KR102373899B1 - Multifunctional cosmetic composition containing complex natural extracts effective for skin wrinkle improvement, whitening, cell regeneration, and wound healing as an active ingredient - Google Patents

Abstract

While the cosmetic activity of extracts prepared by using natural products is being researched, the present invention is completed by confirming skin elasticity enhancement, wrinkle alleviation, skin whitening, anti-inflammation, atopy, skin regeneration, and skin wound recovery effects, etc. of a complex extract using Centella asiatica, Petasites japonicus, Asimina triloba, Portulaca oleracea, Aurea helianthus, Glycine max, Pleurospermum camtschaticum, and others. The present invention provides a multifunctional cosmetic composition containing a complex natural extract having excellent skin elasticity enhancement, wrinkle alleviation, skin whitening, anti-inflammation, atopy, skin regeneration, and skin wound recovery effects.

Description

Multifunctional cosmetic composition containing complex natural extracts effective for skin wrinkle improvement, whitening, cell regeneration, and wound healing as an active ingredient an active ingredient}

The present invention relates to a multifunctional cosmetic composition comprising, as an active ingredient, a complex natural extract having excellent skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, and skin wound healing effects.

The skin is composed of epidermis, dermis, and subcutaneous tissue. The most important role of the skin barrier is to keep our body, which is composed of about 80% water, safely from the external environment, and this barrier function is created and maintained in the epidermis, the outer layer of the skin.

The cells constituting the epidermis are composed of keratinocytes, Langerhans cells, Merkel cells, and melanocytes, which determine the color of the skin. The dermis occupies about 90% of the skin, is 10 to 40 times thicker than the epidermis, and is composed of collagen and elasticity enhancing fibers involved in wrinkles of the skin.

Pigments that affect human skin color include melanin, melanoids, carotene, hemoglobin, and the like. Among them, melanin is a major pigment that determines skin color, and is produced in melanocytes. Specifically, melanin is a black-brown pigment formed by polymerization oxidation reaction by enzymes such as tyrosinase present in melanocytes using tyrosine, an amino acid present in a living body, as a substrate.

The formed melanin moves through the dendrites of the melanocytes to the epidermal cells called keratinocytes. Melanin that has migrated to keratinocytes can be removed by pulling away from the skin as the keratinocytes break away from the epidermis. However, since there is no enzyme that degrades melanin in vivo, melanin once formed is not degraded in vivo. Therefore, in order to brighten the skin, it can be said that suppressing the production of melanin is the key.

The need for whitening cosmetics research is gradually increasing as the recent emotional preference for white skin and the recognition of skin darkening as skin aging. Accordingly, substances exhibiting tyrosinase inhibitory activity, such as ascorbic acid, hydroquinone, kojic acid, glutathione, and arbutin, have been used in combination with cosmetics and pharmaceuticals. , most of these are insufficiently effective or the formulation is unstable, resulting in poor utility.

In particular, compounds such as hydroquinone exhibit a strong bleaching action and have skin sensitization themselves, which can cause skin allergies, etc. its use is limited.

In addition, as the number of external activities increases in recent years, as the skin irritation due to stress and external environments such as UV stimulation and fine dust increases, the induction of skin inflammation is increasing. Accordingly, cosmetics for anti-inflammatory, antioxidant, and collagen synthesis of growing interest.

In addition, recent consumers are increasingly demanding cosmetics that have high-quality functions such as whitening effect according to a special composition and minimize side effects such as irritation and inflammation at the same time. Instead, as consumers' response to cosmetics using natural ingredients increases, the need to develop natural ingredients as raw materials for cosmetics is further raised.

1. Korean Patent Registration No. 10-1746787

Therefore, the present inventor was researching the cosmetic activity of extracts prepared using natural products, and complex extracts using centella asiatica, coltsfoot, paw paw tree, machihyun, geumhwagyu, baektae, and daffodil were used to enhance skin elasticity, improve wrinkles, and whiten skin. , anti-inflammatory, atopy, skin regeneration, skin wound recovery effect, etc. to complete the present invention, the present invention is a complex excellent in skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, skin wound recovery effect An object of the present invention is to provide a multifunctional cosmetic composition containing a natural extract as an active ingredient.

In order to achieve the above technical problem, the present invention

It provides a multifunctional cosmetic composition comprising, as an active ingredient, an extract obtained by extracting a mixture of centella asiatica dry powder, coltsfoot dry powder, paw paw paw tree leaf dry powder, Machihyeon dry powder and Geumhwagyu flower dry powder with water or ethanol as an active ingredient .

In the multifunctional cosmetic composition according to the present invention as described above, the mixture may consist of 1 centella asiatica by weight, 0.8 coltsfoot, 0.7 pawpaw leaves, 1 machihyun 1, and 0.5 geumhwagyu flower.

In the multifunctional cosmetic composition according to the present invention as described above,

The mixture may further include dried Umbrella powder, and the mixture may consist of 1 centella asiatica in weight ratio, 0.8 coltsfoot, 0.7 pawpaw leaves, 1 machihyun, 0.5 geumhwagyu flower, and 1 centella asiatica.

In the multifunctional cosmetic composition according to the present invention as described above,

The mixture further includes dry white powdery powder, and the mixture may consist of 1 centella asiatica by weight, 0.8 coltsfoot, 0.7 pawpaw leaves, 1 machihyeon, 0.5 geumhwagyu flower, and 5 chrysanthemums.

In the multifunctional cosmetic composition according to the present invention as described above,

The multifunctionality is to show remarkable effects on skin elasticity enhancement, skin whitening, skin wrinkle prevention, anti-inflammatory, anti-atopic, and skin regeneration.

According to the present invention, by using a complex extract using Centella asiatica, coltsfoot, pawpaw tree, Machihyun, Geumhwagyu, baektae, and Usan grass as active ingredients, skin elasticity improvement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, skin wound It is possible to manufacture multifunctional cosmetics that can simultaneously exhibit a recovery effect.

Hereinafter, specific contents for carrying out the present invention will be described in detail through Examples and Test Examples.

The present invention provides a multifunctional cosmetic composition containing, as an active ingredient, a complex natural product extract excellent in skin elasticity enhancement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin regeneration, and skin wound recovery effects, centella asiatica, butterbur, paw paw tree, It contains as an active ingredient an extract extracted from a complex natural product using Machihyun, Geumhwagyu, Baektae, and Japanese Umbrella grass.

Specifically, the present invention

First, a multifunctional cosmetic composition comprising, as an active ingredient, an extract obtained by extracting a mixture consisting of centella asiatica dry powder, coltsfoot dry powder, paw paw paw tree leaf dry powder, machihyun dry powder and Geumhwagyu flower dry powder with an extraction solvent as an active ingredient provide,

Here, it provides a multifunctional cosmetic composition, characterized in that it contains the extract as an active ingredient, which is selectively extracted by further including the dried Umbrella grass dry powder, and the dried baektae dry powder.

The extract preparation method in the present invention is as follows.

As an extraction solvent, the mixture of the above-mentioned dry powder of natural products was extracted with one or more solvents selected from the group consisting of water, ethanol, methanol, butanol, ethyl acetate, butylene glycol, hydrous propylene glycol, hydrous glycerin, hydrous ethanol, and hydrous butylene glycol. The active ingredient can be extracted by heating at 50~95℃ for 3~20 hours or by immersion for 1~15 days at 5~37℃ with cooling condenser installed by adding ~20 times the volume to prevent solvent evaporation. can, but is not limited thereto, it is preferable to use water or ethanol as the extraction solvent, and the extraction method using water or ethanol as the extraction solvent is widely known in the art to which the present invention pertains. Do.

In the multifunctional cosmetic composition according to the present invention as described above, the content of the mixture is not particularly limited, but according to various tests of the present invention, when the weight ratio is 1 centella asiatica, butterbur 0.8, paw paw tree leaf 0.7, When it consists of 1 Machihyun and 0.5 Geumhwagyu flower, it shows a remarkable effect. Also, when it contains more dried genitalia powder, the weight ratio is 1 when it is 1 centella asiatica, and when it contains 1 centella asiatica dried powder, the weight is 1 It is preferable that the ratio is 5 white.

As described above, among the natural products used in the present invention, Centella asiatica is a perennial herb commonly found in the mountains or fields of the southern islands of the Korean Peninsula, and the present invention uses the above-ground part of Centella asiatica.

In addition, the coltsfoot (Petasites japonicus) is a perennial plant of the Asteraceae family, and grows well in groups at the foot of a mountain in Korea, where there is some moisture, and the leaves of coltsfoot are used in the present invention.

In addition, the pawpaw tree is a deciduous tree with a small size and opens light yellow to brown fruits, and leaves are used in the present invention.

In addition, the township is an outpost of purslane Portulaca oleracea Linne (purslane and Portulacaceae), and in the present invention, the dried purslane outpost is used.

In addition, the Pleurospermum camtschaticum is a perennial herb that grows in the mountains or fields of the Korean Peninsula, and the present invention uses the above-ground part of the Pleurospermum camtschaticum.

In addition, the baektae is a bean used when making soybeans, and in the present invention, the harvested soybean is dried and used as a powder.

The cosmetic composition in the present invention may include components commonly used in cosmetic compositions, and includes, for example, conventional adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers.

Such cosmetic compositions may be prepared in any formulation conventionally prepared in the art. In one embodiment, it may be any one formulation selected from the group consisting of a high-viscosity emulsifier type, a low-viscosity emulsifier type and a solubilizing formulation, but is not limited thereto. According to the formulation to be prepared, it may include a cosmetic composition formulation component commonly used in the technical field to which the invention pertains. For example, to be formulated as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream, and spray. However, the present invention is not limited thereto. More specifically, flexible lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, sun lotion, sun cream, makeup base, BB cream, cleansing cream, cleansing foam, cleansing water, pack, stick product, balm ( Balm) type product, spray or powder formulation.

Depending on the formulation to be manufactured, additionally, an aqueous phase component, an oil phase component, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a preservative, a fragrance, etc. may be selected and added.

The extract constituting the active ingredient of the cosmetic composition according to the present invention may be included in an amount of 0.001 to 30% by weight relative to the weight of the composition when used alone or in combination of 2 to 3 components. When the extract is less than 0.001% by weight relative to the total weight of the composition, the effect does not appear, and when it exceeds 30% by weight, the degree of increase in the effect for increasing the content is insignificant, there is a problem in formulation stability, and it is not economical.

Hereinafter, the present invention will be described in detail by the following examples and experimental examples. However, the following Examples and Experimental Examples only illustrate the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

< Example One: Centella asiatica , coltsfoot, pawpaw tree , Geumhwagyu Preparation of Ethanol Extract>

Centella asiatica dry powder, coltsfoot dry powder, paw paw paw paw tree leaf dry powder, Machi Prefecture dry powder, and Keumhwagyu flower dry powder were mixed in a weight ratio of 1 centella asiatica, 0.8 coltsfoot, 0.7 pawpaw tree leaf, 1 March flower, 0.5 Keumhwagyu flower. After adding 600 L of 70% ethanol to 6 kg of the mixture, the mixture was extracted at room temperature, filtered and concentrated under reduced pressure to obtain an ethanol extract.

< Example 2: Why Umbrella Grass , Centella asiatica , coltsfoot, pawpaw tree , Geumhwagyu Preparation of Ethanol Extract>

It is the same as in Example 1, except that it is different in that it is extracted using a mixture of the same weight as the Centella asiatica dried powder.

< Example 3: White, Centella asiatica , coltsfoot, pawpaw tree , Geumhwagyu Preparation of Ethanol Extract>

It is the same as in Example 1, except that it is different in that it is extracted using a mixture in which the dried baektae dry powder is mixed 5 times more than Centella asiatica in a weight ratio.

< Example 4: Centella asiatica , coltsfoot, pawpaw tree , Geumhwagyu Preparation of water extract>

It is the same as in Example 1, except that there is a difference in that hot water of 100° C. is used instead of 70% ethanol.

< Example 5: Why Umbrella Grass , Centella asiatica , coltsfoot, pawpaw tree , Geumhwagyu Preparation of water extract>

It is the same as in Example 2, but there is a difference in using hot water at 100° C. instead of 70% ethanol.

< Example 6: White, Centella asiatica , coltsfoot, pawpaw tree , Geumhwagyu Preparation of water extract>

It is the same as in Example 3, except that there is a difference in using hot water at 100° C. instead of 70% ethanol.

< test example One: Elastase Evaluation of skin elasticity enhancement and anti-wrinkle efficacy through inhibitory activity>

Example After adding 40 μl of a solution of proporcine pancreas elastase (2.5 U/ml) dissolved in 50 mM Tris-HCl buffer (pH 8.6) to 40 μl of a test solution diluted in methanol, 50 mM Tris-HCl buffer (pH 8.6) was added as a substrate. 80 μl of N-succinyl-(L-Ala)3-p-nitroanilide (0.5 mg/ml) dissolved in the mixture was added and mixed. After reacting at 37° C. for 30 minutes, absorbance was measured at 410 nm. As a positive control, oleanolic acid was used.

Elastase inhibitory activity was calculated by the absorbance reduction rate of the addition and non-addition groups of the sample solution as shown in the following formula, and the results are shown in Table 1.

Calculation formula: Inhibition rate (%) = 1-(Absorbance of the sample added group / Absorbance of the non-sample group) x 100

Sample Concentration (100 μg/ml) Example 1 Extract 81.3±0.6 Example 2 Extract 90.9±0.7 Example 3 Extract 90.3±0.6 Example 4 Extract 80.2±0.5 Example 5 Extract 89.3±0.6 Example 6 Extract 89.4±0.6 oleanolic acid 70.6±0.5

Elastase, present in the human body's neutrophil granulocytes, is known as an enzyme that is the main cause of reduction of skin elasticity improvement and wrinkle formation by breaking down elastin, an insoluble elastic fiber protein of animal connective tissue, and breaking the network structure of the dermal tissue of the skin. In the case of suppression, it is possible to prevent skin elasticity enhancement maintenance or recovery, and skin wrinkle generation. Looking at the results in Table 1, all of the extracts of the examples of the present invention showed superior effects than the positive control, confirming the salience of the effect. , particularly in Example 2, Example 3, Example 5 and Example 6 showed a very good effect.

< test example 2: Evaluation of whitening efficacy>

One) tyrosinase Inhibitory activity measurement

In 0.5 ml of 0.175M sodium phosphate buffer (pH 6.8), 0.2 ml of 10 mM L-DOPA, a substrate solution, and 0.1 ml of the sample solution were added to a 96 well plate, and 0.2 ml of mushroom tyrosinase (110 U/ml) was added to the mixed solution to 25 After reacting at ℃ for 2 minutes, DOPA chrome generated in the reaction solution was measured at 475 nm, and Arbutin was used as a positive control.

Tyrosinase inhibitory activity was calculated by the absorbance reduction rate of the group with and without the sample solution as shown in the following formula, and the results are shown in Table 2.

Calculation formula: Inhibition rate (%) = 1-(Absorbance in the sample-added group / Absorbance in the no-sample group) x 100

Sample Concentration (100 μg/ml) Example 1 Extract 77.8±0.3 Example 2 Extract 86.5±0.5 Example 3 Extract 85.9±0.6 Example 4 Extract 74.2±0.5 Example 5 Extract 82.9±0.6 Example 6 Extract 82.5±0.8 Arbutin 55.8±0.8

Tyrosinase plays a very important role in the production of melanin in the skin. It is a tyrosine hydroxylase that oxidizes tyrosine in the melanosome to make DOPA. It oxidizes DOPA and acts as a DOPA oxidase that makes DOPA quinone, and acts as an important enzyme for synthesizing melanin polymer. , when this is suppressed, the production of melanin is inhibited, thereby showing a skin whitening effect. Looking at the results in Table 2, all of the extracts of the examples of the present invention showed superior effects than the positive control, confirming the salience of the effect, especially Example 2, Example 3, Example 5 and Example 6 showed a very good effect.

2) Measurement of melanin biosynthesis inhibitory activity

The melanoma cell line, B16F10 cell, was purchased from the Korean Cell Line Bank, and Dulbecco's modified Eagle's containing 1% antibiotic (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA) medium (DMEM) medium was used at 37° C., 5% CO ₂ Incubated in an incubator, and subculture was performed once every 2 days.

B16F10 cells cultured in DMEM medium were inoculated in a 24-well plate at 2.0 X 10 ⁴ cells/well, and then cultured at 37° C., 5% CO ₂ in an incubator for 24 hours, followed by mixing with the sample.

In order to stimulate the sample and cells, 200 nM of α-MSH, a stimulant, was added and incubated in an incubator at 37° C., 5% CO ₂ for 48 hours. After removing the culture medium, wash each well with phosphate buffer (pH 7.4). did

Then, 100 μl of 1N NaOH was added to remove the cells, and then transferred to a 1.5 ml tube. After reacting at 80° C. for 1 hour, absorbance was measured at 405 nm with a spectrophotometer, and the inhibition of melanin biosynthesis was calculated by the absorbance reduction rate of the group with and without the addition of the sample solution, and the results are shown in Table 3.

sample Melanin contents (%) α-MSH(-) - α-MSH (+, 200 nm) 100 α-MSH (+, 200 nM) + Example 1 extract 32.8 ± 1.2 α-MSH (+, 200 nM) + Example 2 extract 25.9 ± 1.5 α-MSH (+, 200 nM) + Example 3 extract 25.1 ± 1.1 α-MSH (+, 200 nM) + Example 4 extract 34.5 ± 1.3 α-MSH (+, 200 nM) + Example 5 extract 26.1 ± 1.2 α-MSH (+, 200 nM) + Example 6 extract 26.2 ± 1.3

Looking at Table 3 above, it was confirmed that melanin synthesis was induced by α-MSH through the α-MSH alone treatment group, and the extracts of the examples of the present invention showed excellent effects compared to the α-MSH alone treatment group. It was possible to confirm the salience, and in particular, Example 2, Example 3, Example 5, and Example 6 showed a very excellent effect.

< test example 3: Evaluation of anti-inflammatory efficacy through nitric oxide (NO) production inhibitory activity>

A murine macrophage cell line, RAW 264.7 cell, was purchased from the Korean Cell Line Bank, and Dulbecco's contains 1% antibiotic (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA). Using modified Eagle's medium (DMEM) medium, it was cultured in an incubator at 37° C., 5% CO ₂ , and subculture was performed once every 2 days.

RAW 264.7 cells were placed in a 24 well plate at 2.0 × 10 ⁵ cells/ml using DMEM medium supplemented with 10% FBS and incubated for 18 hours. and incubated for 24 hours at 37°C, 5% CO ₂ in an incubator.

Then, 100 μl of the cell culture supernatant and 100 μl of Griess reagent were mixed and reacted for 10 minutes in a 96 well plate, and then absorbance was measured at 540 nm.

The amount of NO generated was measured using Griess reagent [1% (w/v) sulfanilamide, 0.1% (w/v) naphylethylenediamine in 2.5% (v/v) phosphoric acid] in the form of NO ₂ ^- present in the cell culture medium. It was measured and confirmed by using sodium nitrite (NaNO ₂ ) as a standard, and the results are shown in Table 4, and 2-amino-4-methylpyridine, a control group, was used as a positive control.

Sample NO production (%) LPS (-) - LPS (+, 1 μg/ml ) 100 2-amino-4-methylpyridine 26.8 ± 1.9 LPS (1 μg/ml) + Example 1 (1000 μg/ml) 19.9 ± 1.3 LPS (1 μg/ml) + Example 2 (1000 μg/ml) 15.7 ± 1.9 LPS (1 μg/ml) + Example 3 (1000 μg/ml) 15.9 ± 1.4 LPS (1 μg/ml) + Example 4 (1000 μg/ml) 20.9 ± 1.2 LPS (1 μg/ml) + Example 5 (1000 μg/ml) 17.1 ± 1.3 LPS (1 μg/ml) + Example 6 (1000 μg/ml) 17.3 ± 1.2

Looking at the results of Table 4, all of the extracts of the examples of the present invention showed superior effects than the positive control, and it was possible to confirm the salience of the effects, particularly in Examples 2, 3, 5 and 6, very showed an excellent effect.

< Sihimye 4: Inflammatory chemokines ( TARC , MDC ) through production inhibitory activity pottery Blood Efficacy Assessment>

HaCaT cells, a human keratinocyte cell line, were purchased from the Korean cell line bank, and were cultured in an incubator at 37°C, 5% CO ₂ using Dulbecco's modified Eagle's medium containing 1% antibiotic and 10% fetal bovine serum. Subculture was performed once.

HaCaT cells 3.0 * 10 ⁵ cells/ml were incubated in a 24 well plate for 18 hours before, and the prepared samples were mixed with IFN-γ and TNF-α at a concentration of 10ng/ml, respectively, and treated simultaneously with the cells at 37°C, 5% CO ₂ incubator incubated for 24 hours.

After that, the culture medium was transferred to a 1.5 ml tube and centrifuged to measure the amount of chemonkines synthesis in the supernatant. The results are shown in Table 5. All samples were frozen until quantitation, and inflammatory chemokines were analyzed by Human enzyme-linked immnunosorbent assay ( ELISA) kit (R&D Systems) was used, and the r ² value of the standard curve for the standard was 0.99 or more.

Sample TARC production (%) - 0 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) 100 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) +
Example 1 (1000 μg/ml) 25.1 ± 1.3 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) +
Example 2 (1000 μg/ml) 17.9 ± 2.1 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) +
Example 3 (1000 μg/ml) 18.3 ± 1.6 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) +
Example 4 (1000 μg/ml) 27.6 ± 1.5 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) +
Example 5 (1000 μg/ml) 19. 4 ± 1.6 TNF-α (10 ng/ml) + IFN-γ (10 ng/ml) +
Example 6 (1000 μg/ml) 20.1 ± 1.2

It is known that the expression of TARC increases in patients with atopic dermatitis. Looking at the results in Table 5, it was confirmed that TARC expression was induced through the TNF-α and IFN-γ alone treatment group, and when treated with the sample of Examples It was confirmed that a high inhibition rate on TARC expression was observed compared to the TNF-α, IFN-γ alone treatment group, and in particular, Example 2, Example 3, Example 5, and Example 6 showed a very good effect.

< Sihimye 5: with fibroblast keratinocytes Evaluation of skin cell regeneration and wound healing effects through mobility evaluation>

HaCaT cells and human dermal fibroblast cells, which are human keratinocytes, were purchased from the Korean cell line bank, and cultured in Dulbecco's modified Eagle's medium medium containing 1% antibiotic and 10% fetal bovine serum at 37°C, 5% CO ₂ incubator. Subculture was performed once every 3 or 4 days.

CytoSelect ^TM 24-Well Wound Healing Assay kit (CELL BIOLABS INC. USA) was used to evaluate fibroblast and keratinocyte mobility. Using sterile forceps, wound field inserts were well fixed in each well of a 24-well plate, and fibroblasts and keratinocytes were inoculated into 24-well plates at 8.0x10 ⁴ cells/well and 3.0x10 ⁵ cells/well, respectively.

Then, after pre-culturing for 18 hours in an incubator at 37° C. and 5% CO ₂ , samples are prepared by concentration. After removing the wound field insert using sterile forceps, the culture medium was removed, washed twice with PBS, and the prepared sample, the composition of Example 1, was treated in each well, and then at 37°C, 5% CO ₂ in an incubator for 2 hours. After culturing, the culture solution was removed, washed with PBS, and replaced with a culture solution containing no sample.

Then, check the cell status hourly for 24 hours, 48 hours, and 72 hours in an incubator at 37 ° C, 5% CO ₂ , remove the culture medium at the time to stop the culture, and wash twice with PBS so that no cell residue remains. Then, 250 μl of cell stain solution was added to each well, reacted at room temperature for 15 minutes, washed 3 times with PBS, and then cell mobility was checked under a microscope with a plate dried at room temperature.

The skin plays a major role in protecting the individual as the primary barrier to the external environment. Keratinocytes are the main cells constituting the epidermis of the skin, and play a role in making the skin barrier through division and differentiation. In skin diseases, the skin barrier is not formed normally or abnormal keratinocyte differentiation occurs. In this case, it can help to form a normal skin barrier by promoting keratinocyte differentiation. It produces collagen that occupies In order to regenerate the dermal layer of the damaged skin, fibroblast regeneration is the most important. In order to check whether the Example extract affects wound healing on keratinocytes and fibroblasts, a wound healing assay was performed, and the untreated control group and Examples The degree of proliferation of the cell group treated with each extract of 1.25 μg/ml was observed, and the results are shown in Table 6.

fibroblast fibroblast sample N.C. N.C. hour 48 hours 72 hours 48 hours 72 hours

sample Example 1 Extract Example 1 Extract Sigin 48 hours 72 hours 48 hours 72 hours

Referring to Table 6, as a result of culturing for 48 hours and 72 hours in fibroblast and 24 hours and 48 hours in Incubation, all of the cell groups treated with the Example 1 extract were fibroblasts and keratinocytes in fibroblasts and keratinocytes as time passed, more cell migration and proliferation than negative control. was observed to be fast, confirming the remarkable effect on skin cell regeneration and wound healing.

As described above, according to the present invention, skin elasticity improvement, wrinkle improvement, skin whitening, anti-inflammatory, atopy, skin whitening, anti-inflammatory, atopy, It is possible to manufacture multifunctional cosmetics that can simultaneously regenerate skin and restore skin wounds.

Claims (7)

In the case of centella asiatica dry powder 1 in weight ratio, an extract obtained by extracting a mixture of coltsfoot dry powder 0.8, paw paw paw tree leaf dry powder 0.7, machihyun dry powder 1 and Geumhwagyu flower dry powder 0.5 with water or ethanol as an active ingredient. Multifunctional cosmetic composition, characterized in that it exhibits effects on skin elasticity enhancement, skin whitening, skin wrinkle prevention, anti-inflammatory, anti-atopic and skin regeneration.
According to claim 1,
The multifunctional cosmetic composition, characterized in that the mixture further comprises a dry powder of Umbrella grass.
3. The method of claim 2,
Multifunctional cosmetic composition, characterized in that when the mixture is 1 centella asiatica in weight ratio, 0.8 coltsfoot, 0.7 pawpaw leaves, 1 machihyun, 0.5 geumhwagyu flower, and 1 honeysuckle.
According to claim 1,
The multifunctional cosmetic composition, characterized in that the mixture further comprises a dry white powder.
5. The method of claim 4,
Multifunctional cosmetic composition, characterized in that when the mixture is 1 centella asiatica in weight ratio, 0.8 coltsfoot, 0.7 pawpaw leaves, 1 machihyun, 0.5 geumhwagyu flower, and 5 baektae.

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