example 5 with Comparative Example 7, the water loss rates are similar, indicating that too high an addition amount cannot effectively improve the moisturizing and repairing effect. Therefore, the essence prepared using the proportions of the components defined in the present invention can maximize its efficacy.

[0139] The embodiments described above are part of the embodiments of the present application, rather than all of the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents the selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present application.

Claims

1. An anti-wrinkle and moisturizing freeze-dried powder composition, characterized in that: The composition is composed of the following mass parts: 0.1-2 parts of Alteromonas fermentation product extract; 0.5-4 parts of soluble collagen; 0.1-0.5 parts of pig umbilical cord extract; Nymphoides callus cell extract 0.1-1 part; Antioxidant 0.1-1 part; The method for preparing the Nymphoides callus cell extract comprises: using its leaves as explant materials, sterilizing, implanting in culture medium, and when the cells are in the division stage, breaking, filtering, and freeze-drying to obtain the extract; The steps of preparing the culture medium are as follows: S1. Weigh MS medium, sucrose, and plant growth hormone; S2. Heat water to boiling under normal pressure, add S1, and stir; adjust pH to 5.5-6; S3. Pack S2, sterilize it, and wait for explant inoculation; The culture conditions are temperature: 26-28°C; time: 3-4 weeks; The ratio of the plant growth hormone NAA to 6-BA is 0.3-0.5; The antioxidant is ascorbic acid.

2. An essence, characterized in that: Comprising the composition of claim 1.

3. The essence according to claim 2, characterized in that The addition ratio of the composition is 1-5wt%.

4. The essence according to claim 2, characterized in that Also included is a vehicle comprising one or more of a thickener, an emulsifier, a skin conditioner, a moisturizer, a pH adjuster, and a preservative.

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