KR20080046830A - Cosmetic composition containing fermented black allium sativum - Google Patents

Abstract

A fermented solution of black allium sativum is provided to promote synthesis of skin cell proteins such as collagen and showing skin improving effect by inhibiting generation of collagenase, thereby being used as a cosmetic composition showing excellent effect of skin damage caused by skin-aging. A cosmetic composition comprises a fermented solution of black allium sativum, which is obtained by culturing black allium sativum powder prepared by ripening allium sativum using Bacillus genus strain and filtering a cultured material. Regarding a dried weight of the cosmetic composition, 0.0001-20.0 wt.% of the fermented solution of the black allium sativum is contained. Further, the fermented solution of black allium sativum promotes biosynthesis of Type I collagen.

Description

Cosmetic composition containing black garlic fermentation broth cultivated using Bacillus microorganisms {Cosmetic composition containing fermented black allium sativum}

 The present invention provides a cosmetic composition capable of effectively improving various skin problems generated by aging by making black garlic fermentation broth made by aging black garlic using microorganisms of Bacillus and containing it in cosmetics.

Skin prevents water from evaporating from our body and prevents harmful ingredients from invading from the outside. For this reason, the skin should always be exposed to external stimuli and have a variety of defenses. In recent years, most cosmetic companies have been conducting a lot of research on skin problems, and many products have been developed. As a person ages, skin aging occurs. The main symptom is wrinkles. The main cause of wrinkles is when collagen, which forms a matrix in the dermis of the skin, breaks down and fails to function. Collagen may be degraded as aging progresses, but collagen is easily broken down to form wrinkles as the activity of the matrix metalloproteinase (MMP-1), which is generated and activated by external stimulation such as ultraviolet rays, increases. There are many increasing clinical reports. Therefore, in recent years, cosmetic products to suppress the production of wrinkles to promote collagen production, but also products that inhibit the degradation of collagen that is essentially produced by inhibiting the production of MMP-1 or inhibiting activity It is developed a lot. In the skin, MMP-1, MMP-2, MMP-9 and other enzymes work. The most fundamental enzyme involved in collagen degradation is MMP-1. In general, conventional substances that promote collagen synthesis include retinoids (RE36068), TGF-β (Transforming growth factor), betulinic acid (JP8-208424), and the like. TGF- inhibits the production of MMP-1. β (Transforming growth factor) is known. Trouble due to skin aging also has the characteristics that skin cells are easily damaged by external stimulation due to pigmentation and deterioration of skin barrier function, and the skin moisturizing power is poor. Therefore, it is ideal to use a substance having such various effects in order to recover skin damage caused by aging. However, most of the raw materials have only one or two effects, and one raw material rarely has various effects. The present invention relates to a cosmetic composition containing a black garlic fermentation broth produced by culturing by adding black garlic using Bacillus sp. Garlic has conventionally been antibacterial, in addition to antithrombotic action, anticancer action, lowering cholesterol and anti-aging, antioxidant activity is known. Garlic has a lot of known problems that the powder is irritating and difficult to use on the skin as it is. In cosmetics, a method of extracting and using water and alcohol using garlic has already been proposed. Cosmetic soap containing an extract extracted with garlic powder or water (Patent No. 1996-4496), garlic extract for cosmetic compositions (Patent No. 2002-191726). However, these extracts can be used as a stimulating ingredient of garlic as it is very likely to irritate the skin.

On the other hand, in recent years there is a case in which the cosmetic composition is provided using a fermentation broth produced using a fermentation product by Bacillus subtilis used in representative fermented foods such as Cheonggukjang and natto (natto). Such fermentation broth has the advantage of degrading the skin irritation by decomposing the stimulus components by the microorganisms or converting to other substances, and the fermentation products by the microorganisms are also widely used as having a good effect on the skin.

The present invention is to provide a cosmetic using a culture medium made by crushing black garlic made by aging garlic for a long time to make fine powder and then cultivating the material using a microorganism of the genus Bacillus. The culture solution used in the present invention was found to have various effects for skin improvement. Black garlic cultures have excellent collagen synthesis effects, have the characteristics of inhibiting the production of collagen degrading enzymes, and are also excellent in inhibiting melanin synthesis. In addition, black garlic culture has a variety of features that relieve skin irritation and repair skin barrier damage.

The present invention was to provide a cosmetic composition using a black garlic culture solution to solve various skin problems damaged by aging.

In the present invention, after making a fermentation broth using Bacillus microorganisms using the black garlic powder, using this active ingredient to effectively improve the skin problems caused by skin aging. First, as a method for inhibiting wrinkle formation, there is generally a method of inhibiting wrinkle generation by promoting collagen synthesis, or inhibiting wrinkle generation by inhibiting the production or activity of collagenase. In addition, in order to inhibit skin pigmentation, it inhibits melanin synthesis and attempts to suppress skin barrier damage.

The black garlic fermentation broth used in the present invention has not only collagen synthesizing effect, but also an effect of inhibiting the production of collagen degrading enzyme, inhibiting pigmentation, and having various characteristics to alleviate skin irritation. Accordingly, the present invention relates to a cosmetic composition showing a very excellent effect on skin damage caused by aging by adding black garlic fermentation broth to cosmetics.

The present invention relates to a cosmetic composition using a fermentation broth using a Bacillus strain using black garlic. Black garlic is garlic made by ripening for about one month at a constant temperature and humidity. It first turns brown and finally turns black. In the present invention, black garlic made by ripening garlic for about one month at a constant temperature and humidity was used.

In addition, Bacillus strains were used to make black garlic fermentation broth. Bacillus strains are Gram-positive microorganisms, capable of forming endospores, and have a rod shape. In the present invention, it was possible to grow in aerobic or anaerobic anaerobic conditions, and as a growing condition, finely made garlic powder (made using 300 mesh), the powder was added to the conventional microbial culture 10g / 1L. Fermentation conditions were grown at about 37 degrees, pH was grown at 5-7.

The black garlic fermentation broth used in the present invention was obtained as follows. After incubation under the same conditions as above for 7 days incubation in a fermentor. After incubation, a culture solution was obtained, and the solution was filtered using a 0.25 uM filter paper. This filtrate was further aged for 10 days at low temperature, filtered and used in the present invention, and the final solid concentration was maintained at 10 g / L before use.

The prepared black garlic fermentation broth is added to conventional basic cosmetics, that is, flexible lotion (skin), nourishing lotion (milk lotion), nourishing cream, massage cream, essence, and pack, and the amount is 0.0001 to 20 based on the dry weight. % By weight, preferably 0.001 to 10% by weight is added to the cosmetic. At this time, the black garlic fermentation broth is less than 0.0001%, the effect is less likely to appear, and if more than 20% is likely to cause irritation to the skin and may have a significant effect on the stabilization of the formulation.

Examples and Experimental Examples below describe the contents of the present invention, but the contents of the present invention are not limited thereto.

Example 1. Preparation of black garlic fermentation broth

After washing with purified water and powdered dry black garlic was made fine using 300 mesh. This was added to Bacillus genus strain culture to 10g / L. To this, glucose was added as a carbon source, and additionally, peptone and NaCl were incubated. Cultivation was carried out using a 5L fermenter for 7 days at 37 degrees, pH 5-7. After incubation, the culture solution was centrifuged to remove the culture cells first, and then filtered by 0.25uM filter paper. The filtrate thus obtained was aged for about 10 days in a low temperature warehouse at 4 degrees, and then the filtrate was filtered using a filter of 0.25 uM and used in the present invention. This fermentation broth used in the present invention was used to maintain a final concentration of 1g / 100mL.

Experimental Example 1. Inhibitory Effect of Black Garlic Fermentation on MMP-1 Production

This experiment was conducted using the sample obtained in the above example, and the experiment proceeded as follows. First, human normal fibroblasts were seeded to 1 × 10 ⁶ cells in each well of a 48-well microplate (Nunc, Denmark) and incubated for 24 hours at 37 ° C. in DMEM medium (Sigma, USA). Subsequently, experiments in which the concentration of the black garlic fermentation broth were changed to DMEM medium without serum at 0.1, 0.5, 1.0, 2.0, and 5.0%, and the control group replaced with the serum-free DMEM medium containing no black garlic fermentation broth were added for 48 hours. Incubated with After incubation, the supernatant of each well was collected and the amount of newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, US), and then the amount of MMP-1 was converted to ng / ml. TGF-β (10 ng / ml, Roche, United States) was used as a positive control. MMP-1 production inhibition was calculated according to the following equation and the results are shown in Table 1.

% Inhibition of MMP-1 production = [1- (MMP-1 amount in experimental group / MMP-1 amount in control group)] 대 100

Table 1 Inhibitory Effects of Black Garlic Fermentation on MMP-1 Production

Test substance (black garlic fermentation broth,%) % Inhibition of MMP-1 production Positive control (TGF-β: 10 ng / ml) 68.5 0.1 12.5 0.5 28.8 1.0 57.3 2.0 68.2 5.0 65.5

Experimental Example 2 Collagen Synthesis Enhancement Effect of Black Garlic Culture

Human normal fibroblasts were seeded to 1 x 10 ⁶ cells in each well of a 48-well microplate and incubated for 24 hours at 37 ° C. in DMEM medium. Subsequently, the experimental group replaced with serum-free DMEM medium with the final concentration of black garlic culture at 0.1, 0.5, 1.0, 2.0, 5.0% and the control group replaced with serum-free DMEM medium without black garlic culture for 24 hours. Incubated with After incubation, the supernatants of each well were collected and the amount of newly synthesized collagen was measured using a kit (Takara, Japan) for the amount of procollagen (procollagen) type I C-peptide (PICP). PICP amount was converted to ng / ml, and TGF-β was also used as a positive control. Collagen biosynthesis increase rate was calculated by the following formula and the results are shown in Table 2.

  Collagen production increase rate (%) = [(test collagen amount / control collagen amount) -1] ㅧ 100

[Table 2] Collagen Synthesis Enhancement Effects of Black Garlic Cultures

Test substance (black garlic fermentation broth,%) Collagen production growth rate (%) Positive control (TGF-β: 10 ng / ml) 71.5 0.1 10.6 0.5 20.5 1.0 33.6 2.0 52.6 5.0 48.9

In the above experimental example, black garlic fermentation broth has the effect of promoting collagen synthesis and at the same time having the effect of inhibiting the biosynthesis of MMP-1, which has the characteristics of collagen degradation.

Experimental Example 3. Experiment to inhibit melanin production in black garlic culture (Melanin assay)

B-16 cells (rat melanoma cells, Korea Cell Line Bank) were seeded in 12-well plates to 10 ⁴ cells / well and cultured for one day. In each well, the black garlic culture solution obtained in Experimental Example was prepared as in Example 1, and the black garlic culture solution was treated according to the concentration, incubated for 3-4 days, and then the supernatant in each well culture solution was discarded, and only melanocytes were centrifuged. Got it. The isolated cells were lysed with 500 μl of 1N NaOH solution dissolved in dimethyl sulfoxide (DMSO) and heated in a 90 ° C. water bath for 10 minutes. It was centrifuged again and the absorbance of the supernatant was measured at 490 nm. As a control, one that was not treated with black garlic culture was used.

Melanin production inhibition was calculated according to the following equation, the results are shown in Table 3.

% Inhibition = [1-absorbance of comparative group (O.D.490) / control absorbance (O.D.490)] ㅧ 100

[Table 3] Melanin production inhibitory effect of black garlic culture

Black garlic fermentation broth (%) Melanin production inhibition rate (%) 0.1 5.6 0.5 12.5 1.0 32.6 2.0 62.6 5.0 78.9

Experimental Example 4. Lactic acid relaxation effect test of black garlic culture

Fibroblasts (Korea Cell Line Bank, South Korea), which are human skin cells, were cultured in T-75 flasks (Falcon, United States) until they grew about 80%. This was again transferred to a 96-well plate (Falcon, USA) to 3 ㅧ 10 ⁴ cells / well and incubated for 24 hours. After culturing, the microscope confirmed whether the cells were completely attached and grew well. The experiment was performed using lactic acid as a source of skin cell stimulation. At this time, lactic acid used 0.2%. That is, 200 μl of DMEM (Sigma, U.S.) medium containing 0.2% lactic acid was added to each of 96 wells and 0.1, 0.5, 1.0, 2.0, 5.0% (v / v) using the black garlic culture solution prepared in Example 1 Adjusted to). At this time, lactic acid and black garlic culture solution were not added to the control group, and only lactic acid was added to the comparison group. In order to compare the viability of the cells 12 hours after the addition of the test substance, the cell viability was measured by adding MTT (Sigma, US) solution (3 mg / ml) at 570 nm using ELISA READER (Molecular Devices, USA). And cell viability (%) was calculated according to the following formula and the experimental results are shown in Table 4 below.

  Cell viability = [(control absorbance-test material absorbance) / control absorbance] ㅧ 100

[Table 4] Cell Stimulation Relaxation Effect of Black Garlic Cultures

   Black garlic culture concentration (%) Cell survival rate (%) 0.0 (without lactic acid) 98.5 0.0% (lactic acid 0.2%) 0.0 0.1 (lactic acid 0.2%) 10.5 0.5 (0.2% lactic acid) 15.7 1.0 (0.2% lactic acid) 25.9 2.0 (0.2% lactic acid) 55.4 5.0 (lactic acid 0.2%) 67.9

 Experimental Example 5. Evaluation of anti-inflammatory effect of black garlic fermentation broth-Carrageenan foot edema

   Inflammation was induced by injecting 10-100 mg / kg of black garlic fermentation broth obtained in Example 1 into the abdominal cavity of rats one hour later, and then injecting 0.5 ml of 0.1% carrageenan solution into the back of the experimental animal. The change in the paw volume of rats immediately after carrageenan injection and 4 hours after injection was measured and shown in Table 5 below.

% Inhibition = (1-ΔV treated group / ΔV control) ㅧ 100 [ΔV: change of foot volume]

[Table 5] Anti-inflammatory effect of black garlic fermentation broth

Black garlic fermentation dose (mg) % Inhibition 10 10.5 20 29.8 50 56.4 100 75.8

Formulation Example 1. Soft Cosmetics

Prescription examples of the flexible cosmetics in cosmetics containing black garlic fermentation broth are as follows.

    ingredient content(%)     Black garlic fermentation broth 0.5     glycerin 5.0     1.3-butylene glycol 3.0 Allantoin 0.1 DL-panthenol 0.3 E.T.A.-2NA 0.02 Benzophenone-9 0.04 Sodium hyaluronate 5.0 Nicole HCO 60 0.4 Methyl paraben 0.2 incense, 0.01 Distilled water Remaining amount Sum 100

Prescription Example 2. Nutritional Cosmetics

Prescription examples of nutrient cosmetics in cosmetics containing black garlic fermentation broth are as follows.

ingredient Content (unit: weight%)     Black garlic fermentation broth      2.0     Glyceryl Stearate SE 1.5 Stearyl alcohol 1.5 lanolin 1.5 Polysorbate 60 1.3 Sorbitan stearate 0.5 Cured Vegetable Oil 1.0 Mineral oil 5.0 Squalane 3.0 Trioctanoine 2.0 Dimethicone     0.8 Tocopherol Acetate 0.5 Carboxy Vinyl Polymer 0.12 glycerin 5.0 1.3-butylene glycol 3.0 Sodium hyaluronate 5.0 Tri ethanolamine 0.12 Methylparaben 0.2 incense 0.02 Distilled water Remaining amount Sum 100

Prescription Example 3. Nutritional Cream

Prescription examples of nutrient creams in cosmetics containing black garlic fermentation broth are as follows.

ingredient Content (unit: weight%)      Black garlic fermentation broth 3.0 Lipophilic monostearate 2.0 Cetearyl Alcohol 2.2 Stearic acid 1.5 Beeswax 1.0 Polysorbate 60 1.5      Sorbitan stearate 0.6 Cured Vegetable Oil 1.0 Squalane 3.0 Mineral oil 5.0 Trioctanoine 5.0 Dimethicone 1.0 Sodium magnesium silicate 0.1 glycerin 5.0 Betaine 3.0 Triethanolamine 1.0 Sodium hyaluronate 4.0 Methylparaben 0.2 incense 0.05 Distilled water Remaining amount Sum 100

Experimental Example 6. Evaluation of wrinkle improvement effect of cosmetics containing black garlic fermentation broth

Wrinkle improvement effect of the cosmetic of the present invention was evaluated through a practical use test. A nutrition cream containing 3% (v / v) of black garlic fermentation broth of Formulation Example 3 and a cream in which black garlic fermentation broth was replaced with purified water in Formulation Example 3 were used. For 20 women, the experimental group was used on one side of the face and the control group on the other side, and the wrinkle improvement after 6 weeks was evaluated by the skin visiometer (C + K, Germany). Evaluated. The result of the wrinkle improvement effect based on this evaluation is as showing in the following table | surface.

[Table 6] Wrinkle improvement effect of nutrition cream containing black garlic fermentation broth (equipment evaluation)

Wrinkle improvement Effectiveness% Great slightly none Cream of Prescription Example 3 13 3 4 80.0 Comparative cream One One 18 10.0

According to the experimental results, the present invention containing the black garlic fermentation broth showed a higher antiwrinkle effect than the comparative example, and skin irritation could not be observed in most of the subjects who applied the cosmetic to the skin.

Experimental Example 7. Skin barrier damage recovery effect of black garlic culture

The cream prepared in Formulation Example 3 and the cream without addition of the black garlic culture solution in Formulation Example 3 were evaluated as follows to evaluate the extent of skin barrier damage recovery as a comparative prescription example. First, categorize hairless mice into groups and select 10 animals in groups 1 and 2 for 5 days. On day six, the skin barrier function of hairless mice is removed using a tape stripping method. Afterwards, apply the product of Formulation Example 3 (containing 3% black garlic culture) to Group 1, and apply the Comparative Formulation Example 3 (product containing no black garlic culture solution) twice a day to visualize the recovery of skin barrier for 1 week. And instrumental analysis. Instrument analysis was compared by measuring the TEWL (TransEpidermalWater Loss) value using a TEWA meter (Corneometer CM820, Conrage + Khazaka, Germany).

[Table 7] Improvement of Skin Barrier by Visual Evaluation

division Skin barrier improvement Excellent good no effect Prescription Example 3 7 3 - Comparative Prescription 3 One One 8

[Table 8] Percutaneous moisture loss rate (%) using TEWA meter

division Percutaneous moisture loss rate (%) 0 days 7th day Prescription Example 3 43.5 5.8 Comparative Prescription 3 44.3 28.5

The present invention relates to the effect of preventing skin aging by effectively improving the skin damage caused by aging using black garlic fermentation broth. The black garlic fermentation broth promotes collagen synthesis and effectively inhibits the biosynthesis of collagen degrading enzyme (MMP-1), which is increased by ultraviolet light. It has the property of suppressing damage. In addition, it has an anti-inflammatory effect and relieves skin cell irritation. In conclusion, the present invention is a cosmetic composition containing the black garlic fermentation broth has the characteristics of improving the skin damage caused by aging and healthy skin.

Claims (9)

   Cosmetic composition characterized by containing a black garlic fermentation broth    [Claim 2] The cosmetic composition according to claim 1, wherein the black garlic fermentation broth contains black garlic fermentation broth made by culturing black garlic powder made by aging general garlic using Bacillus strains.     The cosmetic composition according to claim 1, wherein the black garlic fermentation broth contains 0.0001-20.0% (w / w) based on the dry weight of the silver cosmetic.     The anti-wrinkle cosmetic composition of claim 1, wherein the black garlic fermentation broth promotes biosynthesis of Type I collagen.     The anti-wrinkle cosmetic composition according to claim 1, wherein the black garlic fermentation broth has a feature of inhibiting biosynthesis of collagen degrading enzyme (MMP-1), which is characterized by degrading collagen.     The whitening cosmetic composition according to claim 1, wherein the black garlic fermentation broth has a characteristic of inhibiting melanin production.    The method of claim 1, wherein the black garlic fermentation broth stimulating cosmetic composition, characterized in that to reduce skin cell irritation     The cosmetic composition according to claim 1, wherein the black garlic fermentation broth has a feature of restoring damage to the skin barrier.     The cosmetic composition according to claim 1, wherein the black garlic fermentation broth has an anti-inflammatory effect.