KR101772614B1 - Cosmetic composition with the product fermented by Tremella fuciformis mycelium on medium with endothelial growth factor - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a fermented product obtained by injecting Tremella fuciformis mycelium into a medium comprising an epithelial cell growth factor and fermenting the same, and the fermented product of the present invention has not only stable EGF titer stability and emulsion stability, but also exhibits very excellent efficacy in skin regeneration, skin aging prevention, skin wrinkle reduction, skin barrier function enhancement, skin anti-inflammation, skin moisturizing and skin whitening.

Description

[0001] The present invention relates to a cosmetic composition comprising a fermented product obtained by inoculating a mycelia of white mushroom with a medium containing an epithelial cell growth factor and fermenting the fermented product,

The present invention relates to a cosmetic composition containing a fermented product obtained by inoculating a mycelium of Tremella fuciformis into a medium containing an epithelial cell growth factor and fermenting the fermented product. More particularly, the present invention relates to a cosmetic composition containing EGF titer The present invention relates to a cosmetic composition which is excellent in stability and emulsion stability as well as skin regeneration, skin aging prevention, skin wrinkle improvement, skin barrier function enhancement, skin anti-inflammation, skin moisturizing and skin whitening efficacy.

The skin is an organ that functions as a protective membrane for protecting the living body from the external environment, and prevents the loss of biological components in the human body, that is, water or electrolytes, and prevents harmful substances from entering the human body from the outside.

However, as we age, these functions gradually become weakened, and skin aging proceeds due to ultraviolet rays, stress, disease conditions, environmental factors, activation of free radicals due to scars, and the like. Particularly, as the lipid composition and content of the lipid layer functioning as a skin barrier are changed, the skin function is drastically lowered. When the skin moisture content is decreased, the skin becomes dry, wrinkles are formed and stains, freckles, Skin lesions are induced. When such conditions are intensified, the antioxidant defense network existing in the living body is destroyed, and cells and tissues are damaged, thereby promoting adult diseases and aging. In particular, the oxidation of proteins is caused by the cleavage of collagen, hyaluronic acid, elastin, proteoglycan, fibronectin, which are connective tissues of the skin, If it gets worse and becomes worse, it causes mutation by DNA mutation, cancer induction, immune function deterioration.

In order to solve this problem, a method of supplying epithelial cell growth factor (EGF) to the skin has been used to protect the cell membrane, and already damaged cells are regenerated by active metabolism, Lt; / RTI >

EGF promotes the proliferation of epithelial cells and dermal cells, promotes cell proliferation of fibroblasts that synthesize collagen as a component of dermis, promotes angiogenesis of skin lesion sites and induces branching of other regeneration promoting factors, It plays a key role in skin regeneration, including promoting the synthesis of fibronectin, a substance that forms a network, and minimizing scars on the wound area. Due to this potency of EGF, much research has been focused on techniques for fusing EGF into cosmetics.

However, when EGF is supplied to the skin in a free form, an excessively short half-life is required to effectively exhibit the biological activity. Also, it has been reported that when EGF is added at a certain content or more, it penetrates into the interface to increase the surface tension, thereby reducing the emulsifying power. That is, there is a problem of stability of the cosmetic emulsifier type, so that there is a problem that the content that shows efficacy can not be used. Therefore, there is a desperate need for research for maintaining the skin surface for a long time or improving emulsion stability.

On the other hand, Tremella fuciformis ) is a white throat, a white throat, and a white throat mushroom. It grows in the dead branches of broad-leaved trees from summer to fall and rotates the wood as a by-product. In addition, the diameter of the mushroom is 6 ~ 12cm, height is 3 ~ 6cm, it is split into petal shape, but it shrinks when dried and spreads again when there is moisture. There is gelatinous quality, and the edges are swirled like waves and overlap to form flower shape. It is translucent with light pink or pale yellowish brown, etc. It is almost black when dried and dried. The surface is flat and forms a follicle layer. The size of the spores is 6 ~ 8 ~ 6㎛, and it is a colorless spherical shape with some fine spots and oil droplets. Form consultation is white.

White-throated mushroom is used for edible and medicinal purposes. It has good flavor and delicious soup. In addition, it has attracted attention recently because of its strong anticancer effect, prevention of arteriosclerosis and effect of nourishing tonic. In addition, high nutritional value, protein 11.3%, potassium 1,200 mg, phosphorus 434 mg, iron and calcium are high, and various vitamins content is high. Especially, it has a high fiber content and many colloidal substances, and it acts to wash the esophagus and stomach. In one room, white mushroom was used as a medicinal product to improve the function of the five and to remove the poison of the organ.

Korean Patent No. 10-1369964 (Registered on Feb. 26, 2014) describes an external-use composition for alleviating scalp irritation containing aloe, white throat mushroom and bamboo extract. Korean Patent No. 10-1257993 (registered on Apr. 18, 2013) discloses a method for producing an extract of white mushroom by using an enzyme and a composition for memory enhancement containing the same.

In the present invention, EGF and white mushroom ( Tremella fuciformis ) as a raw material for the cosmetic composition.

The present invention provides a cosmetic composition characterized by containing a fermented product obtained by inoculating a mycelium of Tremella fuciformis into a medium containing epithelial growth factor (EGF), followed by fermentation.

In the cosmetic composition of the present invention, the fermentation is preferably carried out at a temperature of 20 to 40 캜 for 5 to 9 days while maintaining a pH of 5 to 7.

In the cosmetic composition of the present invention, the fermented product is preferably added in an amount of 0.0001 to 80% by weight based on the total weight of the cosmetic composition.

The cosmetic composition of the present invention is preferably used for skin regeneration.

In addition, the cosmetic composition of the present invention is preferably one for prevention of skin aging.

In addition, the cosmetic composition of the present invention is preferably used for improving skin wrinkles.

The cosmetic composition of the present invention is preferably used for enhancing skin barrier function.

In addition, the cosmetic composition of the present invention is preferably used for skin inflammation relief.

The cosmetic composition of the present invention is preferably used for skin moisturizing.

The cosmetic composition of the present invention is preferably a skin whitening agent.

The cosmetic composition of the present invention may be in the form of a solution, suspension, emulsion, paste, lotion, gel, water-soluble liquid, cream, essence, surfactant-containing cleansing oil, oil-in- A basic cosmetic formulation comprising a water (W / O) type; Ointment; A color cosmetic formulation comprising an underwater type and a water-in-oil type make-up base, a foundation, a skin cover, a lipstick, a lip gloss, a face powder, a two-way cake, an eye shadow, a teak color and an eyebrow pencil; And a preparation for scalp; Or the like.

The present invention provides a fermented product obtained by fermenting a mycelium of Tremella fuciformis in a medium containing an epithelial growth factor (EGF) as a cosmetic raw material. Skin barrier enhancement, skin anti-inflammation, skin moisturizing and skin whitening effect were superior to those of EGF alone treatment and other mycelial fermentation.

The present invention also relates to the use of EGF as an anti- fuciformis ), the potency and emulsion stability of EGF could be improved.

In addition, the present invention can reduce consumers' rejection of cosmetic products that contain various conventional chemicals and stimulate skin by using natural materials as raw materials.

FIG. 1 is a graph showing the cell proliferative activity of EGF-white throat mushroom mycelium fermented product and a control group.
FIG. 2 is a graph showing the inhibition rate of MMP-1 activity of the fermented product of EGF-white throat mycelium.
FIG. 3 is a graph showing the transdermal water loss measurement performed in order to examine the skin barrier function enhancing effect of the cosmetic composition containing the fermented product of EGF-white throat mycelium as an active ingredient.

The present invention provides a cosmetic composition characterized by containing a fermented product obtained by inoculating a mycelium of Tremella fuciformis into a medium containing epithelial growth factor (EGF), followed by fermentation.

The fermentation product of the present invention can be produced, for example, by the following steps. (A) inoculating and fermenting the mycelia of white throat to the medium containing EGF; (B) a step of sterilizing or sterilizing after fermentation; And (c) filtrating the fermentation broth to obtain a filtrate.

In the present invention, the medium containing EGF can be produced by adding EGF to a mushroom mycelium, more preferably a conventional medium capable of culturing mycelia of white throat.

In the present invention, the fermentation is preferably carried out at a temperature of 20 to 40 캜 for 5 to 9 days while maintaining a pH of 5 to 7. Under such fermentation conditions, the fermentation of the mycelia of white throat can be maximized, and fermentation products can be obtained with optimum yield and productivity.

On the other hand, in the cosmetic composition of the present invention, the fermented product is preferably added in an amount of 0.0001 to 80% by weight based on the total weight of the cosmetic composition. It is difficult to exhibit skin regeneration, prevention of skin aging, improvement of skin wrinkles, enhancement of skin barrier function, skin anti-inflammation, skin moisturizing and skin whitening effect upon addition of fermented product, When added, the increase in the effect of the addition amount is insignificant, which is not economical.

On the other hand, the cosmetic composition of the present invention can be preferably used for regenerating skin, preventing skin aging, improving skin wrinkles, enhancing skin barrier function, alleviating skin irritation, moisturizing skin, and whitening skin.

On the other hand, the cosmetic composition of the present invention can be used as a cosmetic composition such as a solution, a suspension, an emulsion, a paste, a lotion, a gel, a water-soluble liquid, a cream, an essence, a surfactant-containing cleansing oil, A basic cosmetic formulation comprising a water (W / O) type; Ointment; A color cosmetic formulation comprising an underwater type and a water-in-oil type make-up base, a foundation, a skin cover, a lipstick, a lip gloss, a face powder, a two-way cake, an eye shadow, a teak color and an eyebrow pencil; And a preparation for scalp; Or the like.

In addition, the cosmetic composition of the present invention may be applied selectively to the skin in the form of an aerosol, or may be applied to the skin in the form of a solid, for example, a stick.

On the other hand, the cosmetic composition according to the present invention may preferably contain, besides the above-mentioned fermentation product, other components capable of synergizing the main effect within a range not impairing the intended main effect of the present invention . Such as antioxidants, stabilizers, solubilizers, vitamins, customary adjuvants such as pigments and flavoring agents, and carriers.

When the formulation of the cosmetic composition of the present invention is a suspension, the carrier component may be a liquid diluent such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester , Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or draagacant, and the like can be used.

When the formulation of the cosmetic composition of the present invention is an interfacial active agent-containing cleansing agent, the carrier component may be aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionide, imidazolinium derivative, methyltaurate , Fatty acid amide ether sulfide, alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

According to the experiment of the present invention as described below, the fermentation product of the present invention is excellent in stability of EGF and stability of emulsification, and is excellent in skin regeneration, prevention of skin aging, improvement of skin wrinkles, enhancement of skin barrier function, It was confirmed that skin whitening effect was superior to that of conventional EGF treatment and fermentation with other mycelium.

Hereinafter, the present invention will be described in more detail with reference to the following Examples and Experimental Examples. However, the scope of the present invention is not limited to the following examples and experimental examples, but includes modifications of equivalent technical ideas.

[ Example  One: EGF - White throat mushroom  mycelium Fermented  Produce]

The white mushroom ( Tremella fuciformis strains were isolated directly from fresh fruiting bodies. White mushroom mycelia were isolated from yeast-malt juice agar medium (yeast extract 3 g, malt juice 3 g, glucose 10 g, peptone 5 g, agar 20 g, distilled water 1 L) The slides were incubated at 4 ° C and subcultured every month. The mycelia grown on the slope medium were aseptically homogenized and inoculated in a liquid medium adjusted to pH 6 to 5% (v / v) as a simple complex medium containing 3% glucose and 1.0% yeast extract, EGF 0.3% were mixed and then incubated in a fermenter at a temperature of 30 ° C, a revolution of 150 rpm, and aeration of 1.5 vvm for 7 days.

The mycelia were removed by centrifugation, sterilized at 125 ° C, 1.5 atm for 15 min, and filtered at 0.45 μm to obtain a filtrate (hereinafter referred to as EGF-fermented white mushroom mycelium).

[ Comparative Example  One: White throat mushroom  mycelium Fermented  Produce]

In Example 1, a fermented white mushroom mycelium was obtained in the same manner without mixing EGF.

[ Comparative Example  2: EGF - Mushroom mycelium Fermented  Produce]

In this Comparative Example 2, a fermentation product was prepared by fermenting EGF using mycelia of pine mushroom instead of mycelium of white mushroom. Morchella esculenta was directly isolated from fresh fruiting bodies and the mycelia of mung bean were grown in yeast-malt juice agar medium (3 g yeast extract, 3 g malt juice, 10 g glucose, 5 g peptone, 20 g agar g, and 1 L of distilled water), stored at 4 ° C, and subcultured every month. The mycelia grown on the slope medium were aseptically homogenized and inoculated with 5% (v / v) in a liquid medium adjusted to a pH of 6 as a simple complex medium containing 3% glucose and 1.0% yeast extract , And 0.3% EGF were mixed and then cultured in a fermenter at a temperature of 30 ° C, a rotation speed of 150 rpm, and aeration of 1.5 vvm for 7 days.

After completion of the cultivation, the mycelium was removed by centrifugation, and the culture filtrate was sterilized at 125 ° C, 1.5 atm, and 15 min, and filtered at 0.45 μm to obtain a filtrate (hereinafter referred to as "EGF-mushroom mycelium fermentation product").

[ Comparative Example  3: EGF - Tricolored mushroom  mycelium Fermented  Produce]

In this Comparative Example 3, EGF was fermented by using mycelia of three colors of mushrooms, instead of mycelium of white mushroom, to prepare a fermented product. Daedaleopsis tricolor strains were directly isolated from fresh fruiting bodies. Yeast - malt juice agar medium (3 g yeast extract, 3 g malt juice, 10 g glucose, 5 g peptone, 20 g of agar, 1 L of distilled water), stored at 4 ° C, and subcultured every month. The mycelia grown on the slope medium were aseptically homogenized and inoculated in a liquid medium adjusted to pH 6 to 5% (v / v) as a simple complex medium containing 3% glucose and 1.0% yeast extract, EGF 0.3% were mixed and then incubated in a fermenter at a temperature of 30 ° C, a revolution of 150 rpm, and aeration of 1.5 vvm for 7 days.

After completion of the cultivation, the mycelium was removed by centrifugation, and the culture filtrate was sterilized at 125 ° C., 1.5 atm for 15 min, and filtered at 0.45 μm to obtain a filtrate (hereinafter referred to as EGF-tricolor coated mushroom mycelium fermentation product).

[ Comparative Example  4: EGF - Avi mushroom  mycelium Fermented  Produce]

In this Comparative Example 4, a fermented product was prepared by fermenting EGF with mycelia of mackerel rather than mycelium of white mushroom. The mushroom used in the experiment ( Pleurotus Ferulea ) was isolated directly from fresh fruiting bodies. The mushroom mycelia were harvested in yeast-malt juice agar medium (3 g yeast extract, 3 g malt juice, 10 g glucose, 5 g peptone, 20 g agar, 1 L distilled water) The slides were incubated at 4 ° C and subcultured every month. The mycelia grown on the slope medium were aseptically homogenized and inoculated in a liquid medium adjusted to pH 6 to 5% (v / v) as a simple complex medium containing 3% glucose and 1.0% yeast extract, EGF 0.3% were mixed and then incubated in a fermenter at a temperature of 30 ° C, a revolution of 150 rpm, and aeration of 1.5 vvm for 7 days.

After completion of the cultivation, the mycelium was removed by centrifugation, and the culture filtrate was sterilized at 125 ° C, 1.5 atm, and 15 min, and then filtered at 0.45 μm to obtain a filtrate (hereinafter, referred to as an EGF-adiomyocyte fermentation product).

[ Experimental Example  1: Evaluation of cell activity effect]

In Experiment 1, 0.025% (w / w), 0.05% (w / w), 0.1% (w / w) and 0.2% (w / w) of the fermented product of the EGF- w / w)) skin cell activity. At this time, as a control group, the samples prepared in Comparative Examples 1 to 4 and EGP were used.

Human dermal fibroblast (HDF) was inoculated in a 96-well plate (5 x 10 4 cells / well) and stabilized for 24 hours. After that, the fermented mycelia of EGF-white mushroom mycelia was treated with DMEM medium containing 0.5% FBS (0.025%, 0.05%, 0.1%, 0.2%) and cultured in a 37 ° C CO 2 incubator for 48 hours . Cell activity was then measured using an MTT assay.

As a result of the experiment, it was confirmed that the EGF-white agaricus fermented product of Example 1 was further increased in cell activity in a concentration-dependent manner compared with the control of Comparative Examples 1 to 4 (FIG. 1)

[ Experimental Example  2: protease ( MMP -1) Measurement of inhibition rate of activity]

In Experimental Example 2, the in vitro metalloproteinase (MMP-1) inhibition rate of the EGF-fermented white mushroom mycelium obtained in Example 1 was measured.

Human dermal fibroblast (HDF) was inoculated into a 24-well plate at a concentration of 5 x 10 < 4 > cells / well, and then stabilized in a 37 DEG C incubator for 24 hours. Then, it was rinsed twice with PBS, and UVA was irradiated with 6 J / cm 2 in the presence of PBS. After the irradiation, the medium was replaced with a medium supplemented with 0.2% FBS. The EGF-white mushroom mycelium fermentation (0.025%, 0.05%, 0.1%, 0.2%) and the fermented white mushroom mycelium (0.2% And 100 nM of adenosine, respectively. After 48 hours, the supernatant was measured at 450 nm using a human total MMP-1 ELISA kit (human total MMP-1 Cat. No. DY901, R & D systems).

As a result of the experiment, it was confirmed that the fermented product of EGF-mushroom mycelia inhibited MMP-1 up to about 25% from 0.025% as low as the positive control adenosine (AD) (FIG. 2).

Based on this, it was found that the composition for external application for skin of the present invention effectively inhibited the proteolytic enzyme, thereby preventing skin elasticity and wrinkle formation.

[ Experimental Example  3: Type 1 Procollagen  Biosynthesis promoting effect measurement]

In Experimental Example 3, the effect of promoting the biosynthesis of type 1 procollagen, which is a skin matrix component, in the fermentation product of EGF-white mushroom mycelium obtained in Example 1 was examined. For this, TGF-beta was used as a positive control, and procollagen assay was performed as follows.

Human dermal fibroblast (HDF) was purchased from American Type Culture Collection (ATCC, USA), and 10% fetal bovine serum (FBS) was added to DMEM / F-12 cm < 2 >. When 70 ~ 80% of the cells were grown, the cells were subcultured at a ratio of 1: 3, and the cells cultured in the third to fourth passages were used for the experiment.

For the measurement of the amount of procollagen, the fibroblasts were cultured in a 48-well plate at a concentration of 90% or more, and then the fermented product of EGF-white mushroom mycelia was added at concentrations of 0.2%, 0.1% and 0.05% After 24 hours, the amount of procollagen liberated in the medium was measured using a procollagen type-1 C-peptide EIA kit (MK101, TaKaRa, Japan).

Name of sample Promoting effect of procollagen biosynthesis (%) EGF 0.006 wt% 13 0.2% by weight of fermented mycelium of white throat, 9 EGF-white throat mushroom Mycelial fermentation added 0.2% by weight 37 0.1% by weight of fermented product of EGF-white throat mushroom mycelium 19 EGF-white throat mushroom Mycelium fermentation 0.05 wt% added 14 Addition of 0.001% by weight of TGF-β 36

The results of measuring the promoting effect of procollagen biosynthesis (Table 1), 13% when treated with 0.006% by weight of EGF, 9% with 0.2% by weight of the fermented white mushroom mycelium, 0.2% by weight of the fermented broth of EGF- The biosynthesis of procollagen was promoted by 37%, which was similar to that of 0.001% by weight of the cell signaling substance TGF-β.

From the above results, it was confirmed that the fermented product of EGF-white throat mushroom had excellent promoter biosynthesis promoting activity, and it was found that the present invention has excellent effect of improving skin wrinkles.

[ Manufacturing example  One: EGF - White throat mushroom  mycelium The fermented product  Containing Cosmetic  Produce]

A cosmetic composition containing 10.0% by weight of a fermented product of EGF-white throat mycelia of Example 1 was prepared according to the composition shown in the following Table 4 and referred to as Prescription Example 1.

On the other hand, cosmetic compositions containing each of the EGF-mushroom mycelial fermentation product of Comparative Examples 2 to 4, the EGF-3 mushroom mycelia fermentation product or the EGF-mushroom mycelial fermentation product were respectively prepared and referred to as Comparative Prescription Examples 2 to 4, Other cosmetic compositions treated with EGF alone were referred to as Comparative Prescription Example 5 (Table 2).

ingredient Prescription Example 1
content
(weight%) Comparative Prescription Example 1
content
(weight%) Comparative Prescription Example 2
content
(weight%) Comparative Prescription Example 3
content
(weight%) Comparative Prescription Example 4
content
(weight%) Comparative Prescription Example 5
content
(weight%) Example 1 10.0 - - - - - Comparative Example 1 - 10.0 - - - - Comparative Example 2 -  - 10.0 - - - Comparative Example 3 - - - 10 - - Comparative Example 4 - - - - 10 - EGF - - - - - 1.8 μg / ml 1,3-BG 10.0 10.0 10.0 10.0 10.0 10.0 glycerin 5.1 5.1 5.1 5.1 5.1 5.1 Propylene glycol 4.2 4.2 4.2 4.2 4.2 4.2 Tocopheryl acetate 3.0 3.0 3.0 3.0 3.0 3.0 Liquid paraffin 4.6 4.6 4.6 4.6 4.6 4.6 Triethanolamine 1.0 1.0 1.0 1.0 1.0 1.0 Squalane 3.1 3.1 3.1 3.1 3.1 3.1 Macadamia nut oil 2.5 2.5 2.5 2.5 2.5 2.5 Polysorbate 1.6 1.6 1.6 1.6 1.6 1.6 Sorbitan sesquiterate 1.6 1.6 1.6 1.6 1.6 1.6 Propylparaben 0.6 0.6 0.6 0.6 0.6 0.6 Carboxyl vinyl polymer 1.5 1.5 1.5 1.5 1.5 1.5 incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water Balance Balance Balance Balance Balance Balance system 100.0 100.0 100.0 100.0 100.0 100.0

[ Experimental Example  4: EGF Potency  Stability evaluation]

The content of EGF was confirmed by HPLC using the fermented product prepared in Prescription Example 1 and Comparative Prescription Examples 1 to 5 as a test substance. The EGF components contained in each test substance were dissolved in purified water and the solution filtered through a 0.45 μm PVDF syringe filter (Watman, England) was quantitatively analyzed using an HPLC apparatus. Columns were C18 column (4.6 x 250 nm), PDA (200-400 nm) detector, mobile phase HPLC grade acetonitrile (A) and water (B) for 0-10 minutes, 10% A; 10 to 30 minutes 10 to 90% A; The flow rate was 1.0 mL / min, and the sample injection amount was 10.0 μl. The results are shown in Table 3 below (Table 3).

Prescription Example 1 Comparative Prescription Example 1 Comparative Prescription Example 2 Comparative Prescription Example 3 Comparative Prescription Example 4 Comparative Prescription Example 5 cold storage 2.2 μg / ml 0.0 μg / ml 1.6 μg / ml 1.5 μg / ml 1.3 μg / ml 1.8 μg / ml Room temperature 2.2 μg / ml 0.0 μg / ml 1.6 μg / ml 1.5 μg / ml 1.2 μg / ml 1.8 μg / ml Window 2.1 μg / ml 0.0 μg / ml 1.5 μg / ml 1.4 μg / ml 1.2 μg / ml 1.6 μg / ml High temperature 2.0 μg / ml 0.0 μg / ml 1.4 μg / ml 1.4 μg / ml 1.1 μg / ml 1.4 μg / ml

As can be seen from the above results, it was found that the potency stability of EGF was maintained only in the case of Prescription Example 1 in which fermentation with white mushroom was carried out. It was thought that the fermentation component produced during the fermentation of white mushroom caused EGF stability.

[ Experimental Example  5: Evaluation of emulsion stability]

The stability of the formulation was measured by using the fermented product prepared in Prescription Example 1 and Comparative Prescription Examples 1 to 5 as a test substance under refrigeration, normal temperature, window, and high temperature for 4 weeks. Table 4 shows the results according to appearance and viscosity reduction (Table 4).

Prescription Example 1 Comparative Prescription Example 1 Comparative Prescription Example 2 Comparative Prescription Example 3 Comparative Prescription Example 4 Comparative Prescription Example 5 cold storage Good Viscosity decrease Good Good Viscosity decrease Good Room temperature Good Viscosity decrease Good Good Viscosity decrease Viscosity decrease Window Good Viscosity decrease Viscosity decrease Viscosity decrease Viscosity decrease Viscosity decrease High temperature Good Viscosity decrease Viscosity decrease Viscosity decrease Viscosity decrease Viscosity decrease

As shown in Table 4, although the emulsions were not visually separated, all of the comparative formulations 1 to 5 showed a decrease in viscosity and thus affected the formulation. However, in the case of Formulation Example 1, it was confirmed that the formulation stability was excellent.

[ Experimental Example  6: EGF - White throat mushroom  mycelium Fermented  Skin barrier function enhancement effect]

In Experimental Example 6, the effect of enhancing the skin barrier function of Prescription Example 1 was investigated, which is a cosmetic containing the fermented product of the white mushroom mycelium prepared in Preparation Example 1 as an active ingredient.

Transepidermal water loss (TEWL) means that water inside the skin is evaporated when the barrier function of the skin is lost. The evaluation of moisture evaporation is expressed in g / m², which evaporates at unit area per hour.

The skin of the inner part of the arm was peeled off using a tape to remove the keratin of the skin artificially, and after applying the above-described Prescription Example 1 and Comparative Prescription Examples 1 to 4 respectively, 1 hour, 2 hours, 4 hours, 6 hours After a period of time, the moisture evaporation of the skin was measured using a transdermal water loss meter (Tewameter TM 210, Courage and Khazaka, Germany). The untreated skin was used as a control.

As a result of the experiment, Comparative Example 1, which is a cosmetic preparation containing the fermented product of mycelia of white mushroom mycelium as an active ingredient, and Comparative Example 1, which is a cosmetic containing the fermented product of mycelia of white mushroom mycelium as an active ingredient, Comparative Preparation Example 2, which is a cosmetic composition containing an active ingredient of a mushroom mycelium as an active ingredient, Comparative Preparation Example 3, which is a cosmetic composition containing a fermented product of mushroom mycelia of three colors as an active ingredient, In comparative formulation example 4, it was confirmed that both of them had an effect of inhibiting transdermal water loss. However, Prescription Example 1 of the present invention showed a better effect of inhibiting transdermal water loss than Comparative Prescription Examples 1 to 4. Thus, it was found that the present invention has a more excellent effect on enhancing the skin barrier function (FIG. 3).

[ Experimental Example  7: Measurement of inhibitory effect of inflammatory cytokine expression by ultraviolet irradiation]

Experimental Example 7 measures the anti-inflammatory effect of the fermented mycelia of EGF-mushroom mycelia obtained in Example 1 by measuring the degree of suppression of inflammatory cytokine expression expressed by ultraviolet irradiation.

Human dermal fibroblast (HDF) was added to a 24-well test plate in an amount of 5x10 4 cells and adhered for 24 hours. Each well was washed once with PBS and 500 占 퐇 of PBS was added to each well. After irradiating the fibroblasts with 10 mJ / cm2 of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki Co., Japan), PBS was removed and the cell culture medium ≪ / RTI > supplemented medium) was added.

The EGF-white rot fungus fermented product to be evaluated was treated and cultured for 5 hours. After that, 150 [mu] l of the culture supernatant was taken to quantitate IL-1 [beta], thereby determining the effect of inhibiting the expression of inflammatory cytokines in the EGF-mycelia fermentation product. The amount of IL-1 [beta] was quantitated using an enzyme-linked immunosorbent assay (ELISA), and the production rate of IL-1 [beta] was calculated by the following formula (1). As a positive control, nordihydroguaiaretic acid (NDGA) was used.

[ Equation 1 ]

Inhibitory rate of inflammatory cytokine expression (%) = [1- (St-Bo) / (Bt-Bo)] x 100

Bo: IL-1? Production in wells not subjected to ultraviolet irradiation and not treated

Bt: IL-1? Production in wells irradiated with ultraviolet rays and not treated with the sample

St: IL-1? Production in well irradiated with ultraviolet light

The inhibitory effect of inflammatory cytokine expression by ultraviolet irradiation was measured (Table 5). When treated with 0.006% by weight of EGF, 3.1%, and with 0.2% by weight of fermented white mushroom mycelium, the inflammatory cytokine IL-1β Lt; / RTI > EGF-white rot fungi fermented mycelium produced the inflammatory cytokine IL-1β It was confirmed that the generation of inflammation due to ultraviolet rays can be effectively prevented at a low concentration by controlling 18.7% at 0.1% concentration and 32.5% at 0.2% concentration.

Name of sample Inhibitory rate of inflammatory cytokine expression (%) EGF 0.006 wt% 3.1 0.2% by weight of fermented mycelium of white throat, 4.3 EGF-fermented white mushroom mycelial body 0.1% 18.7 EGF-fermented white mushroom mycelia 0.2% 32.5 NDGA 10 μM 27.8

From the above results, it could be deduced that the cosmetic composition containing the fermented product of EGF-white throat mycelia of the present invention as an active ingredient shows excellent anti-inflammatory effect.

[ Experimental Example  8: EGF - White throat mushroom  mycelium Fermented  skin Moisture power  Improvement effect]

In Experimental Example 8, the effect of improving the skin moisturizing power of the cosmetic preparation containing the fermented product of the white mushroom mycelium prepared in Preparation Example 1 as an active ingredient, Prescription Example 1 and Comparative Prescription Examples 1 to 5 was examined. The cosmetic composition of Prescription Example 1 and Comparative Prescription Examples 1 to 5 prepared in Preparation Example 1 was divided into two groups of 20 persons per one set of 20 to 40 persons without skin disease, And applied to the face and forearm for a month.

The skin conductivity was measured using a moisture analyzer (CM820 courage Khazaka electronic GmbH, Germany) under constant temperature and humidity conditions (24 ° C, 40% humidity) before the application and the skin conductivity increase rate after 1 week, 2 weeks, (%) Were measured and the rate of increase was evaluated. The results are shown in Table 6 below.

sample After 1 week after 2 weeks After 4 weeks Prescription Example 1 51 57 62 Comparative Prescription Example 1 5 8 8 Comparative Prescription Example 2 26 33 38 Comparative Prescription Example 3 18 22 26 Comparative Prescription Example 4 7 10 13 Comparative Prescription Example 5 23 36 35

(unit: %)

As shown in the results of Table 6, in the case of Formulation Example 1, which is a cosmetic containing the fermented product of mycelium of mycelium of the present invention as an active ingredient, the rate of skin conductivity increase was much superior to Comparative Formulation Examples 1 to 5.

In general, the skin conductivity is in proportion to the skin moisture content. Thus, the above results show that the cosmetic composition containing the fermented product of the present invention as the active ingredient maintains the skin moisture content higher than that of the cosmetic composition not containing it.

[ Experimental Example  9: B16F1  Measurement of inhibitory effect on melanin synthesis using melanocytes]

In Experimental Example 9, albutin known as a whitening agent was used as a comparative sample and B16F1 melanocyte was used for confirming the melanin synthesis inhibitory effect of the EGF-white throat mushroom fermentation product obtained in Example 1 above. The B16F1 melanocyte used in Experimental Example 9 is a cell line derived from a mouse, and is a cell that secretes a melanin pigment called melanin.

The inhibitory effect of B16F1 melanin on melanin synthesis was measured as follows. B16F1 melanocytes were plated at a concentration of 2x10 per well in a 6-well plate, and the cells were adhered, treated at a concentration not causing toxicity, and cultured for 72 hours. After culturing for 72 hours, cells were detached with trypsin-EDTA, the number of cells was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out with a slight modification of the method of Rotan (R Lotan and D Lotan, Cancer Res, 40: 3345-3350, 1980). Cell pellets were washed once with PBS and then 1 ml of homogenization buffer (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. After centrifugation (3,000 rpm, 10 min), 1N NaOH (10% DMSO) was added to the cell filtrate to dissolve the extracted melanin, and the absorbance of melanin was measured at 405 nm with a microplate reader. Melanin was quantified The percent inhibition of melanin formation in the sample was measured.

The inhibition rate (%) of melanin formation of B16F1 melanocytes was calculated by the following equation (2).

& Quot; (2 ) & quot ;

Melanin synthesis inhibition rate (%) = [(A-B) / A] x 100

A: Amount of melanin in wells to which no sample was added

B: Amount of melanin in the well to which the sample was added

Inhibition of melanin synthesis by B16F1 melanocytes was measured (Table 7). When treated with 0.006% by weight of EGF and 11% by 0.2% by weight of fermented broth of white mushroom mycelium, inhibition of melanin synthesis was 1% 0.2% by weight of fermented broth of white throat mushroom showed melanin synthesis inhibition rate similar to that of 0.2% by weight of arbutin.

Name of sample  Melanin synthesis inhibition (%) EGF 0.006 wt% 11 0.2% by weight of fermented mycelium of white throat, One 0.2% by weight of EGF-fermented mycelia of white throat, 30 EGF-fermented white mushroom mycelia 0.01 wt% 21 EGF-fermented white mushroom mycelia 0.005 wt% 8 Arbutin 0.2 wt% 32

From the above results, it was possible to confirm the excellent inhibition of melanin synthesis of the fermented product of EGF-white throat mycelia, and the excellent whitening effect of the present invention was confirmed.

Claims (11)

To the medium containing epithelial cell growth factor (EGF), white marrow mushroom ( Tremella fuciformis ), and then fermenting the fermented product.
The method according to claim 1,
The above-
Characterized in that it is carried out at a temperature of 20 to 40 占 폚 while maintaining a pH of 5 to 7 for 5 to 9 days.
The method according to claim 1,
The fermented product may contain,
Is added in an amount of 0.0001 to 80% by weight based on the total weight of the cosmetic composition.
The method according to claim 1,
In the cosmetic composition,
Wherein the cosmetic composition is for skin regeneration.
The method according to claim 1,
In the cosmetic composition,
The cosmetic composition is characterized in that it is for preventing skin aging.
The method according to claim 1,
In the cosmetic composition,
Wherein the cosmetic composition is for improving skin wrinkles.
The method according to claim 1,
In the cosmetic composition,
Wherein the composition is for enhancing skin barrier function.
The method according to claim 1,
In the cosmetic composition,
Wherein the cosmetic composition is for skin inflammation relief.
The method according to claim 1,
In the cosmetic composition,
Wherein the composition is for moisturizing the skin.
The method according to claim 1,
In the cosmetic composition,
Wherein the cosmetic composition is for skin whitening.
The method according to claim 1,
In the cosmetic composition,
(Cosmetics), cosmetics, gels, water-soluble liquids, creams, essences, surfactant-containing cleansing, oils, oil-in-water (O / W) ; Ointment; A color cosmetic formulation comprising an underwater type and a water-in-oil type make-up base, a foundation, a skin cover, a lipstick, a lip gloss, a face powder, a two-way cake, an eye shadow, a teak color and an eyebrow pencil; And a preparation for scalp; And the cosmetic composition of the present invention.

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