CN115120550A

CN115120550A - Hair cosmetic composition comprising a culture of streptococci providing beneficial effects on the hair

Info

Publication number: CN115120550A
Application number: CN202111152017.5A
Authority: CN
Inventors: 朴慧琳; 金珍英; 李康爀; 申松锡
Original assignee: Hyundai Bioland Co Ltd
Current assignee: Hyundai Bioland Co Ltd
Priority date: 2021-03-26
Filing date: 2021-09-29
Publication date: 2022-09-30
Also published as: KR102315133B1

Abstract

The present invention relates to a hair cosmetic composition comprising a culture solution of streptococcus cultured in a manner that hyaluronic acid is not formed, and more particularly, to a hair cosmetic composition for preventing hair loss and hair growth comprising a culture solution of streptococcus strain cultured under acidic pH conditions in a manner that hyaluronic acid is not formed as an effective ingredient, and a culturing method thereof. According to the present invention, the effects of hair loss alleviation, hair growth, hair root strengthening and hair coating of a culture solution of Streptococcus cultured under acidic pH conditions in such a manner that hyaluronic acid is not formed are better than those of a culture solution of Streptococcus cultured under neutral pH conditions in such a manner that hyaluronic acid is formed in the past. Therefore, the culture solution of Streptococcus cultured according to the present invention can be effectively used as a raw material for a hair cosmetic composition for preventing alopecia and hair growth.

Description

Hair cosmetic composition comprising a culture of streptococci providing beneficial effects on the hair

Technical Field

The present invention relates to a hair cosmetic composition comprising a culture solution of streptococcus cultured in a manner that hyaluronic acid is not formed, and more particularly, to a hair cosmetic composition for preventing hair loss and hair growth comprising a culture solution of streptococcus strain cultured under acidic pH conditions in a manner that hyaluronic acid is not formed as an effective ingredient, and a culturing method thereof.

Background

In modern society, the number of people who feel troubled by hair loss is increasing, and the age group thereof is also gradually becoming lower. For these reasons, alopecia is currently a major social problem in modern society where beauty is important.

The main causes of alopecia include a functional disorder of hair-forming cells, excessive secretion of male hormones, excessive secretion of sebum, excessive production of dandruff, and insufficient nutrient intake. These problems are not only caused by hereditary and aging phenomena, but also are caused by a combination of dietary habits, mental stress, hygiene, and the like, but the clear cause of alopecia has not been clarified.

Recently, there is a growing demand for women or young people who use anti-hair loss products in their daily lives from the viewpoint of care, and in response to such a trend, shampoo compositions which are used daily in a usual manner contain active ingredients contributing to the anti-hair loss to prevent hair loss.

Currently, typical hair loss preventive agents include finasteride, minoxidil, and the like, which are approved by the FDA in the united states. Although these drugs have clinically significant effects, they have disadvantages such as not only side effects such as sexual dysfunction and skin irritation, but also female unavailability or limitation. Researches for developing a natural substance or a culture of a microorganism usable as a raw material which can improve safety of the disadvantages of these drugs without being restricted by sex are actively being conducted recently.

Hair cosmetic compositions for hair growth comprising a fermented product obtained by fermenting streptococcus strains are mentioned in korean patent laid-open publication No. 10-1098526, and hair cosmetic compositions comprising a complex lactobacillus fermented liquid are mentioned in korean patent laid-open publication No. 10-2001-0107152. Further, korean laid-open patent No. 10-2020-0126920 has mentioned a pharmaceutical composition for preventing or treating alopecia containing hyaluronic acid and proteoglycan-coupled protein as active ingredients, korean granted patent No. 10-1516445 has mentioned a hair modifier using cationized hyaluronic acid and/or a salt thereof, and a final report of development industry of characteristic (leading) industry in 2019 has published "development of a latest alopecia reliever using hyaluronic acid particles containing hair-related signaling polypeptide".

As for the culture conditions of the Streptococcus strains, previously published papers "Effect of pH, a positioning and incubation on hyaluronic acid production by Streptococcus zoepidemicus" (Biotechnology Letters volume 16, pages507-512(1994)) revealed that pH 6.7. + -. 0.2 is the optimum pH for the hyaluronic acid production yield of Streptococcus, Korean patent granted No. 10-1784864 revealed that the pH is preferably maintained to 6.5-7.5 in order to prevent the inhibition of the strain culture due to the lactic acid produced when culturing Streptococcus, and Korean patent granted No. 10-1322227 revealed that the culture conditions for Streptococcus optimally producing hyaluronic acid are the conditions of pH 7.0 and 34 ℃. That is, it was revealed that the culture conditions of the conventional hyaluronic acid-producing streptococci were only neutral pH.

In contrast, the present inventors have combined the above-mentioned prior art techniques and have conducted experiments to culture streptococcus strains in a manner that enables the production of more hyaluronic acid having an effect on preventing hair loss, and have occasionally found that a streptococcus culture solution cultured under acidic pH conditions without forming hyaluronic acid is rather superior in hair loss alleviation, hair growth, hair root strengthening and hair coating effects than a conventional streptococcus culture solution cultured under neutral pH conditions in a manner that forms hyaluronic acid, thereby completing the present invention.

Disclosure of Invention

Accordingly, a primary object of the present invention is to provide a hair cosmetic composition for preventing hair loss and hair growth, which comprises a culture solution of streptococcus strain cultured so as not to form hyaluronic acid.

Another object of the present invention is to provide a method for culturing a Streptococcus strain in such a manner that hyaluronic acid is not formed.

According to an embodiment of the present invention, there is provided a hair cosmetic composition comprising, as an active ingredient, a culture solution of a streptococcus strain cultured under acidic pH conditions in such a manner that hyaluronic acid is not formed.

The present invention provides a hair cosmetic composition characterized in that the acidic pH condition is pH 2 to 4. In the conventional culture of Streptococcus strains, the strains are usually cultured under neutral pH conditions of pH 6.5 to 7.5 for the optimal culture of the strains and the production of hyaluronic acid. The acidic pH condition of the present invention is a severe condition that can inhibit the culture of Streptococcus strains and the production of hyaluronic acid, as compared with the conventional neutral pH condition. In the experimental example 1 of the present invention, it was initially confirmed that hyaluronic acid was hardly formed in the examples cultured under acidic conditions of pH 2 to 4, unlike the neutral pH conditions.

The present invention provides a hair cosmetic composition characterized in that any Streptococcus strain known to produce hyaluronic acid can be used as the Streptococcus strain, and preferably any one selected from the group consisting of Streptococcus zooepidemicus (Streptococcus zoepidemicus), Streptococcus Thermophilus (Streptococcus Thermophilus), and Streptococcus salivarius (Streptococcus salivarius).

In the present invention, the culture solution includes not only a culture stock solution but also a purified culture product obtained by purifying the culture stock solution. The present invention provides a hair cosmetic composition characterized in that any purification method known in the art of culturing microorganisms can be used for the purification of the culture solution, and preferably, the culture solution is purified by a centrifugal separator, a filter and activated carbon.

The present invention provides a hair cosmetic composition characterized in that the above-mentioned culture solution has hair loss alleviation, hair growth, hair root strengthening and hair-spreading effects better than a culture solution of a streptococcus strain cultured under neutral pH conditions so as to form hyaluronic acid. In the present invention, the effect of the present invention was demonstrated by evaluating the tensile strength of hair in experimental example 2, measuring the average roughness of hair by AFM in experimental example 3, evaluating the strengthening of hair roots in experimental example 4, evaluating the hair growth in experimental example 5, and evaluating the alleviation of male pattern alopecia in experimental example 6.

Since hyaluronic acid has been known to have a hair loss preventing effect, it is expected that a culture solution of streptococcus strain cultured in a manner capable of producing more hyaluronic acid is more effective in hair loss alleviation, hair growth, and the like, however, the present inventors have unexpectedly and initially found that a culture solution of streptococcus strain cultured under acidic pH conditions in a manner such that hyaluronic acid is not formed is more effective in hair loss alleviation, hair growth, and the like, thereby completing the present invention. This is presumed to be due to the harsh culture conditions of acidic pH, resulting in the formation of better components in the culture solution that contribute to alopecia alleviation, hair growth.

In the present invention, there is provided a hair cosmetic composition characterized in that a culture solution of streptococcus strain cultured under acidic pH conditions in such a manner that the above hyaluronic acid is not formed is contained in an amount of 0.01 to 100% by weight.

The present invention provides a hair cosmetic composition in a form selected from the group consisting of a shampoo, a hair tonic, a hair conditioner, a hair treatment cream, a hair lotion, a gel water, a hair mask, a hair spray, a mousse, a shampoo, a hair powder, a hair oil, and the like.

According to another embodiment of the present invention, the present invention provides a method for culturing a Streptococcus strain in a manner that does not form hyaluronic acid, comprising the steps of:

a) inoculating a Streptococcus strain to a sterilized production medium, and culturing at a culture temperature of 20-40 ℃ and a pH of 2-4 at 100-250 rpm for 2-8 hours; and the number of the first and second groups,

b) and then culturing at 0-100 rpm for 20-30 hours at a culture temperature of 20-30 ℃ and a pH of 2-4.

In the present invention, an inoculum culture step for propagating the Streptococcus strain prior to the inoculation in the above-mentioned step a) may also be included. Preferably, the step of culturing the inoculum includes the step of inoculating the Streptococcus strain in a sterilized liquid medium and culturing at pH 6-8 and 27-35 ℃ at 100-250 rpm for 15-20 hours.

The present invention provides a method for culturing a Streptococcus strain, wherein any production medium containing nutrients such as glucose and inorganic salts for culturing the Streptococcus strain can be used as the production medium in step a), but preferably, glucose is contained in an amount of 3 to 7% per 1L of water, yeast concentrate is contained in an amount of 0.3 to 0.7% per 1L of water, potassium phosphate is contained in an amount of 0.1 to 0.3% per 1L of water, magnesium sulfate is contained in an amount of 0.03 to 0.07% per 1L of water, and polypeptone is contained in an amount of 1 to 3% per 1L of water.

In the present invention, in the above step a) and step b), lactic acid is produced by culturing Streptococcus, and the pH is lowered, so that the pH can be maintained at 2 to 4 without using an alkaline pH adjuster such as NaOH as in the prior art.

In the present invention, in the step b), not only the acidic pH condition but also the culture temperature is adjusted to 20 to 30 ℃ lower than the proper culture temperature, and the rpm is also adjusted to 0 to 100rpm lower than the proper rpm, so that the culture is performed under more severe conditions, thereby more surely ensuring that hyaluronic acid is not formed. In experimental example 1 of the present invention, it was confirmed that hyaluronic acid was hardly formed in the example cultured under the severe conditions as described above.

According to the present invention, the streptococcal culture fluid cultured under acidic pH conditions in such a manner that hyaluronic acid is not formed has hair loss alleviation, hair growth, hair root strengthening and hair coating effects better than the streptococcal culture fluid cultured under neutral pH conditions in such a manner that hyaluronic acid is formed in the past. Therefore, the culture solution of Streptococcus cultured according to the present invention can be effectively used as a raw material for a hair cosmetic composition for preventing alopecia and hair growth.

Drawings

Fig. 1 is a photograph showing a color change after a carbazole reaction based on the generation of hyaluronic acid.

FIG. 2 is a graph showing the results of real-time polymerase chain reaction (HGF Realtime PCR) of hepatocyte growth factor for evaluation of hair root enhancement.

FIG. 3 is a graph showing the results of VEGF Realtime PCR for evaluating hair root enhancement.

Fig. 4 is a graph showing the results of Wnt10b real-time polymerase chain reaction (Wnt10b realtome PCR) for evaluating hair growth.

Fig. 5 is a graph showing the results of Androgen receptor real-time Polymerase Chain Reaction (PCR) in order to evaluate the alleviation of male pattern baldness.

FIG. 6 is a graph showing the results of the TGF-. beta.2 real-time polymerase chain reaction (TGF-. beta.2 real PCR) for the evaluation of the alleviation of male pattern baldness.

Detailed Description

The present invention will be described in more detail below with reference to examples. These examples are only for illustrating the present invention, and thus the scope of the present invention should not be construed as being limited thereto.

Example 1 procedure for culturing Streptococcus strains in a manner that does not form hyaluronic acid

Streptococcus strain (S.zoopeptidimicus HDB5021) was cultured at 30 ℃ for 17 hours using Todd-Hewitt sterilized liquid medium (BD Co., U.S. pH 7). The cultured inoculum solution was inoculated into a sterilized production medium (5% glucose, 0.5% yeast concentrate, 0.2% potassium phosphate, 0.05% magnesium sulfate, 1.5% polypeptone per 1L of water), and then cultured under conditions of 1vvm, 35 ℃, pH 2 to 4, and 200rpm for 6 hours. Then, the cells were cultured at 50rpm at a culture temperature of 25 ℃ and a pH of 2 to 4 for 24 hours.

Example 2 procedure for purifying the culture broth obtained in example 1

The culture solution obtained in example 1 was diluted with 100 times of purified water. The precipitate was allowed to settle by running at 4,000rpm for 5min using a centrifugal separator (Centrifuge MF-80, Hanil, Korea), and only the supernatant was removed and filtered using a 0.3Fm filter. And (3) treating the filtered filtrate by using 1-10% of activated carbon to remove color and smell.

Comparative example 1 culture solution cultured in a manner to produce hyaluronic acid

Streptococcus strain (S.zoopeptidimicus HDB5021) was cultured at 30 ℃ for 17 hours using Todd-Hewitt sterilized liquid medium (BD Co., U.S. pH). After the completion of the culture, the inoculum solution was inoculated into a sterilized production medium (5% glucose, 0.5% yeast concentrate, 0.2% potassium phosphate, 0.05% magnesium sulfate, 1.5% polypeptone per 1L of water), cultured at 1vvm, 35 ℃ and 200rpm for 6 hours, and the pH was adjusted to 7 using a 10% NaOH solution. Subsequently, the incubation temperature was maintained at 35 ℃ and the pH was further adjusted to 7 using 10% NaOH solution, and incubation was continued at 600rpm for 24 hours. The culture broth was also purified as in example 2.

Experimental example 1 hyaluronic acid comparative confirmation experiment of example 2 and comparative example 1

Hyaluronic acid productivity was quantitatively analyzed by the carbazole method (z.disco, j.biol.chem.167,189 (1947)). 100mL of the culture solution obtained in Experimental example 2 and 100mL of the culture solution obtained in comparative example 1 were precipitated in 10-fold ethanol, and the precipitate obtained in the 250 mesh filter cloth was freeze-dried and used in the analysis. Fig. 1 is a photograph showing a color change after a carbazole reaction based on the generation of hyaluronic acid.

After the carbazole reaction, the absorbance was measured by a Spectrophotometer (Spectrophotometer V-730, JASCO, UK), the hyaluronic acid content was calculated, and the experiment was repeated 3 times, and the average value thereof is shown in table 1.

TABLE 1

Distinguishing	Hyaluronic acid concentration (g/L)
		Example 2	0
Comparative example 1	5.51

Experimental example 2 measurement of tensile Strength of Hair

As the hair used in the experiment, the hair of a female 25 years old who had not undergone chemical treatment such as permanent wave, hair dyeing, bleaching and dyeing for 18 months or more was used. Even if the hair is subjected to the same conditions, the hair is exposed to different degrees in daily life (shampoo, hair drying, ultraviolet exposure, etc.), and therefore, the hair is cut with scissors at a position 2cm away from the root of the hair. The collected hair was washed with neutral shampoo to remove residual contaminants, and then naturally dried, and about 1g each of them was used to prepare a hair bundle.

In order to produce a damaged hair bundle, the previously produced hair bundle was left to stand in a solution composed of 6% hydrogen peroxide, 0.5% ammonium hydroxide and 0.1% EDTA for 3 hours, rinsed with water for 1 minute or more, and then dried with a hair dryer for use.

In normal hair tresses and damaged hair tresses, after example 2 and comparative example 1 were immersed for 2 hours, washed with neutral shampoo, naturally dried, and then tensile strength was measured by the following method.

Tensile strength was measured using a Universal Testing Machine (Instron 5584, USA). Evaluation was conducted by the korean textile research institute (FITI), and the tensile test was performed in accordance with the tensile strength and elongation of korean industrial standard fibers for measuring the single fiber yarn (KS K ISO 5079: 2007). The test was conducted at a tensile rate of 30mm/min, and for the reliability of the measured values, the average value was obtained after 10 measurements per sample, and the results are shown in Table 2.

TABLE 2

Experimental example 3 measurement of average roughness of hair by AFM

In order to quantitatively compare and analyze the fine structure of the hair surface, Line profiles (Line profiles) were shown in the groups treated with normal hair in example 2 and comparative example 1, respectively, and the average roughness was determined. The surface fine structure of normal hair was very rough and there were many impurities, but when treated with example 2, the surface was very smooth, showing the result that impurities were removed. The width of 5 μm x 5 μm was designated at 3 points, and the average roughness Ra of the entire area was determined after topographies were measured, and the results are shown in the following table.

TABLE 3

Distinguishing	Average roughness Ra
		Normal Hair + example 2	60.83±23.41nm
Normal Hair + comparative example 1	10.64±4.84nm

Experimental example 4 evaluation of Hair root strengthening

To evaluate the hair root strengthening efficacy, Hepatocyte Growth Factor (HGF), Vascular Endothelial Growth Factor (VEGF) gene expression experiments were performed. Growth factors expressed from hair follicles include insulin-like growth factor 1 (IGF-1), Vascular Endothelial Growth Factor (VEGF), Hepatocyte Growth Factor (HGF), Keratinocyte Growth Factor (KGF), and the like, and these growth factors form and grow hairs to maintain the growth phase of the hairs [1,2 ]. HGF is known to stimulate the growth of hair follicles in hair papilla cells and regulate the hair growth cycle [3], and VEGF is known to supply nutrients to form hair roots and regulate the size of hairs through the regulation of the hair cycle and the vasodilation around the hair roots [2,4 ].

In 6well multiwell plates (6well multiplate) (corning), at 2.5X10 ⁵ cells/well were inoculated with DMEM medium containing 10% bovine serum and dermal papilla cells (DPCs, cell biology) each 2 , and after 24 hours of culture, each well (well) was replaced with serum-free (serum free) medium. After treating serum-free (serum free) medium with the sample, the medium was cultured for 24 hours. After washing (washing) with Phosphate Buffered Saline (PBS), cultured cells were treated with QIAzol (qiagen, USA) to perform Cell lysis (Cell lysis), and then RNA was isolated using the protocol (protocol) provided by the manufacturer (qiagen). After washing with PBS (washing), the cultured cells were treated with QIAzol Lysis Reagent (QIAzol Lysis Reagent) (QIAGEN)1 , and (prep) RNA was prepared using the protocol provided by QIAGEN, and the RNA was prepared using Qubit ^TM RNA BR Assay kit, in which cDNA is synthesized and Real-time polymerase chain reaction (Real-time PCR) is performed after quantification of the isolated RNA. cDNA Synthesis Using the cDNA Synthesis Kit (Kit) (qPCRBIO cDNA Synthesis Kit, PCRBIOS, London, UK), experiments were performed according to the Kit method. Real-time PCR after amplifying genes using Real-time PCR Kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOSYSTEMS, London, UK), the amplified products were quantitatively analyzed.

Primers (primers) for HGF, VEGF, and β -actin (β -actin) used in PCR were synthesized by cosmo gene tech (Korea) and used, and the primer sequences (primer sequences) are shown in [ Table 4 ].

TABLE 4

The cDNA synthesis reaction conditions were that HGF, VEGF, and β -actin were all at 95 ℃ for 5 seconds and 60 ℃ for 25 seconds for 40 cycles. The Control group (Control) used purified water treated as a dissolution solvent, and Minoxidil (Minoxidil) used as a positive Control group was known to have an effect of promoting hair growth through improvement of blood circulation and opening of potassium channels. The results of expression levels are shown in tables 5 and 6 and FIGS. 2 and 3. As shown in fig. 2, the HGF gene expressed in the growth phase in the dermal papilla cell line was increased in concentration-dependently as compared to the Control (Control) in example 2, thereby having the efficacy of strengthening the hair root at the cellular level. As shown in fig. 3, example 2 showed that the effect of strengthening hair roots at the cellular level was exhibited by inducing new blood vessel formation in hair follicles, increasing the supply of nutrients to hair roots and hairs, and increasing the concentration of VEGF gene expression that induces cell proliferation and migration in a manner dependent on the Control (Control).

TABLE 5

TABLE 6

Experimental example 5 evaluation of Hair growth

To evaluate the hair growth efficacy, the hair was heldLine Wnt10b (Wireless-related model tumor tissue integration site 10b) gene expression experiment. It is known that the Wnt signaling pathway (Wnt signalling) is one of the most important pathways for Hair follicle development, regulates the proliferation and migration of Hair cells (germinal cell line, outer root sheath cell, Hair papilla cell), and is associated with the regeneration of Hair follicles [5,6 ]]. Also, it is known that Wnt10 acts to promote epithelial cell division and hair growth [7]. In 6well multiwell plates (6well multiwell) (corning), at 2.5X10 ⁵ cells/well were inoculated with 2 each of MSCM medium containing 5% bovine serum and Germinal cells (Human Hair stromal cells, HHGMC, ScienCell), and cultured for 24 hours, after which each well (well) was treated with a sample and cultured for 24 hours. After washing (washing) with Phosphate Buffered Saline (PBS), cultured cells were treated with QIAzol (qiagen, USA) to perform Cell lysis (Cell lysis), and then RNA was isolated using the protocol (protocol) provided by the manufacturer (qiagen). Using qubits ^TM RNA BR Assay kit, quantification of the RNA isolated, cDNA synthesis, Real-time polymerase chain reaction (Real-time PCR). cDNA Synthesis Using the cDNA Synthesis Kit (Kit) (qPCRBIO cDNA Synthesis Kit, PCRBIOS, London, UK), experiments were performed according to the Kit method. Real-time PCR after amplifying genes using Real-time PCR Kit (2x qPCRBIO SyGreen Blue mix Lo-ROX, PCRBIOSYSTEMS, London, UK), the amplified products were quantitatively analyzed.

Wnt10b, primer for beta-actin (primer) used in PCR was synthesized by cosmo genetech (Korea) and used, and the primer sequence (primer sequence) is shown in [ Table 7 ].

TABLE 7

The conditions for the cDNA synthesis were Wnt10b and β -actin 5 seconds at 95 ℃ and 25 seconds at 60 ℃ for 40 cycles. The Control group (Control) used purified water treated as a dissolution solvent, and Caffeine (Caffeine) used as a positive Control group was known as a substance inducing hair growth. The results of expression levels are shown in table 8 and fig. 4. As shown in fig. 4, in example 2, the expression of Wnt10b, which is one of Wnt signaling pathways (Wnt signaling) involved in the proliferation of hair cells and the production of hair follicles (hair follicles), was increased as compared to the Control group (Control), and thus the effect of promoting hair growth was exhibited at a cellular level.

TABLE 8

Experimental example 6 evaluation of alleviation of Male alopecia

Male pattern alopecia converts testosterone (testosteron) present in hair root cells into the stronger metabolite Dihydrotestosterone (DHT), which binds to androgen receptors (androgen receptors) of hair follicles, inhibits the activity of adenylate cyclase (adenylyl cyclase) to activate intracellular metabolism, decreases the concentration of intracellular Cyclic AMP, and decreases sugar metabolism of hair cells, obstructs energy supply, delays protein synthesis, shortens the growth phase of hair follicles, and so on, and this process is repeated to increase the ratio of resting-stage hair follicles, which indicates that hairs are tapered short [8-13 ]. To evaluate the male pattern alopecia alleviating efficacy, gene expression experiments of Transforming growth factor-beta 2 (TGF-beta 2), Androgen Receptor (AR) were performed. The Androgen receptor (Androgen receptor) is a transporter of DHT, which functions to transfer DHT to hair cells, induce apoptosis of the hair cells, and cause alopecia, and TGF- β 2 is known to promote the growth phase of hair to shift to the catagen phase [8-13 ].

In 6-well multiwell plates (6well multiplate) (corning), at 2.5X10 ⁵ cells/wells, seeded hair papilla cells (DPCs, cell biology) 2 , after 24 hours of culture, wells were replaced with serum free (serum free) medium and then treated with 5 α -Dihydrotestosterone (5 α -Dihydrotestosterone) (DHT, seleckchem korea) solution (solution) inducing male pattern alopecia exceptThe test group of the non-treated group was incubated for 48 hours after the treatment with the test sample. After washing (washing) with Phosphate Buffered Saline (PBS), the cultured cells were treated with QIAzol (qiagen, USA) to perform Cell lysis (Cell lysis), and then RNA was isolated using the protocol (protocol) provided by the manufacturer (qiagen). Using Qubit ^TM RNA BR Assay kit, in which cDNA is synthesized and Real-time polymerase chain reaction (Real-time PCR) is performed after quantification of the isolated RNA. cDNA Synthesis Using the cDNA Synthesis Kit (Kit) (qPCRBIO cDNA Synthesis Kit, PCRBIOS, London, UK), experiments were performed according to the Kit method. Real-time PCR after amplifying genes using Real-time PCR Kit (2x qPCRBIO SyGreen Blue mixLo-ROX, PCRBIOSYSTEMS, London, UK), the amplified products were quantitatively analyzed. Primers (primers) for transforming growth factor-beta 2 (TGF-. beta.2), androgen receptor (androgen receptor), and beta-actin (beta-actin) used in PCR were synthesized by cosmo gene tech, Inc. (Korea), and the primer sequences (primer sequences) were shown in [ Table 9 ]]In (1).

TABLE 9

The cDNA synthesis reaction conditions were 40 cycles of TGF-. beta.2, AR, and β -actin at 95 ℃ for 5 seconds and 60 ℃ for 25 seconds. The Control group (Control) used purified water treated as a dissolution solvent, and Minoxidil (Minoxidil) used as a positive Control group was known to have an effect of promoting hair growth through improvement of blood circulation and opening of potassium channels. The results of expression levels are shown in tables 10 and 11 and FIGS. 5 and 6. As shown in fig. 5, in example 2, the effect of alleviating male pattern baldness at a cellular level is achieved by blocking the expression of androgen receptor (androgen receptor) which is a transport receptor of DHT that induces male pattern baldness, inhibiting the transfer of DHT to hair follicle cells, and blocking baldness. As shown in fig. 6, TGF- β 2 increased by DHT inducing male pattern alopecia in example 2 promoted the shift from the anagen phase to the catagen phase in the hair cycle and inhibited the proliferation of epithelial cells, thereby promoting alopecia, compared to Control (Control), and it has the effect of relieving male pattern alopecia at the cellular level by inhibiting the expression of these TGF- β 2 genes.

TABLE 10

TABLE 11

Dosage form example 1: preparation of hair lotion

The hair spray was prepared by a conventional method using the following ingredients listed in the following table.

TABLE 12

	Composition (I)	Content (percentage by weight)
			1	Example 2 culture purification	5.0
2	Stearic acid	5.0
			3	Cetyl alcohol	0.5
4	Polyoxyethylene sorbitan sesquioleate	0.8
			5	Triethanolamine	0.4
6	Glycerol	5.0
			7	Ethanol	8.0
8	Perfume	Proper amount of
			9	Purified water	To 100

Dosage form example 2: preparation of shampoo

The shampoo was prepared by a conventional method according to the following composition ingredients described in the table.

Watch 13

	Composition (A)	Content (percentage by weight)
			1	Culture purification of example 2	5.0
2	Polyquaternary ammonium salts	0.3
			3	Ethylenediaminetetraacetic acid disodium salt	0.05
4	Hydroxy phenyl methyl ester	0.3
			5	Polyoxyethylene sorbitan monooleate	0.5
6	Sodium laureth sulfate	20.0
			7	Cocoyl amphocarboxyglycinate	20.0
8	Cocoamidopropyl betaine	5.0
			9	Coconut oil fatty acid diethanolamide	3.0
10	Methylchloroisothiazolinone/methylisothiazolinone	0.04
			11	Hydrolyzed silk	0.5
12	Sodium chloride	0.7
			13	Citric acid	0.3
14	Perfume	Proper amount of
			15	Purified water	To 100

Dosage form example 3: preparation of hairdressing gel

The hairdressing gel was prepared by a conventional method according to the composition described in the following table.

TABLE 14

	Composition (I)	Content (percentage by weight)
			1	Culture purification of example 2	5.0
2	Dialkyl methyl ammonium chloride	1.0
			3	Monostearyltrimethyl amine chloride	1.0
4	Hexadecanol and octadecanol	2.0
			5	Isostearoyl lactyl lactate	1.0
6	Paraffin wax	3.0
			7	Polypeptides	5.0
8	Cationized cellulose	3.0
			9	Polyoxyethylene oleyl ether	0.5
10	Hydroxy phenyl methyl ester	0.2
			11	Perfume	Proper amount of
12	Purified water	To 100

While the invention has been described with reference to the above embodiments, this is by way of example only, and various modifications and equivalent other embodiments will be apparent to those skilled in the art from this disclosure. Therefore, the true technical scope of the present invention should be determined by the technical matters of the appended claims.

Claims

1. A hair cosmetic composition characterized by containing, as an active ingredient, a culture solution of a Streptococcus strain cultured under acidic pH conditions so as not to form hyaluronic acid.

2. The hair cosmetic composition according to claim 1, wherein the acidic pH condition is pH 2 to 4.

3. The hair cosmetic composition according to claim 1, wherein the streptococcus strain is any one selected from the group consisting of streptococcus zooepidemicus, streptococcus thermophilus and streptococcus salivarius.

4. The hair cosmetic composition according to claim 1, wherein the culture solution is purified by a centrifugal separator, a filter and activated carbon.

5. The hair cosmetic composition according to claim 1, wherein the culture solution has better hair growth, hair root strengthening, and hair application effects than a culture solution of Streptococcus strain cultured under neutral pH conditions in such a manner as to form hyaluronic acid.

6. The hair cosmetic composition according to claim 1, wherein the formulation of the hair cosmetic composition is any one selected from the group consisting of shampoo, hair tonic, hair conditioner, hair treatment, hair cream, hair lotion, gel, hair mask, hair spray, mousse, hair soap, hair powder, and hair oil.

7. A method for culturing a Streptococcus strain in a manner that does not form hyaluronic acid, comprising the steps of:

step a), after inoculating a streptococcus strain into a sterilized production culture medium, culturing for 2-8 hours at a culture temperature of 20-40 ℃ and a pH value of 2-4 at a speed of 100-250 rpm; and the number of the first and second groups,

and b), then culturing for 20-30 hours at 0-100 rpm under the conditions that the culture temperature is 20-30 ℃ and the pH value is 2-4.

8. The method of claim 7, wherein the production medium contains 3-7% of glucose, 0.3-0.7% of yeast concentrate, 0.1-0.3% of potassium phosphate, 0.03-0.07% of magnesium sulfate, and 1-3% of polypeptone per 1L of water.