Composition for inducing epidermal damage regeneration

Abstract

The present invention relates to: a composition for regenerating or repairing skin damage and / or promoting epidermal cell differentiation, comprising calcium lactate as an active ingredient; and a method therefor, wherein the skin damage includes that caused by ultraviolet rays. The present invention induces differentiation of epidermal keratinocytes so as to form a biological barrier and allows the skin to maintain moisture on its own, and, as a temporary effect, plays a role in protecting the skin from UV by forming nanoparticles on the surface thereof.

Description

Composition for inducing regeneration of damaged skin epidermis

[0001] The present invention relates to a composition that induces the regeneration of skin epidermal damage, and more specifically, to a composition for inducing the regeneration of skin epidermal damage including ultraviolet light damage to the skin epidermal skin.

[0002] Keratinocytes originate and divide from the epidermal layer (Stratum basale) bordering the dermis, moving to the outside of the skin through the Stratum spinosum, Stratum granulosum, and Stratum lucideum. Ultimately, they form the Stratum corneum, which acts as a physical barrier for the skin.

[0003] In this process, each layer of the epidermis undergoes a specific differentiation process, and to finally form the Stratum corneum, a stratum corneum is formed through terminal differentiation, which creates a stratum corneum of keratinized keratinocytes.

[0004] Differentiation markers exist for each stage and are as follows. The differentiation markers for Stratum basal are K5, K14, and K15. The differentiation markers for Stratum spinosum are K1 and K10. The differentiation markers for Stratum granulosum are transglutaminase and involucrin. The differentiation markers for Stratum corneum are filaggrin, loricrin, and caspase 14.

[0005] Calcium is known to be a representative substance that influences the differentiation of keratinocytes, along with vitamin D and retinoic acid. In particular, calcium concentration decreases as one moves from the stratum corneum to the stratum basal. Keratinocytes proliferate at low calcium concentrations within the skin, while differentiation is induced by the increasing calcium concentrations as one moves upward. This is referred to as skin barrier homeostasis, and if this homeostasis is disrupted, it can lead to diseases such as inflammation or itching, as seen in atopic dermatitis.

[0006] As a result, it creates a barrier against physicochemical hazards exposed from the outside, biologically prevents infection, or maintains moisture within the skin.

[0007] While there are various causes that disrupt skin homeostasis, a representative factor is skin damage caused by high-energy UV wavelengths. The range of UV wavelengths that penetrate the skin is roughly the dermis for UVA and the epidermis for UVB.

[0008] Under normal circumstances, the skin is not exposed to strong UV rays in the short term. However, it is inevitably exposed to high-energy UV wavelengths from sunlight for extended periods. This not only disrupts the skin's homeostasis but also causes changes in the extracellular matrix, which can lead to the formation of wrinkles or, in severe cases, inflammation or cancer.

[0009] TiO2 and ZnO are primarily used to protect the skin from the environment described above. When sunscreens containing these two compounds are applied to the skin, they protect the skin by reflecting or scattering UV wavelengths. However, as the amount of these two inorganic sunscreen compounds increases, the whitening (white cast) phenomenon becomes severe. Therefore, there is a need to develop high-performance inorganic sunscreens that can maintain UV protection while reducing the amount of added compounds.

[0010] Furthermore, the differentiation of keratinocytes in damaged skin epidermis is crucial for the functional regeneration of the epidermis. To promote keratinocyte differentiation, a representative method involves supplying calcium, a differentiation inducer, to the epidermis. However, since inorganic calcium preparations have limitations for human application due to their low stability, the development of safe calcium must be prioritized.

[0011] [Prior Art]

[0012] Republic of Korea Patent Registration No. 101965202

[0013] The present invention was devised to solve the above-mentioned problems and address the aforementioned needs, and the objective of the present invention is to provide a composition capable of regenerating and repairing skin damage caused by environmental factors such as ultraviolet rays, temperature, and humidity.

[0014] Another objective of the present invention is to provide a composition having a skin protection effect against keratinocyte differentiation and / or UV rays, etc.

[0015] To achieve the above objective, the present invention provides a composition for regenerating or repairing skin damage comprising calcium lactate as an active ingredient.

[0016] In one embodiment of the present invention, the composition is preferably prepared by adding calcium lactate to a mixed solution of phosphate and carbonate, but is not limited thereto.

[0017] In another embodiment of the present invention, the effective amount of calcium lactate in the composition is preferably 0.125 mM or more, and the effective amount of calcium lactate in the composition is more preferably 0.125 mM or more to 10 mM, but is not limited thereto.

[0018] In one embodiment of the present invention, the skin damage is characterized by being caused by ultraviolet rays, but is not limited thereto.

[0019] The present invention also provides a method for regenerating or repairing skin damage, comprising administering an effective amount of calcium lactate to a subject requiring skin damage regeneration or repair.

[0020] In one embodiment of the present invention, the effective amount of calcium lactate is preferably prepared by adding calcium lactate to a mixed solution of phosphate and carbonate, but is not limited thereto.

[0021] In another embodiment of the present invention, the effective amount of calcium lactate is preferably 0.125 mM or more, and the effective amount of calcium lactate is more preferably 0.125 mM or more to 10 mM, but is not limited thereto.

[0022] In another embodiment of the present invention, the skin damage is preferably caused by ultraviolet rays, but is not limited thereto.

[0023] In addition, the present invention provides a composition for promoting skin epidermal cell differentiation comprising calcium lactate as an active ingredient.

[0024] In one embodiment of the present invention, the effective amount of calcium lactate in the composition is preferably 2.5 mM or more, and the effective amount of calcium lactate in the composition is more preferably 2.5 mM or more to 5.0 mM or less, but is not limited thereto.

[0025] The present invention will be described below.

[0026] The term 'METI' as used in the specification and / or drawings of the present invention refers to calcium lactate.

[0027] In the composition of the present invention, other ingredients commonly used for external use in cosmetics or quasi-drugs, such as water-soluble alcohols and thickeners, may be incorporated within a range that does not impair the effects of the present invention, and furthermore, powders such as inorganic pigments and extenders, moisturizers, chelating agents, preservatives, colorants, fragrances, etc., may be appropriately incorporated as desired.

[0028] Various water-soluble alcohols may be incorporated into the composition of the present invention.

[0029] Water-soluble alcohols are one or more selected from lower alcohols, polyhydric alcohols, polyhydric alcohol aggregates, dihydric alcohol alkyl ethers, dihydric alcohol alkyl ethers, dihydric alcohol ether esters, glycerin monoalkyl ethers, sugar alcohols, monosaccharides, oligosaccharides, polysaccharides, and derivatives thereof.

[0030] Examples of lower alcohols include ethanol, propanol, isopropanol, isobutyl alcohol, t-butyl alcohol, etc.

[0031] Polyhydric alcohols include, for example, dihydric alcohols (e.g., dipropylene glycol, 1,3-butylene glycol, ethylene glycol, trimethylene glycol, 1,2-butylene glycol, tetramethylene glycol, 2,3-butylene glycol, pentamethylene glycol, 2-butene-1,4-diol, hexylene glycol, octylene glycol, etc.), and trihydric alcohols (e.g., glycerin, trimethylolpropane, etc.).

[0032] Tetrahydric alcohols (e.g., diglycerin, 1,2,6-hexanetriol, pentaerythritol, etc.), pentahydric alcohols (e.g., xylitol, triglycerin, etc.), hexahydric alcohols (e.g., sorbitol, mannitol, etc.), polyhydric alcohol aggregates (e.g., diethylene glycol, dipropylene glycol-triethylene glycol, polypropylene glycol, tetraethylene glycol, diglycerin-triglycerin, tetraglycerin, polyglycerin, etc.), dihydric alcohol alkyl ethers (e.g., ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, ethylene glycol monophenyl ether, ethylene glycol monohexyl ether, ethylene glycol mono2-methylhexyl ether, ethylene glycol isoamyl ether, ethylene glycol benzyl ether, ethylene glycol isopropyl ether, ), dihydric alcohol alkyl ethers (e.g., diethylene glycol dimethyl ether, ethylene glycol diethyl ether, ethylene glycol dibutyl ether, etc.), dihydric alcohol alkyl ethers (e.g., diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol butyl ether, diethylene glycol methyl ethyl ether, triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, propylene glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monobutyl ether, propylene glycol isopropyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, dipropylene glycol butyl ether, etc.), dihydric alcohol ether esters (e.g., ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, ethylene glycol monobutyl ether) Acetate, ethylene glycol monophenyl ether acetate, ethylene glycol diazibate, ethylene glycol disaxinate, diethylene glycol monoethyl ether acetate, diethylene glycol monobutyl ether acetate, propylene glycol monomethyl ether acetate, propylene glycol monoethyl ether acetate,Examples include propylene glycol monopropyl ether acetate, propylene glycol monophenyl ether acetate, etc.), glycerin monoalkyl ethers (e.g., xyl alcohol, cerakyl alcohol, batyl alcohol, etc.), sugar alcohols (e.g., malttriose, mannitol, sucrose, erythritol, glucose, fructose, starch hydrolyzed sugar, maltose, starch hydrolyzed sugar reduced alcohol, etc.), glycolides, tetrahydropulpril alcohol, POE-tetrahydropulpril alcohol, POP-butyl ether, POP / POE-butyl ether tripolyoxypropylene glycerin ether, POP-glycerin ether, POP-glycerin ether phosphoric acid, POP / POE-pentane erythritol ether, polyglycerin, etc.

[0033] As monosaccharides, for example, 3-oses (e.g., D-glycerylaldehyde, dihydroxyacetone, etc.), 4-oses (e.g., D-erythose, D-erytholos, D-threose, erythritol, etc.), 5-oses (e.g., L-arabinose, D-xylose, L-lyxose, D-arabinose, D-ribose, D-ribulose, D-xylose, L-xylose, etc.), 6-oses (e.g., D-glucose, D-tharose, D-busicose, D-galactose, D-fructose, L-galactose, L-mannose, D-tagatose, etc.), 7-oses (e.g., aldoheptose, heprose, etc.), 8-oses (e.g., octrose, etc.), deoxysugars (e.g., 2-deoxy D-ribose, Examples include 6-deoxy L-galactose, 6-deoxy L-mannose, etc.), amino sugars (e.g., D-glucosamine, D-galactosamine, sialic acid, aminouronic acid, muramic acid, etc.), and uronic acids (e.g., D-glucuronic acid, D-mannuronic acid, L-glucuronic acid, D-galacturonic acid, L-isulonic acid, etc.).

[0034] Examples of oligosaccharides include sucrose, kunchianos, unberiferose, lactose, planteos, isoricoses, α,α-trehalose, raffinose, ricoses, unbiricin, starchos, verbascososes, etc.

[0035] Examples of polysaccharides include cellulose, quince seed, starch, galactan, delmatan sulfate, glycogen, gum arabic, heparan sulfate, traganthate, keratan sulfate, chondroitin, xanthan gum, guagam, dextran, keratan sulfate, locust bean gum, saxinoglucan, etc.

[0036] Other polyols include polyoxyethylene methylglucoside (E-10) and polyoxypropylene methylglucoside (glucam P-10).

[0037] Various thickeners may be incorporated into this sunscreen composition of the present invention.

[0038] Examples of thickening agents include gum arabic, carrageenan, karaya gum, tragacanth gum, carob gum, quince seed (marmero), casein, dextrin, gelatin, sodium pectinate, sodium arginate, methyl cellulose, ethyl cellulose, CMC, hydroxyethyl cellulose, hydroxypropyl cellulose, PVA, PVM, PVP, sodium polyacrylate, carboxyvinyl polymer, locust bean gum, gouagam, tamarint gum, cellulose dialkyldimethylammonium sulfate, xanthan gum, magnesium aluminum silicate, bentonite, hetolite, Al-Mg silicate (bi gum), laponite, anhydrous silica, etc.

[0039] Examples of natural water-soluble polymers include plant-based polymers (e.g., gum arabic, tragacanth gum, galactan, guar gum, carob gum, karaya gum, carrageenan, pectin, agar, quince seed (Marmero), algae colloid (brown algae extract), starch (rice, corn, potato, wheat), glycyrrhizic acid), microbial-based polymers (e.g., xanthan gum, dextran, saxinoglucan, pullulan, etc.), and animal-based polymers (e.g., collagen, casein, albumin, gelatin, etc.).

[0040] Examples of semi-synthetic water-soluble polymers include starch-based polymers (e.g., carboxymethyl starch, methyl hydroxypropyl starch, etc.), cellulose-based polymers (methyl cellulose, ethyl cellulose, methyl hydroxypropyl cellulose, hydroxyethyl cellulose, sodium cellulose sulfate, hydroxypropyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, crystalline cellulose, cellulose powder, etc.), and alginate-based polymers (e.g., sodium alginate, propylene glycol alginate ester, etc.).

[0041] Examples of water-soluble polymers for synthesis include vinyl polymers (e.g., polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, carboxyvinyl polymer, etc.), polyoxyethylene polymers (e.g., polyethylene glycol 20,000, 40,000, 60,000, etc.), acrylic polymers (e.g., sodium polyacrylate, polyethyl acrylate, polyacrylamide, etc.), polyethyleneimine, cationic polymers, etc.

[0042] As powder components, for example, inorganic powders (e.g., talc, kaolin, mica, cirrus mica (seric mica), muscovite, phlogopite, synthetic mica, boron mica, biotite, pyroclasticite, magnesium carbonate, calcium carbonate, aluminum silicate, barium silicate, calcium silicate, magnesium silicate, strontium silicate, metal tungstate salts, magnesium, silica, zeolite, barium sulfate, calcined calcium sulfate (bovine gypsum), calcium phosphate, fluoroapatite, hydroxyapatite, ceramic powder, metal soaps (e.g., zinc myristate, calcium palmitate, aluminum stearate), boron nitride, etc.), organic powders (e.g., polyamide resin powder (nylon powder), polyethylene powder, polymethyl methacrylate powder, polystyrene powder, copolymer resin powder of styrene and acrylic acid, benzoguanamine resin powder, polytetrafluoroethylene powder, Cellulose powder, etc.), inorganic white pigments (e.g., titanium dioxide, zinc oxide, etc.), inorganic red pigments (e.g., iron oxide (red iron oxide), iron titanate, etc.), inorganic brown pigments (e.g., γ-iron oxide, etc.), inorganic yellow pigments (e.g., iron sulfate, ochre, etc.), inorganic black pigments (e.g., black iron oxide, low-order titanium oxide, etc.), inorganic purple pigments (e.g., mango violet, cobalt violet, etc.), inorganic green pigments (e.g., chromium oxide, chromium hydroxide, cobalt titanate, etc.), inorganic blue pigments (e.g., ultramarine, indigo, etc.), pearl pigments (e.g., titanium oxide-coated mica, titanium oxide-coated bismuth oxychloride, titanium oxide-coated talc, colored titanium oxide-coated mica, bismuth oxychloride, irrind, etc.), metal powder pigments (e.g., aluminum powder, copper powder, etc.). ), organic pigments such as zirconium, barium or aluminum lakes (e.g., organic pigments such as Red 201, Red 202, Red 204, Red 205, Red 220, Red 226, Red 228, Red 405, Orange 203, Orange 204, Yellow 205, Yellow 401, and Blue 404, Red 3, Red 104,Examples include Red 106, Red 227, Red 230, Red 401, Red 505, Orange 205, Yellow 4, Yellow 5, Yellow 202, Yellow 203, Green 3, and Blue 1, etc.), and natural pigments (e.g., chlorophyll, β-carotene, etc.).

[0043] Examples of moisturizers include chondroitin sulfate, hyaluronic acid, mucoitin sulfate, caronic acid, atherocollagen, cholesteryl 12-hydroxy stearate, sodium lactate, bile salt, DL-pyrrolidone carboxylate, short-chain soluble collagen, diglycerin (EO) PO adduct, chestnut rose extract, yarrow extract, melilot extract, etc.

[0044] Examples of metal ion chelating agents include 1-hydroxyethane-1, 1-diphosphonic acid, 1-hydroxyethane-1, 1-diphosphonic acid tetrasodium salt, disodium edetate, trisodium edetate, tetrasodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, phosphoric acid, citric acid, ascorbic acid, succinic acid, edetic acid, ethylenediamine hydroxyethyl triacetate trisodium, etc.

[0045] Examples of amino acids include neutral amino acids (e.g., threonine, cysteine, etc.) and alkaline amino acids (e.g., hydroxylysine, etc.). Also, examples of amino acid derivatives include acyl sarcosine sodium (lauroyl sarcosine sodium), acyl glutamate, acyl β-alanine sodium, glutathione, etc.

[0046] Examples of organic amines include monoethanolamine, diethanolamine, triethanolamine, morpholine, tetrakis(2-hydroxypropyl) ethylenediamine, triisopropanolamine, 2-amino-2-methyl-1,3-propanediol, 2-amino2-methyl-1-propanol, etc.

[0047] Examples of polymer emulsions include acrylic resin emulsion, polyethyl acrylate emulsion, acrylic resin liquid, polyacrylate alkyl ester emulsion, polyvinyl acetate resin emulsion, natural rubber latex, etc.

[0048] Examples of pH adjusters include buffers such as lactic acid-sodium lactic acid, citrate-sodium citrate, and succinate-sodium succinate.

[0049] Examples of vitamins include vitamins A, B1, B2, B6, C, E and their derivatives, pantothenic acid and its derivatives, and biotin.

[0050] Examples of antioxidants include tocopherols, dibutyl hydroxy toluene, butyl hydroxy anisole, gallic acid esters, etc.

[0051] Examples of antioxidants include phosphoric acid, citric acid, ascorbic acid, maleic acid, maronic acid, succinic acid, fumaric acid, caefalin, hexametaphosphate, phytic acid, ethylenediamine tetraacetic acid, etc.

[0052] Other possible ingredients include, for example, preservatives (methylparaben, ethylparaben, butylparaben, phenoxyethanol, etc.), anti-inflammatory agents (e.g., glycyrrhetinic acid derivatives, salicylic acid derivatives, hinoxyol, zinc oxide, allantoin, etc.), whitening agents (e.g., Saxifraga extract, arbutin, etc.), various extracts (e.g., Phellodendron amurense, Coptis japonica, Lithospermum erythrorhizon, Paeonia lactiflora, Angelica dahurica, birch, sage, loquat, carrot, aloe, Malva verticillata, iris, grape, Coix lacryma-jobi, loofah, lily, saffron, Cnidium officinale, ginger, Capsicum sarmentosum, Ononis, garlic, chili pepper, Citrus peel-Angelica gigas, seaweed, etc.), stimulants (e.g., royal jelly, photosensitizers, cholesterol derivatives, etc.), and blood circulation promoters (e.g., nonyl acid arenilamide, nicotinate benzyl ester, Examples include nicotinic acid β-butoxyethyl ester, capsaicin, zingerone, canthalistinche, ichtamol, tannic acid, α-borneol, nicotinic acid tocopherol, inositol hexanicotinate, secrandelate, cinnarizine, trazoline, acetylcholine, verapamil, cepharanthin, γ-oryzanol, etc.), anti-seborrheic agents (e.g., sulfur, thianthol, etc.), and anti-inflammatory agents (e.g., tranexamic acid, thiotaurine, hypotaurine, etc.).

[0053] According to one embodiment, the above-mentioned ingredient, which is generally used for external use in cosmetics or quasi-drugs, may be included in an amount of 50% by weight or less relative to the total weight of the composition, for example, 45% by weight or less, 40% by weight or less, 35% by weight or less, 30% by weight or less, 25% by weight or less, 20% by weight or less, 15% by weight or less, 10% by weight or less, 5% by weight or less, 4% by weight or less, 3% by weight or less, 2% by weight or less, 1% by weight or less, 0.1% by weight or less, or 0.01% by weight or less.

[0054] Since the composition of the present invention is basically applied to the skin, it can be formulated by referring to cosmetic compositions in the art, for example, as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation, spray, etc., and known ingredients or known combinations necessary for this purpose may be used.

[0055] In one embodiment, the formulation of the composition may be in a liquid state. The liquid state may be low viscosity or non-viscous.

[0056] The composition of the present invention can be prepared in a group of formulations suitable for low-viscosity or non-viscosity formulations, and more specifically, can be prepared in a formulation suitable for a product in which a low-viscosity or non-viscosity liquid is total-packaged in a high concentration, for example, a skin toner, a mist, a cosmetic lotion, or a toner pad.

[0057] In addition, the composition of the present invention may be used by applying it alone or in combination, or by applying it in combination with other cosmetic compositions other than the present invention. Furthermore, the cosmetic composition according to the present invention may be used according to conventional methods of use, and the frequency of use may be varied depending on the user's skin condition or preference.

[0058] As can be seen from the present invention, the invention induces the differentiation of keratinocytes in the epidermis to form a biological barrier and maintain moisture within the skin itself, and as a temporary effect, forms nanoparticles on the surface to protect the skin from UV rays.

[0059] Figure 1 is a graph showing the amount of nanoparticles produced by absorbance when calcium lactate is added to a liquid solution containing phosphate and carbonate.

[0060] Figures 2 to 4 are, respectively, a graph showing the diameter of the total nanoparticles generated over time, a graph showing the diameter of the nanoparticles generated over time, and a graph showing the measured zeta potential of the nanoparticles generated over time.

[0061] Figure 5 is a graph showing the absorbance values ​​of nanoparticles produced after a 10-minute reaction in the UV wavelength range, and Figure 6 is a graph showing the absorbance values ​​of nanoparticles produced after a 120-minute reaction in the UV wavelength range.

[0062] Figures 7 and 8 are, respectively, immunoblot images of differentiation-inducing proteins quantified in keratinocytes and microscopic images of keratinocytes before and after differentiation.

[0063] Figures 9 and 10 are, respectively, an immunoblot showing the differentiation-inducing protein of fibroblasts and myofibroblasts according to the concentration of calcium lactate and an immunoblot showing the protein indicating intercellular adhesion of fibroblasts and myofibroblasts according to the concentration of calcium lactate.

[0064] Figures 11 and 12 are graphs showing the transglutaminase activity of keratinocytes cultured for 1 hour in RPMI1640 medium without serum and the transglutaminase activity of keratinocytes cultured for 24 hours in RPMI1640 medium with serum.

[0065] The present invention will be described in more detail below through non-limiting examples. However, the following examples are described for the purpose of illustrating the present invention, and the scope of the present invention shall not be interpreted as being limited by the following examples.

[0066] Example 1: Nanoparticle-forming ability of calcium lactate

[0067] 1-1. Formation of Nanoparticles

[0068] To synthesize nanoparticles, a solution containing dissolved phosphate and carbonate was added to a 96-well plate at a volume of 200 μl / well. Subsequently, 200 mM calcium lactate stock solution was added to achieve concentrations of 0, 0.625, 1.25, 2.5, 5.0, and 10.0 mM, respectively, and the reaction was carried out. Afterward, absorbance was measured at a wavelength of 660 nm at 0, 10, 30, and 60 minutes. In the above experiment, the phosphate was added at a concentration of 4800 mg / L (sodium dihydrogen phosphate, NaH2PO), and the carbonate was added at a concentration of 2000 mg / L (sodium bicarbonate, NaHCO3).

[0069] The reaction began immediately upon the addition of calcium lactate, and the absorbance value rose to a constant level at 0 minutes. It was confirmed that after 10 minutes or more, the reaction was complete and the slope of the graph did not increase. When calcium lactate was reacted at a concentration of 0.625 mM, it reacted completely immediately upon addition and maintained a constant absorbance value starting from 0 minutes; however, when reacted at a concentration of 1.25 mM or higher, it was confirmed that the amount of nanoparticles produced increased proportionally to the concentration, indicating that the absorbance value increased proportionally to the concentration. (Fig. 1)

[0070] 1-2. Characteristics of Nanoparticles

[0071] The size of the nanoparticles prepared by the above method was measured at different time intervals. 1 ml of a solution containing phosphate and carbonate was placed in an EP tube, and 12.5 µl of a 200 mM calcium lactate solution was added to make the concentration 2.5 mM. After leaving the mixture at room temperature for the reaction time, the size and surface charge of the nanoparticles were measured using dynamic light scattering analysis with a cuvette.

[0072] Looking at the graph of the average size of all particles, it can be seen that they grow to an average of 6,000 nm. However, the average size of the most numerous particles ultimately increases to an average of 4,000 nm. Additionally, it was observed that nanoparticles of various sizes are formed before 10 minutes, and that their sizes become somewhat similar when the reaction is carried out for 30 to 60 minutes or longer (Figs. 2 to 3).

[0073] When measuring the surface charge of these particles, it can be seen that the zeta value is -10mV around 10 minutes, when small-sized particles predominate, but decreases as time passes and the particle size increases (Fig. 4).

[0074] Example 2: UVA and UVB blocking activity of nanoparticles

[0075] For the UV blocking experiment, nanoparticles were prepared by adding 2.5 mM calcium lactate to a solution of dissolved phosphate and carbonate in a 96-well plate. After adding calcium lactate, absorbance was measured using a plate reader at wavelengths in the UVA and UVB ranges 10 and 120 minutes later.

[0076] It can be observed that absorbance increases in the UV range when nanoparticles are formed. Particles reacted for 10 minutes exhibited the highest absorbance in the 300–350 nm wavelength range (Fig. 5). As time passed, sufficient nanoparticle formation occurred, and absorbance increased by approximately 1.5 to 2 times after 120 minutes (Fig. 6).

[0077] Example 3: Terminal differentiation induction activity of epidermal keratinocytes

[0078] 3-1. Quantification of Differentiation-Inducing Proteins in Undifferentiated Keratinocytes

[0079] Using human-derived HaCaT cells, a type of keratinocyte, differentiation markers were identified via immunoblotting to determine whether keratinocytes differentiate in response to calcium lactate. HaCaT cells were 4.5 x 10⁶ 5The cells were seeded into 6-well plates and cultured in DMEM medium for 24 hours. Once the cells stabilized, the medium was changed to FBS-free RPMI1640 medium, and then cultured for 1 hour under hypoxic conditions with calcium lactate at concentrations of 0, 1.5, 2.5, and 5.0 mM. Cells were collected from each group, and proteins were extracted using Lysis buffer. The extracted proteins were quantified, and immunoblotting was performed according to the concentrations.

[0080] The experimental results confirmed that the protein concentrations of Involucrin and Filaggrin, keratinocyte differentiation markers, increased with increasing calcium lactate concentration. At 1.5 mM calcium lactate, there was no difference compared to untreated cells, but starting from 2.5 mM calcium lactate, the protein concentrations of Involucrin and Filaggrin increased. Under these conditions, it was confirmed that calcium lactate can differentiate keratinocytes within one hour. Furthermore, it was observed that differentiation into corneocytes present on the skin surface occurs due to the decrease in cadherin. (Fig. 7)

[0081] 3-2. Induction of telogenesis in undifferentiated keratinocytes

[0082] As in Example 3-1, the HaCaT cells were 4.5 x 10 5 The cells were dispensed into a 6-well plate and cultured in DMEM medium for 24 hours. After cell stabilization, the cells were cultured for 1 hour under hypoxic conditions in FBS-free RPMI 1640 medium with calcium lactate at concentrations of 0, 1.5, 2.5, and 5.0 mM.

[0083] The morphology and changes of cells treated with calcium lactate were observed using an optical microscope. While cells not treated with calcium lactate maintained a normal cell morphology, it was observed that as the concentration of calcium lactate increased, the cells differentiated and their morphology changed.

[0084] In particular, cell morphology changed significantly starting from 2.5 mM calcium lactate, and at 5 mM, it was observed that the overall cell size decreased. This appears to be the formation of a cell layer similar to the stratum corneum. (Fig. 8)

[0085] Example 4: Induction of pluripotency and EMT inhibitory activity in fibroblasts and fibroblasts

[0086] Fibroblasts and fibroblasts 4.5 x 10 5 Cells were seeded into 60 mm plates and stabilized by incubating in RPMI1640 medium for 24 hours. After stabilization, the medium was replaced with FBS-free RPMI1640, and calcium lactate was added at concentrations of 0, 0.5, 1, and 2.5 mM, respectively. Fibroblast and fibroblast differentiation was induced by incubating under hypoxic conditions for 1 hour, while the EMT inhibition experiment involved incubation under hypoxic conditions for 5 minutes. The calcium lactate-treated cells were collected by group, and proteins were extracted using Lysis buffer. The extracted proteins were quantified, and immunoblotting was performed according to the concentration.

[0087] It was confirmed that fibroblasts and fibroblasts, which may be exposed due to skin damage, undergo differentiation into keratinocytes within one hour (Fig. 9). Additionally, E-cadherin increases and N-cadherin decreases within five minutes in fibroblasts and fibroblasts, suggesting that EMT is inhibited rapidly (Fig. 10).

[0088] Example 5: Transglutaminase activity in keratinocytes

[0089] To measure the transglutaminase enzyme activity of keratinocytes, HaCaT 4.5 x 10 5The cells were seeded into 6-well plates and cultured in DMEM medium for 24 hours. After stabilizing the cells, they were divided into two groups of four. One group was cultured for 1 hour in serum-free RPMI1640 medium under hypoxic conditions, treated with calcium lactate at concentrations of 0, 1.5, 2.5, and 5.0 mM, while the other group was cultured for 24 hours in RPMI1640 medium containing 10% serum under hypoxic conditions, treated with calcium lactate at concentrations of 0, 1.5, 2.5, and 5.0 mM. After the designated culture time for each group was completed, cells were collected and proteins were extracted using Lysis buffer. Protein analysis was performed using a Transglutaminase activity assay kit (Sigma Aldrich, cs1070). Subsequent experimental procedures followed the instructions provided in the kit.

[0090] Under conditions without serum, no changes in the transglutaminase enzyme were observed due to rapid differentiation (Fig. 11). However, in keratinocytes cultured for 24 hours in the presence of serum, the activity of transglutaminase appears to increase due to the calcium in calcium lactate (Fig. 12). It was found that keratinocytes differentiate into Stratum granulosum by calcium lactate through the increased activity of transglutaminase, a representative differentiation marker, which binds structural proteins to enable the stratum corneum to function as a skin barrier. Furthermore, as shown in Example 3, the increase in Filaggrin creates a cell layer similar to Stratum corneum, which will enable rapid formation of a skin barrier.

Claims

1. A composition for regenerating or repairing skin damage containing calcium lactate as an active ingredient.

2. A composition for skin damage regeneration or repair according to claim 1, characterized in that the composition is prepared by adding calcium lactate to a mixed solution of phosphate and carbonate.

3. A composition for regenerating or repairing skin damage, characterized in that, in claim 1 or 2, the effective amount of calcium lactate in the composition is 0.125 mM or more.

4. A composition for regenerating or repairing skin damage, characterized in that, in claim 1 or 2, the effective amount of calcium lactate in the composition is 0.125 mM or more to 10 mM.

5. A composition for regenerating or repairing skin damage according to claim 1 or 2, characterized in that the skin damage is caused by ultraviolet rays.

6. A method for regenerating or repairing skin damage, comprising administering an effective amount of calcium lactate to a subject requiring skin damage regeneration or repair.

7. A method for regenerating or repairing skin damage according to claim 6, characterized in that the effective amount of calcium lactate is prepared by adding calcium lactate to a mixed solution of phosphate and carbonate.

8. A method for regenerating or repairing skin damage according to claim 6 or 7, characterized in that the effective amount of calcium lactate is 0.125 mM or more.

9. A method for regenerating or repairing skin damage according to claim 6 or 7, characterized in that the effective amount of calcium lactate is 0.125 mM or more to 10 mM.

10. A method for regenerating or repairing skin damage according to claim 6 or 7, wherein the skin damage is caused by ultraviolet rays.

11. A composition for promoting skin epidermal cell differentiation containing calcium lactate as an active ingredient.

12. A composition for promoting skin epidermal cell differentiation, characterized in that, in claim 11, the effective amount of calcium lactate in the composition is 2.5 mM or more.

13. A composition for promoting skin epidermal cell differentiation, characterized in that, in claim 11 or 12, the effective amount of calcium lactate in the composition is 2.5 mM or more to 5.0 mM or less.

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