KR102260314B1 - Cosmetic composition for wrinkle improvement - Google Patents

Abstract

The present invention relates to a cosmetic composition containing fermented aloe as an active ingredient and a manufacturing method thereof. The manufacturing method of the present invention comprises: a first step of fermenting aloe with lactic acid bacteria; a second step of post-processing the fermented aloe obtained after fermentation with ultrasonic waves at room temperature; and a third step of concentrating the fermented aloe and then freeze-drying the fermented aloe to manufacture the fermented aloe in the form of powder. The existing cosmetic compositions using aloe mainly remove the aloe skin and extract and use the gel existing in the endothelium, or use only aloe vera gel and fermented products using specific lactic acid bacteria, mushrooms fungus, basidiomycetes, etc. Since more than 95% of the gel is water and the content of active ingredients is low, a large amount of the extract must be used to achieve the desired effect as a functional improvement material. In order to solve the above disadvantage, the destruction of various active ingredients contained in aloe is minimized by fermenting by-products such as the skin, etc. remaining after removing the entire ground part or gel of aloe. Through its high antioxidant effect, the present invention can provide effects such as wrinkle reduction, whitening effects, promotion of skin cell activity, anti-atopic dermatitis, etc.

Description

Wrinkle improvement cosmetic composition {Cosmetic composition for wrinkle improvement}

The present invention relates to a cosmetic composition containing fermented aloe as an active ingredient, and a method for preparing the same, and more particularly, to the whole above-ground part of Aloe aborescens or Aloe verana, rather than using a gel component usually obtained after peeling aloe. The present invention relates to a cosmetic composition containing, as an active ingredient, a fermented aloe product having an wrinkle-improving effect of a fermented product obtained by using lactic acid bacteria that is edible only in the skin part remaining after obtaining a gel from Saponaria, and a method for manufacturing the same.

There are about 500 different types of aloe belonging to the genus Aloe, among which there are three species used for food and medicinal purposes: Aloe Vera and Aloe Aborescens. Among them, aloe vera is widely known and is used in various fields from food to cosmetic materials, but it is confirmed that there is no use as a cosmetic composition for fermented products of aloe vera.

It is confirmed that most of the papers using fermented aloe vera are used as food additives, and there is no research as a cosmetic composition. A paper using an aloe vera extract inhibited prostaglindan, an inflammatory mediator, and confirmed an anti-inflammatory effect (Beatriz V et al., J Ethnopharmacol, 55(1): 69-75, 1996). Antibacterial activity has been reported by inhibiting bacteria such as Staphylococcus aureus, Pseudomonas aeruginosa, and Trichophyton mentagraphytes (Olaleye MT et al., Afr J Biotechnol, 4(12): 1413-1414, 2005). In addition, it was confirmed that collagen synthesis and proliferation were promoted in human skin cell lines using aloe vera gel extract, confirming its value as a cosmetic material. In addition, many studies on aloe vera extracts in various fields have been reported.

As a thesis on the fermented product of Aloe aborecens, it was confirmed that the fermented product of Aloe aborecens was not toxic to organs such as liver, kidney, and heart as a result of single and repeated administration toxicity test in rats (Cho JH et al. Lab Anim Res, 27(3): 235-244, 2011) and other papers using fermented products of Aloe aborescens have not been reported. Papers on various efficacy effects of Aloe aborescens extract have been reported, and the cytoprotective effect from oxidative stress was confirmed using the antioxidant activity of Aloe aborescens (Beppu H et al., J Ethnopharmacol, 89(1)). : 37-45, 2003), it was confirmed that when Aloe aborescens juice was applied to the affected area, it promoted skin regeneration and was effective in wound healing (Jia Y et al., J Ethnopharmacol, 120(2): 181-9, 2008). ). The reason why it is effective in wound healing is due to the antifungal and anti-inflammatory effects of Aloe aborescens extract. The study on the antifungal effect effectively inhibiting the proliferation of Trichophyton mentagrophytes conducted by Fujita K et al. 14(1): 132-6, 1978) reported an anti-inflammatory study by Masatoshi et al. (Masatoshi YM et al., Agric Biol Chem, 1627-1629, 1991). As in the above studies, several papers have been previously reported on Aloe Aborescens, but studies on skin anti-aging and anti-wrinkle effects using Aloe Aborescens are insufficient.

Conventional technology includes fermented aloe vera with a patent for fermented aloe for whitening cosmetic material and whitening cosmetics using the same (Korean Patent Registration No. 10-11032830000), as well as aloe fermented and hydrolyzed extracts that prevent and improve atopic dermatitis. manufacturing method and manufactured cosmetics (Korea Patent Registration No. 10-12050970000). However, all of these aloe fermentation-related patents use only aloe vera gel, and some of the fermented bacteria use specific lactic acid bacteria or mushrooms or basidiomycetes, etc. it is about

In particular, it is understood that there is no patent application for fermented aborescens technology at present, and a patent related to aborescens extract has been applied for a food composition having a function of improving and preventing constipation (Korean Patent Application No. 10-2008-0122550), A patent for a composition for external application for skin containing aloe extract and capsaicin as active ingredients, which has effects such as moisturizing the skin, improving blood circulation, and decomposing cellulite, has been registered (Korean Patent Registration No. 10-17641310000).

That is, functional cosmetic compositions for providing anti-oxidation, skin cell activity promotion, wrinkle improvement, and whitening effects as well as atopy efficacy using fermented aloe have not been provided so far.

Republic of Korea Patent Registration No. 10-11032830000 "The Manufacturing Method of Fermented Aloe for Whitening Effect in skin, and the Functional Whitening Cosmetics Containing Fermented Aloe" Republic of Korea Patent Registration No. 10-12050970000 "Method for producing fermented and hydrolyzed aloe extract showing atopic dermatitis prevention and improvement effect and cosmetics manufactured using the same" Korean Patent Application No. 10-2008-0122550 (December 4, 2008) No. "Constipation releaving Health Food composition containing Aloe vera and Aloe vera and Aloe having the function of improving and preventing constipation containing Aloe vera and Aloe borescens as main ingredients aborescens)" Republic of Korea Patent Registration No. 10-17641310000 "EXTERNAL PREPARATION FOR SKIN COMPRISING ALOE EXTRACTS AND CAPSAICIN containing aloe extract and capsaicin as active ingredients"

The present invention is to solve the above problems, and as a result of diligent efforts to find cosmetic compositions having various functions, high antioxidant effect by minimizing the destruction of useful substances through the lactic acid bacteria fermentation process of aloe outpost or aloe by-product remaining after obtaining the gel. Provided is a cosmetic composition containing fermented aloe as an active ingredient for providing effects such as anti-atopic function through skin cell activation and promotion and skin immunity enhancement along with wrinkle improvement and whitening effect, and a manufacturing method thereof it is to do

Another object of the present invention is to provide a cosmetic composition containing fermented aloe as an active ingredient, and a method for preparing the same.

Another object of the present invention is to provide a cosmetic composition containing fermented aloe as an active ingredient, and a method for preparing the same.

However, the objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, according to an embodiment of the present invention, there is provided a method for producing a cosmetic composition containing fermented aloe as an active ingredient, comprising: a first step of fermenting aloe with lactic acid bacteria; a second step of post-processing the aloe fermented product obtained after fermentation with ultrasonic waves at room temperature; and a third step of concentrating the fermented aloe and then freeze-drying to prepare a powder; It may be characterized in that it consists of.

In this case, in the second step, the aloe fermented product is subjected to ultrasonic post-treatment at room temperature to extract the aloe fermented product, and the post-sonicated treatment of the fermented product is performed by fermenting the aloe fermented product under ultrasonic conditions of 20 to 120 kHz at room temperature for 30 minutes It may be characterized by extraction for 6 hours.

In addition, the lactic acid bacteria used in the fermentation of aloe in the first step are: Leuconostoc mesenteroides, Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei, Saccharomyces exiguus, Lactobacillus acidophilus, Bifidobacterium animalis, Bifidobacterium bifidum, Lobactobactillus casei delbrueckii, Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Saccharomyces cerevisiae, Rhodopseudomonas palustris, Saccharomyces fragilis, Aspergilus niger, Bacillus stearothermophilus, and Acetobacter can be characterized in that it comprises at least one selected from the group consisting of xylinum have.

In addition, when fermenting aloe in the first step, the entire above-ground part of aloe or the remaining by-products except for the aloe gel are fermented as an active ingredient, and the aloe powder obtained by drying in the third step is used in an amount of 1% to 90% based on the total weight of the cosmetic composition. It can be characterized by adding.

In addition, the kind to be fermented in the first step may include any one or more of Aloe aborescens, Aloe vera, and Aloe saponaria.

To achieve the above object, a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention may include fermented aloe as an active ingredient.

In this case, the kind to be fermented in the present invention may include any one or more of aloe aborescens, aloe vera, and aloe saponaria.

Further, in the present invention, the fermented aloe obtained by fermenting aloe with lactic acid bacteria and then post-treating the fermented aloe with ultrasonic waves at room temperature is used. It may be characterized in that the fermented aloe extracted at room temperature for 30 minutes to 6 hours is used.

In addition, the lactic acid bacteria used for the fermentation of aloe in the present invention are: Leuconostoc mesenteroides, Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei, Saccharomyces exiguus, Lactobacillus acidophilus, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacactii casei Any one or more selected from the group consisting of Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Saccharomyces cerevisiae, Rhodopseudomonas palustris, Saccharomyces fragilis, Aspergilus niger, Bacillus stearothermophilus, and Acetobacter xylinum may be selected from the group consisting of.

In addition, during aloe fermentation in the present invention, the whole above-ground part of aloe or by-products other than aloe gel are fermented as an active ingredient, and then dried aloe powder is added in an amount of 1% to 90% based on the total weight of the cosmetic composition. can be done with

In the cosmetic composition containing fermented aloe as an active ingredient, and the manufacturing method thereof, according to an embodiment of the present invention, the cosmetic composition using aloe is mainly used by removing the aloe skin and extracting the gel present in the endothelium as it is, or Only aloe vera gel has been used with fermented products using specific lactic acid bacteria, mushrooms, or basidiomycetes, but the gel contains more than 95% water and has a low content of active ingredients, so it is necessary to use a large amount of extract to achieve the desired effect as a functional cosmetic material. In order to solve many shortcomings, the destruction of various active ingredients contained in aloe is minimized by fermenting by-products such as the skin remaining after removing the entire above-ground part or gel of aloe, improving wrinkles, whitening effect, and promoting skin cell activity through high antioxidant effect. and anti-atopic effects.

In addition, the cosmetic composition containing the Aloe aborescens extract as an active ingredient, and the preparation method thereof, according to another embodiment of the present invention have improved wrinkle improvement, whitening effect, skin cell activity promotion and anti-atopic dermatitis through antioxidant effect. As a novel functional cosmetic composition, it provides an effect that can be applied to various cosmetic formulations.

1 is a flowchart illustrating a method for preparing a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention.
2 is a graph showing the results of analysis of cell activity of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention.
3 is a graph showing the results of analysis of free radical scavenging ability of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention.
4 is a graph showing the results of analysis of collagen synthesis ability of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention.
5 shows the results of analysis of the inhibition rate of Matrix metalloproteinase (MMP-1) in fermented aloe, aloe vera gel and gel fermented product used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention; It is a graph.
6 is a graph showing the results of analyzing the tyrosinase enzyme activity inhibition rate of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention. .
7 is an aloe fermented product used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention, aloe vera gel, and the degree of inhibition of nitrooxide (NO) production from macrophages of the gel fermented product; It is a graph showing the analysis result.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, detailed description of preferred embodiments of the present invention will be described with reference to the accompanying drawings. In the following description of the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description thereof will be omitted.

The present invention provides a functional cosmetic composition comprising, as an active ingredient, a fermented product obtained by fermenting lactic acid bacteria with an aloe plant or a by-product after obtaining a gel with ultrasonic waves at room temperature. Therefore, in an embodiment of the present invention, when aloe fermented product was treated on human skin cell line (CCD-986sk), the present invention based on the results of experiments such as increased cellular metabolic activity, wrinkle improvement, whitening and skin immunity enhancement effect in the cells was completed.

Hereinafter, the present invention will be described in detail.

1 is a flowchart illustrating a method for preparing a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention. Referring to FIG. 1 , the fermented product of aloe aborescens is prepared by (a) fermenting aloe with lactic acid bacteria (S11); (b) post-processing the fermented product obtained after fermentation with ultrasonic waves at room temperature (S12); (c) concentrating the fermented product and then freeze-drying to prepare it in the form of a powder (S13). The present invention provides a functional cosmetic composition comprising fermented aloe as an active ingredient. to provide.

The aloe used in step S11 is not limited to aloe arborescens, and may be a functional cosmetic composition comprising at least one of aloe vera, aloe arborescens, and aloe saponaria.

In addition, the aloe used in step S11 may be a functional cosmetic composition comprising at least one of aloe vera, aloe aborescens, and aloe saponaria as an edible aloe species. However, it is not limited to the above three types of aloe, and various aloe species may be targeted.

In addition, the aloe used in step S11 may provide a functional cosmetic composition including not only the aloe gel, but the whole above-ground part of aloe or a fermented by-product excluding the aloe gel as an active ingredient. In addition, it may be fermented by adding 1% to 90% (by weight) of aloe powder obtained by drying the by-product remaining after removing the whole above-ground part of aloe or aloe gel during fermentation, based on the total weight of the cosmetic composition.

In another embodiment of the present invention, the extract prepared by refluxing the whole above-ground part of aloe during fermentation or the by-product remaining for aloe gel with hot water at 100° C. for 6 to 24 hours is filtered with a filter paper, and the resulting extract is used as the total volume of the cosmetic composition. It may be fermented by adding 10% to 90% (volume ratio).

In another embodiment of the present invention, the types of lactic acid bacteria that can be used for fermentation are Leuconostoc mesenteroides, Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus paracasei, Saccharomyces exiguus, Lactobacillus acidophilus, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacillus acidophilus, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacillus , characterized in that it comprises the Lactobacillus delbrueckii, Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Saccharomyces cerevisiae, Rhodopseudomonas palustris, Saccharomyces fragilis, Aspergilus niger, Bacillus stearothermophilus, and at least one selected from the group consisting of Acetobacter xylinum It may be a functional cosmetic composition.

Meanwhile, it is possible to provide a functional cosmetic composition comprising, as an active ingredient, a fermented aloe obtained after post-treatment of the fermentation broth obtained through lactic acid bacteria fermentation of aloe at room temperature with ultrasonic waves in step (S12).

And in another embodiment of the present invention, the post-sonication treatment of the fermented product comprises extracting the fermented Aloe aborescens of the above example under ultrasonic conditions of 20 to 120 kHz at room temperature for 30 minutes to 6 hours. It may be a functional cosmetic composition comprising

In another embodiment of the present invention, when the cosmetic composition is mixed with a radical such as a DPPH solution of a preset weight ratio (eg, 0.001 to 0.0005 parts by weight of the total weight), antioxidant efficacy can be confirmed.

In another embodiment of the present invention, after administering the cosmetic composition to human skin fibroblasts (CCD-986sk), using 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) solution The effect of promoting the growth of human skin cells can be confirmed.

In another embodiment of the present invention, when the cosmetic composition is administered to the cells, the wrinkle improvement effect can be confirmed by increasing the amount of collagen synthesis.

In another embodiment of the present invention, when the cosmetic composition is administered to the cells, the wrinkle improvement effect can be confirmed by inhibiting the expression of MMP-1.

In another embodiment of the present invention, the whitening effect can be confirmed when the cosmetic composition is mixed with tyrosinase.

In another embodiment of the present invention, it is possible to confirm the anti-atopic effect through skin immunity enhancement by suppressing the amount of NO produced after administration of the cosmetic composition to macrophages.

[Example] Aloe fermentation and post-treatment

1-1. Manufacture of ingredients for aloe fermentation

For the preparation of the material for the fermentation of aloe of the present invention, the entire above-ground part of Aloe aborescens is dried and then powdered and used as a fermentation sample. Alternatively, after removing the shell of Aloe Vera or Aloe Saponaria, the gel is taken, and the remaining peel by-product is dried to prepare a powder and used for fermentation. In addition, an aloe vera gel obtained after removing the aloe vera shell or a fermented product obtained by fermenting the gel is used as a control sample.

1-2. Aloe Fermentation

To prepare the fermented aborescens of the present invention, the samples obtained in 1-1 were added to the MRS medium, which is a medium for culturing lactic acid bacteria, in a ratio of 1:10 (w/v or v/v)), and then at 37°C. Cultured L actobacillus plantarum (ATCC®14917) 1-10% (v/v) of the bacteria were inoculated. After the inoculation was completed, fermentation was carried out at a temperature of 37° C. at a stirring speed of 120 rpm for 4 days to obtain a fermented aloe.

1-3. Post-treatment of aloe fermented broth

For the ultrasonic post-treatment of the fermented iloe of the present invention, the fermentation broth obtained in 1-2 above was treated at room temperature under ultrasonic conditions of 20 to 120 kHz for 30 minutes to 6 hours.

The fermented aloe obtained above is filtered through a reduced pressure filtration device. Then, it was concentrated under reduced pressure and then freeze-dried to prepare a powder.

2 is a graph showing the results of analysis of cell activity of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention. 3 is a graph showing the results of analysis of free radical scavenging ability of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention. 4 is a graph showing the results of analysis of collagen synthesis ability of fermented aloe, aloe vera gel, and fermented gel used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention. 5 shows the results of analysis of the inhibition rate of Matrix metalloproteinase (MMP-1) in fermented aloe, aloe vera gel and gel fermented product used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention; It is a graph. 6 is a graph showing the results of analyzing the tyrosinase enzyme activity inhibition rate of an aloe fermented product, an aloe vera gel, and a gel fermented product used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention. . 7 is an aloe fermented product used in a cosmetic composition containing fermented aloe as an active ingredient according to an embodiment of the present invention, aloe vera gel, and the degree of inhibition of nitrooxide (NO) production from macrophages of the fermented gel; It is a graph showing the analysis result.

Hereinafter, with reference to FIGS. 2 to 7, cell activity and efficacy evaluation of a cosmetic composition containing fermented aloe as an active ingredient will be described with specific experimental examples.

[Evaluation of cell activity and efficacy]

2-1. cell culture

Human skin cell lines (CCD-986sk) were each cultured in Dulbecco's modified Eagle's medium supplemented with 10% Fetal bovine serum. Each of the cells was cultured in a culture dish at 37° C., 5% CO ₂ while exchanging the medium every 2 days after culturing in a culture dish.

2-2. Assessment of cytotoxicity and cellular activity

^{1×10 4} human skin cell line (CCD-986sk) in a 48-well plate After inoculation at a cell/well concentration, aloe fermented products and control groups were administered at each concentration (1%, 0.5%, 0.1%, 0.05% and 0.01%), and then cultured for 24 hours without substance administration. Cell viability analysis (MTT, thiazolyl blue tetrazolium bromide) was performed as a negative control group.

As a result, as shown in FIG. 2 , even at a concentration of 1% of the aloe fermented product administered group, the cell viability of the negative control group that was not treated with anything was significantly increased to activate cell metabolism, and cell metabolism was activated at other concentrations as well. It was confirmed that the cell viability was increased compared to the negative control group according to the present invention, so it was predicted that the fermented iloe did not simply induce cytotoxicity, but also significantly promoted the activity of mitochondria in the cell to exhibit the desired cosmetic activity. On the other hand, the aloe vera gel and fermented ivera gel used as a control showed no significant difference in cell viability and did not induce cytotoxicity compared to the untreated negative control at all concentrations. However, it was difficult to expect high activity because the degree of promoting cell activity was not higher than that of abo essence fermented products or vera bark fermented products.

2-3. Efficacy evaluation

2-3-1. Antioxidant evaluation

The antioxidant activity of aloe fermented products used in the experiment was confirmed by measuring DPPH scavenging activity. Trolox (0.05%) was used as a positive control, and 0.2 mM DPPH solution was added to the samples prepared by each concentration (0.5%, 0.25%, 0.125%) of fermented aloe, followed by reaction at room temperature for 30 minutes. After the reaction was completed, the absorbance was measured at 520 nm, and the DPPH radical scavenging ability according to the concentration was confirmed by Equation 1 below. The free radical scavenging activity was expressed as a percentage compared to the negative control to which the sample was not added.

As a result, as shown in FIG. 3 , when aloe fermented products were administered, the DPPH radical scavenging ability was significantly increased in a concentration-dependent manner in all experimental groups compared to the negative control group, and at a concentration of 0.5%, more than 95% of the radical scavenging ability was exhibited. This is considered to be a fairly high antioxidant effect close to 100% of Trolox used as a control. On the other hand, the aloe vera gel used as a control had very little antioxidant effect, and the fermented aloe vera also showed a low free radical scavenging ability of about 60% compared to the fermented aloe of the two types, demonstrating that the two fermented products had excellent antioxidant efficacy.

2-3-2. Evaluation of wrinkle improvement effect by collagen synthesis ability

Fibroblast lines were placed in a 48-well plate in a dish at 1×10 ^{4 per well.} After inoculation at a cell/well concentration, aloe fermented products and controls were administered at each concentration (1%, 0.5%, 0.1%, 0.05% and 0.01%), and cultured for 24 hours. As a positive control, TGF-beta1 was used at a concentration of 0.1 ng/ml. The culture medium was collected and used for the experiment, and the amount of collagen synthesis in the cell culture medium was measured using a procollagen type 1 peptide EIA kit (Takara biomedical). 100ul of the antibody-POD conjugate solution was put into each well, and then 20ul of the sample and standard were added and reacted at 37℃ for 3 hours. After the reaction was completed, the culture medium was removed from each well and washed 4 times with wash buffer. 100ul of the substrate solution was added and reacted at room temperature for 15 minutes. After stopping the reaction by adding 100 ul of Stop solution (1N H2SO4), absorbance was measured at 450 nm with an ELISA reader.

As a result, as shown in FIG. 4 , when the aloe fermented product was administered, collagen synthesis was significantly increased in all experimental groups compared to the negative control group. In particular, at a concentration of 0.5% fermented product added, the collagen synthesis increased by 65% or more compared to the negative control group. On the other hand, aloe vera gel and gel fermented products, which are widely used in conventional cosmetics as a control group, showed only about a 45% increase in collagen synthesis compared to the negative control group at the highest concentration of 0.5%. It was confirmed that this high collagen synthesis ability.

2-3-3. Wrinkle improvement effect evaluation by MMP-1 inhibitory ability

Fibroblast lines were placed in 48-well plate dishes 1×10 ^{4 per hole.} After inoculation at a cell/well concentration, aloe fermented products and controls were administered at each concentration (1%, 0.5%, 0.1%, 0.05%, and 0.01%), and cultured for 24 hours. As a positive control, TGF-b was used at a concentration of 0.1 ng/ml. The culture medium was collected and used for the experiment, and the amount of collagen synthesis in the cell culture medium was measured using the Human MMP-2 ELISA kit (Abcam). 100ul of the antibody-POD conjugate solution was put into each well, and then 50ul of the sample and standard were added and reacted at 37℃ for 3 hours. After the reaction was completed, the culture medium was removed from each well and washed 4 times with wash buffer. 100ul of the substrate solution was added and reacted at room temperature for 15 minutes. After stopping the reaction by adding 100 ul of Stop solution (1N H2SO4), absorbance was measured at 450 nm with an ELISA reader.

As a result, as shown in FIG. 5 , when the aloe fermented product was administered, the amount of MMP-1 production was significantly decreased in all experimental groups compared to the negative control group. In particular, the production of MMP-1 at the maximum added concentration of 0.5% was higher than that of the negative control group. Compared to the decrease of about 40% or more, the amount of MMP-1 decreased by 20-30% in aloe vera gel and fermented gels in a similar manner, indicating that the fermented product had a very high MMP-1 production inhibitory effect. Even at a low concentration of 0.05%, aloe fermented products inhibited production by more than 20%, whereas vera gel or fermented gels showed a slight decrease of less than 10%.

2-3-4. Whitening effect evaluation

To verify the whitening effect of the aloe fermented product, the tyrosinase inhibitory effect was analyzed. To this end, first, mushroom-derived tyrosinase (sigma aldrich) was dissolved in 0.1 M phosphate buffer at 2000 units/ml and cryopreserved, and L-3,4-dihydroxylphenyl alanine (L-3,4-dihydroxylphenyl alanine, L -DOPA) was prepared immediately before analysis.

Tyrosinase inhibitory activity was preincubated with tyrosinase (78 units/mL) and 1/15M phosphate buffer at 25°C for 5 minutes, and aloe fermented products and controls were administered at 0.125%, 0.25%, and 0.5%, A positive control was used as arbutin. The L-DOPA was added thereto, and the absorbance was measured at 475 nm for 10 minutes, and then the enzyme inhibition rate (%) was calculated from Equation 2 below.

As a result, as shown in FIG. 6 , when the aloe fermented products were administered, the tyrosinase inhibition rate increased in a concentration-dependent manner depending on the treatment concentration. In particular, at the highest concentration of 0.5%, the tyrosinase inhibition rate was measured to be about 30% or more. On the other hand, aloe vera gel or fermented gel showed a low inhibition rate of enzyme activity of about 5-6% at the same maximum concentration of 0.5%, and showed a significantly lower inhibition rate than the two types of fermented aloe.

2-3-5 Measurement of skin immunity enhancement ability

In order to confirm the anti-atopic effect of aloe fermented product, the degree of inhibition of NO production in macrophages, which is most commonly used, was measured. For this purpose, first, murine macrophage RAW 264.7 cells were cultured. The culture and differentiation medium is Dulbecco' modified Eagle' medium (DMEM) by adding 10% Fetal bovine serum to high glucose, 5% CO ₂ , After sufficiently incubating in an incubator at 37°C, the cells are centrifuged to collect 1.5×10 ^{5 cells.} Cells/ml were dispensed into 96-well plates and cultured for 24 hours. After 24 hours, 0.15, 0.25, and 0.5% aloe samples were divided into 1ug/mL Lipolysaccharides (LPS)-treated and untreated groups, and 200 μl was added, and incubated for another 24 hours. After 24 hours, was transferred to a new 96 well plate as the supernatant of the cultured samples 50 ul were added to Griess reagent 50 μl, and measuring the absorbance at 540 nm wavelength after allowed to stand for 20 minutes at room temperature once again, a determination method using NaNO ₂ was used to quantify the amount of NO production.

As a result, as shown in FIG. 7 , when the NO production amount of all samples was checked, it was confirmed that the aloe fermented products significantly suppressed NO production compared to the two control groups at all treatment concentrations, thereby effectively suppressing skin inflammation. In particular, it was confirmed that the production rate of NO was significantly reduced by more than 50% even after LPS treatment of Arobescens and aloe peel fermented products, and thus had a high skin immunity effect, which is consistent with the antioxidant activity described above in 2-3-1. From the results, it was confirmed that the excellent cosmetic effect of the fermented aloe was due to the high antioxidant activity.

So far, the present invention has been looked at with respect to preferred embodiments thereof. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in modified forms without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. In particular, fermented products using various types of aloe and bacteria that can be used to obtain a fermented product have the same function, and thus are not excluded from the scope of this embodiment. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (10)

It contains fermented aloe as an active ingredient,
The type to be fermented is a wrinkle-improving cosmetic composition comprising aloe arborescens.
The method according to claim 1,
After fermenting aloe with lactic acid bacteria, the fermented aloe obtained by ultrasonically post-treating the fermented aloe at room temperature is used, and the post-sonication treatment of the fermented product is performed by fermenting the fermented aloe under ultrasonic waves of 20 to 120 kHz at room temperature for 30 minutes A wrinkle-improving cosmetic composition comprising fermented aloe extracted for 6 hours.
The method according to claim 1,
The lactic acid bacteria used in the fermentation of aloe are,
Leuconostoc mesenteroides, Bifidobacterium longum, Lactobacillus rhamnosus , Lactobacillus paracasei, Saccharomyces exiguus, Lactobacillus acidophilus, Bifidobacterium animalis, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus plantarum, Lactococcus diacetylactis, Lactococcus lactis, Streptococcus thermophilus, Saccharomyces cerevisiae , Rhodopseudomonas palustris, Saccharomyces fragilis, Aspergilus niger, Bacillus stearothermophilus, and Acetobacter xylinum wrinkle improvement cosmetic composition comprising any one or more selected from the group consisting of.
The method according to claim 1,
Wrinkle improvement cosmetic, characterized in that it is prepared by fermenting the whole above-ground part of aloe or by-products other than aloe gel as an active ingredient during aloe fermentation, and then adding 1% to 90% of aloe powder obtained by drying based on the total weight of the cosmetic composition. composition.
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