Cosmetic composition for alleviating hair loss symptoms and improving skin elasticity comprising a Bacillus subtilis Bs5C fermentation product and a complex marine extract

Abstract

The present invention relates to a cosmetic composition comprising, as active ingredients, a concentrated fermented supernatant obtained from a fermentation culture of the Bacillus subtilis Bs5C strain (accession number: KCTC 18585P), a complex marine extract comprising microalgae extract and seaweed extract, Ectoine, and Spermidine. The microalgae extract may include Chlorella vulgaris extract, plankton extract, and Spirulina platensis extract, and the seaweed extract may include Laminaria japonica extract and Sargassum fulvellum extract. The cosmetic composition of the present invention can provide effects such as alleviating hair loss, suppressing scalp inflammation, and improving skin elasticity, and can be usefully used to comprehensively improve the condition of the skin and scalp.

Description

Technology Field

[0001] The present invention relates to a cosmetic composition, specifically, Bacillus subtilis ( Bacillus subtilis The present invention relates to a cosmetic composition containing a fermented product derived from the Bs5C strain and a complex marine extract that can provide effects of alleviating hair loss, suppressing scalp inflammation, and improving skin elasticity. Background Technology

[0002] Recently, with the progression of population aging and changes in living environments, interest in skin and hair-related issues such as hair loss, scalp troubles, and decreased skin elasticity has been increasing. In particular, as cases of deteriorating scalp and skin health due to various factors—including external environmental pollution, UV exposure, stress, and changes in lifestyle—are on the rise, the industrial demand for functional cosmetics to address these issues is continuously expanding. Consequently, there is active development of cosmetic materials and compositions capable of simultaneously improving the condition of the skin and scalp, and further enhancing the self-regenerative and restorative capabilities of skin and scalp cells to improve long-term health status, or "skin longevity."

[0003] Meanwhile, the cosmetics industry has recently been focusing on the development of technologies utilizing bio-derived materials based on microbial fermentation, natural extracts derived from marine biological resources, and bioactive substances related to skin protection or cell activation. In particular, marine-derived materials and microbial-derived ingredients have been reported to exhibit various physiological activities, such as antioxidant, anti-inflammatory, moisturizing, and cell activation promotion, leading to ongoing research to apply them to cosmetic compositions.

[0004] However, cosmetic compositions based on conventional technology often focus on specific functions or single mechanisms of action, which limits their ability to effectively and simultaneously address complex skin and scalp issues such as hair loss, scalp condition improvement, and skin elasticity enhancement. Furthermore, simply mixing various functional materials makes it difficult to reliably secure the expected level of biological activity resulting from the interactions between the ingredients; therefore, there is a need for the development of new cosmetic compositions capable of providing enhanced functionality. Prior art literature

[0005] Republic of Korea Published Patent Application No. 10-2021-0155987 The problem to be solved

[0006] The present invention aims to provide a cosmetic composition that overcomes the limitations of conventional cosmetic compositions, which are centered on a single function and have difficulty simultaneously improving various skin and scalp problems, and provides a complex effect—such as alleviating hair loss, suppressing scalp inflammation, and improving skin elasticity—in a stable and continuous manner, while enhancing the self-regenerative and restorative power of skin and scalp cells to contribute to skin longevity.

[0007] In addition, the present invention has another objective of providing a cosmetic composition that can contribute to improving scalp health and skin elasticity by providing a composition capable of simultaneously exhibiting physiological activities related to enhancing the activity of skin and scalp cells, inhibiting inflammatory responses, and improving skin structure.

[0008] However, the problems that this invention seeks to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by a person skilled in the art from the description below. means of solving the problem

[0009] To achieve the above objectives, the present invention (Bacillus subtilis ( Bacillus subtilisA cosmetic composition comprising, as active ingredients: a concentrated fermented supernatant obtained from a fermentation culture of the Bs5C strain (accession number: KCTC 18585P); a complex marine extract comprising a microalgae extract and a seaweed extract; Ectoine; and Spermidine, wherein the microalgae extract is Chlorella vulgaris ( Chlorella vulgaris ) extract, plankton extract and Spirulina platensis( Spirulina platensis It includes an extract, and the seaweed extract is kelp ( Laminaria japonica ) extract and Sargassum ( Sargassum fulvellum A cosmetic composition is provided that includes an extract, wherein the cosmetic composition is intended for alleviating hair loss, suppressing scalp inflammation, or improving skin elasticity, and the weight ratio of the microalgae extract to the seaweed extract is 3:2.

[0010] In one embodiment of the present invention, the microalgae extract and the seaweed extract may act complementarily to simultaneously exhibit cell activity enhancement, antioxidant, and anti-inflammatory effects, and for example, it may be most preferable to include the microalgae extract and the seaweed extract in a weight ratio of 3:2. This composition ratio may exhibit excellent effects in terms of promoting the proliferation of dermal papilla cells and inhibiting inflammatory responses.

[0011] On the other hand, if the weight ratio of the microalgae extract and the seaweed extract deviates from the above range, the balance between the physiologically active components of each ingredient is impaired, which may result in a relative decrease in the cell activity-promoting effect or the anti-inflammatory effect. For example, if the proportion of the microalgae extract is excessively high, antioxidant and cell activity-promoting effects may be observed, but the anti-inflammatory and scalp environment improvement effects caused by seaweed-derived polysaccharides and mineral components may not be sufficiently manifested; conversely, if the proportion of the seaweed extract is excessively high, anti-inflammatory effects may be observed, but the cell activity and hair papilla cell proliferation-promoting effects caused by microalgae-derived nutrients may be relatively reduced.

[0012] In one embodiment of the present invention, the cosmetic composition may further comprise 0.005% to 0.015% by weight of fucoxanthin relative to the total weight of the composition. Fucoxanthin is a bioactive substance of the carotenoid class that is abundant in brown algae, and can exhibit excellent antioxidant activity and anti-inflammatory effects, and can help protect skin and scalp cells from oxidative stress. In addition, fucoxanthin can contribute to alleviating scalp inflammatory responses by inhibiting the generation of reactive oxygen species (ROS) and reducing the expression of inflammatory cytokines, and can also help improve skin elasticity by promoting collagen synthesis in skin cells and inhibiting the expression of collagenases. Furthermore, the fucoxanthin is the Bacillus subtilis of the present invention ( Bacillus subtilis When used in combination with a fermented supernatant concentrate derived from the Bs5C strain and a complex marine extract, the antioxidant and anti-inflammatory effects work complementarily to maintain the activity of hair papilla cells and improve the scalp environment, which can have a more favorable effect. Effects of the invention

[0013] The cosmetic composition according to the embodiments of the present invention can provide an effect of alleviating hair loss by improving the hair growth environment through promoting the proliferation of hair papilla cells and increasing the expression of growth factors, and can also contribute to enhancing the self-regenerating and restorative capabilities of scalp cells. In addition, it can exhibit an effect of improving the condition of the scalp by alleviating the inflammatory response of the scalp through the inhibition of the expression of inflammatory cytokines.

[0014] Furthermore, the cosmetic composition according to the embodiments of the present invention can provide an effect of improving skin elasticity by enhancing the activity of skin cells, promoting collagen production, and inhibiting the expression of collagen-degrading enzymes. In addition, by strengthening the self-regenerating and restorative power of skin cells, it can contribute to maintaining long-term skin health, i.e., improving skin longevity. Accordingly, the cosmetic composition of the present invention can stably provide a complex improvement effect on the scalp and skin. Specific details for implementing the invention

[0015] The present invention relates to Bacillus subtilis ( Bacillus subtilis A cosmetic composition comprising, as active ingredients: a concentrated fermented supernatant obtained from a fermentation culture of the Bs5C strain (accession number: KCTC 18585P); a complex marine extract comprising a microalgae extract and a seaweed extract; Ectoine; and Spermidine, wherein the microalgae extract is Chlorella vulgaris ( Chlorella vulgaris ) extract, plankton extract and Spirulina platensis( Spirulina platensis It includes an extract, and the seaweed extract is kelp ( Laminaria japonica ) extract and Sargassum ( Sargassum fulvellum A cosmetic composition is provided that includes an extract, wherein the cosmetic composition is intended for alleviating hair loss, suppressing scalp inflammation, or improving skin elasticity, and the weight ratio of the microalgae extract to the seaweed extract is 3:2.

[0017] In one embodiment of the present invention, the fermented supernatant concentrate is Bacillus subtilis ( Bacillus subtilis It may be obtained by concentrating the supernatant after removing cells from the fermentation culture medium of the Bs5C strain. The fermentation supernatant may contain enzymes, peptides, amino acids, and various metabolites generated during the fermentation process, and these bioactive components may contribute to promoting cell metabolism and improving the scalp environment. In addition, the fermentation supernatant concentrate may help enhance the activity of dermal papilla cells, increase the expression of cell growth factors, and stabilize the scalp environment.

[0018] In one embodiment of the present invention, the above Bacillus subtilis ( Bacillus subtilis The Bs5C strain may be a microorganism capable of producing various physiologically active metabolites during the fermentation process as well as producing proteolytic enzymes.

[0019] In one embodiment of the present invention, the above Bacillus subtilis ( Bacillus subtilis The enzyme derived from the supernatant of the Bs5C strain fermentation may include a protease, for example, a neutral protease, a serine protease, a metalloprotease, or a combination thereof.

[0021] In one embodiment of the present invention, the microalgae extract is Chlorella vulgaris ( Chlorella vulgaris ), plankton and Spirulina platensis ( Spirulina platensis It may include an extract obtained from ). The microalgae are rich in proteins, amino acids, vitamins, minerals, and antioxidant components, which can help promote the activity of skin and scalp cells. For example, the microalgae extract may have the effect of promoting the proliferation of dermal papilla cells and activating the metabolism of skin cells.

[0022] In one embodiment of the present invention, the seaweed extract is kelp ( Laminaria japonica ) and Sargassum ( Sargassum fulvellum It may include an extract obtained from ). The seaweed contains fucoidan, alginic acid, polyphenols, and various minerals, which may exhibit anti-inflammatory and antioxidant activities, and may help improve the scalp environment and maintain skin elasticity.

[0024] In one embodiment of the present invention, the fermented supernatant concentrate may be included in an amount of 0.7% to 1.3% by weight relative to the total weight of the composition. If the content is lower than the above range, the concentration of the physiologically active substance contained in the fermented supernatant is insufficient, so the effect of promoting dermal papilla cell activity or increasing growth factor expression may not be sufficiently manifested. On the other hand, if it exceeds the above range, the concentration of the proteinaceous component in the composition increases, which may lead to a decrease in formulation stability or a tendency for the effect of increasing cell activity to become saturated.

[0025] In one embodiment of the present invention, the complex marine extract may be included in an amount of 0.3% to 0.7% by weight relative to the total weight of the composition. If the content is lower than the above range, the concentration of antioxidant and anti-inflammatory active components derived from microalgae and seaweed may not be sufficient, and thus the effect of enhancing cell activity or inhibiting inflammation may not be sufficiently exhibited. On the other hand, if it exceeds the above range, an increase in formulation viscosity or a decrease in physical stability may occur due to the polysaccharide components derived from seaweed.

[0027] In one embodiment of the present invention, the ectoine is a low-molecular-weight protective substance derived from microorganisms in extreme environments that can exhibit cell protection and moisturizing effects, and can play a role in stabilizing the skin barrier and protecting the skin from external environmental stress. The ectoine may be included in an amount of 0.05% to 0.15% by weight relative to the total weight of the composition. If the content is lower than the above range, the cell protection effect, moisturizing effect, or skin barrier stabilization effect by the ectoine may not be sufficiently expressed. On the other hand, if it exceeds the above range, formulation stability may be reduced due to an increase in solute concentration within the composition, or the effect of increasing efficacy may tend to become saturated due to interactions between ingredients.

[0028] In one embodiment of the present invention, the spermidine is a polyamine-based bioactive substance that may be involved in cell proliferation and the maintenance of cell homeostasis, and may help enhance the activity of hair follicle cells. The spermidine may be included in an amount of 0.001% to 0.003% by weight relative to the total weight of the composition. If the content is lower than the above range, the effect of promoting cell proliferation or maintaining cell homeostasis may not be sufficiently expressed. On the other hand, if it exceeds the above range, the solute concentration in the composition increases, which may cause changes in the physical properties of the formulation or a tendency for the increase in effect to saturate.

[0030] In one embodiment of the present invention, Bacillus subtilis included in the cosmetic composition ( Bacillus subtilisThe fermented supernatant concentrate derived from the Bs5C strain, a complex marine extract containing microalgae and seaweed, ectoin, and spermidine act complementarily to exhibit complex improvement effects on the scalp and skin. For example, the fermented supernatant concentrate can contribute to enhancing cell activity and promoting protein synthesis, while the complex marine extract can play a role in stabilizing the scalp and skin environment through antioxidant and anti-inflammatory actions. Additionally, ectoin and spermidine can help maintain cellular physiological functions by contributing to cell protection and stress relief. When these ingredients are included together, effects such as enhanced dermal papilla cell activity, alleviation of inflammatory responses, and improvement of skin structure may manifest more efficiently; consequently, they can simultaneously provide complex effects such as hair loss relief, suppression of scalp inflammation, and improvement of skin elasticity.

[0031] In one embodiment of the present invention, the cosmetic composition may be a composite biomaterial-based composition containing both a bioactive ingredient derived from microorganisms and a bioactive ingredient derived from marine sources. For example, Bacillus subtilis ( Bacillus subtilis The fermented supernatant derived from the Bs5C strain can act as a bioactive component derived from microorganisms, and a complex marine extract containing microalgae and seaweed extracts can act as a bioactive component derived from marine organisms. This combination of microorganism-derived and marine-derived materials can simultaneously provide effects such as enhanced cell activity, anti-inflammatory action, and improvement of skin structure through different biological mechanisms, thereby exhibiting complex improvement effects on the scalp and skin.

[0032] In one embodiment of the present invention, the cosmetic composition may be used alone or in combination with other cosmetic ingredients. For example, the other cosmetic ingredients may include moisturizers, emulsifiers, surfactants, thickeners, skin conditioning agents, antioxidants, vitamins, plant extracts, fragrances, colorants, preservatives, sunscreens, pH adjusters, or combinations thereof.

[0033] More specifically, the moisturizer may include glycerin, butylene glycol, propylene glycol, hyaluronic acid, or panthenol, and the thickening agent may include carbomer, carbopol, xanthan gum, hydroxyethylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, or a combination thereof. Additionally, the pH adjuster may include tromethamine, triethanolamine, sodium hydroxide, potassium hydroxide, citric acid, lactic acid, or a combination thereof. Meanwhile, the preservative may include 1,2-hexanediol, phenoxyethanol, ethylhexylglycerin, benzyl alcohol, sodium benzoate, potassium sorbate, or a combination thereof. Meanwhile, the above antioxidant may include tocopherol, ascorbic acid, green tea extract, or coenzyme Q10, etc., and the above plant extract may include aloe, green tea, centella asiatica, licorice, or rosemary extract, etc. The above cosmetic composition may be used in combination with these ingredients to improve the stability, moisturizing properties, or skin conditioning effect of the cosmetic formulation.

[0035] The present invention relates to Bacillus subtilis ( Bacillus subtilis A cosmetic composition comprising, as active ingredients: a concentrated fermented supernatant obtained from a fermentation culture of the Bs5C strain (accession number: KCTC 18585P); a complex marine extract comprising a microalgae extract and a seaweed extract; Ectoine; and Spermidine, wherein the microalgae extract is Chlorella vulgaris ( Chlorella vulgaris ) extract, plankton extract and Spirulina platensis( Spirulina platensis It includes an extract, and the seaweed extract is kelp ( Laminaria japonica ) extract and Sargassum ( Sargassum fulvellum A method for alleviating hair loss, improving scalp condition, or improving skin elasticity is provided, comprising the step of applying a cosmetic composition to the skin or scalp of an individual, wherein the cosmetic composition is for alleviating hair loss, suppressing scalp inflammation, or improving skin elasticity, and the weight ratio of the microalgae extract to the seaweed extract is 3:2.

[0036] In one embodiment of the present invention, the method may include the step of applying the cosmetic composition directly to the scalp or skin of an individual. The individual may be, for example, a human, and may be, in particular, a subject who exhibits symptoms of hair loss or is concerned about scalp inflammation or a decrease in skin elasticity. The cosmetic composition may be applied to the scalp or skin once or several times a day. For example, the cosmetic composition may be prepared in the form of a scalp tonic, hair serum, ampoule, lotion, cream, or essence and applied to the scalp or skin. In addition, it may be prepared in various formulations such as a scalp essence, hair mist, hair treatment, hair conditioner, scalp scaler, hair pack, mask pack, gel, spray, or foam. Furthermore, the cosmetic composition may be utilized in various forms, not only as a product applied directly to the scalp and skin, but also as a functional cosmetic for scalp care or a cosmetic for skin care.

[0037] By applying the cosmetic composition to the scalp or skin by the above method, it may help increase the activity of dermal papilla cells and alleviate scalp inflammatory reactions, and at the same time promote collagen synthesis in skin cells to demonstrate an effect of improving skin elasticity. In addition, the above method may be a non-therapeutic method for cosmetic purposes.

[0039] The present invention relates to Bacillus subtilis ( Bacillus subtilis A cosmetic composition comprising, as active ingredients: a concentrated fermented supernatant obtained from a fermentation culture of the Bs5C strain (accession number: KCTC 18585P); a complex marine extract comprising a microalgae extract and a seaweed extract; Ectoine; and Spermidine, wherein the microalgae extract is Chlorella vulgaris ( Chlorella vulgaris ) extract, plankton extract and Spirulina platensis( Spirulina platensis It includes an extract, and the seaweed extract is kelp ( Laminaria japonica ) extract and Sargassum ( Sargassum fulvellum A cosmetic composition comprising an extract, wherein the cosmetic composition is intended for alleviating hair loss, suppressing scalp inflammation, or improving skin elasticity, and wherein the weight ratio of the microalgae extract to the seaweed extract is 3:2, is provided for use in manufacturing a cosmetic or quasi-drug for alleviating hair loss, improving scalp condition, or improving skin elasticity.

[0040] In one embodiment of the present invention, the cosmetic or quasi-drug may be manufactured for the purpose of alleviating hair loss, improving scalp condition, or improving skin elasticity. The cosmetic or quasi-drug may be manufactured in various formulations applicable to the scalp or skin, for example, in the form of a scalp tonic, hair essence, hair serum, shampoo, conditioner, lotion, cream, ampoule, or serum. The cosmetic or quasi-drug may exhibit scalp environment improvement, anti-inflammatory action, and antioxidant action through the combined action of the fermented supernatant concentrate, complex marine extract, ectoin, and spermidine, thereby helping to improve the hair growth environment and maintain skin elasticity. Additionally, the use may be for non-therapeutic cosmetic purposes.

[0042] Hereinafter, the structure of the present invention and the resulting effects are to be explained in more detail through specific embodiments and comparative examples. However, these embodiments are intended to explain the present invention more specifically, and the scope of the present invention is not limited to these embodiments.

[0044] [Example]

[0045] Examples 1 and 2 were prepared according to the weights in Table 1 below:

[0046] [Table 1]

[0047]

[0048] Example 1. Preparation of a cosmetic composition for alleviating hair loss, suppressing scalp inflammation, or improving skin elasticity

[0049] (1) Bacillus subtilis ( Bacillus subtilis Preparation of Bs5C strain fermentation supernatant concentrate

[0050] Bacillus subtilis Bacillus subtilis The Bs5C strain (KCTC 18585P) was inoculated into LB liquid medium and pre-cultured at 30°C for 24 hours. 5 mL of the pre-culture solution was then inoculated into 1 L of fermentation medium (10 g peptone, 10 g yeast extract, 10 g glucose, 40.5 g MgSO4, 20.5 g CaCl2, 42 g K2HPO4, 40.5 g KH2PO4, remainder purified water) and cultured with shaking at 180 rpm at 30°C for 72 hours. After cultivation was complete, the culture medium was centrifuged at 8,000 rpm for 20 minutes to remove the cells, and the culture supernatant was recovered. The recovered supernatant was filtered through a 0.45 μm membrane filter and concentrated using a vacuum concentrator to obtain Bacillus subtilis ( Bacillus subtilis A concentrated supernatant of Bs5C strain fermentation was prepared.

[0051] (2) Preparation of microalgae extract

[0052] Chlorella vulgaris powder, plankton powder, and Spirulina platensis powder were each dispersed in purified water. Cellulase was added to Chlorella, and lysozyme was added to plankton and Spirulina, and the mixture was stirred at 50°C for 3 hours to perform a cell wall decomposition reaction. Subsequently, ultrasonic treatment was performed for 30 minutes using a 30 kHz ultrasonic disperser, and low-temperature extraction was carried out at 40°C for 6 hours. The extracts were filtered and freeze-dried to obtain Chlorella vulgaris extract, plankton extract, and Spirulina platensis extract, which were then mixed in equal weights to prepare microalgae extracts.

[0053] (3) Preparation of seaweed extract

[0054] Dried kelp and Sargassum were washed with purified water, cut into pieces approximately 2 cm in size, and steamed at 75°C for 20 minutes. Subsequently, primary drying was performed at 70°C for 2 hours, followed by secondary drying at 55°C for 8 hours. The dried raw materials were frozen at -45°C for 4 hours and aged at room temperature for 6 hours. The aged raw materials were then ground into powder, purified water was added, and reflux extraction was performed at 55°C. The extracts were filtered and concentrated under reduced pressure to obtain kelp extract and Sargassum extract, which were then mixed in equal weights to prepare the seaweed extract.

[0055] (4) Preparation of cosmetic composition

[0056] After adding 50 g of glycerin and 30 g of butylene glycol to purified water and dissolving them, the Bacillus subtilis prepared in (1) above ( Bacillus subtilis10 g of the Bs5C strain fermentation supernatant concentrate, 3 g of the microalgae extract prepared in (2), and 2 g of the seaweed extract prepared in (3) were added sequentially and stirred. Then, 1 g of Ectoine and 0.02 g of Spermidine were added and completely dissolved, followed by the addition of 2 g of the thickening agent Carbomer and the neutralization with 1 g of Tromethamine to adjust the viscosity, and the addition of 5 g of 1,2-Hexanediol as a preservative to be uniformly mixed to prepare a cosmetic composition for alleviating hair loss and improving the scalp.

[0057] Example 2. Additional addition of fucoxanthin

[0058] A cosmetic composition for alleviating hair loss and improving the scalp was prepared in the same manner as in Example 1, except that 0.10 g of fucoxanthin was additionally added in step (4) of Example 1.

[0060] Comparative Examples 1 to 10 were prepared according to the weights in Table 2 below:

[0061] [Table 2]

[0062]

[0063] Comparative Examples 1 and 2. Low and high content of Bs5C strain fermentation supernatant concentrate

[0064] Comparative Examples 1 and 2 are Bacillus subtilis ( Bacillus subtilis A cosmetic composition was prepared in the same manner as in Example 1, except that the Bs5C strain fermentation supernatant concentrate was included in amounts of 5 g (0.5 wt%) and 15 g (1.5 wt%), respectively.

[0065] Comparative Examples 3 and 4. Low and high content of complex marine extract

[0066] Comparative Examples 3 and 4 were prepared using the same method as Example 1, except that the weight ratio of the microalgae extract and the seaweed extract was maintained at 3:2, while the total weight of the two extracts was adjusted to 2 g (0.2 wt%) and 8 g (0.8 wt%), respectively.

[0067] Comparative Examples 5 to 7. Change in weight ratio of microalgae extract and seaweed extract

[0068] Comparative Examples 5 to 7 were prepared using the same method as Example 1, except that the total weight of the microalgae extract and seaweed extract was maintained at 5 g, while the weight ratio of each extract was adjusted to 1:1, 4:1, and 1:4, respectively.

[0069] Comparative Examples 8 and 9. Low and high content of echtoin

[0070] Comparative Examples 8 and 9 were prepared using the same method as Example 1, except that they contained 0.3 g (0.03 wt%) and 1.8 g (0.18 wt%) of echtoin, respectively.

[0071] Comparative Example 10. No addition of echtoin and spermidine

[0072] Comparative Example 10 was prepared by the same method as Example 1, except that echtoin and spermidine were not added.

[0074] Experimental Example 1. Evaluation of hair growth promotion effects through dermal papilla cell proliferation and growth factor expression

[0075] (1) Experimental method

[0076] In order to evaluate the effect of cosmetic compositions according to Example 1, Example 2, and Comparative Examples 1 to 10 on the activity of dermal papilla cells, which play an important role in hair growth, cell proliferation and growth factor expression tests were performed using human dermal papilla cells (HDPC).

[0077] Human dermal papilla cells were cultured in DMEM medium (containing 10% FBS and 1% penicillin-streptomycin) at 37°C under 5% CO2 conditions, and the cells were placed in a 96-well plate at a ratio of 1 × 10⁶ 4After dispensing at a density of cells / well, the cells were stabilized for 24 hours. Subsequently, each well was treated with the cosmetic compositions prepared in Example 1, Example 2 and Comparative Examples 1 to 10 at the same concentration and cultured for 48 hours. Cell proliferation rate was measured using an MTT assay; an MTT solution (5 mg / mL) was added to each well and reacted for 4 hours, after which the resulting formazan was dissolved in DMSO and the absorbance was measured at 570 nm.

[0078] In addition, the supernatant of cells cultured under the same conditions was collected, and the expression levels of VEGF (Vascular Endothelial Growth Factor) and IGF-1 (Insulin-like Growth Factor-1) were measured using an ELISA kit. Cell proliferation rate was calculated as a relative proliferation rate (%) with the control group, which was not treated with the sample, set as 100%, and the expression levels of VEGF and IGF-1 were converted into relative values ​​(%) compared to the control group.

[0079] (2) Experimental results

[0080] [Table 3]

[0081]

[0082] Examples 1 and 2 showed a significantly increased dermal papilla cell proliferation rate compared to the control group and all comparative examples, and the expression levels of VEGF and IGF-1 were also the highest. In particular, Example 2 was confirmed to have superior dermal papilla cell proliferation rate, VEGF expression, and IGF-1 expression levels compared to Example 1. On the other hand, Comparative Examples 1 to 10 all showed lower values ​​compared to Examples 1 and 2.

[0084] Experimental Example 2. Evaluation of scalp improvement effects through inflammation suppression

[0085] (1) Experimental method

[0086] In order to evaluate the effect of cosmetic compositions according to Example 1, Example 2, and Comparative Examples 1 to 10 on scalp inflammatory response, an inflammatory cytokine expression test using human keratinocytes (HaCaT keratinocytes) was performed.

[0087] HaCaT cells were cultured in DMEM medium (containing 10% FBS and 1% penicillin-streptomycin) at 37°C under 5% CO2 conditions, and the cells were placed in a 96-well plate at a ratio of 1 × 10⁶ 4 After seeding at a density of cells / well, the cells were stabilized for 24 hours. Subsequently, each well was treated with the cosmetic compositions prepared in Example 1, Example 2, and Comparative Examples 1 to 10 at the same concentration and pretreated for 2 hours. Then, LPS (Lipopolysaccharide) was applied to induce an inflammatory response, and the cells were cultured for 24 hours. After culture, the cell culture supernatant was collected, and the expression levels of the inflammatory cytokines IL-6 (Interleukin-6) and TNF-α (Tumor Necrosis Factor-α) were measured using an ELISA kit. The expression levels of each cytokine were calculated as a relative expression rate (%) based on the inflammation-induced group treated only with LPS as 100%.

[0088] (2) Experimental results

[0089] [Table 4]

[0090]

[0091] In Examples 1 and 2, the expression of inflammatory cytokines IL-6 and TNF-α was significantly reduced. In particular, Example 2 showed a lower expression rate than Example 1, demonstrating the most superior anti-inflammatory effect. On the other hand, Comparative Examples 1 to 10 showed higher expression rates of inflammatory cytokines than in Examples 1 and 2.

[0093] Experimental Example 3. Evaluation of skin improvement effects through the promotion of collagen production

[0094] (1) Experimental method

[0095] To evaluate the effects of cosmetic compositions according to Example 1, Example 2, and Comparative Examples 1 to 10 on skin elasticity and skin improvement, a collagen production and degradation enzyme expression test was performed using human dermal fibroblasts.

[0096] Human dermal fibroblasts were cultured in DMEM medium (containing 10% FBS and 1% penicillin-streptomycin) at 37°C under 5% CO2 conditions, and the cells were plated in a 96-well plate at a ratio of 1 × 10⁶ 4 After dispensing at a density of cells / well, the cells were stabilized for 24 hours. Subsequently, each well was treated with the cosmetic compositions prepared in Example 1, Example 2, and Comparative Examples 1 to 10 at the same concentration and cultured for 48 hours. After culture, the cell culture supernatant was collected, and the production of Pro-collagen type I was measured using an ELISA kit, while simultaneously measuring the expression level of MMP-1 (Matrix Metalloproteinase-1), a collagenase. The production of Pro-collagen and the expression level of MMP-1 were calculated as relative expression rates (%) based on the control group as 100%.

[0097] (2) Experimental results

[0098] [Table 5]

[0099]

[0100] Examples 1 and 2 showed a significant increase in the production of pro-collagen compared to the control group and comparative examples, while simultaneously showing a decrease in the expression of MMP-1, a collagen-degrading enzyme. In particular, Example 2 exhibited the most superior skin improvement effect, showing a higher collagen production effect and lower MMP-1 expression than Example 1. On the other hand, Comparative Examples 1 to 10 showed a relatively lower collagen production effect and a higher MMP-1 expression rate compared to the examples.

[0102] The various embodiments described above may be embodied in other specific forms without departing from the technical idea and essential features. Accordingly, the above detailed description should not be interpreted restrictively in all respects but should be considered exemplary. The scope of the various embodiments shall be determined by a reasonable interpretation of the appended claims, and all modifications within the equivalent scope of the various embodiments are included within the scope of the various embodiments. Furthermore, embodiments may be constructed by combining claims that do not have an explicit citation relationship in the patent claims, or they may be included as new claims through amendments made after filing.

Claims

Claim 1 Bacillus subtilis with accession number KCTC 18585P ( Bacillus subtilis A cosmetic composition comprising a concentrated fermentation supernatant obtained from a fermentation culture medium of a Bs5C strain; a complex marine extract comprising a microalgae extract and a seaweed extract; Ectoine; and Spermidine as active ingredients, wherein the microalgae extract is Chlorella vulgaris ( Chlorella vulgaris ) extract, plankton extract and Spirulina platensis( Spirulina platensis It includes an extract, and the seaweed extract is kelp ( Laminaria japonica ) extract and Sargassum ( Sargassum fulvellum A cosmetic composition comprising an extract, wherein the cosmetic composition is intended for alleviating hair loss, suppressing scalp inflammation, or improving skin elasticity, and wherein the weight ratio of the microalgae extract to the seaweed extract is 3:

2. Claim 2 A cosmetic composition according to claim 1, wherein the cosmetic composition further comprises 0.005% to 0.015% by weight of fucoxanthin based on the total weight of the composition. Claim 3 A cosmetic composition according to claim 1, wherein the fermented supernatant concentrate is included in an amount of 0.7% to 1.3% by weight relative to the total weight of the composition, and the complex marine extract is included in an amount of 0.3% to 0.7% by weight relative to the total weight of the composition. Claim 4 A cosmetic composition according to claim 1, wherein the echtoin is included in an amount of 0.05% to 0.15% by weight relative to the total weight of the composition, and the spermidine is included in an amount of 0.001% to 0.003% by weight relative to the total weight of the composition.

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