KR20160092569A - A Cosmetic Composition Having An Antioxidant Activity, Anti-wrinkle, And Moisturizing Effects Contaning A Two-stage Fermentation Product Of Yeast And Lactic Acid Bacteria - Google Patents
A Cosmetic Composition Having An Antioxidant Activity, Anti-wrinkle, And Moisturizing Effects Contaning A Two-stage Fermentation Product Of Yeast And Lactic Acid Bacteria Download PDFInfo
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- A—HUMAN NECESSITIES
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A61Q19/08—Anti-ageing preparations
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Abstract
The present invention relates to a method for producing a two-stage fermentation extract and a cosmetic composition comprising the same as an active ingredient. More specifically, the present invention relates to a method for fermenting two or more stages using lactic acid bacteria after completion of a first stage fermentation using yeast, comprising the fermentation extract prepared by the above method and antioxidants containing the same as an active ingredient, To a cosmetic composition for moisturizing.
Description
The present invention relates to a cosmetic composition containing a fermented extract having two or more stages, more specifically, not only has an effect of enhancing a physiological activity through fermentation, but also exhibits an antioxidative effect and a moisture content enhancing effect associated with prevention of aging, Or two-stage or two-stage fermentation extract having excellent skin wrinkle and moisturizing effect induced by light damage, and a cosmetic herbal composition containing the same as an active ingredient.
Recently, as the economic level of modern people has improved and the participation of social life has increased, the interest in skin and beauty health has increased to a high level in various age groups. Many beauty cosmetics and foods have been developed for the purpose of preventing skin aging and delaying the aging process of skin to maintain healthy skin. In particular, researches for alleviation and improvement of skin wrinkles using fermented extracts are actively under way.
Fermentation is the process of decomposing organic matter by enzymes secreted by microorganisms. It is also fermentation that microorganisms produce useful substances in the process of obtaining energy. Traditional Korean fermented foods such as doenjang and cheonggukjang are typical examples of kimchi, as well as familiarity with Koreans. This fermentation process is an anaerobic respiration microorganism that completely decomposes organic matter and produces other kinds of organic matter, thus generating a small amount of energy. These kinds of fermentation are lactic acid fermentation, alcohol fermentation, propionic acid fermentation, and methane fermentation depending on microorganisms.
Lactic acid bacteria, which are representative of microorganisms involved in such fermentation, decompose sugars such as glucose or lactose to produce organic acids such as lactic acid and acetic acid, and produce enzymes such as amylase, cellulase, lipase and protease as byproducts, Thereby facilitating skin penetration.
Therefore, the present inventors have made intensive researches on fermentation products having wrinkle, moisturizing effect and anti-aging effect. As a result, it has been confirmed that the fermented extract has the effect of improving wrinkles, moisturizing and anti-aging without cytotoxicity, And the function is further strengthened. Based on this finding, the present invention has been completed.
The object of the present invention is to provide a cosmetic composition comprising a two-stage fermentation extract having anti-aging effect, skin wrinkle improvement and moisturizing effect as a functional raw material.
To this end, the present invention provides a cosmetic composition for anti-aging, moisturizing effect and skin wrinkle improvement comprising a fermentation extract of two or more stages by yeast and lactic acid bacteria.
The present invention relates to a fermented extract of two or more stages, which exhibits excellent fermentation efficiency through two or more stages of fermentation than the conventional one-stage fermentation and exhibits antioxidative, wrinkle-improving and moisturizing effects associated with anti-aging . Therefore, it has the effect of providing a cosmetic composition having enhanced skin collagen synthesis promotion, inhibition of collagenase activity, skin wrinkle, improvement and prevention of skin aging, and skin moisturizing ability, while exhibiting low side effects.
Hereinafter, the present invention will be described in more detail with reference to Examples. The following examples are provided for the purpose of more clearly illustrating the present invention and are not intended to limit the scope of the present invention.
<Example 1> Preparation of Saccharomyces-Lactobacillus two-stage fermentation extract
Saccharomyces cerevisiae was activated in a YM solid medium at 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized YM liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours. do.
Lactobacillus paracasei is activated in an MRS solid medium at 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized MRS liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours to be used as a seed culture .
YM liquid medium so that the concentration of Saccharomyces cerevisiae is 10 7 to 10 8 CFU / ml, and the mixture is incubated at 25 ° C to 50 ° C for 18 to 120 hours to perform one-stage fermentation.
The Lactobacillus paracasei bacteria previously cultured in the first stage fermentation product is added so as to have a concentration of 10 7 to 10 8 CFU / ml, and cultured at 25 ° C to 50 ° C for 18 to 120 hours to perform two-stage fermentation Respectively.
The fermentation was terminated by heating at 100 ° C for 20 minutes to induce loss of function of the bacteria. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.
Example 2: Preparation of Galactomyces-Lactobacillus two-stage fermentation extract
Galactomyces is activated in a YM solid medium at a temperature of 20 ° C to 40 ° C for 24 hours, inoculated into a sterilized YM liquid medium after one colony, and cultured in a 20 ° C to 40 ° C incubator for 24 hours. .
Lactobacillus paracasei was activated in an MRS solid medium at a temperature of 20 ° C to 40 ° C for 24 hours, inoculated into a sterilized MRS liquid medium after one colony, and cultured in a 20 ° C to 40 ° C incubator for 24 hours. do.
YM liquid medium to a concentration of 10 7 to 10 8 CFU / ml, and then cultured at 20 ° C to 40 ° C for 18 to 120 hours to carry out one-stage fermentation.
The Lactobacillus paracasei bacteria previously cultivated in the first stage fermentation product is added so as to have a concentration of 10 7 to 10 8 CFU / ml and cultured at a temperature of 20 ° C to 40 ° C for 18 to 120 hours to perform a two-stage fermentation, Respectively.
The fermentation was terminated by heating at 100 ° C for 20 minutes to induce loss of function of the bacteria. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.
<Example 3> Preparation of Saccharomyces-Lactobacillus-Lactobacillus three-stage fermentation extract
Saccharomyces cerevisiae was activated in a YM solid medium at 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized YM liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours. do.
Lactobacillus paracasei was activated in an MRS solid medium at a temperature of 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized MRS liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours. do.
Lactobacillus bifidus was activated in an MRS solid medium at a temperature of 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized MRS liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours, .
YM liquid medium so that the concentration of Saccharomyces cerevisiae is 10 7 to 10 8 CFU / ml, and the mixture is incubated at 25 ° C to 50 ° C for 18 to 120 hours to perform one-stage fermentation.
The Lactobacillus paracasei bacteria preliminarily cultured in the first-stage fermentation product is added to a concentration of 10 7 to 10 8 CFU / ml, followed by culturing at 25 ° C to 50 ° C for 18 to 120 hours to perform two-stage fermentation.
The Lactobacillus bifidus bacteria previously cultivated in the two-stage fermentation product is added so as to have a concentration of 10 7 to 10 8 CFU / ml, and cultured at 25 ° C to 50 ° C for 18 to 120 hours to perform three-stage fermentation Respectively.
The fermentation was terminated by heating at 100 ° C for 20 minutes to induce loss of function of the bacteria. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.
Example 4 Preparation of Galactomyces-Lactobacillus-Lactobacillus Three-Stage Fermented Extract
Galactomyces is activated in a YM solid medium at a temperature of 20 ° C to 40 ° C for 24 hours, inoculated into a sterilized YM liquid medium after one colony, and cultured in a 20 ° C to 40 ° C incubator for 24 hours. .
Lactobacillus paracasei was activated in an MRS solid medium at a temperature of 20 ° C to 40 ° C for 24 hours, inoculated into a sterilized MRS liquid medium after one colony, and cultured in a 20 ° C to 40 ° C incubator for 24 hours. do.
Lactobacillus bifidus was activated in an MRS solid medium at a temperature of 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized MRS liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours, .
YM liquid medium to a concentration of 10 7 to 10 8 CFU / ml, and then cultured at 20 ° C to 40 ° C for 18 to 120 hours to carry out one-stage fermentation.
The Lactobacillus paracasei bacteria previously cultivated in the first-stage fermentation product is added to a concentration of 10 7 to 10 8 CFU / ml, followed by culturing at 20 ° C to 40 ° C for 18 to 120 hours to perform two-stage fermentation.
The Lactobacillus bifidus bacteria previously cultivated in the two-stage fermentation product is added so as to have a concentration of 10 7 to 10 8 CFU / ml, and cultured at 25 ° C to 50 ° C for 18 to 120 hours to perform three-stage fermentation Respectively.
The fermentation was terminated by heating at 100 ° C for 20 minutes to induce loss of function of the bacteria. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.
≪ Comparative Example 1 > Preparation of Saccharomyces 1 stage fermentation extract
Saccharomyces cerevisiae was activated in a YM solid medium at 25 ° C to 50 ° C for 24 hours, inoculated into a sterilized YM liquid medium after one colony, and cultured in a 25 ° C to 50 ° C incubator for 24 hours. do.
Saccharomyces cerevisiae bacteria previously cultivated in a YM liquid medium were added so as to have a concentration of 10 7 to 10 8 CFU / ml, and cultured at 25 ° C to 50 ° C for 18 to 120 hours to effect a one-stage fermentation.
The fermentation was terminated by heating at 100 ° C for 20 minutes to induce loss of function of the bacteria. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.
≪ Comparative Example 2 > Preparation of Galactomyces single step fermentation extract
Galactomyces is activated in a YM solid medium at a temperature of 20 ° C to 40 ° C for 24 hours, inoculated into a sterilized YM liquid medium after one colony, and cultured in a 20 ° C to 40 ° C incubator for 24 hours. .
YM liquid culture medium to a concentration of 10 7 to 10 8 CFU / ml and incubated at a temperature of 20 ° C to 40 ° C for 18 to 120 hours to obtain a fermented product.
The fermentation was terminated by heating at 100 ° C for 20 minutes to induce loss of function of the bacteria. The precipitate was removed using a centrifuge and the supernatant was collected, or the filtered supernatant was used.
<Experimental Example 1> Antioxidant activity measurement
DPPH (2.2-diphenyl-1-picryl hydrezyl) radical scavenging activity was used to measure the antioxidant activity of the two-stage fermentation extract.
0.4 mM DPPH (2.2-diphenyl-1-picryl hydrezyl) solution was dissolved in ethanol and filtered. The prepared DPPH solution and the sample solution were mixed at a ratio of 1: 1 and vigorously mixed. Then, the mixture was allowed to stand at room temperature for 10 minutes. After the absorbance at 515 nm was measured using a microplate reader (BioTek, USA), DPPH radical removal rate (%) was calculated by the following equation (1). At this time, 0.1 mg / ml of ascorbic acid was used as a positive control to remove radicals.
As shown in Table 1, the Saccharomyces 1-stage fermentation extract of Comparative Example 1 and the Galactomyces 1-stage fermentation extract of Comparative Example 2 exhibit similar antioxidative activities. It was also confirmed that the two-stage fermentation extract of Example 1 and Example 2 and the three-stage fermentation extract of Example 3 and Example 4 showed an increase in antioxidative activity compared to Comparative Example 1 and Comparative Example 2. The fermented extracts of two or more stages showed higher antioxidant ability than the fermented extract of one stage.
[Equation 1]
DPPH radical scavenging rate (%) = (A - B) / A x 100
Where A is the absorbance of the blank test solution and B is the absorbance of the test solution
<Experimental Example 2> Effect of improving skin wrinkles
<2-1> Evaluation of collagen biosynthesis
A procollagen assay was performed to measure the amount of newly formed collagen in Fibroblast cells. Fibroblast, one of the cell lines involved in collagen production, was inoculated into Dulbecco's modified essential medium (DMEM Welgene, Korea) containing gentamicin (50 ug / mL) and 10% fetal bovine serum (FBS, Welgene, Korea) incubator. 2 × 10 5 Fibroblast cells were inoculated on a 6-well plate and incubated for 24 hours. The medium was removed, washed with PBS, and irradiated at 12.5 mJ at a wavelength of 312 nm. Thereafter, the sample was treated with DMEM medium at different concentrations and further cultured for 48-72 hours. Ascorbic acid 200 μM was used as a positive control, the UV (+) group as a negative control, and the control group as a control. After 48-72 hours of incubation, the medium was collected and the collected collagen was collected using a Procollagen Type-I C-Peptide (PIP) EIA kit (Takara Bio Inc. MK101) Respectively. Add 100ul of Antibody-POD conjugate solution to each well, add 20ul of sample or standard, mix well and leave at 37 ℃ for 3 hours. After the reaction was completed, the contents were removed and washed 4 times with 400 μl of PBS. After removing 100 μl of the substrate solution, react at room temperature for 15 minutes, add 100 μl of stop solution and mix gently. Using a microplate reader (BioTek, USA), measure the absorbance at 450 nm, (Table 2).
As shown in Table 2, in order to examine the anti-wrinkle activity of the two-stage fermentation extract by ultraviolet stimulation, the two-stage fermentation extract was compared with the first stage fermentation extract for comparison. The two-stage fermentation extract of Example 1 and Example 2 and the three-stage fermentation extract of Example 3 and Example 4 exhibited a stronger collagen synthesis performance than the one-stage fermentation extract of Comparative Example 1 and Comparative Example 2, And the effect of anti-aging and wrinkle improvement
(-)
(+)
<2-2> Evaluation of inhibition of MMP-1
MMP-1 (Matrix metalloproteinase I, collagenase), which degrades collagen type I, is the most widely known enzyme that degrades collagen. A procollagen assay was performed to measure the amount of newly formed collagen in Fibroblast cells. Fibroblast, one of the cell lines involved in collagen production, was cultured in Dulbecco's modified essential medium (DMEM Welgene, Korea) containing gentamicin (50 ug / mL) and 10% fetal bovine serum (FBS, Welgene, Korea) CO2 incubator. 2 × 10 5 Fibroblast cells were inoculated on a 6-well plate and incubated for 24 hours. The medium was removed, washed with PBS, and irradiated at 12.5 mJ at a wavelength of 312 nm. Thereafter, the sample was treated with DMEM medium for each concentration and further cultured for 48 hours. Ascorbic acid was compared with the results of the test using 200uM as positive control, UV (+) as negative control, and control group as control. The medium was collected after 48 hours of incubation after the sample treatment. The collected medium was measured for absorbance at 450 nm with an ELISA reader using an MMP-1 measurement kit (RPN 2610, Amersham Bioscience, UK). Add a buffer to a 96-well plate coated with anti-MMP1 antibody as a blank, add 100 μl of the sample and the standard solution in the medium, and incubate for 2 hours at room temperature without shaking. Fill the wells with Washing buffer, wash 3 times and wash thoroughly. Add 100 μl peroxide conjugate and incubate at room temperature for 2 hours without shaking. Fill the wells with washing buffer and incubate for 30 minutes. After stopping the reaction with 100 μl 1M H 2 SO 4, the absorbance was measured at 450 nm using a microplate reader (BioTek, USA) (Table 3).
As shown in Table 3, in order to examine anti-wrinkle activity of the two-stage fermentation extract by ultraviolet stimulation, the two-stage fermentation extract was compared with the one-stage fermentation extract for comparison. In the case of the two-stage fermentation extract of Example 1 and Example 2, and the three-stage fermentation extract of Example 3 and Example 4, MMP-1 activity was inhibited more strongly than the one-stage fermentation extract of Comparative Example 1 and Comparative Example 2, This is consistent with the results of collagen biosynthesis (Table 2).
(+)
Example 5: Preparation of emulsion-type cosmetic composition containing two-stage fermentation extract
Cosmetic preparations containing two-stage fermentation extracts were prepared in the same manner as the cosmetic preparations containing the two-stage fermentation extracts of Examples 1 and 2, respectively (Formulations 1 and 2). For comparison of efficacy, glycerin (Comparative Formulation Example 1) containing 1,3-butylene glycol, a cosmetic composition containing glycerin and sorbitol as a moisturizing agent (Comparative Formulation Example 2), a cosmetic composition containing neither an extract nor a moisturizer (Comparative Formulation Example 3 ) Are shown in Table 4 below.
(Example 1)
(Example 2)
<Experimental Example 3> Moisturizing effect of the two-stage fermentation extract
The clinical moisturizing effects of the cosmetic compositions of Formulation Examples 1 and 2 were measured as follows. A, B and C groups were divided into 6 groups (A, B, C, D, E, F) by randomly dividing 150 healthy men and women who felt skin dry through a questionnaire. (Comparative Formulation Example 1) containing glycerin and 1,3-butylene glycol as moisturizing agents instead of the two-stage fermentation extract, and glycerin and sorbitol as moisturizing agents were added to D, E, and F groups, respectively A cosmetic composition (Comparative Formulation Example 2) and a cosmetic composition containing no extract or moisturizer (Comparative Formulation Example 3) were applied to the face for 2 weeks, twice a day for 8 weeks. The moisturizing effect was evaluated by Corneometer CM 820 (Corage + Khazaka, Germany) for every 2 weeks and after the start of the test. The experimental results are shown in Table 5 below.
As shown in Table 5, it was found that the moisturizing effect of the cosmetic compositions (Formulation Examples 1 and 2) containing the two-stage fermentation extract of the present invention was superior to the cosmetic compositions containing other moisturizing agents (Comparative Formulation Examples 1 and 2) I could confirm.
≪ Example 6 > Flexible longevity (skin lotion)
The number of softening times containing the above two-stage fermentation extraction was prepared according to a conventional method as shown in Table 6 below.
Example 7 Milk Lotion (Milky Lotion)
Milk lotions containing the above two-stage fermentation extraction were prepared by heating the water phase and the oil phase at 75 DEG C and emulsifying them for 5 minutes, respectively, as shown in Table 7 below.
≪ Example 8 >
As shown in Table 8 below, the nutrient cream containing the two-stage fermentation extraction was prepared by emulsifying the aqueous phase and the oil phase at 75 ° C for 5 minutes, followed by cooling and defoaming.
<Example 9> Preparation of cosmetic composition of essence formulation containing two-stage fermentation extract
Cosmetic formulations containing the two-stage fermentation extracts were prepared in the same manner as the cosmetic formulations (Formulation examples 3 and 4) containing the two-stage fermentation extracts of Examples 1 and 2, respectively, except for the two-stage fermentation extract (Comparative Formulation Example 4) are shown in Table 9 below.
<Experimental Example 4> Effect of wrinkle improvement, sensation of use and satisfaction of the two-stage fermentation extract
Tests were conducted using the essences prepared in Formulation Examples 3 and 4 and Comparative Formulation Example 4 to evaluate skin wrinkle-improving effect, feeling and feeling of satisfaction with the cosmetic compositions of Formulation Examples 3 and 4. [ For the wrinkle improvement effect items of the skin, the wrinkle improvement effect of the essence of Comparative Formulation Example 4 was relatively evaluated based on the essence of Formulation Examples 3 and 4, and the feeling and satisfaction were relatively evaluated.
The effect of wrinkle improvement, feeling of use, and satisfaction after using the essence prepared for 100 people twice a day for 4 weeks were surveyed. The evaluation was carried out on the basis of very good, excellent, medium, and poor. The results are shown in Tables 10 and 11 below.
As shown in Table 10, the effect of improving the wrinkles of the skin relative to the cosmetic compositions of Formulation Examples 3 and 4 according to the present invention was superior to that of Formulation Example 3, and that of Formulation Example 4 was 85, The degree of improvement of the skin was very excellent.
As shown in Table 11, the comparative feeling for the cosmetic compositions of Formulation Examples 3 and 4 according to the present invention was excellent, 78 for Formulation Example 3, 79 for Formulation Example 4, and 87% It is found that the feeling and the satisfaction are very excellent as compared with Example 4.
Claims (5)
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Cited By (7)
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WO2020245797A1 (en) * | 2019-06-05 | 2020-12-10 | Lac2biome S.r.l. | Compositions comprising a bacterial strain lactobacillus paracasei and hyaluronic acid and the use thereof for the treatment of the skin |
WO2021112298A1 (en) * | 2019-12-05 | 2021-06-10 | 주식회사 네이처인랩 | Method for producing complex fermentation product of lichen-derived minerals and galactomyces |
CN113151067A (en) * | 2021-03-31 | 2021-07-23 | 绽妍生物科技有限公司 | Extraction and separation process of lactobacillus fermentation extract |
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CN113924357A (en) * | 2019-04-02 | 2022-01-11 | 株式会社爱托健 | Bifidobacterium longum strain or cosmetic composition containing the same |
WO2020245797A1 (en) * | 2019-06-05 | 2020-12-10 | Lac2biome S.r.l. | Compositions comprising a bacterial strain lactobacillus paracasei and hyaluronic acid and the use thereof for the treatment of the skin |
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CN113151067A (en) * | 2021-03-31 | 2021-07-23 | 绽妍生物科技有限公司 | Extraction and separation process of lactobacillus fermentation extract |
WO2024014816A1 (en) * | 2022-07-14 | 2024-01-18 | 주식회사 라비오 | Cosmetic composition for improving scalp comprising fermented microorganism as active ingredient, and method for preparing same |
KR102627236B1 (en) * | 2023-04-14 | 2024-01-19 | 인코스(주) | A sea squirt shell ferment made by using bioconversion process, composition including the same and cosmetic composition including the same for anti-inflammatory, anti-oxidation, anti-wrinkle, skin regeneration, skin elasticity and skin soothing |
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