Skin preparation containing lactic acid bacteria fermentation product of aloe vera leaves and its manufacturing method

Abstract

【assignment】 The present invention provides a new skin topical agent and a method for producing the same, which contains, as an active ingredient, a lactic acid bacteria fermentation product of the juice of Aloe vera leaves, by using the TY-610 strain, a new strain of Lactiplantibacillus plantarum. [Solution] The present invention provides an external skin preparation containing a lactic acid bacteria fermentation product of aloe vera leaves, The lactic acid bacteria is Lactiplantibacillus plantarum TY-610 strain ( Contract Number: NITE B P-03877) and a method for producing the same.

Description

Cross References

[0001] This application claims priority in Japan based on Patent Application No. 2023-070742 filed on April 24, 2023 and Patent Application No. 2023-218768 filed on December 26, 2023, all of which are incorporated herein by reference in their entirety. [Technical field]

[0002] The present invention relates to an external skin preparation containing a lactic acid bacteria fermentation product of aloe vera leaves, a method for producing the same, and the like. [Background technology]

[0003] Lactic acid bacteria fermentation products are generally used in skin lotions and skin care products because they are effective in improving wrinkles and / or sagging skin. Among them, moisturizers using aloe fermentation products produced by various lactic acid bacteria (Patent Document 1) and cleansing cosmetics containing aloe fermentation products (Patent Document 2) have been reported as using the aloe plant. However, these products used existing strains such as standard strains of the lactic acid bacteria Lactiplantibacillus plantarum. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 3795011 [Patent Document 2] JP 2022-164406 A Summary of the Invention [Problem to be solved by the invention]

[0005] As a result of intensive research, the present inventors have found a new strain of Lactiplantibacillus plantarum, TY-610. Therefore, the present invention aims to provide a new skin care product containing, as an active ingredient, a lactic acid fermentation product of the juice of Aloe vera leaves, using the said strain, and a method for producing the same. [Means for solving the problem]

[0006] (1) In one embodiment of the present invention, a skin topical preparation for achieving the above object comprises: Contains lactic acid fermentation product of aloe vera leaves, The lactic acid bacteria is Lactiplantibacillus plantarum TY-610 strain ( Contract Number: NITE B P-03877). (2) In another embodiment of the topical skin preparation, the preparation preferably has hyaluronic acid production promoting activity. (3) In another embodiment of the topical skin preparation, it is preferable to prevent and / or improve dry skin. (4) In another embodiment of the topical skin preparation, the balance of the resident skin flora is preferably improved. (5) In one embodiment for achieving the above object, a method for producing an external skin preparation containing a lactic acid bacteria fermentation product produced from aloe vera leaf juice comprises the steps of: In the process of producing a lactic acid bacteria fermentation product, The juice was treated with Lactiplantibacillus plantarum TY-610 strain ( Contract Number: NITE B and a fermentation step of fermenting the mixture by a method comprising the steps of: The step of producing the lactic acid bacteria fermentation product preferably includes the steps of: an aloin removal step for removing aloin from the juice; The juice after the aloin removal process was treated with Lactiplantibacillus plantarum TY-610 strain ( Contract Number: NITE B and a fermentation step of fermenting the mixture by a method comprising the steps of: Effect of the Invention

[0007] According to the present invention, by using the TY-610 strain, which is a novel strain of Lactiplantibacillus plantarum, it is possible to provide a novel skin topical agent containing, as an active ingredient, a lactic acid bacteria fermentation product of Aloe vera leaf juice, and a method for producing the same. [Brief description of the drawings]

[0008] [Figure 1] FIG. 1 shows the measurement sites in a human skin monitoring test (Test Example 8). [Diagram 2] Figure 2 shows a micrograph of a specimen obtained as a result of a human skin monitoring test (Test Example 9, Evaluation of multilayer peeling of the stratum corneum in a human monitoring test). The line in the figure indicates 100.0 μm. When the peeled stratum corneum approaches a monolayer, the staining pigment appears faint. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Next, each embodiment of the present invention will be described. Note that each embodiment described below does not limit the invention according to the claims, and all of the elements and combinations thereof described in each embodiment are not necessarily essential to the solution of the present invention.

[0010] (Aloe vera leaves) The Aloe vera leaves used in the present invention are composed of an outer skin consisting of a thick cuticle layer and mesophyll present inside the outer skin. In the present invention, the "Aloe vera leaves" may be used, for example, raw or dried, or may be crushed. The amount of Aloe vera leaves used as a raw material in the present invention is not particularly limited, but is preferably 0.01% or more, more preferably 0.1% or more, and particularly preferably 1% or more of the total weight of the raw material used in the fermentation process.

[0011] (TY-610 strain) In the present invention, the lactic acid bacteria fermentation of aloe vera leaves is performed using Lactiplantibacillus plantarum TY-610 strain (hereinafter, simply referred to as TY-610 strain). The TY-610 strain is a new strain of Lactiplantibacillus plantarum isolated from pickled cabbage. The TY-610 strain is different from the standard strain in that it has a spinnability, and therefore is thought to produce and release mucilaginous polysaccharides outside the bacterial cell. The TY-610 strain has been deposited at the National Institute of Technology and Evaluation (NPMD) (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan) on April 5, 2023 under the accession number NITE P-03877. Subsequently, the TY-610 strain was requested to be transferred from domestic deposit (original deposit date: April 5, 2023) to international deposit under the Budapest Treaty on March 13, 2024 at the international depository authority, the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (NPMD) (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan). Contract Number NITE B Has been granted P-03877.

[0012] (Lactic acid fermentation product) In the present invention, the term "lactic acid bacteria fermentation product" refers to a product obtained by fermenting aloe vera leaves with the above-mentioned TY-610 strain. The form of the lactic acid bacteria fermentation product is not particularly limited, and may be a liquid or a solid such as a powder, and is selected as desired depending on the form of use of the present invention. In addition, the lactic acid bacteria fermentation product according to the present invention may contain additives such as preservatives, antibacterial agents, and chelating agents. Preservatives, antibacterial agents, and chelating agents include hydroxybenzoic acids and their salts or esters, such as methylparaben, ethylparaben, propylparaben, and butylparaben; salicylic acid; sodium benzoate; phenoxyethanol; 1,2-diols, such as 1,2-pentanediol and 1,2-hexanediol; isothiazolinone derivatives, such as methylchloroisothiazolinone and methylisothiazolinone; imidazolinium urea; dehydroacetic acid and its salts; phenols; halogenated bisphenols, such as triclosan, acid amides, and quaternary ammonium salts; trichlorocarbanide, zinc pyrithione, benzalkonium chloride, benzethonium chloride, sorbic acid, chlorhexidine, chlorhexidine gluconate, halocarban, hexachlorophene, and hinokitiol; phenol, isopropyl pheno Examples of additives include other phenols such as ethanol, cresol, thymol, parachlorophenol, phenylphenol, and phenylphenol sodium; phenylethyl alcohol, photosensitizers, antibacterial zeolite, silver ions; edetates (ethylenediaminetetraacetates) such as EDTA, EDTA2Na, EDTA3Na, and EDTA4Na; hydroxyethylethylenediaminetriacetates such as HEDTA3Na; pentetates (diethylenetriaminepentaacetates); phytic acid; phosphonic acids such as etidronic acid and salts thereof such as sodium salts; sodium oxalate; polyamino acids such as polyaspartic acid and polyglutamic acid; sodium polyphosphate, sodium metaphosphate, phosphoric acid; sodium citrate, citric acid, alanine, dihydroxyethylglycine, gluconic acid, ascorbic acid, succinic acid, and tartaric acid. The content of additives in the lactic acid bacteria fermentation product is not particularly limited, but is preferably 1% or less, more preferably 0.5% or less, and particularly preferably 0.1% or less, based on the total weight of the lactic acid bacteria fermentation product.

[0013] (Form of topical skin preparation) The topical skin preparations according to the present invention are in suitable forms for use, such as ampoules, capsules, powders, granules, liquids, gels, foams, emulsions, sheets, mists, sprays, etc., and include 1) medicines, 2) quasi-drugs, and 3) topical or whole-body topical skin preparations (for example, basic cosmetics such as skin lotions, milky lotions, creams, ointments, lotions, oils, and packs, face washes and skin cleansers such as solid soaps, liquid soaps, and hand washes, massage preparations, cleansing preparations, hair removal preparations, depilatories, shaving treatments, aftershave lotions, preshave lotions, shaving creams, foundations, lipsticks, blushers, eyeshadows, and eyeliners). , make-up cosmetics such as mascara, perfumes, nail beautifying products, nail beautifying enamel, nail beautifying enamel remover, poultices, plasters, tapes, sheets, patches, aerosols, etc.), 4) medicinal and / or cosmetic preparations applied to the scalp and hair (e.g. shampoos, rinses, hair treatments, pre-hair treatments, permanent solutions, hair dyes, hair styling products, hair tonics, hair growth and tonic products, poultices, plasters, tapes, sheets, aerosols, etc.), 5) bath products used by adding them to the bath, 6) other products such as underarm odor prevention agents, deodorants, antiperspirants, sanitary products, sanitary cotton, wet tissues, etc.

[0014] (Components of topical skin preparations) In addition, in the production of such agents, the following components and additives can be selected and used in combination as necessary within the range that does not impair the effects of the present invention. The amount of these to be added to the formulation is not particularly limited, but it is generally considered preferable to add about 0.0001 to 50%.

[0015] (1) Various oils and fats Avocado oil, almond oil, fennel oil, perilla oil, olive oil, orange oil, orange roughage oil, sesame oil, cacao butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, kukui nut oil, safflower oil, shea butter, liquid shea butter, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, Japan wax, coconut oil, beef tallow, lard, squalene, squalane, pristane, or hydrogenated products of these oils and fats (hardened oils, etc.).

[0016] (2) Waxes Beeswax, carnauba wax, spermaceti, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax, etc.

[0017] (3) Mineral oil Liquid paraffin, Vaseline, paraffin, ozokeride, ceresin, microcrystalline wax, etc.

[0018] (4) Fatty acids Natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and lanolin fatty acids; synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid, and isopentanoic acid.

[0019] (5) Alcohol Natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, and phenoxyethanol; and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, and 2-octyldodecanol.

[0020] (6) Polyhydric alcohols Ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1,3-butylene glycol, pentyl glycol, glycerin, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, maltitol, and the like.

[0021] (7) Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, etc.

[0022] (8) Metal soaps Examples of such stearate include aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, and zinc undecylenate.

[0023] (9) Gums, sugars or water-soluble polymeric compounds Gum arabic, gum benzoin, gum dammar, guaiac butter, Irish moss, gum karaya, gum tragacanth, carob gum, quince seed, agar, casein, lactose, fructose, sucrose or its ester, trehalose or its derivative, dextrin, gelatin, pectin, starch, carrageenan, carboxymethylchitin or chitosan, hydroxyalkyl (C2-C4) chitin or chitosan with an alkylene (C2-C4) oxide such as ethylene oxide added thereto, low molecular weight chitin or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or its salt, hyaline Examples of suitable cellulose acetate hydrochloride include hydroxypropyl cellulose, ...

[0024] (10) Surfactants Anionic surfactants (alkyl carboxylates, alkyl sulfonates, alkyl sulfates, alkyl phosphates), cationic surfactants (alkyl amine salts, alkyl quaternary ammonium salts), amphoteric surfactants: carboxylate type amphoteric surfactants (amino type, betaine type), sulfate ester type amphoteric surfactants, sulfonate type amphoteric surfactants, phosphate ester type amphoteric surfactants, nonionic surfactants (ether type nonionic surfactants, ether ester type nonionic surfactants, ester type nonionic surfactants, block polymer type nonionic surfactants, nitrogen-containing nonionic surfactants), other surfactants (natural surfactants, derivatives of protein hydrolysates, polymeric surfactants, surfactants containing titanium and silicon, fluorocarbon surfactants), etc.

[0025] (11) Various vitamins Vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), Vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, inositol, Vitamin C group: vitamin C acid or its derivatives, Vitamin D group: ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotachysterol, vitamin E group: vitamin E or its derivatives, ubiquinones, vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3), menadiol (vitamin K4), others, essential fatty acids (vitamin F), carnitine, ferulic acid, γ-oryzanol, orotic acid, vitamin P group (rutin, eriocitrin, hesperidin), vitamin U, etc.

[0026] (12) Various amino acids Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine, histidine, and the like, and amino acid derivatives thereof such as sulfates, phosphates, nitrates, citrates, or pyrrolidone carboxylic acid.

[0027] (13) Various additives derived from plant or animal sources These may be added by subjecting them to processing that is conventionally performed according to the type and form of the product to which they are to be added (e.g., any combination of processing such as crushing, milling, washing, hydrolysis, fermentation, refining, squeezing, extraction, fractionation, filtration, drying, powderization, granulation, dissolution, sterilization, pH adjustment, deodorization, and decolorization), and then arbitrarily selecting from various materials.

[0028] The upper limit of the content of the lactic acid bacteria fermentation product in the topical skin preparation is not particularly limited, and may be, for example, 50% by mass, 10% by mass, 5% by mass, or 1% by mass of the total weight of the topical skin preparation. The lower limit of the content of the lactic acid bacteria fermentation product in the topical skin preparation is not particularly limited, and is preferably 0.001% by mass, more preferably 0.01% by mass, even more preferably 0.05% by mass, and particularly preferably 0.1% by mass of the total weight of the topical skin preparation.

[0029] (Hyaluronic acid production promoting activity) In the present invention, "promotion of hyaluronic acid production" refers to promoting hyaluronic acid production in the skin, and "hyaluronic acid production-promoting activity" refers to activity that causes the above-mentioned promotion of hyaluronic acid production.

[0030] (Improves the balance of the skin's resident flora) In the present invention, "skin flora" refers to microorganisms present on the human epidermis, scalp, mucous membrane, etc., which are possessed by many people. Human skin flora is broadly divided into two types: good bacteria and bad bacteria. Good bacteria refers to bacteria that have a beneficial effect on the skin. An example of good bacteria is Staphylococcus epidermidis. Staphylococcus epidermidis keeps the skin weakly acidic by breaking down sebum into fatty acids and glycerin. On the other hand, bad bacteria refers to bacteria that have a harmful effect on the skin. An example of bad bacteria is Staphylococcus aureus. Staphylococcus aureus causes skin diseases such as atopic dermatitis. In the present invention, "improving the balance of the skin flora" refers to not inhibiting or promoting the growth of good bacteria and / or inhibiting the growth of bad bacteria among the above-mentioned skin flora, and preferably promoting the growth of Staphylococcus epidermidis and / or suppressing the growth of Staphylococcus aureus.

[0031] (Dry skin) In the present invention, "dry skin" refers to a state in which moisture is lost from the stratum corneum of the skin. When moisture is lost from the stratum corneum, the barrier function of the skin is weakened, making it easier for external stimuli to penetrate. When inflammatory cytokines are produced in association with the penetration of external stimuli, inflammation is induced, resulting in itching and redness. Scratching the affected area, etc., causes eczema, and the skin turnover period is shortened. As a result, the number of stratum corneum cells with immature moisturizing functions increases, making the skin more prone to dryness. Examples of inflammatory cytokines include IL-1α, which is associated with a decrease in epidermal barrier function and contributes to rough skin, and IL-6, which induces redness. In the present invention, "preventing and / or improving dry skin" refers to suppressing the production of the above-mentioned inflammatory cytokines. The type of inflammatory cytokines to be suppressed is not particularly limited, but is preferably IL-1α and / or IL-6.

[0032] In another aspect of the present invention, a method for producing an external skin preparation is provided.

[0033] (Aloin removal process) Aloin is a medicinal ingredient contained in the outer layer of Aloe vera leaves, and has bitterness and laxative activity. Aloin is usually removed during the manufacturing process of cosmetics. In the present invention, the manufacturing method of a skin topical agent preferably includes an aloin removal step. The aloin removal step is not limited as long as it can remove aloin, and any known method such as adsorption to activated carbon can be used.

[0034] (Fermentation process) The fermentation step in the present invention will now be described in detail.

[0035] In the present invention, the fermentation process is carried out by mixing the juice component of the aloe vera leaves after the aloin removal process with the TY-610 strain. The juice component of the aloe vera leaves after the aloin removal process is in powder form in the examples described below, but is not limited thereto and may be, for example, liquid. The juice component is suspended or added to a fermentation medium and then subjected to the fermentation process. The fermentation medium is water or a mixture of water and lower alcohols (methanol, ethanol, propanol, etc.) or glycols (ethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, etc.) in an appropriate amount relative to the juice component. Sugars such as glucose, fructose, sucrose, sugar, granulated sugar, and white sugar may be added to the fermentation medium. Yeast extract may also be added to the fermentation medium.

[0036] The liquid component suspension is sterilized to remove unwanted bacteria that may hinder fermentation before being subjected to the fermentation process. In this case, the sterilization and removal method may be a method in which the fermentation material is washed and sterilized in advance with sterilizing ethanol or the like, and then suspended in a sterile medium such as sterile water, or a method in which the fermentation material is suspended in a medium and then the suspension is heat sterilized. As the heat sterilization method, an autoclave sterilization method in which the suspension is heated at 120 to 130°C for 10 to 20 minutes, or an intermittent sterilization method in which the suspension is kept at 80 to 90°C for 60 to 120 minutes and this is repeated once a day for 2 to 3 days, is generally used.

[0037] Fermentation is carried out by adding a medium containing the TY-610 strain to the fermentation medium after heat sterilization. The time required for fermentation is not particularly limited, but is preferably 12 to 36 hours, more preferably 15 to 30 hours, and particularly preferably 18 to 24 hours. The temperature condition during fermentation is preferably 15 to 37°C, more preferably 20 to 35°C, and particularly preferably 25 to 32°C. After the fermentation is completed, the TY-610 strain cells are removed, but the method for removing the cells is not particularly limited, and known methods such as centrifugation and filtration can be used. The sugar content (Brix) of the lactic acid bacteria fermentation product is not particularly limited, but is preferably 0.1 to 20°, more preferably 0.5 to 15°, and particularly preferably 1 to 10°.

[0038] A preservative may be added to the lactic acid bacteria fermentation product after the removal of the bacteria. Here, the preservative is not particularly limited as long as it is a preservative that is added to the above-mentioned skin external preparation. However, such preservatives may cause skin irritation and stickiness when added to a skin external preparation, so it is desired to reduce the amount used as much as possible. Therefore, there is no limit to the lower limit concentration when adding a preservative, and it is sufficient to exceed 0% by mass. The upper limit concentration is 1% by mass, preferably 0.5% by mass, and more preferably 0.1% by mass. Preservatives that have a preservative effect at these low concentrations are, for example, parabens, benzoic acid and its salts, and more preferably sodium benzoate.

[0039] The pH of the mixed solution containing the suspension and a medium containing the TY-610 strain (hereinafter simply referred to as the mixed solution) before fermentation is preferably about 4 to 5, more preferably about 4.5 to 5, and particularly preferably about 4.7. The pH after fermentation is preferably about 3 to 4, more preferably about 3.5 to 4, and particularly preferably about 3.7. The acidity of the mixed solution before fermentation is preferably about 0.2 to 0.3, more preferably about 0.21, in terms of lactic acid. The acidity of the mixed solution after fermentation is preferably about 0.7 to 0.8, more preferably about 0.76, in terms of lactic acid.

[0040] When the lactic acid bacteria fermentation product produced in the above fermentation step is provided in the form of an external skin preparation, it can be provided, for example, together with a pharma- ceutical or cosmetically acceptable carrier (water, oily components, etc.) in addition to the lactic acid bacteria fermentation product. Furthermore, if necessary, for example, the above-mentioned commonly used components and additives can be added and prepared into a desired form by a conventional method.

[0041] The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. In the following examples, the unit % of the numerical values ​​showing the amount of each component added means % by mass. EXAMPLES

[0042] The experimental materials used in the experiments described below in the test examples are as follows: Human epidermal keratinocytes: Normal human epidermal keratinocytes, Kurabo, KK-4009, Frozen NHEK (NB) neonatal derived, KURABO Keratinocyte culture medium (1): HuMedia-KG2 specially ordered GC supplementary medium, Kurabo, KK-2170S. This specially ordered GC supplementary medium is supplemented with calcium (final concentration 0.06 mM). Keratinocyte culture medium (2): Medium (1) minus BPE (Bovine Pituitary Extract) and EGF (Epidermal Growth Factor). Staphylococcus epidermidis: Standard strain of Staphylococcus epidermidis (ATCC12228) ·NB medium: Nutrient Broth No.2 liquid medium (Oxoid Limited, CM0067B)

[0043] (Preparation of lactic acid bacteria fermentation product) The lactic acid bacteria fermentation product (aloe vera juice component (aloe powder) that has undergone a fermentation process) used in the following test examples was produced according to the following process.

[0044] The unfermented material used in the following test examples was aloe powder (a juice component of aloe vera leaves that has undergone an aloin removal process, Organic SQ Freeze Dried Aloe Vera Leaf Powder 100X (manufactured by Aroe Laboratories, Inc.)).

[0045] 1. Pre-culture of TY-610 strain The TY-610 strain was added to the medium having the composition shown in Table 1 below at 1 × 10 6 The bacteria were inoculated to a concentration of cfu / mL and cultured at 30°C for 18 hours.

[0046] [Table 1]

[0047] 2. Preparation of aloe vera leaf juice component (aloe powder) after aloin removal process As the juice component of aloe vera leaves that has been subjected to the aloin removal process, we used Organic SQ Freeze Dried Aloe Vera Leaf Powder 100X (manufactured by Aroe Laboratories, Inc.), which is an aloe powder. Note that the unfermented material shown in the following test examples is this aloe powder.

[0048] 3. Fermentation process of aloe powder 6.0 kg of the above aloe powder (about 1% of the total weight of the mixed liquid) was suspended in a fermentation medium having the composition shown in Table 2 below to obtain a total of 594.0 kg of suspension.

[0049] [Table 2]

[0050] The sugar content (Brix) of the suspension was 2.0°.

[0051] The suspension was heat sterilized at 90° C. for 1 minute. The suspension after heat sterilization was cooled to about 30° C. 6.0 kg of a preculture medium containing the TY-610 strain was added to the suspension after cooling.

[0052] The suspension after the addition was subjected to a fermentation process for 18 hours under the condition of 30° C. The sugar content (Brix) of the suspension after the fermentation process was 1.9°.

[0053] 4. Removal of TY-610 strain As described below, the TY-610 strain was removed from the suspension after the fermentation process. The lactic acid bacteria fermentation product after the fermentation process was centrifuged (8,200×g) using a continuous centrifuge to collect the supernatant. The supernatant was filtered by a precoating method to obtain a filtrate.

[0054] (Test Example 1) Evaluation of hyaluronic acid production promoting activity In this test example, a non-fermented product or a lactic acid bacteria fermented product was added to human epidermal keratinocytes, and the change in the amount of hyaluronic acid produced was evaluated (quantified).

[0055] (Preparation of human epidermal keratinocytes) Human epidermal keratinocytes were cultured at 5 × 10 4 The cells were seeded at 1000 cells / mL onto a 24-well culture plate. The seeded cells were cultured in keratinocyte culture medium (1) under conditions of 5% CO2 and 37°C for 24 hours until the cells reached 75% confluence. The cells used in Test Example 1 were prepared by this culture.

[0056] (Addition of sample to epidermal keratinocytes) The prepared cells were cultured for 72 hours under the conditions of 5% CO2 and 37°C as shown in Table 3 below. After the culture, the supernatant was collected. The hyaluronic acid in the collected supernatant was measured using QnE Hyaluronic Acid (HA) ELISA Assay Kit (BTP, BTP-96200). The measurement results are shown in Table 3.

[0057] [Table 3]

[0058] The measurement results shown in Table 3 are values ​​(relative values) calculated from the average values ​​measured for each group (n=3), with the control group set at 100. In Table 3, an * indicates a significant difference of p<0.05 compared to the control group in a Dunnett test with n=3. In Table 3, a † indicates a significant difference of p<0.05 compared to the group to which a final concentration of 0.1% non-fermented material was added in a Dunnett test with n=3. As shown in Table 3 etc., an increase in hyaluronic acid production was observed by adding a specified amount of lactic acid bacteria fermentation product, compared to the control group etc.

[0059] (Test Example 2) Dryness Irritation Evaluation Test In this test example, a lactic acid bacteria fermentation product was added to human epidermal keratinocytes, and changes in the amounts of IL-1α and IL-6 produced due to dryness stimulation were evaluated.

[0060] (Preparation of human epidermal keratinocytes) Human epidermal keratinocytes were cultured at 5 × 10 4The cells were seeded at 1000 cells / mL onto a 24-well culture plate. The seeded cells were cultured in keratinocyte culture medium (1) under conditions of 5% CO2 and 37°C for 24 hours until the cells reached 75% confluence. The cells used in Test Example 2 were prepared by this culture.

[0061] (Addition of sample to epidermal keratinocytes) The prepared cells were cultured for 24 hours under the conditions of 5% CO2 and 37°C under the conditions shown in Table 4 below. After the culture, the supernatant was removed. After this removal, the following actions were performed. For the groups other than the control group listed in Table 4, dry stimulation (standing at 25°C) was performed for 5 minutes with the top lid of the 24-well culture plate open. For the control group shown in Table 4, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the 24-well culture plate was left to stand for 5 minutes at 25°C without opening the top lid. After the standing, the control group was cultured for 24 hours under the following conditions of 5% CO2 and 37°C.

[0062] For the dry stimulation group, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After the culture, the IL-1α production amount and IL-6 production amount in the collected supernatant were measured for the dry stimulation group and the control group. The IL-1α production amount was measured using a human IL1-alpha ELISA Kit (PGI, KE00123). The IL-6 production amount was measured using a human IL-6 Quantikine ELISA Kit (R&D, D6050). The measurement results are shown in Table 4. The measurement results shown in Table 4 show values ​​(relative values) shown from the average values ​​measured for each group (n=3) when the control group was set to 100. * in Table 4 indicates a significant difference of p<0.05 compared to the non-treated group in the Dunnett test for n=3. As shown in Table 4 etc., it was confirmed that the addition of the lactic acid bacteria fermentation product reduced the amount of IL-1α produced and the amount of IL-6 produced, compared to the non-addition group.

[0063] [Table 4]

[0064] (Test Example 3) Evaluation test of dry irritation to cells in an inflamed state In this test example, a lactic acid bacteria fermentation product was added to cells in an inflamed state, and the change in IL-1α production due to dryness stimulation was evaluated.

[0065] (Preparation of epidermal keratinocytes) Human epidermal keratinocytes were cultured at 5 × 10 4 The cells were seeded at 1000 cells / mL onto a 24-well culture plate. The seeded cells were cultured in keratinocyte culture medium (1) under conditions of 5% CO2 and 37°C for 24 hours until the cells reached 75% confluence. The cells used in Test Example 3 were prepared by this culture.

[0066] (Preparation of inflamed and non-inflamed cells) Using the prepared cells, cells in an inflammatory state and cells in a non-inflammatory state were prepared as follows.

[0067] The following two types of inflammatory cells were prepared. In Groups 1 and 5 shown in Table 5 below, inflammatory cell type 1 was used. In Groups 2 and 6 shown in Table 5 below, inflammatory cell type 2 was used. Inflammatory cells #1: The cells were seeded in a keratinocyte culture medium (2) containing TNF-α (final concentration 20 ng / mL) and INF-γ (final concentration 20 ng / mL). The seeded cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After the culture, the cells were recovered. The recovered cells were designated as inflammatory cells No. 1. Inflammatory cells #2: The cells were seeded in a keratinocyte culture medium (2) containing TNF-α (final concentration 20 ng / mL). The seeded cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After the culture, the cells were recovered. The recovered cells were designated as inflamed cells No. 2.

[0068] Non-inflammatory cells were prepared as follows. The cells were seeded in keratinocyte culture medium (2) containing purified water (30 μL). The seeded cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After the culture, the cells were recovered. The recovered cells were used as non-inflammatory cells. Non-inflammatory cells were used in Groups 3, 4, 7, and 8 shown in Table 5 below.

[0069] (Addition of sample to inflamed or non-inflamed cells) The cells were cultured for 24 hours under the conditions (addition of samples to cells) shown in Table 5 below, at 5% CO2 and 37°C. After the culture, the supernatant was removed. After this removal, the following actions were performed. Groups 1, 2, 3, 5, 6, and 7 shown in Table 5 were subjected to dry stimulation (standing at 25°C) for 5 minutes with the top lid of the 24-well culture plate open. For Groups 4 and 8 in Table 5, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the 24-well culture plate was left to stand for 5 minutes at 25°C without opening the top lid. After the standing, Groups 4 and 8 were cultured for 24 hours under the following conditions of 5% CO2 and 37°C.

[0070] After the dry stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After the culture, the amount of IL-1α produced in the collected supernatant was measured for the groups in which the dry stimulation was performed, group 4, and group 8. TM The amount of IL-1α produced was measured using an IL-1 alpha ELISA Kit (PGI, KE00268). The results are shown in Table 5. The results shown in Table 5 are the values ​​(relative values) shown from the average values ​​measured for each group (n=3) with Group 8 (control group) set at 100. * and other marks in Table 5 indicate the following, based on Student's t-test with n=3. As shown in Table 5 and other tables, a decrease in the amount of IL-1α produced was confirmed by the addition of lactic acid bacteria fermentation product. ·*: Comparison between Group 1 and Group 5 indicates a significant difference of p<0.05 compared to Group 5. ·**: Comparison between Group 2 and Group 6 indicates a significant difference of p<0.05 compared to Group 6. ·***: Comparison between group 3 and group 7 indicates a significant difference of p<0.05 compared with group 7. ·****: Comparison between Group 4 and Group 8, indicates a significant difference of p<0.05 compared with Group 8.

[0071] [Table 5]

[0072] (Test Example 4) Staphylococcus epidermidis growth test In this test example, a lactic acid bacteria fermentation product was added to cultured Staphylococcus epidermidis, and the effect on the proliferation of Staphylococcus epidermidis was evaluated.

[0073] (Staphylococcus epidermidis culture) Staphylococcus epidermidis was cultured (cultured at 37° C. for 20 hours) under the conditions shown in Table 6. The number of Staphylococcus epidermidis after the culture (the concentration of Staphylococcus epidermidis contained in the medium, CFU / mL) was calculated according to the following formula 1.

[0074] X = (C / S) x 10 3 ×N (Formula 1) The concentration of Staphylococcus epidermidis contained in the culture solution cultured for 20 hours is X (CFU / mL). Since the culture solution was diluted 1 / C, the concentration of Staphylococcus epidermidis in the diluted culture solution is expressed as X / C (CFU / mL). The diluted Staphylococcus epidermidis culture solution is expressed as S (μL), that is, S x 10 -3 Since the culture was inoculated onto a 10 mL plate, the number of Staphylococcus epidermidis in the inoculated culture was (X / C) × S × 10 -3 It is expressed as CFU. If N colonies are obtained on the plate after inoculation, it is (X / C) × S × 10 -3 =N.

[0075] The calculated results are shown in Table 6. In Table 6, * indicates a significant difference of p<0.05 in Student's t-test (n=3) between the number of cocci in the group containing 0.10% unfermented material (group 2) and the number of cocci in the group containing 0.10% lactic acid bacteria fermented product (group 1). As shown in Table 6, it was confirmed that the number of Staphylococcus epidermidis was higher with the addition of a lactic acid bacteria fermented product (group 1) than with the addition of an unfermented material (group 2).

[0076] [Table 6]

[0077] (Test Example 5) Staphylococcus aureus growth test Staphylococcus aureus was cultured (cultured at 37° C. for 20 hours) under the conditions shown in Table 7. The number of Staphylococcus aureus after the culture (concentration of Staphylococcus aureus contained in the medium, CFU / mL) was calculated according to the following formula 2.

[0078] X = (C / S) x 10 3 ×N (Formula 2) The concentration of Staphylococcus aureus in the culture solution cultured for 20 hours is X (CFU / mL). Since the culture solution was diluted 1 / C, the concentration of Staphylococcus aureus in the diluted culture solution is expressed as X / C (CFU / mL). The diluted Staphylococcus aureus culture solution is expressed as S (μL), that is, S x 10 -3 Since the culture was inoculated onto a 10 mL plate, the number of Staphylococcus aureus in the inoculated culture was (X / C) × S × 10 -3 It is expressed as CFU. If N colonies are obtained on the plate after inoculation, it is (X / C) × S × 10 -3 =N.

[0079] The calculated results are shown in Table 7. * in Table 7 indicates a significant difference of p<0.05 in Student's t-test (n=3) between the number of cocci in the group (group 2) to which the lactic acid bacteria fermentation product was not added and the number of cocci in the group (group 1) to which 0.10% of the lactic acid bacteria fermentation product was added. As shown in Table 7, it was confirmed that the number of Staphylococcus aureus was lower with the addition of the lactic acid bacteria fermentation product (group 1) compared to the group to which the lactic acid bacteria fermentation product was not added (group 2). From the results shown in Table 6 (results showing an increase in the number of Staphylococcus epidermidis due to the addition of the lactic acid bacteria fermentation product) and the results shown in Table 7 (results showing a decrease in the number of Staphylococcus aureus due to the addition of the lactic acid bacteria fermentation product), it is considered that the balance of the resident skin flora can be improved by adding a certain lactic acid bacteria fermentation product.

[0080] [Table 7]

[0081] (Test Example 6) Dryness Irritation Evaluation Test In this test example, a lactic acid bacteria fermentation product was added to human epidermal keratinocytes, and the amount of IL-1α receptor antagonist (IL-1RA) produced by dry stimulation was evaluated.

[0082] (Preparation of human epidermal keratinocytes) Human epidermal keratinocytes were cultured at 5 × 10 4 The cells were seeded at 1000 cells / mL onto a 24-well culture plate. The seeded cells were cultured in keratinocyte culture medium (1) under conditions of 5% CO2 and 37°C for 24 hours until the cells reached 75% confluence. The cells used in Test Example 6 were prepared by this culture.

[0083] (Addition of sample to epidermal keratinocytes) The prepared cells were cultured for 24 hours under the conditions of 5% CO2 and 37°C under the conditions shown in Table 8 below. After the culture, the supernatant was removed. After this removal, the following actions were performed.

[0084] Seven dry stimulations: In groups 1 and 5, the following actions were performed: Dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After the stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed.

[0085] Five dry stimulations: In groups 2 and 6, the following actions were performed: Dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After the stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed.

[0086] Three dry stimulations: In groups 3 and 7, the following actions were performed: Dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After the stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed. After this removal, dry stimulation (standing at 25°C) was performed for 5 minutes with the lid of the 24-well culture plate open. After this stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After this addition, the wells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After this culture, the supernatant was removed.

[0087] No dry irritation: The following actions were performed in Groups 4 and 8. 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the 24-well culture plate was left to stand for 5 minutes at 25°C without opening the top lid. After the standing, the control group was cultured for 24 hours under the following conditions: 5% CO2, 37°C.

[0088] For the group subjected to dry stimulation, 1 mL of keratinocyte culture medium (2) was added to each well. After the addition, the cells were cultured for 24 hours under conditions of 5% CO2 and 37°C. After the 24-hour culture, the amount of IL-1RA produced in the collected supernatant was measured for the group subjected to dry stimulation and the group without dry stimulation. The amount of IL-1RA produced was measured using an IL-1ra / IL-1F3, Human, ELISA Kit (R&D Systems, DRA00B). The results of the measurement are shown in Table 8. The results of the measurement shown in Table 8 are values ​​(relative values) shown from the average values ​​measured for each group (n=3) with Group 8 (control group) set at 100. * in Table 8 indicates a significant difference of p<0.05 compared to Group 5 in a Student's t-test of n=3. ** in Table 8 indicates a significant difference of p<0.05 compared to Group 6 in a Student's t-test of n=3. In Table 8, *** indicates a significant difference of p<0.05 compared to Group 7 in Student's t-test (n=3). As shown in Table 8, a decrease in the amount of IL-1RA produced was confirmed by the addition of the lactic acid bacteria fermentation product.

[0089] [Table 8]

[0090] (Test Example 7) Evaluation of skin condition improvement effect by human monitor test This monitoring study was conducted between January 2023 and March 2023, with 12 healthy men and women aged 20 to 69 years (average age: 44.6 years (average age for men: 42.2 years, average age for women: 45.4 years)) as subjects.

[0091] (Test Method) A lotion having the composition shown in Table 9 (Table 9 lotion) was applied to the left half of the face of the subject for a specified period, and a lotion shown in Table 10 (Table 10 lotion) was applied to the right half of the face of the subject for a specified period. This specified period was 4 weeks, twice a day (morning and evening). After 4 weeks, 42 mg of Table 9 lotion and 42 mg of Table 10 lotion were applied to each subject. Then, measurements (measurement of skin moisture content and skin moisture evaporation) were performed on day 0 before application of these lotions and 4 weeks after application of the lotions. The moisture content and the moisture evaporation (TEWL) of the skin were measured using a Corneometer (CM825, Courage+Khazaka). The skin texture density was measured using a skin moisture distribution and shape measurement device "MoistureMapMM100 (Courage+Khazaka)".

[0092] [Table 9]

[0093] [Table 10]

[0094] (Measurement results) The results shown below are calculated as the average value of the 12 subjects (average value of the measurements), and the value 0 days after the calculation (before the application) is set as 100 (relative value). The moisture content of the skin was measured after 4 weeks and was 103.36 in the group treated with lotion in Table 10, while it was 110.62 in the group treated with lotion in Table 9 (p<0.01 (Wilcoxon test) compared to the value of the group treated with lotion in Table 9 on day 0). The transepidermal water loss (TEWL) was measured. After 4 weeks, the group treated with lotion in Table 10 had a value of 102.35, while the group treated with lotion in Table 9 had a value of 109.80 (p<0.05 (Wilcoxon test) compared to the value of the group treated with lotion in Table 9 on day 0). The skin texture density was measured. After 4 weeks, the group treated with the lotion in Table 10 had a texture density of 92.47, while the group treated with the lotion in Table 9 had a texture density of 106.47 (p<0.05 (Wilcoxon test) compared to the value of the group treated with the lotion in Table 10 after 4 weeks). These measurement results confirmed that applying a lotion containing lactic acid bacteria fermentation products (lotion listed in Table 9) to the skin maintained the skin's moisture content, compared to applying a lotion not containing lactic acid bacteria fermentation products (lotion listed in Table 10).

[0095] (Test Example 8) Evaluation of nasolabial fold length by human monitor test This monitoring study was conducted from late January 2023 to early March 2023 on 12 healthy men and women aged 20 to 69 (those who answered in a pre-study questionnaire that "my skin condition changes easily depending on the season and / or I am concerned about rough or dry skin"; average age was 44.6 years (average age for men: 42.2 years, average age for women: 45.4 years)).

[0096] A lotion having the composition shown in Table 9 (Table 9 Lotion) was applied to the left half of the face of the subject for a specified period of time, and a lotion shown in Table 10 (Table 10 Lotion) was applied to the right half of the face of the subject for a specified period of time. This specified period was twice a day (morning and evening) for four weeks. After four weeks, 14 g of the Table 9 Lotion and 14 g of the Table 10 Lotion were applied to each subject. Measurements (measurement of nasolabial folds) were then taken on day 0 before application of these lotions and four weeks after application of the lotions. Antera3DTM The measurement (measurement of nasolabial folds) was performed using a Miravex (Miravex, Dublin, Ireland). The measurement site (nasolabial folds) is shown in Figure 1.

[0097] (Measurement results) The results shown below are the average value of the 12 subjects (average value of the measurements), and the value 0 days after the calculation (before application) is set to 100 (relative value). After 4 weeks, the value was 101.8 for the group applied with the lotion in Table 10, and 100.2 for the group applied with the lotion in Table 9.

[0098] Of the 12 people, 6 people were diagnosed with hay fever by a doctor (these 6 people did not receive medication for the prevention or treatment of hay fever), and the results are also shown below. The results shown below are the average value of the 6 people (average value of the measurement), and the value 0 days after the calculation (before the application) is set to 100 (relative value). After 4 weeks, the value of the Table 10 lotion application group was 103.5, while the value of the Table 9 lotion application group was 100.0 (p<0.05 in a paired t-test compared with the value of the Table 10 lotion application group after 4 weeks).

[0099] (Test Example 9) Evaluation of Stratum Corneum Lamellar Delamination in Human Monitor Tests The evaluation was carried out with reference to the method described in the literature (Journal of the Japanese Society of Cosmetic Science, Vol. 38, No. 2, p. 87-91, 2014, Evaluation and Study of Layered Peeling Using Image Analysis). This monitor test was conducted from late January 2023 to early March 2023 on 12 healthy men and women aged 20 to 69 years (those who answered "My skin condition changes easily depending on the season" in a questionnaire before the test, with an average age of 44.6 years (average age of men: 42.2 years, average age of women: 45.4 years)).

[0100] A lotion having the composition shown in Table 9 (Table 9 Lotion) was applied to the left half of the subject's face for a specified period of time, and a lotion shown in Table 10 (Table 10 Lotion) was applied to the right half of the subject's face for a specified period of time. This specified period was twice a day (morning and evening) for four weeks. After four weeks, 14 g of the Table 9 Lotion and 14 g of the Table 10 Lotion were applied to each subject.

[0101] (Analysis by tape stripping method) The stratum corneum of the cheek was sampled as follows: Using a keratin checker AST-01 (Nippon Ash Co., Ltd.), samples were taken twice, one from each side, before application (day 0) and four weeks after application, for a total of four samples per subject.

[0102] The stratum corneum was stained as follows. First, the stratum corneum checker to which the stratum corneum had been transferred was lightly washed in a water bath. After the washing, the checker was immersed for 10 minutes in a staining solution in which 1% gentian violet (Kanto Chemical) and 0.5% brilliant green (Merck) were dissolved. After the immersion, the checker was rinsed in a water bath and then air-dried.

[0103] After air drying (after the stratum corneum was dry), the stratum corneum was attached to a mount for a stratum corneum checker. The stratum corneum (stratum corneum cells) attached to the mount was photographed using a digital microscope VHX-5000 (Keyence). The color adjustments for photography were set to R583, G265, and B452, and the size was set to the standard "1600 x 1200", and the stratum corneum images were taken. In order to evaluate the size of the stratum corneum and the degree of delamination, a standard scale for each was prepared, and the stratum corneum images were evaluated visually. Figure 2 shows a micrograph of a specimen from one randomly selected person (a man in his 50s) out of the 12 people.

[0104] (Measurement results) As shown in the micrograph of FIG. 2, in the photograph taken four weeks later, it was confirmed that the group applied with the lotion shown in Table 9 tended to show less peeling of the layer compared to the group applied with the lotion shown in Table 10.

[0105] Exfoliation of keratinocytes means that the adhesive mechanism between epidermal keratinocytes weakens, causing old keratinocytes to peel off from the skin surface. Exfoliation is promoted by decomposing a protein called desmosome, which adheres keratinocytes to each other (J.Soc.Cosmet.Chem.Jpn., Mechanism of Adhesion and Exfoliation of the Stratum Corneum and its Role in Skin Care, p16-p26). However, when the activity of this proteolytic enzyme decreases, the decomposition of desmosomes is inhibited, making it difficult for keratin to peel off smoothly, and stratification occurs. This is thought to cause skin problems such as dullness, roughness, and uneven texture. Therefore, it is thought that promoting the exfoliation of keratinocytes and suppressing stratification will improve dullness and make the skin smoother. [Industrial Applicability]

[0106] The present invention can be used for external skin preparations and methods for producing the same.

Claims

1. A skin external preparation containing a lactic acid bacteria fermentation product of aloe vera leaves, The skin external preparation, wherein the lactic acid bacterium is Lactiplantibacillus plantarum TY-610 strain (accession number: NITEBP-03877).

2. The skin topical preparation according to claim 1 , which has hyaluronic acid production promoting activity.

3. The external skin preparation according to claim 1 for preventing and / or improving dry skin.

4. The external skin preparation according to claim 1 for improving the balance of the normal skin flora.

5. A method for producing a skin care product containing a lactic acid bacteria fermentation product produced from the juice of aloe vera leaves, comprising: The step of producing a lactic acid bacteria fermentation product comprises: The method for producing an external skin preparation includes a fermentation step of fermenting the juice with Lactiplantibacillus plantarum TY-610 strain (Accession No.: NITEBP-03877).

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