KR101267321B1 - Cosmetic composition comprising earthworm mucus and mud as an effective components for regenerating skin and improving skin wrinkle - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin regeneration and wrinkle improvement including earthworm mucus and mud as an active ingredient, in particular the cosmetic composition of the present invention has a high cation substitution capacity and electrical conductivity, humic acid, fulvic acid, humin or organic matter It has high content of, and contains Jangheung mud with high content of calcium and magnesium ions and earthworm slime extracted from living state without stress, and shows remarkably excellent skin regeneration effect and wrinkle improvement. Through the functional cosmetic composition of the public, excellence of natural products in Jangheung region, Jeollanam-do can be promoted to increase local incomes and revitalize the local economy.

Description

Cosmetic composition comprising earthworm mucus and mud as an active ingredient {Cosmetic composition comprising earthworm mucus and mud as an effective components for regenerating skin and improving skin wrinkle}

The present invention relates to a functional cosmetic composition based on a natural material capable of skin regeneration and wrinkle improvement.

Mud (MUD) is a natural sea mud that emits large amounts of far-infrared rays and is rich in minerals. It also contains beneficial ingredients such as germanium and bentonite, which help prevent skin aging, maintain moisture balance, clean the skin, and treat skin waste. It is known to have an excellent effect such as removal.

According to the Korea International Trade Association statistics, most of the mud in Korea depends on imports. Among them, Israel's Dead Sea Mud, Glacier Mud on the Canadian Columbia coast, Russia's Baikal Mud, California Clay Mud and New Zealand Volcanic Mud are well known around the world. It is partly manufactured and commercialized in Boryeong and North Korea.

Cosmetic related technology using mud is a cosmetic raw material mud manufacturing composition emitted by far-infrared energy of Korea Patent Publication No. 2002-0087590, the composition of the mud pack of Korea Patent Publication No. 2001-0090063, the mud of Korea Patent Publication No. 2003-0060426 Color cosmetic composition for body painting containing, mud color cream composition containing mud of Korea Patent Publication No. 2004-0022868, cosmetic composition containing mud and herbs and black rice powder of Korean Patent Registration No. 10-0515956, Korean Patent Publication 2004 Numerous patented technologies have been disclosed, including mud extracts and cosmetic compositions containing herbal oils of -0067377.

In addition, the mud of a particular region is used, and the technology using the boryeong acid mud is a method of extracting a cosmetic raw material having skin moisturizing effect and cell growth effect from Boryeong acid mud of Korea Patent Publication No. 2001-0079027, and Korea Patent Publication No. Skin cosmetic composition containing Boryeong acid mud of 1998-0034570, extraction method of Boryeong acid mud extract of Korea Patent Publication No. 2010-90533, whitening functional cosmetic composition containing the extract, and Youngil Mansan mud There is a skin cosmetic composition using the Youngilman acid rock mud of the Patent Publication No. 2005-48792.

Jeonnam Jangheung area has a natural environment of uncontaminated nature, and undeveloped natural resources are scattered everywhere, and it is adjacent to the coast where mud can be generated due to the characteristics of the region, but in order to industrialize Changheung Mud in the past There was no effort.

On the other hand, protease is a generic term for proteolytic enzymes having proteolytic activity such as papain and trypsin, and when these substances are applied to the epidermis, they break down proteins such as keratin, which are exfoliation or keratin of the epidermis, and are digested and absorbed as a kind of induced protein. It can be used as cosmetics and additives by performing peptone, a nutritious ingredient, which can be easily hydrated, thereby facilitating epidermal metabolism and maintaining normalization of skin function as well as excellent moisture control as a moisturizing additive. Substance.

In an attempt to effectively extract such protease components from earthworms, Korean Patent No. 390760 discloses a method of centrifugation after drying, pulverizing, and dissolving earthworms, and Korean Patent No. 301306 discloses earthworms in a solvent that the earthworm does not like. A method of simultaneously extracting body fluids and mucus by dipping is disclosed. In Korean Patent No. 578952, a method of crushing earthworms in the roots and ocher of herbs used as a medicinal herb, and in Korean Patent No. 833465, the earthworms are alive. A method of extracting with water in a state, restoring the freshness of the earthworm, and reusing the same is disclosed.

However, the earthworm extract or earthworm mucus prepared by the above method only mentions the normalization or moisturizing effect of skin function by the protease component, and does not mention the effect of skin regeneration or skin wrinkle improvement.

The present inventors compared and analyzed Jangheung-san mud and Boryeong-san mud, but they are similar in color and far-infrared radiation, but have high cation substitution capacity and electrical conductivity, high humic acid, fulvic acid, humin or organic matter, and natural The high content of calcium ions in detoxification and stress suppression, and the magnesium ions in promoting waste excretion and collagen binding, making them suitable for cosmetic compositions. When used with the extracted earthworm mucus was confirmed that the skin regeneration effect and skin wrinkle improvement effect is significantly improved to complete the present invention.

It is an object of the present invention to provide a functional cosmetic composition comprising earthworm mucus and mud as an active ingredient having excellent skin regeneration effect and skin wrinkle improvement effect.

The cosmetic composition for skin regeneration and skin wrinkle improvement of the present invention includes earthworm mucus and mud as an active ingredient.

The earthworm mucus in the cosmetic composition for skin regeneration and skin wrinkle improvement of the present invention, the pretreatment step of excretion of the stool in the earthworm body and then washed with water; And the earthworm slime obtained through the step of extracting the slime from the earthworm by immersing the pretreated earthworm in a container containing water for 20 to 60 minutes.

Earthworm mucus of the present invention, unlike the earthworm dry ground powder or organic solvent extract does not contain toxins that occur when the earthworm is stressed to die, and also has an excellent antioxidant activity about 20 times compared to the normal earthworm dry ground powder.

If the pre-treated earthworm is an earthworm used for extracting the earthworm slime, put it in the earthworm recuperator and supply air at 5-20 ° C. for 2 to 10 minutes at intervals of 30 to 60 minutes to recover the earthworm's freshness. After undergoing a refreshing step, it is preferable to extract the mucus to discharge the mucus from the earthworm by placing the refreshed earthworm in a container containing water and soaking for 20 to 60 minutes.

When the earthworm is immersed in a container containing water, the earthworm is stressed and discharges a large amount of mucus as one of self-defense mechanisms. The ratio of earthworm to water should be about 1: 1.5. The longer the earthworm is immersed in water, the greater the amount of mucus discharged. However, if the earthworm is immersed in the water for a certain time, the earthworm will die due to epidermal respiration. Therefore, the immersion time of the earthworm is good not to exceed 60 minutes, preferably immersed for 20 to 60 minutes. In addition, what is necessary is just to use the thing of the temperature of water that the earthworm is suitable for growth, ie, the range of 10-30 degreeC.

In addition, the inlet is preferably formed in the upper portion for extracting the earthworm mucus, the interior is provided with a receiving space, the insertion groove is formed in the upper periphery of the inlet, the side connection is connected to the pressure reducing pump to induce pressure reduction inside Body is provided with; An o-ring inserted into the insertion groove of the body; A separating member which is inserted into the storage space of the body while the upper end is over the inlet of the body, and has a filtering net at the bottom thereof to separate only the liquid from the solution including the earthworm and the slime flowing into the bottom; It is preferable to use an earthworm mucus extraction device consisting of.

The particle diameter of the strainer does not necessarily need to be limited, but it is preferable to have a size of about 0.05 to 0.5 mm because it should satisfy the size of the foreign matter including the earthworm.

In addition, in extracting the earthworm slime by using the earthworm slime extracting device, after connecting the connector of the earthworm slime extractor with a decompression pump while operating the decompression pump separating the solution containing the earthworm and slime obtained in the slime extracting step It is preferable that the solution is poured through the strainer to separate the solution, and the solution obtained in the separation step is centrifuged to collect only the supernatant to recover the earthworm mucus.

In addition, the earthworm mucus obtained is preferably used after diluting to a constant concentration after drying in order to ensure uniformity of quality as a cosmetic material. Although the drying method is not specifically limited, It is preferable to carry out lyophilization in order to suppress the denaturation by the heat of the active ingredient of an earthworm mucus, without requiring an excipient.

In the cosmetic composition for skin regeneration and skin wrinkle improvement of the present invention, the mud is taken after drying the mud sample.

Mud can be any of domestic mud, Israel's Dead Sea Mud, glacier mud on the coast of Columbia, Canada, Baikal mud, California clay mud, New Zealand's volcanic mud, but preferably mud, particularly preferably Changheung's mud. Mud and color and far-infrared radiation are similar, but have high cation substitution capacity and electric conductivity, high humic acid, fulvic acid, humin and organic matter, detoxification and stress suppression among natural minerals and waste excretion It is suitable for the cosmetic composition of the present invention because the content of magnesium ions that promotes and promotes collagen binding is high.

In the cosmetic composition for skin regeneration and skin wrinkle improvement of the present invention, the earthworm slime based on a solid content of 0.001 to 0.1% by weight, preferably 0.01 to 0.5% by weight. If the earthworm mucus content is less than the lower limit, sufficient skin regeneration effect and skin wrinkle improvement effect may not be achieved. If the earthworm mucus content is exceeded, the solubility of the earthworm mucus may be oversaturated and the earthworm mucus may not be diluted.

In addition, in the cosmetic composition for skin regeneration and skin wrinkle improvement of the present invention, mud contains 0.5 to 10% by weight, preferably 1 to 5% by weight, based on solids, and below the lower limit, detoxification and stress inhibiting action in natural minerals. The content of calcium ions and magnesium ions that promote waste excretion and collagen binding are lowered, so that sufficient skin regeneration and skin wrinkle improvement effects cannot be achieved. As with the solubility of the mud, there is a problem that useful ingredients such as calcium and magnesium are not eluted anymore.

The cosmetic composition for skin regeneration and skin wrinkle improvement of the present invention may be prepared in any one form selected from lotion (skin), lotion, essence, nutrition cream and pack.

Cosmetic composition of the present invention may further comprise a suitable carrier, excipient or diluent commonly used in the manufacture of cosmetics.

Carriers, excipients or diluents which may be included in the cosmetic composition include purified water, oils, waxes, fatty acids, fatty acid alcohols, fatty acid esters, surfactants, humectants, thickening agents, antioxidants, viscosity stabilizers ), Chelating agents, buffers, lower alcohols, and the like. Whitening agents, moisturizers, vitamins, sunscreens, perfumes, dyes, and antimicrobials may be included as needed.

As the oil, hydrogenated vegetable oil, perm oil, cottonseed oil, olive oil, palm oil, jojoba oil, avocado oil may be used, and waxes, beeswax, carnauba, candelilla, montan, ceresin, liquid paraffin, and lanolin are used as waxes. Can be used. Stearic acid, linoleic acid, linolenic acid, oleic acid may be used as fatty acids, and cetyl alcohol, octyl dodecanol, oleyl alcohol, pantenol, lanolin alcohol, stearyl alcohol, hexadecanol may be used as fatty acid alcohol. Isopropyl myristate, isopropyl palmitate, butyl stearate may be used as the ester. Examples of the surfactant include anionic surfactants such as sodium stearate, sodium cetyl sulfate, polyoxyethylene laurylether phosphate, sodium N-acyl glutamate; Cationic surfactants such as stearyldimethylbenzylammonium chloride and stearyltrimethylammonium chloride; Amphoteric surfactants such as alkylaminoethylglycine hydrochloride and lecithin; Glycerin monostearate, sorbitan monostearate, sucrose fatty acid ester, propylene glycol monostearate, polyoxyethylene oleyl ether, polyethylene glycol monostearate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene coconut fatty acid monoethanol Nonionic surfactants such as monoethanol arnide, polyoxypropylene glycol, polyoxyethylene castor oil, polyoxyethylene lanolin, and the like. Glycerin, 1,3-butylene glycol, propylene glycol may be used as the hygroscopic agent, and ethanol or isopropanol may be used as the lower alcohol. Examples of thickeners include sodium alginate, sodium caseate, gelatin agar, xanthan gum, starch, cellulose ethers (e.g. hydroxyethyl cellulose, methyl cellulose, carboxymethyl cellulose, hydroxy propylmethyl cellulose), polyvinylpyrrolidone, Polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose and the like. As antioxidants, butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, citric acid, ethoxyquin can be used, and chelating agents include disodium edetate and ethane hydroxy diphosphate. Citric acid, sodium citrate, boric acid, borax, disodium hydrogen phosphate are available as buffers. However, it is not limited to the above mentioned carriers, excipients or diluents.

The present invention has a high cation substitution capacity and electrical conductivity, high content of humic acid, fulvic acid, humin or organic matter, and the earthworm mud extracted in a live state without stress with Jangheung acid mud having a high content of calcium and magnesium ions. It is possible to provide a high value-added functional cosmetic composition exhibiting a remarkably excellent skin regeneration effect and skin wrinkle improvement effect, and through this to promote the excellence of natural products in Jangheung, Jeollanam-do, and to increase local income and activate the local economy.

1 is a graph of MTT analysis results confirming that Jangheung-san mud and earthworm mucus used as raw materials of the cosmetic composition of the present invention exhibit cytotoxicity to human keratinocytes.
FIG. 2 shows cell photographs at 1 and 50 μl in each experimental group of FIG. 1.
3 is a graph showing the free radical scavenging ability according to the concentration of earthworm mucus.
4a and 4b are graphs showing the free radical scavenging ability of the earthworm ground powder in different concentration ranges.
5 is a photograph of a state in which only one side of the HR-1 mouse was scratched in Experimental Example 4. FIG.
6 is a skin thickness comparison graph (left) and a skin cross-sectional photograph (right) for comparing the skin regeneration effect of the lotion of Example 2 and the comparative example.
7 is a skin thickness comparison graph (left) and a skin cross-sectional photograph (right) for comparing skin regeneration effects of essences of Example 3 and Comparative Example.
8 is a graph (left) and a Western blot photograph (right) showing collagen expression levels for comparing skin wrinkle improvement effects of skins of Example 1 and Comparative Example.
9 is a graph (left) and a Western blot photograph (right) showing collagen expression levels for comparing skin wrinkle improvement effects of lotion of Example 2 and Comparative Example.

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following examples are provided only to aid understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Preparation Example 1 Preparation of Mud Raw Material

After harvesting clean rainwater mud from Jukcheong-ri, Gwansan-eup, Jeollanam-do (GPS information: N34 ° 34'27.1 ″ E126 ° 58′47.4 ″), mud raw materials were manufactured using a wet soil maker in the Jeollanam-do Natural Resources Research Institute. Firstly, the wet muds were removed by primary screening, stirred for at least 4 hours, and then dispersed in a settling tank designed to naturally precipitate from dense particles, while particles were normally dispersed according to Stokes law. Sufficient settling time was allowed for a while, and high purity mud raw materials were collected. The collected mud raw material was sufficiently dried for 1 day or more with a far-infrared dryer, and then the raw materials were pulverized and only particles of 325 mesh or less were used for preparing the cosmetic composition.

In addition, for comparison with Jangheung mud, Boryeong mud raw materials were prepared from mud collected from Boryeong area in the same way as Jangheung mud.

Preparation Example 2 Preparation of Earthworm Raw Material

The earthworms were left for 3 days to excrete all the stools in the body, and then 1 kg of earthworms washed with water were placed in a container containing 1.5 kg of water and immersed for 40 minutes to discharge mucus from the earthworms. Put the discharged mucus and earthworm in a decompression funnel equipped with a strainer and operate the decompression pump to collect only the earthworm mucus that passed through the strainer, and centrifuge to remove the supernatant and obtain 50 g as a supernatant. Earthworm mucus powder was prepared.

In addition, the earthworms were sufficiently washed with distilled water for comparison with the earthworm mucus powder, and then put in a grinder and pulverized at 18,000 rpm for 5 minutes, pre-frozen at -80 ° C, and lyophilized for 2 days. At this time, the freeze-dried portion such as blood was removed and then lyophilized for 1 day to prepare an earthworm crushed powder.

Experimental Example 1: Cytotoxicity of Jangheung Mountain Mud and Earthworm Mucus

The MTT assay uses 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT) to form a pigment product called formazan that is insoluble in water in living cells. It is a method to measure cytotoxicity against.

It was confirmed whether the Changheung-san mud and earthworm mucus of the present invention is toxic to human keratinocytes. Human keratinocytes were counted in 1 x 10 ⁵ cells / well in 96-well plates using a heamacytometer and then aliquoted. 1 ~ 50 μl of the extract of the preparation of the present invention in each well (0.7 / w)% was incubated in DMEM medium at 37 ° C., 5% carbon dioxide for 24 hours. The supernatants were then collected separately on a plate and 50 to 150 μl of a solution of 2-5 mg / ml 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT, Sigma M5655, USA) Addition and incubation for 3 hours under the same conditions. Subsequently, the cell culture medium was removed, 200 μl of dimethyl sulfoxide (DMSO, Sigma D2650, USA) was treated in each well, wrapped in foil, and stirred for 1 hour. Then, 200 μl was taken in 96 wells, followed by Enzyme-Linked Immunosorbent Assay (ELISA). The absorbance was measured at 570 nm. Cell viability is shown in FIG. 1 as a percentage based on the absorbance intensity of the control group using pure water, and the cell photographs at 1 and 50 μl of each experimental group are shown in FIG. 2.

From the above results, Jangheung-san mud and earthworm mucus intended to be used as a raw material of the cosmetic composition of the present invention did not exhibit cytotoxicity in 1 ~ 50 μl, especially when the mud and earthworm mucus were added together, unlike when only the earthworm mucus was added. It was confirmed that rather enhance the cell growth.

Experimental Example 2: Comparison of the properties of Jangheungsan Mud and Boryeong Mountain Mud

Table 1 shows the analysis of the moisture and ash content of Jangheung acid mud and Boryeong acid mud of Preparation Example 1.

division Jangheungsan Mud Boryeong Mountain Mud Water (% by weight) 2.5 4.8 Ash content (% by weight) 92.7 90.1

From the results of Table 1, it can be seen that Jangheung mud may be somewhat superior in purity due to low moisture content and high ash content.

Far-infrared emissivity, pH, cation substitution capacity, and electrical conductivity of Jangheung acid mud and Boryeong acid mud were analyzed and shown in Table 2.

division Jangheungsan Mud Boryeong Mountain Mud Far-infrared emissivity 0.94 0.94 pH 8.3 8.1 Cationic Substitution Capacity (cmol ⁺ / kg) 20.76 11.25 Electrical Conductivity (μS / cm) 78.0 56.5

Jangheung-san mud showed far-infrared emissivity similar to Boryeong-san mud, but Changheung-san mud showed higher activity than that of Boryeong-san mud in cation substitution capacity and electrical conductivity.

Organic content, humic acid, fulvic acid, and humin content of Jangheung Mountain Mud and Boryeong Mountain Mud were analyzed and shown in Table 3.

division Jangheungsan Mud Boryeong Mountain Mud Organic matter (% by weight) 1.67 1.18 Humic acid (% by weight) 1.08 0.77 Fulvic acid (% by weight) 0.29 0.21 Humin (% by weight) 0.29 0.21

Jangheungsan mud showed higher levels of organic content as well as humic acid, fulvic acid and humin, which are useful for skin stabilization.

The potassium, sodium, calcium and magnesium ion contents of the Jangheung and Mud Boryeong mud are shown in Table 4.

division Jangheungsan Mud Boryeong Mountain Mud Potassium Ion (cmol ⁺ / kg) 2.46 2.20 Sodium ion (cmol ⁺ / kg) 1.25 1.25 Calcium ion (cmol ⁺ / kg) 41.3 5.17 Magnesium ion (cmol ⁺ / kg) 29.8 21.1

Jangheung Mountain Mud did not show any significant difference between Boryeong acid Mud and potassium and sodium ions, but the calcium and magnesium contents were high, especially calcium showed that Changheung Mud was 8 times higher than Boryeong Mud.

Experimental Example 3: Comparison of Antioxidant Activity between Earthworm Mucus and Earthworm Ground Powder

The free radical scavenging ability of DPPH was confirmed in comparison with the earthworm pulverized powder whether the earthworm mucus of the present invention exhibited antioxidant activity.

1 mM DPPH (1,1-diphenyl-2-picryl hydrazyl, Sigma D9132-1G, USA) generates free radicals in ethanol. Ethanol was added to 0.5 ml of 0.1 mM DPPH solution prepared with ethanol in 0.4 ml and the earthworm slime powder and earthworm crushed powder of Preparation Example 2 were diluted by concentration so that the final concentration was 10%. The mixture was stirred vigorously for 10 seconds, stored in a cool dark place for 30 minutes, and the absorbance was measured at 517 nm using an ELISA. The degree of antioxidant activity was expressed as a percentage based on the absorbance of the control group using pure water.

% Free radical scavenging capacity = [100- (reaction absorbance of each sample / control absorbance of control)] x 100

Comparing the antioxidant activity of the earthworm slime and earthworm crushed powder in the range of 0.1 to 0.5% by weight, the earthworm crushed powder showed little antioxidant activity, while the earthworm slime showed strong antioxidant activity depending on the dose (FIG. 3 and FIG. 4a). In addition, even if the concentration of the earthworm crushed powder is increased by more than 10 times, the antioxidant activity was less than half of the earthworm mucus (Fig. 4b).

Example 1 Preparation of Skin (Cosmetic Water)

Earthworm Mucus Powder (0.5% Diluent) 3.00

Jangheung acid mud powder (50% diluent) 6.00

Jangheungsan Cypress Oil 0.60

Hydroxyethylene Cellulose (2% aqueous solution) 12.00

Xanthan gum (2% aqueous solution) 2.00

1,3-butylene glycol 6.00

Glycerin 4.00

Sodium Hyaluronate (1% aqueous solution) 5.00

Purified water level

(Unit: wt%)

Example 2: Preparation of Lotion

Earthworm Mucus Powder (0.5% Diluent) 3.00

Jangheung acid mud powder (50% diluent) 6.00

Jangheungsan Cypress Oil 0.60

Ascorbic Acid-2-Magnesium Phosphate 1.00

Water Soluble Collagen (1% Aqueous Solution) 1.00

Sodium Citrate 0.10

Citric Acid 0.05

1,3-butylene glycol 3.00

Purified water level

(Unit: wt%)

Example 3: Preparation of Essence

Earthworm Mucus Powder (0.5% Diluent) 2.00

Jangheung Mountain Mud Powder (50% Diluent) 1.00

Jangheungsan Cypress Oil 0.20

Glycerin 8.00

Sodium hyaluronate (1% aqueous solution) 2.50

Lecithin 0.60

Macadamia Nut Oil 10.00

Purified water balance

Example 4: Preparation of Nutritional Cream

Earthworm Mucus Powder (0.5% Diluent) 2.50

Jangheung acid mud powder (50% diluent) 1.25

Jangheungsan Cypress Oil 0.20

Glycerin 5.00

Triethanolamine 0.50

Liquid Paraffin 5.00

Squalene 5.00

Macadamia Nut Oil 15.00

Polysorbate 60 1.00

Solbitan Sesquioleate 1.00

Carboxyl Vinyl Polymer 0.20

Purified water level

(Unit: wt%)

Example 5: Preparation of Pack

Earthworm Mucus Powder (0.5% Diluent) 2.50

Jangheung acid mud powder (50% diluent) 1.25

Jangheungsan Cypress Oil 0.20

Polyvinyl Alcohol 13.00

L-ascorbic acid-2-magnesium phosphate salt 1.00

Lauroylhydroxyproline 1.00

Water-soluble collagen (1% aqueous solution) 2.00

1,3-butylene glycol 3.00

Ethanol 5.00

Purified water level

(Unit: wt%)

Comparative example

To compare the effect with the cosmetic composition of the present invention, a skin, lotion, essence, nourishing cream, pack (manufactured by Kolmar Korea, product name mudrang) containing boryeong acid mud as a raw material was used as a comparative example.

In addition, since cosmetic compositions containing earthworm powder or earthworm extract are not commercially available, skins, lotions, essences (manufactured by Dassault & Company, product name Swanicoco) and nourishing cream (manufactured by Ezekiel) , Product name Secret Key), and pack (manufactured by Hanul Herb, product name Happy Snail) were used as comparative examples.

Experimental Example 4: Skin Regeneration Effect

After stabilizing indoors for 1 week in HR-1 mouse, the skin was scratched as shown in FIG. 5 to compare with normal skin, and the skin regeneration effect was compared after 1 week after 100 μl of each cosmetic composition was applied.

Take the skin tissue and let it stand in sucrose 10, 20, 30% for 1 day, take it out, slice the tissue by 20 μm, stain with H / E (hematoxylin and eosin) staining method, and measure the thickness of the regenerated tissue under an optical microscope. Observed.

6 is a comparison of the skin regeneration effect of the lotion of Example 2 and Comparative Example, the skin regeneration effect was the most excellent in Example 2, the next lotion containing snail mucus, followed by lotion containing Boryeong acid mud.

Figure 7 is a comparison of the skin regeneration effect of the essence of Example 3 and Comparative Example was the most excellent skin regeneration effect in Example 2, next was the essence containing the Boryeong acid mud, essence containing snail mucus is skin regeneration effect Not shown.

Experimental Example 5: Skin wrinkle improvement effect

In order to confirm the skin wrinkle improvement effect, it was confirmed through an animal test for 30 days. Take skin tissue and take RNA sample from tissue using Easy-blue kit. The collected RNA samples are made into cDNA using RT-PCR (reverse transcriptase), and the collagen gene is expressed using Col1A1 primer, and the cDNA is amplified by PCR. Samples obtained after PCR were electrophoresed on a gel, and the expression level of the genes was confirmed using an image analyzer.

Compared with the UV-treated control group, the wrinkle improvement effect of the skins of Example 1 and Comparative Example was unique in the skin of Example 1 (FIG. 8).

In addition, among the lotions of Example 2 and Comparative Example, the wrinkle improvement effect was observed in the lotion containing Example 2 and Boryeong acid mud, and the product containing snail mucus did not show any wrinkle improvement effect (FIG. 9).

Claims (4)

Earthworm mucus and mud as an active ingredient,
The earthworm slime is a pre-treatment step to excrete the stool in the earthworm body and then washed with water; And extracting the mucus from the earthworm by putting the pretreated earthworm in a container containing water and immersing it for 20 to 60 minutes at 10 to 30 ° C. The cosmetic composition for skin regeneration.
delete The cosmetic composition for skin regeneration according to claim 1, wherein the mud is Jangheung San Mud.
The method according to claim 1 or 3, which contains 0.001 to 0.1% by weight of the earthworm mucus and 0.5 to 10% by weight of the mud, based on a solid content, prepared in any one form selected from skins, lotions, essences, nourishing creams and packs. Cosmetic composition for skin regeneration characterized in that it is.

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