KR20100131613A - Skin composition for external application containing tomato callus and preparing the same - Google Patents

Abstract

PURPOSE: An external use skin composition containing tomato callus extract is provided to ensure skin cell activation, anti-wrinkling, skin whitening and anti-oxidation. CONSTITUTION: An external use skin composition for improving skin condition contains 0.1-10 weight% of tomato callus extract as an active ingredient. A method for manufacturing the external use skin composition comprises: a step of inducing callus form rice plant seeds and culturing; and a step of preparing the composition having the callus.

Description

Skin composition for external application containing tomato callus extract and method for preparing the same {Skin Composition for External Application Containing Tomato Callus and Preparing the Same}

The present invention relates to a composition for external application for skin containing tomato callus extract and a method for producing the same.

Modern man is always threatened from the diseases associated with it by various internal and external stresses, and the skin is not free from such stresses. Various studies have been attempted to keep the skin healthy from such a skin stress, and in recent years, interest in cosmetics using natural extracts obtained from natural substances having little skin irritation is increasing.

On the other hand, plants have long been a source of various natural substances that help human well-being. Most of these substances are secondary metabolites and are used for pharmaceuticals, food additives, cosmetic raw materials, fragrances, and pigments. Despite the synthesis of numerous new compounds by the development of the field of organic synthesis, plants are still recognized as the source of new substances. Considering that there are more than a thousand new substances with new chemical structures found in plants every year, many of them are bioactive, which makes them an important ingredient.

 Many chemicals produced by plants are classified as primary metabolites because they are essential for cellular function. Primary plant metabolites are universally present in plant populations and constitute the majority of the chemicals produced by plants, while other compounds are classified as secondary metabolites and because of their many uses, both scientific research and commercial use are the focus of both. It is.

In some cases secondary metabolites are produced by plants, for example in response to stress induced by microbial infection, UV radiation, mechanical damage and also by treatment with organic and inorganic materials. The variety of secondary metabolites produced under such stress conditions is broad and includes alkaloids, terpenoids, prebonoids, carotenoids, phenolic compounds and glycosides. However, not all secondary metabolites are produced in large quantities by all plants, and the kinds of metabolites depend largely on the type of plant.

Plants have an excellent totipotency in which the entire individual is formed even when only a part of the leaves, stems, and roots constituting the plant are transplanted. Plant cells are callus derived from plant tissues. Callus is an amorphous tissue or cell mass that has lost its ability to cause normal organogenesis or tissue differentiation.Auxin or Cytokinins Occurs when cultured in a medium containing or wounding the plant, or treated with the auxin of plants.

Callus can be derived from any part of the plant such as leaves, stems, roots, rhizomes, and fruits through plant cell culture, especially tissue culture, and is characterized by being continuously produced by subculture. The basic principle of the cultivation of plant cells takes advantage of the biological fact that every plant cell has the ability to build the whole plant from which the cell is derived. This pluripotency is comparable to the pluripotency of embryonic stem cells in animals. Therefore, it can be accepted that plant cells have a positive effect on the protection and activation of skin stem cells.

On the other hand, callus is an oily tissue in which a wound cell recovers division ability and prevents and enlarges when a wound is caused to a plant, and a special mass of tissue generated by culturing the tissue cut from the plant in an auxin-containing medium or the like. In recent years, interest in the use of such callus has been increasing and is being studied in various ways. Further, as a public patent related to callus, there is a 10-2008-0038068 or the like.

Meanwhile, as a cosmetic composition related to tomatoes, a cosmetic composition for preventing skin aging containing a supercritical fluid extract of tomatoes, grapes and safflower seeds (Korean Patent Registration No. 10-0803924), a cosmetic composition containing a pomegranate extract and a tomato extract ( Korean Patent Publication No. 10-2007-0008150), a cosmetic composition containing a tomato extract (Korean Patent Publication No. 10-2005-0076307), etc. Inducing callus from tomato seeds, callus extract was used, or there was no example of preparing a cosmetic composition for inducing a useful substance to the tomato callus.

Thus, the inventors of the present invention, while trying to prepare a skin external preparation composition having skin improvement ability, confirmed that the tomato callus has a skin cell activation, skin wrinkle improvement, skin whitening and antioxidant effect, and completed the present invention.

Accordingly, it is an object of the present invention to provide a skin external preparation composition for improving skin containing tomato callus extract as an active ingredient.

Still another object of the present invention is to provide a method for preparing a skin external composition for skin improvement containing the tomato callus extract as an active ingredient.

In order to achieve the above object, the present invention provides a skin external composition for skin improvement containing tomato callus extract as an active ingredient.

The present invention also comprises the steps of (a) inducing callus from the tomato seed culture; And (b) provides a method for producing a skin external composition for skin improvement containing tomato callus extract comprising the step of preparing a composition containing the callus.

The tomato callus-containing external composition for skin according to the present invention activates skin cells due to skin cell growth and proliferation, has a moisturizing effect, and has an excellent effect on skin regeneration and skin soothing.

The present invention relates to a skin external preparation composition for improving skin containing the extract of tomato callus as an active ingredient and a method for preparing the same. Specifically, the present invention induces callus from the tomato seeds using tissue culture technology, and the skin A topical composition, for example, relates to a use as a cosmetic raw material and a method for producing the same.

The present invention relates to a skin external composition for skin improvement containing tomato callus extract as an active ingredient in one aspect. Specifically, the tomato callus extract may contain 0.01 to 30% by weight, preferably 0.1 to 10% by weight based on the total weight of the composition. If it is contained in less than 0.1% by weight, it is difficult to exhibit the desired skin improvement effect, when containing more than 10% by weight there is a disadvantage that shows a problem in the formulation.

In the present invention, "extract" refers to an extract obtained by pulverizing the tomato callus itself or tomato callus, or by hot water extraction, ethanol extraction, or the like.

In the present invention, the meaning of "contained as an active ingredient" means that the composition for external application of the skin can contain the skin improving effect, for example, can be improved in the proliferation activity of the skin cells and is contained to an extent that can be activated. do. In addition, it means that it contains an effective amount that can exhibit a moisturizing effect, skin soothing effect and the like as the external preparation for skin.

In the present invention, the tomato (Solanum Lycopersicum MILL.) Is a yearly perennial plant of the dicotyledon plant moss-tree branch, also known as a yearly persimmon, tomato fruit is about 90% water and vitamins A, B, C, E, K and potassium, calcium Contains a lot of minerals, carotene and lycopene. Beta-carotene and lycopene in tomatoes are known to prevent cancer, and tomatoes are also known to play a role in relieving constipation and making elastic and fair skin. It also controls the amount of moisture in the body to suppress overeating. It is known to function to promote digestion and to promote the action of the stomach, pancreas, liver and the like. Tomatoes are known to be effective in improving the function of the large intestine, which lowers the level of cholesterol in the blood and prevents obesity. Tomatoes are known to be particularly effective in preventing vitamins from aging and preventing osteoporosis and senile dementia, especially among vitamin C. These vitamins contain anti-aging ingredients that help to rejuvenate your body, prevent osteoporosis, and vitamin C in tomatoes is more potent in inhibiting carcinogens than vitamin C in other fruits and vegetables. (Herb illustration, Ahn Deok-gyun, 1998, 699p), also promotes metabolism and regulates the amount of urine, and is good for febrile diseases such as hepatitis (熱 性病). In addition, it is described in agreement that the customary skin disease is effective by drinking a little water boiled tomato root, stem, and leaf, washing the body frequently with this water, and drinking a little foreign body.

In the present invention, the tomato has a variety of varieties, including wind, young water. Among these, the gust of wind is very strong and leafy phenomenon is easy to cultivate, good cold and overheating and low malformation and less swelling is known as a large species. The cultivation type is semi-tactile, and in the case of semi-tactile cultivation, it is advantageous that the formal seedling is planted in the first flowering period, and in suppressing cultivation, 4-5 seedlings of main leaves are planted. The minimum temperature must be maintained above 10 ℃ during the seedling period.

On the other hand, Young-soo has good lightness and hypertrophy even at low temperatures, and especially because of its small leaves, it has good light transmission. Cultivation is palpable. In the following examples, callus induction was used using a varietal varieties, but in addition to this, seeds derived from various tomato varieties may also be callus induction.

 In the present invention, callus refers to a special tissue or a mass of cells generated when a tissue cut out of a plant is cultured in an auxin-containing medium, a certain kind of plant is wounded, or an upper mouth is treated with auxin. Normally, callus is an amorphous tissue or cell mass that has lost its ability to cause normal organogenesis or tissue differentiation, and is mostly composed of flow cells. In a broad sense, it may also include plant tumor tissue caused by infections such as agrobacterium.

When the undifferentiated cell mass callus is formed through tissue culture, that is, the cells in the plant are oriented immediately upon proliferation, and are assembled regularly into specific tissues and organs. However, when various plant growth hormones are given from the outside, the control that has been maintained until then is released, and cells are interpreted to proliferate arbitrarily. When the callus obtained by this dedifferentiation is put in a liquid medium having the same composition and shaken for culture, the cells are separated and continue to proliferate as a suspended state. The callus or cultured cells are subdivided and proliferated permanently by passage in fresh medium, which is essentially different from the differentiated cells in which complete plants age and die. Callus or cultured cells, which have been passaged for several generations, are applied to a solid medium from which various plant growth hormones have been removed. In other words, the plant is restored in one somatic cell that makes up the plant. Plant tissue cells isolated from the parent plants have the potential to be regenerated into complete plants when cultured in an appropriate culture environment. This morphogenic ability is of great value for the practical use of cosmetic raw materials, as well as for academic importance such as plant development.

In the present invention, the plant tissue for inducing callus may be any part of the seed or the plant. For example, germinated or sterilized seeds can be germinated to cut away parts of the plant, or to cut and sterilize parts of the plant. In this way callus is induced through sterile tissue culture methods. Preferably, the plant fragments are cultured in a tissue culture medium. The medium includes MS (Murashige & Skoog, 1962) medium, N6 (Chu et al. 1975) medium, B5 (Gamborg et al., 1968), NN (Nitsch and Nitsch, 1967) medium, and the basic medium of various plants Available at Plant growth regulators should be added to this basal medium. The most commonly used plant growth regulators are 2,4-D (2,4-dichlorophenoxyacetic acid), 2,4,5-T (2), which are auxins. , 4,5-trichlorophenoxyacetic acid, Dicamba (2-methoxy-3,6-dichlorobenzoic acid), IAA (Indole-3-acetic acid), IBA (Indole-3-butyric acid), MCPA (2-methyl-4 -chlorophenoxyacetic acid), NAA (1-naphthylacetic acid), NOA (2-naphthyloxyacetic acid), Picloram (4-amino-2,5,6-trichloropicolinic acid) and Cytokinin BAP (6-benzylaminopurine) , 2iP (N ^6- (2-isopentyl) adenine), Kinetin (6-furfurylaminopurine), Thidiazuron (1-phenyl-3- (1,2,3-thiadiazol-5-yl) urea), Zeatin (4-hydroxy -3-methyl-trans-2-butenylaminopurine is used at a concentration suitable for the plant, taking into account the species, age, site, and ripening of the plant. In addition, secondary metabolites such as alkaloids, terpenoids, prebonoids, carotenoids, phenolic compounds, and glycosides can be added to plant types to enhance the various physiological activities of callus induced by growth regulators. Can be added accordingly.

A summary of the preferred callus induction and culture method is as follows. A part of the tissue of the plant is separated and sterilized, and then cultured in a medium containing growth regulators of auxin or cytokinin in the plant basic medium. After incubation for 2 to 3 weeks, when the cells of the tissue divide and the callus, which is a mass of cells, is separated, only the callus is separated and placed on a fresh medium, and then passaged by observing a constant time interval or callus state.

Conventional studies on tomato callus, biotechnological perspectives on the callus itself, such as confirming the peroxidase activity of the transformed tomato callus, inducing callus to examine the morphological characteristics, and the effect on plant regeneration The study was only attempted. The present inventors newly confirmed that the tomato callus has an antioxidant activity and has a proliferative activity of skin cells, and manufactured an external preparation composition for skin improvement comprising the same.

In another aspect, the present invention, (a) inducing callus from the tomato seed cultured; And (b) relates to a method for producing a skin external composition for skin improvement containing a tomato callus extract comprising the step of preparing a composition containing a tomato callus.

In the step (a), specifically, it is possible to select an appropriate medium to induce callus by germinating tomato seeds. If there is a medium generally used in the callus culture in the art, it can be used without limitation.

In the present invention, as a medium for inducing callus from tomato seeds, including the medium used in the callus induction step in the production of GMO tomato, etc., all that is conventionally available as a tomato callus induction medium can be applied. Medium added with 2.0 mg / L BA, 0.01 mg / L IBA, 30 g / L sucrose, and 8 g / L agar in MS medium (Murashige and Skoog 1962, Duchefa, Cat No. M0221) can be used. In addition to this, all plant basal medium may be applicable.

When the callus is cultured in a medium containing useful substances, the callus absorbs the useful substances added to the medium and thus contains the useful substances in the callus. It can exhibit physiological activity. The introduction of the useful substance in the callus may introduce the useful substance into the cultured callus by additionally performing the step of densifying the induced callus in the medium containing the useful substance.

In the present invention, as an example of the useful substance, there may be mentioned Lycopene, which is a kind of phytochemical. The lycopene is a carotenoid having about 40 carbon chains, and is a natural pigment that makes red color contained in tomatoes, guavas, watermelons, and the like, and is particularly contained in tomatoes. Lycopene is known to have a physiological activity that prevents cell damage caused by free radicals generated in the body through excellent antioxidant activity.

 In addition, since lycopene has the highest antioxidant activity among carotenoids, it has been reported to prevent cell damage caused by reactive oxygen species and reactive nitrogen species, which is very effective in preventing and treating various diseases (DiMascio P., Kaiser S. and H., Sies. Lycopene as the most efficient biological carotenoid singlet oxygen quencher.Arch.Biochem.Biophys. 1989.274: 532-538; Bohm F., Tinkler JH and TG, Truscott.Carotenoids protect against cell membrane damage by the nitrogen dioxide radical.Nature Medicine. 1995. 2: 98-99 etc.). In other words, lycopene has a strong antioxidant function that prevents oxidative damage of intracellular DNA and cell membranes to inhibit the development of cancer cells and has a direct proliferation inhibitory effect on cancer cells. It has been suppressed. In addition, it improves lipid metabolism in the blood to prevent cardiovascular diseases such as angina pectoris, myocardial infarction and stroke, and increases the immune cell catcher to increase the body's immune function. It has also been reported to be effective in improving impotence. The physiological activity or efficacy of such lycopene reduces the risk of cancer, heart disease, etc., the US FDA has already approved the health claim (Health claim) and confirmed as a safe food (GRAS) (GRN no 156).

In step (b), the callus obtained in step (a) may be powdered and included in the composition. In addition to this, it can be prepared by containing the extract obtained by hot water extraction method, ethanol extraction method and the like in an appropriate concentration.

Specifically, in the composition for external application for skin containing tomato callus extract according to the present invention, in addition to the aqueous phase component and the oil phase component, additives may be included. The additives include alcohols, colorants, fragrances and the like.

In addition, the topical skin preparation composition according to the invention can be prepared in various forms, for example emulsions, lotions, creams (water-in-oil, water-in-oil, multiphase), solutions, suspensions (anhydrous and aqueous), anhydrous products (Oil and glycol based), gels, masks, packs, powders and the like formulations.

The method of preparing the tomato callus extract-containing skin external preparation composition according to the present invention is not limited to the above-described preparation method. Tomato callus extract-containing skin external preparation composition according to the invention can be prepared.

In addition, the topical skin composition according to the present invention may be a cosmetic composition or a pharmaceutical composition.

When the external skin composition is a pharmaceutical composition, it may further include a pharmaceutically acceptable carrier that is already known and used.

In the cosmetic composition, an acceptable carrier may be included in the cosmetic formulation. Herein, "acceptable carrier in cosmetic preparation" means a compound or composition already known and used that may be included in a cosmetic preparation or a compound or composition which will be developed in the future, and which has no toxicity, instability or irritation that can be adapted to the human body upon contact with the skin. Say nothing.

The carrier may be included in the topical skin composition of the present invention in an amount from about 1% to about 99.99% by weight, preferably from about 90% to about 99.99% by weight of the composition. However, since the ratio depends on the formulation of the above-described skin external preparation composition of the present invention and its specific application site (face, neck, etc.) or its preferred application amount, the ratio may be viewed in any aspect. It should not be understood as limiting the scope of the invention.

Meanwhile, examples of the carrier include alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosity modifiers, emulsions, stabilizers, sunscreen agents, colorants, fragrances, and the like. The alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosity modifiers, emulsions, stabilizers, sunscreens, colorants, compounds / compositions that can be used as fragrances and the like are already known in the art, Appropriate materials / compositions may be selected and used.

As an embodiment of the present invention, the external preparation composition for skin according to the present invention may include glycerin, butylene glycol, propyleneglycol, polyoxyethylene hardened castor oil, ethanol, triethanolamine, etc., in addition to the above-mentioned seaweed extract, A trace amount may be included if necessary such as a medicine, a pigment, purified water, and the like.

In particular, the external preparation composition for the skin may be prepared in the form of general emulsion formulations and solubilized formulations, using a conventionally known manufacturing method, in addition to the manufacturing method specifically disclosed in the present invention.

When the cosmetic composition is prepared, cosmetics of the emulsion formulations include nutrient cosmetics, creams, essences, etc., and cosmetics of the solubilized formulations include soft cosmetics. In addition, by containing a dermatologically acceptable medium or base, it can be prepared in the form of topical or systemic adjuvants commonly used in the field of dermatology.

In addition, formulations of suitable cosmetics include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionics obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase, for example. It may be provided in the form of a vesicle dispersant in the form of a cream, skin, lotion, powder, ointment, spray or concealed stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the topical skin composition of the present invention may further comprise fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ions Any commonly used in type or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics It may contain adjuvants conventionally used in the cosmetic or dermatology field as other ingredients. In addition, the above components may be introduced in an amount generally used in the dermatology field.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Example 1 Preparation of Tomato Callus According to the Present Invention

(1) Preparation of tomato callus according to the present invention

Among the various varieties of tomato (Lycopersicon esculentumMill), 'young water' was used as a material, and seed germination was immersed in 70% ethanol for 30 seconds, washed with sterile water and again with disinfectant solution (30% Lax + Tween20) for 20 minutes. After disinfection, the cells were washed three times with sterile water and seeded by adding 8 g / L of agar and 30 g / L of sucrose to MS medium (Murashige and Skoog, 1962) having reduced salt concentration to 1/2.

Seeds sown were incubated for 7 days at 25 ° C. and 70% humidity to induce germination, cut the cotyledons of germinated seeds, and cut the cotyledons into MS basal medium (Murashige and Skoog 1962, Duchefa, Cat No. M0221) added BA 2.0 mg / L, IBA 0.01 mg / L, sucrose 30 g / L, agar 8 g / L and was suspended in a solid medium adjusted to pH 5.8. After about 20-25 weeks, callus was induced from the cut cotyledons dentified in the medium. The derived callus was finely ground in liquid nitrogen, dissolved in purified water, and used in the following experimental example.

(2) Preparation of Tomato Callus Derived from Lycopene-Containing Medium

After sterilization and washing in the same manner as in (1), sucrose 30g / L and agar 8g / L are added to the MS medium, Lycopene 100mg / L is added thereto, and then adjusted to pH5.6˜5.7, BA Callus was induced at 25 ° C. using a solid medium containing 2.0 mg / L and IBA 0.01 mg / L.

Experimental Example 1 Test of Determination of Tyrosinase Inhibition Effect of a Composition According to the Present Invention

In order to confirm the whitening effect of the callus extract according to the present invention prepared in Example 1, Tyrosinase (Tyrosinase) which is an enzyme that helps melanin production by promoting the oxidation process of a substance called Tyrosine in vivo The extent to which the function was suppressed was examined. Specifically, a method of measuring the degree of inhibition of the formation of a black polymer called melanin by oxidizing tyrosine by inhibiting the function of tyrosinase (Pomerantz SH, J. Biochem., 24: 161-168, 1966) is applied. Determined.

 Into the tomato callus extract of the present invention in 96 well-plate to 0.5%, 1%, 5% by weight, respectively, 220µl of 100mM sodium phosphate buffer (pH 6.8), 40µl of 1.5mM L-tyrosine solution After adding 20 μl of mushroom tyrosinase (1,500 units / ml, Sigma) and reacting at 37 ° C. for 20 minutes, 0.2 ml of the solution was transferred to a 96 well plate, and the microplate reader was placed. The absorbance at 490 nm was used to determine the inhibition rate for tyrosinase. The degree of inhibition of tyrosinase function was expressed as a percentage based on the absorbance intensity of the control using pure water.

[Equation 1]

% Inhibition = 1-[(Experimental absorbance-Blank absorbance) / control absorbance] X 100

As shown in FIG. 1, 0.5% of tomato callus extract showed 5% oxidation inhibition, 1% of 48% oxidation inhibition and 5% compared to the control group using purified water. In one case, it showed 50% oxidation inhibition. In case of containing 0.5% of tomato callus extract derived from lycopene-added medium, it showed 30% oxidation inhibition, 1%, 65% oxidation inhibition, and 5% 67.6% showed oxidation inhibition. Therefore, the composition according to the present invention was also possible to inhibit the oxidation mediated by the enzyme, it was confirmed that it can be applied as a whitening material.

Experimental Example 2: Antioxidant Test Using DPPH

Compound 1,1-diphenyl-2-picryl hydrazyl (DPPH, Sigma D9132-1G, USA) generates free radicals in ethanol, the amount of free radicals produced by mixing with the composition according to the invention in a proportion By checking whether the degree of decrease (free radical scavenging activity test) was checked whether the antioxidant effect. The free radical scavenging activity was modified from the method of Kim et al. picryl hydrazyl, Sigma D9132-1G, USA) reagent was used.

150 μl of 0.2 mM DPPH solution (ethanol in the case of Blanc) was prepared to contain various extracts at various concentrations, and 150 μl of these mixtures were added and mixed, and the absorbance was measured at 517 nm after being allowed to stand at room temperature for 30 minutes. The control group was set to the case using purified water. After absorbance measurement for the experimental group and the control group, the degree of antioxidant capacity was expressed as a percentage based on the absorbance intensity of the control group using pure water.

Using the following Equation 2 to obtain the free radical scavenging effect was as shown in FIG.

[Equation 2]

Free radical scavenging effect (%) = 1- [(Experimental absorbance-Blank absorbance) / control absorbance] X 100

As shown in Figure 2, compared to the control using purified water, 5% of the tomato callus extract, 15% of the DPPH free radicals of the reaction system containing 39%, 32% and 0.5% when containing 1% When 5% of the tomato callus extract derived from the lycopene-added medium contained 5% of the DPPH free radicals in the reaction system, it was found that the amount of the DPPH free radicals decreased by 63%, 1%, 60% and 0.5%, respectively. Could.

Experimental Example 3: Effect test on the cell growth and proliferation of tomato callus according to the present invention

In order to examine the proliferative activity of the skin cells of the tomato callus extract prepared in Example 1, human skin fibroblasts (Human Skin Fibroblast, CCD-986Sk) was cultured by aliquoting from the American Type Culture Collection (ATCC). Each cell was inoculated in a 24 well culture plate at a concentration of 1 × 10 ⁴ cells / ml. Medium was used DMEM (Dubelcco'S Modified Eagle Medium, BRL, USA) containing 10% FBS. After 48 hours of incubation in DMEM containing 10% FBS and 25-30% of the surface of the culture vessel, the tomato callus extract prepared in Example 1 was replaced with FBS-free DMEM containing 0.5-5% for 24 hours. It was further cultured. After incubation, 50 µl of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT, Sigma M5655, USA) solution (2.5 mg / ml) was added and 3 hours After further incubation, the supernatant was removed, 200 μl of Dimethylsulfoxide (DMSO, Sigma D2650, USA) solution was added to each well, followed by stirring for 20 minutes to dissolve the formazan crystals formed. The absorbance was measured at 570 nm with an Enzyme-Linked Immunosorbent Assay (ELISA). The proliferative activity of the skin cells was expressed as a percentage calculated according to the following equation based on the absorbance intensity of the control group using pure water, and the results were as shown in FIG.

&Quot; (3) "

% Cell proliferation effect = [(experimental absorbance-control absorbance) / control absorbance] × 100

As shown in Figure 3, 115% when containing 5% tomato callus extract, 110% when containing 1% tomato callus extract, 101% when containing 0.5% tomato callus extract, lycopene When 5% of the tomato callus extract derived from the added medium contained 123%, the growth rate was 123%, and in the case of 1% and 0.5%, the growth was 120% and 105%, respectively. Therefore, it was confirmed that the composition containing the tomato callus extract according to the present invention or the composition containing the tomato callus extract derived from the lycopene-added medium has an effect on the activation of skin cell growth and proliferation.

Experimental Example 4: Collagen Synthesis Enhancement Effect Test

Human Skin Fibroblast (DH) containing 10% FBS, Penicillin (50 U / ml) and Streptomycin (50 µg / ml) in a cell incubator maintained at 37 ° C. in a 5% CO ₂ incubator. Dulbecco's Modified Eagle's Medium, Gibco, USA), 500 μl was dispensed into 24 well plates at 1 × 10 ⁵ / ml and incubated for 24 hours. Fresh medium containing a control extract and a mixed extract of various concentrations of 0.05 ~ 5% was added and incubated in 37 ℃, 5% CO ₂ incubator for 48 hours. After 48 hours, 20 μl of the supernatant of the medium was taken and measured using a Procollagen Type I C-Peptide EIA Kit (PICP, Takara, Cat No. MK101) to determine the amount of newly synthesized collagen. PICP amount was converted into ng / 1 × 10 ⁵ cells, and the results are shown in FIG. 4.

As shown in FIG. 4, when the collagen synthesis rate related to wrinkle improvement contains 5%, 1%, and 0.5% of tomato callus extract, respectively, compared to the control group using purified water, 135%, 127%, and 120, respectively, are concentration-dependent. In the case of compositions containing 5%, 1%, and 0.5% of tomato callus extract derived from lycopene-added medium, respectively, 145%, 142%, and 132% of collagen synthesis, respectively. The effect was shown.

1 is a graph showing the test results of tyrosinase inhibitory effect of the composition containing tomato callus extract according to the present invention.

Figure 2 is a graph showing the results of testing the antioxidant effect of the composition containing tomato callus extract according to the present invention.

Figure 3 is a graph showing the results of human fibroblast growth and proliferation test of the composition containing tomato callus extract according to the present invention.

Figure 4 is a graph showing the results of the collagen synthesis enhancement test of the composition containing tomato callus extract according to the present invention.

Claims (5)

Skin external composition for improving skin containing tomato callus extract as an active ingredient. According to claim 1, wherein the tomato callus extract is a skin external composition for skin improvement, characterized in that containing 0.1 to 10% by weight relative to the total weight of the composition. Method for preparing a skin external composition for skin improvement containing tomato callus extract comprising the following steps: (a) inducing and incubating callus from rice seeds; And (b) preparing a composition containing the callus. The method of claim 3, wherein the callus is powdered and contained in the composition. The method according to claim 3, wherein in step (a), the callus is induced from rice seeds and cultured in a medium containing epigallocatechinallate.

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