KR100903283B1 - Hair growing material which contains a deer antler ingredient and its manufacturing method - Google Patents

Abstract

A hair growth stimulator containing a Cervi Parvum Cornu component is provided to prevent hair loss and promote hair growth. A hair growth stimulator containing a Cervi Parvum Cornu component comprises medium of Cervi Parvum Cornu-derived cell as an active ingredient. A manufacturing method of hair growth stimulator comprises: a step of treating Cervi Parvum Cornu tissue with DMEM-collagenase medium and culturing in a culture room having 5% of CO2 at 34-37°C for six hours; a step of centrifuging the medium containing the cells of Cervi Parvum Cornu; a step of culturing the cells in a culture room having 5% of CO2 at 34-37°C for 24-72 hours; and a step of collecting medium containing the Cervi Parvum Cornu component.

Description

Hair Growing Material Containing Deer Antler and Its Manufacturing Method {Hair Growing material which contains a deer antler ingredient and Its manufacturing method}

The human hair is about 100,000 to 150,000 pieces, each hair has a different cycle and grow and drop out through the anagen, catagen, and telogen. This cycle is repeated over three to six years, with the result that on average 50-100 hairs fall out normally. In general, alopecia is a condition in which a hair grows shorter in growth stages, particularly in the head of a hair, in particular, in the head where the hair is kept or at a reduced size.
Many studies have been conducted to treat hair loss in recent decades, but the cause of hair loss is still unknown. Looking at the information on the hair loss factors that have been identified so far, the inhibition of the growth or function of dermal papilla associated with hair cycle regulation, abnormal hair cycle due to the action of male hormones, hair cycle due to blood flow to the scalp Abnormal changes, anticancer drugs, mental stress, physical stimulation, and environmental pollution.
Alopecia is classified into the causes and forms of alopecia, genotyping alopecia, seborrheic alopecia, alopecia areata, alopecia areata due to drug side effects, alopecia due to aquatic cells, alopecia due to environmental changes, and alopecia areata. In particular, a growing number of people are suffering from hair loss due to excessive stress, dietary changes, pollution, etc., and their age is also decreasing and female hair loss population is increasing.
Currently, medically proven methods for treating hair loss are drug treatment and hair transplantation. In the case of drugs used for treatment, currently prescribed in domestic hospitals, there are two types of drugs, 'finasteride' and 'minoxidil'. FDA approval. In addition, there are hormonal agents for inhibiting the action of male hormones such as estrogens and estradiol, and testosterone activity inhibitors such as pentacatecanoic acid and fenasteride.

Deer antler refers to the deer's horns, which are cut before being hardened into keratin and used as medicine. Traditionally, ginseng is the best prosthetic tonic along with ginseng. Therefore, the purpose of deer breeding in Korea is mainly for the production of antler and the efficacy of antler as a medicinal has been recorded from a long time ago.
Dongbogam: Neural breakdown, fatigue recovery, sexual function is good substance.
Herbal cost: blood and anemia is effective, and protects yanggi.
Herbal tree: protects tablets, water, yin and blood, and is good for restoring after illness, weak people, pulmonary tuberculosis and pulmonary function.
Prosthetic smell: All weakness, malnutrition, pain in the lower back or legs, skin feelings, etc.
Herbal tablets: uterine bleeding, cold hypothalamus, etc. are good for all kinds of women.
Cheongeumbang: to treat insomnia, heart palpitations.
Religious herbaceous tree: Take care after birth, it is the best protection.
Pharmacokinetics, hereditary minor theory: Covers nerves, relaxes the fetus. Strengthen the weakness and strengthen the bones.
The main component of the deer antler is composed of lipids, proteins, carbohydrates, minerals and trace elements, etc., and these antlers are considered to have various physiological activities due to these components. On the other hand, studies on the physiological function of antler are actively conducted in the field of Korean traditional medicine, anti-fatigue effect, immune activity increase action, protein synthesis promotion action, endocrine function promotion action, hematopoietic action, immune activity increase action, analgesic action, cholesterol lowering Action, liver function improvement, growth promoting action and diabetes treatment effect and the like have been reported.

Deer Antler is an expensive medicine and its field of use is limited compared to its excellent efficacy. An object of the present invention relates to a culture solution containing the antler component and the use thereof through the culture of antler-derived cells with less side effects, effectively activate the hair root to prevent hair loss, promote hair growth and provide a hair growth composition that lasts the effect It is in that. The present invention was to investigate the growth of the antler cell and the antler cell containing the antler cell culture in vitro test method, to provide a basis for the antler component of the antler culture medium can be used as a hair growth treatment.

The present invention proposes a method for producing antler components through culturing antler-derived cells, and cultures the antler-derived cells to have the effect of promoting hair growth and preventing hair loss through the activity of hair roots. Said.

The cultivation of the antler-derived cells was carried out, and thus the hair growth experiment was carried out through the culture medium containing the antler component, the medium before culturing the antler cell (regular medium) and minoxidil sulfate (minoxidil sulfate). Hair follicles of Wistar rats were isolated and cultured in 37 ° C, 5% CO ² incubator with one follicle in each well of a 24-well plate. Dog hair follicles were used. During the culture, the medium was changed every 3 days, and the culture medium containing the medium and the antler component was treated at concentrations of 1, 10 and 100 ug / ml, and the minoxidil sulfate (Sigma, USA), a positive control, at a concentration of 1 uM. Treated with. The hair follicle length of each culture was measured using an image analyzer (DP analyzer; Olympus, Japan), and the average value of hair follicle length change was obtained, and the growth degree was measured by comparing with the average length of the control group.

Example 1
1-1 Extraction and Culture of Deer Antler-Derived Cells
Three- to seven-year-old healthy and breeding ages were chosen to produce deer antler, which is more than 4.5 kilograms of deer antler per year. The extracted antler was removed from the skin using a sterile surgical instrument, and cultured in phosphate buffered saline (PBS) for about 2 hours to remove blood from the antler tissue. Deer antler tissue 1 to 3 cm from the upper end of the antler was separated using a surgical scissors and a knife. Since collagen was used for the chemical degradation (Collagenase). First, DMEM (Dulbecco's Modified Eagle's Medium) medium was subjected to type 1-S collagenase (Collagenase Type IS) or type 4 collagenase (Collagenase Type IV) or type 3 collagenase. (Collagenase type III) was prepared at a concentration of 0.5 mg / ml. Thus prepared DMEM-collagenase medium was treated to the antler tissue and incubated for about 6 hours in a 34 ~ 37 ℃, 5% CO ₂ incubator. The medium containing the antler cells was centrifuged at 1200 rpm for 5 minutes to remove collagenase and impurities. The isolated cells were inoculated in a flask containing a cell culture medium containing 5% FBS (fetal bovaine serum) and incubated for 24 to 72 hours in an incubator at 34 to 37 ° C and 5% CO ₂ . Only cells that adhere to the vessel surface to produce a monolayer are passaged.
1-2 Composition of Deer Antler Medium
D-media (# 78-5470EB, Gibco) was used as a medium for culturing antler-derived cells, and 5% to 10% of FBS (fetal bovaine serum) was added to form a medium.
1-3 Culture medium containing antler component
1 x 10 ⁵ antler-derived cells Dogs were inoculated in 25T flasks containing antler medium (5% FBS + D-media), incubated for 24 to 72 hours in an incubator at 34-37 ° C and 5% CO ₂ , and the cells grown to 80% on the surface of the culture vessel. When the culture medium was collected, the foreign material was removed by centrifugation at 1200 rpm for 5 minutes in a centrifuge.
Example 2
2-1 materials
The culture medium containing the test medium and the antler component were lyophilized, and each lyophilized powder was dissolved in dimethyl sulfoxide (DMSO) and used for the experiment.
2-2 In Vitro Hair Growth Efficacy
2-2-1 Isolation and Culture of Rat Hair Follicles
Three-week-old male rats (Wistar rat) were purchased from a central laboratory animal (Japan SLC, Hamamatsu, Janpan) Co., Ltd. and anesthetized with ethyl ether and cervical spine was also killed. Isolation of rat left and right beard pads (mystacial pads) to isolate E / P buffer containing 100 units / ml penicillin-100 µg / ml streptomycin (Gibco Inc, NY, USA) Earle's balanced salts solution (EBSS, Sigma MO, USA) + phosphate buffered solution (PBS, phosphate-buffered saline; Sigma MO, USA). The hair follicles were carefully separated while observing under a dissecting microscope. Until the hair follicles were separated, the separated hair follicles were placed in a Petri dish containing E / P buffer solution and incubated for 1 hour at 37 ° C and 5% CO ₂ incubator. 2 mM L-glutamine (Gbco Inc, NY, USA), 10 μg / ml insulin (Sigma MO, USA), 50 nM in each well of a 24 well plate. 500 μl William medium (Williams' medium E; Gibco Inc, NY, USA) containing hydrocortisone (Sigma MO, USA) and 100 units / ml penicillin-100 μg / ml streptomycin Put, and put one follicle in one well (37) and incubated in 37 ° C, 5% CO ₂ thermostat. 7-9 hair follicles were used in one experimental group, and the culture medium was changed every 3 days. Normal medium and antler-containing cultures were treated at concentrations of 1, 10 and 100 ug / ml, and minoxidil sulfate (Sigma, USA), a positive control, was treated at a concentration of 1 uM.
2-2-2 Measurement of hair follicle growth
The morphology of the hair follicles in culture was photographed using a microscope (Olympus, Japan). Hair follicle length was measured at 0, 7, 10, 14 and 21 days using an image analyzer (DP controller; Olympus, Japan). The average value of the hair follicle length change was calculated and compared with the average length of the control group.
Example 3:
3-1 In Vitro Hair Growth Efficacy of Normal Medium
Growth hair follicles of three-week-old experimental rats were isolated and cultured for three weeks, and hair follicle length was measured to examine the hair growth efficacy of the antler medium (FIG. 1). After comparing the length difference (%) of hair follicles between the antler medium treatment group and the control group 21 days after culture, the control group (100 ± 6%) increased the growth of hair fibers to 111.4 ± 6.1% and showed no increase at concentrations of 10 and 100 ug / ml. The positive control 1uM minoxidil sulphate showed 111.8 ± 6.7% growth (FIG. 2).
3-2 In Vitro Hair Growth Efficacy of Antler-Containing Cultures
Growth hair follicles of the three-week-old experimental rats were isolated and cultured for three weeks, and hair follicle length was measured to examine the hair growth efficacy of the antler component-containing culture solution (FIG. 3). After comparing the length difference (%) of the hair follicles between the treated group containing the antler component and the control group at 21 days after the culture, the concentrations of 10 ug / ml when treated with 1, 10 and 100 ug / ml of the antler component were respectively. Hair fiber growth was more increased to 141.3 ± 8.2% than the control (100 ± 6%) and there was no increase effect at concentrations of 1 and 100 ug / ml. The positive control 1uM minoxidil sulphate showed 111.8 ± 6.7% growth (FIG. 4).
result
After incubation, each hair follicle length was measured using an image analyzer (DP controller; Olympus, Japan), and the average value of hair follicle length change was determined, and the growth degree was measured by comparing with the average length of the control group. The results show that the growth effect of the antler component-containing medium is excellent (Table 1).
Control Growth Effect Plain badge 111.4 ± 6.1% Deer Antler-Containing Culture 141.3 ± 8.2% minoxidil sulphate 111.8 ± 6.7%

1 is a photograph of hair growth efficacy of general medium.
Figure 2 is a graph of the hair growth efficacy of the normal medium.
3 is a photograph of hair growth efficacy of the antler component-containing culture solution.
Figure 4 is a graph of the hair growth efficiency of the antler component-containing culture solution.

Claims (4)

Hair growth promoting composition comprising a culture solution of antler-derived cells as an active ingredient The hair growth promoting composition according to claim 1, wherein the formulation of the hair growth promoting composition of claim 1 is a liquid, a spray, a gel, a paste, an emulsion, a cream, a conditioner, or a shampoo. delete delete

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