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

Hair growing material which contains a deer antler ingredient and its manufacturing method Download PDF

Info

Publication number
KR100903283B1
KR100903283B1 KR1020080088645A KR20080088645A KR100903283B1 KR 100903283 B1 KR100903283 B1 KR 100903283B1 KR 1020080088645 A KR1020080088645 A KR 1020080088645A KR 20080088645 A KR20080088645 A KR 20080088645A KR 100903283 B1 KR100903283 B1 KR 100903283B1
Authority
KR
South Korea
Prior art keywords
hair
antler
medium
hair growth
growth
Prior art date
Application number
KR1020080088645A
Other languages
Korean (ko)
Inventor
이성민
Original Assignee
이성민
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 이성민 filed Critical 이성민
Priority to KR1020080088645A priority Critical patent/KR100903283B1/en
Application granted granted Critical
Publication of KR100903283B1 publication Critical patent/KR100903283B1/en
Priority to PCT/KR2009/004972 priority patent/WO2010030092A2/en

Links

Images

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/32Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00Preparations for affecting hair growth
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9728Fungi, e.g. yeasts
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/06Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/10Dispersions; Emulsions

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Birds (AREA)
  • Botany (AREA)
  • Dermatology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Dispersion Chemistry (AREA)
  • Rheumatology (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • Zoology (AREA)
  • Cosmetics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

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

녹용 성분을 함유한 발모촉진물 및 그의 제조방법 {Hair Growing material which contains a deer antler ingredient and Its manufacturing method}Hair Growing Material Containing Deer Antler and Its Manufacturing Method {Hair Growing material which contains a deer antler ingredient and Its manufacturing method}

인체의 모발은 약 10만~15만개 정도이며, 각각의 모발은 서로 다른 주기를 가지며 성장기(anagen), 퇴행기(catagen), 휴지기(telogen)를 거쳐 성장, 탈락하게 된다. 이러한 주기는 3~6년에 걸쳐 반복되는데, 이 결과 일일 평균 50~100개의 모발이 정상적으로 탈락하게 된다. 일반적으로 탈모증이라 함은 성장기가 짧아져서 정상적으로 존재해야할 유모부, 특히 머리카락이 있는 두부에 모발이 휴지기 상태에서 지속되거나, 또는 그 크기 수가 감소하여 생기게 되는 상태이다.
최근 몇 십년동안 탈모 치료를 위한 많은 연구가 진행되고 있지만, 아직도 탈모의 원인이 무엇인지는 정확히 알려져 있지는 않다. 지금까지 밝혀진 탈모요인에 대한 내용을 살펴보면, 모발 주기 조절과 관련된 모유듀(dermal papilla)의 증식억제 또는 기능저하, 남성호르몬의 작용에 의한 모발주기의 비정상화, 두피로의 혈류량 저하로 인한 모발주기의 비정상적 변화, 항암제, 정신적 스트레스, 물리적 자극, 및 환경오염 등 거론되고 있다.
탈모증은 탈모의 원인과 형태로 구분되는데 유전형탈모, 지루성탈모, 원형탈모증, 영양결핍탈모증 약물 부작용에 의한 탈모증, 진구류에 의한 탈모증, 환경변화 부적응에 의한 탈모증, 신경성 탈모증 등 그 수가 대단히 많다. 특히 최근 들어 과다한 스트레스의 증가와 식생활의 변화, 공해 등으로 인한 탈모로 고민하는 인구가 늘어나고 있는 추세로 그 연령 또한 낮아지고 여성의 탈모 인구도 늘어나고 있다.
현재 탈모 치료와 관련하여 의학적 효능이 입증된 방식은 약물치료와 모발이식술이다. 치료에 사용되는 약물의 경우, 현재 국내 병원에서 처방되는 것은 피나스레리드(finasteride) 계열의 ‘먹는 약’과 미녹시딜(minoxidil) 계열의 ‘바르는 약’ 등 두 가지로 판단 기준은 미국 식품의약국(FDA)의 승인 여부다. 이외에 에스테로겐, 에스트라지올 등의 남성호르몬 작용을 억제하기 위한 호르몬제 및 펜타테칸산, 휘나스테리드 등의 남성 호르몬 활성 억제제 등이 있다.
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.

녹용이란 사슴의 뿔이 딱딱하게 각질화되기 전에 잘라서 약으로 사용하는 것을 말하며 전통적으로 인삼과 더불어 가장 우수한 보형강장제로 한방에서 수침액으로 빈번히 사용되어 왔다. 따라서, 국내에서 사슴사육의 목적은 주로 녹용생산을 위한 것이고 녹용의 약용으로서의 효능은 한방에서 오래전부터 기록되어 있다.
동의보감 : 신경쇠약, 피로회복, 성기능 저하에 좋은 물질이다.
본초비용 : 보혈, 빈혈에 특효가 있고, 양기를 보호해 준다.
본초강목 : 정과 수, 음과 혈을 보호하며, 병후 원기회복, 허약한 사람, 폐결핵, 폐기능 저하에 좋다.
의방유취 : 남녀의 모든 허약증, 영양실조, 허리나 다리의 통증, 피부소양감 등을 낫게 한다.
본초정 : 자궁출혈, 냉대하증 등 여성의 온갖 병에 좋다.
천금방 : 불면증, 가슴 두근거림을 치료한다.
종교본초강목 : 산전 후 몸조리, 가장 보호에 탁월하다.
약성론, 본초경소론 : 신경을 덮혀 준다, 태아를 편안하게 한다. 약기를 보강하며 뼈를 튼튼하게 한다.
이러한, 녹용의 주성분은 지질, 단백질, 탄수화물, 무기질 및 미량원소 등으로 구성되어 있으며 이런 성분들에 기인하여 녹용이 다양한 생리활성이 발형하는 것으로 받아들여지도 있다. 한편, 녹용의 생리기능에대한 연구는 국내 한의학분야에서 활발히 진행되어, 항피로효과, 면역활성 증가작용, 단백질 합성촉진작용, 내분비 기능촉진작용, 조혈작용, 면역활성 증가작용, 진통작용, 콜레스테롤 저하 작용, 간기능개선작용, 성장촉진작용 및 당뇨병치료효과 등이 보고되고 있다.
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.

녹용은 고가의 약재로 뛰어난 효능에 비하여 그 사용 분야가 한정되어 있다. 본 발명의 목적은 녹용유래세포의 배양을 통해 녹용성분이 함유된 배양액과 이의 사용법에 관한 것으로 부작용이 적고, 효과적으로 모근을 활성화하여 탈모를 방지하고 발모를 촉진하며 그 효과가 지속되는 발모 조성물로 제공된다는 것에 있다. 본 발명은 녹용 배지 및 녹용성분이 함유된 녹용세포 배양 후 배지의 육모 효능을 시험관 내 실험방법으로 조사하여, 녹용 배양액의 녹용성분이 발모치료제로 이용할 수 있는 근거를 마련하고자 하였다.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.

본 발명은 녹용유래세포의 배양을 통하여 녹용성분 생산 방법을 제시하고, 녹용유래세포를 배양하여 녹용성분이 함유되어있는 배양액이 모근의 활성을 통하여 모발의 발모를 촉진하고 탈모를 방지하는 효과가 있다는 것을 밝혔다.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.

녹용유래세포의 배양을 실시하고 이에 따라 녹용성분이 함유된 배양액과 대조물질인 녹용 세포 배양 전의 배지(일반 배지) 그리고 황산 미녹시딜(minoxidil sulfate)을 통하여 발모실험을 실시하였다. 실험용 쥐(Wistar rat)의 모낭을 분리하고 24 웰 플레이트(24 well plate)의 각 웰(well)에 하나씩의 모낭을 넣어서 37°C, 5% CO2 항온기에서 배양하였으며, 한 실험군에 7-9개의 모낭을 이용하였다. 배양 중에 배지는 3 일마다 교환하였으며 일반 배지 및 녹용 성분 함유 배양액을 1, 10 및 100 ug/㎖의 농도로 처리하고, 양성 대조물질인 미녹시딜 황산 (minoxidil sulfate ; Sigma, USA)는 1 uM의 농도로 처리하였다. 배양 각각의 모낭 길이를 이미지 분석장치(image analyzer,DP controller; Olympus, Japan)를 사용하여 측정하고 모낭 길이 변화의 평균값을 구하고 대조군의 평균길이와 비교하여 성장 정도를 측정하였다. 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.

실시예1 :
1-1 녹용유래세포의 추출 및 배양
3~7세의 건강하고 번식 적령기며, 연간 4.5킬로그램 이상의 녹용을 생산하는 우량종 숫사슴을 선택하여, 녹용의 적출을 시도하였다. 적출한 녹용을 멸균된 수술 도구를 사용하여 피부를 제거하고, 인산완충용액(PBS; Phosphate Buffered Saline)에 약 2시간 동안 배양하여 녹용 조직에 있는 혈액을 제거하였다. 녹용 윗 말단에서 아래로 1~3cm 되는 녹용조직을 수술용 가위 및 칼를 이용하여 분리하였다. 이후 화학적 분해를 위해 교원질분해 효소(Collagenase)를 이용하였다. 먼저 DMEM(둘베코의 개질된 이글 배지; Dulbecco's Modified Eagle's Medium) 배지에 제1-S형 교원질분해효소(Collagenase Type I-S) 또는 제4형 교원질분해효소(Collagenase Type IV) 또는 제3형 교원질분해효소(Collagenase type III)를 0.5mg/ml의 농도로 조제하였다. 이렇게 준비된 DMEM-collagenase 배지를 녹용 조직에 처리하여 34~37℃,5% CO2의 배양기에 약 6시간 정도 배양하였다. 녹용 세포가 함유된 배지는 원심분리기에서 1200 rpm로 5분간 원심분리하여 교원질분해효소(collagenase) 및 불순물을 제거하였다. 이렇게 분리된 세포는 5%의 FBS(우태혈청; fetal bovaine serum)를 함유하는 세포배양용 배지가 담긴 플라스크에 접종하고, 34~37℃,5% CO2의 배양기에서 24~72시간 배양하여 배양용기 표면에 부착하여 세포단층을 생성하는 세포만을 계대배양하였다.
1-2 녹용 배지의 구성
녹용유래세포의 배양을 위한 배지는 D-media(#78-5470EB, Gibco)를 사용하였고 5%~10%의 FBS(우태혈청; fetal bovaine serum)를 추가하여 배지를 구성하였다.
1-3 녹용 성분 함유 배양액
녹용유래세포를 1x 105 개씩 녹용 배지(5% FBS + D-media)가 담긴 25T 플라스크에 접종하여, 34~37℃, 5% CO2의 배양기에서 24~72시간 배양하고, 세포가 배양용기 표면에 80%가 되도록 자랐을 때 배양된 배지를 수거하였으며, 원심분리기에서 1200 rpm로 5분간 원심분리하여 이물질을 제거하였다.
실시예2 :
2-1 재료
본 실험재료인 일반 배지 및 녹용 성분 함유 배양액은 동결건조하고, 각각의 동결건조 파우더를 디메틸 술폭시드(DMSO; dimethyl sulfoxide)로 녹여 실험에 사용하였다.
2-2 시험관 내 육모 효능 측정
2-2-1 Rat 모낭의 분리 및 배양
생후 3주령인 실험용 쥐(Wistar rat) 수컷을 (Japan SLC, Hamamatsu, Janpan) ㈜ 중앙실험동물로부터 구입하여 에틸에테르(ethyl ether)로 마취 후 경추도살 하였다. Rat 왼쪽과 오른쪽 수염 부위(mystacial pads)를 분리하여 100 units/㎖ 페니실린(penicillin) - 100 ㎍/㎖ 스트렙토마이신(streptomycin) (Gibco Inc, NY, USA)이 함유된 E/P 완충용액 [이글의 평형염용액(Earle's balanced salts solution; EBSS, Sigma MO, USA) + 인산완충용액(PBS,phosphate-buffered saline; Sigma MO, USA)]에 넣었다. 해부 현미경으로 관찰하며 모낭을 조심스럽게 분리하였다. 모낭이 모두 분리될 때까지 E/P 완충용액을 넣은 페트리 접시에 분리된 모낭을 넣고 37°C, 5% CO2 항온기에서 1시간 정도 배양하였다. 24웰 배양용기(24 well plate)의 각 월(well)에 2 mM L-글루타민(L-glutamine; Gibco Inc, NY, USA), 10 ㎍/㎖ 인슐린(insulin; Sigma MO, USA), 50 nM 하이드로코르티손(hydrocortisone; Sigma MO, USA)과 100 units/㎖ 페니실린(penicillin) - 100 ㎍/㎖ 스트렙토마이신(streptomycin)을 포함하는 500 ㎕의 윌리엄 배지(Williams' medium E; Gibco Inc, NY, USA)을 넣고, 하나의 웰(well)에 하나씩의 모낭을 넣어서 37°C, 5% CO2 항온기에서 배양하였다. 한 실험군에 7-9개의 모낭을 이용하였으며, 배양 중에 배지는 3 일마다 교환하였다. 일반 배지 및 녹용 성분 함유 배양액을 1, 10 및 100 ug/㎖의 농도로 처리하였으며, 양성 대조물질인 미녹시딜 황산(minoxidil sulfate; Sigma, USA)은 1 uM의 농도로 처리하였다.
2-2-2 모낭 성장의 측정
배양중인 모낭의 형태는 현미경 (Olympus, Japan)을 사용하여 촬영하였다. 모낭 길이는 이미지 분석장치 (image analyzer; DP controller; Olympus, Japan)를 사용하여 0, 7, 10, 14 및 21일에 측정하였다. 모낭 길이 변화의 평균값을 구하고 대조군의 평균길이와 비교하여 성장 정도를 측정하였다.
실시예 3 :
3-1 일반 배지의 시험관 내 육모 효능
생후 3 주령 실험용 쥐의 성장기 모낭을 분리하여 3주간 배양하면서 모낭 길이를 측정하여 녹용 배지의 육모 효능을 조사하였다 (도 1). 배양 후 21일째 녹용 배지 처리군과 대조군과의 모낭의 길이 차이 (%)를 비교한 결과, 1, 10 및 100 ug/ml 녹용 배지를 각각 처리하였을 때 1 ug/ml의 농도에서 대조군 (100± 6%)보다 모섬유의 성장이 111.4± 6.1%로 증가하였으며 10 및 100 ug/ml의 농도에서는 증가효과가 없었다. 양성대조군인 1uM 미녹시딜 황산(minoxidil sulphate)는 111.8± 6.7% 성장을 나타내었다 (도 2).
3-2 녹용 성분 함유 배양액의 시험관 내 육모 효능
생후 3 주령 실험용 쥐의 성장기 모낭을 분리하여 3주간 배양하면서 모낭 길이를 측정하여 녹용 성분 함유 배양액의 육모 효능을 조사하였다 (도 3). 배양 후 21일째 녹용 성분 함유 배양액 처리군과 대조군과의 모낭의 길이 차이 (%)를 비교한 결과, 1, 10 및 100 ug/ml 녹용 성분 함유 배양액을 각각 처리하였을 때 10 ug/ml의 농도에서 대조군 (100± 6%)보다 모섬유(hair fiber)의 성장이 141.3± 8.2%로 더 증가하였으며 1 및 100 ug/ml의 농도에서는 증가효과가 없었다. 양성대조군인 1uM 미녹시딜 황산(minoxidil sulphate)는 111.8± 6.7% 성장을 나타내었다.(도 4).
결과
배양 21후 각각의 모낭 길이를 이미지 분석장치(image analyzer; DP controller; Olympus, Japan)를 사용하여 측정하고 모낭 길이 변화의 평균값을 구하고 대조군의 평균길이와 비교하여 성장 정도를 측정하였다. 결과 녹용 성분 함유 배지의 성장효과가 뛰어난 것으로 보여진다.(표 1)
대조군 비교 성장효과 일반 배지 111.4± 6.1% 녹용 성분 함유 배양액 141.3± 8.2% minoxidil sulphate 111.8± 6.7%
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%

도1은 일반 배지의 육모 효능 사진이다.
도2은 일반 배지의 육모 효능을 그래프화한 것이다.
도3은 녹용 성분 함유 배양액의 육모 효능 사진이다.
도4은 녹용 성분 함유 배양액의 육모 효능 그래프화한 것이다.
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)

녹용유래세포의 배양액을 유효성분으로 포함하는 발모 촉진 조성물Hair growth promoting composition comprising a culture solution of antler-derived cells as an active ingredient 제 1항에 있어서, 상기항의 발모 촉진 조성물의 제형이 액상, 스프레이, 겔, 페이스트, 유제, 크림, 콘디셔너 또는 샴푸인 것을 특징으로 하는 발모 촉진 조성물The 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. 삭제delete 삭제delete
KR1020080088645A 2008-09-09 2008-09-09 Hair growing material which contains a deer antler ingredient and its manufacturing method KR100903283B1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
KR1020080088645A KR100903283B1 (en) 2008-09-09 2008-09-09 Hair growing material which contains a deer antler ingredient and its manufacturing method
PCT/KR2009/004972 WO2010030092A2 (en) 2008-09-09 2009-09-03 Hair growth stimulating composition containing a deer antler ingredient, and method for preparing same

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
KR1020080088645A KR100903283B1 (en) 2008-09-09 2008-09-09 Hair growing material which contains a deer antler ingredient and its manufacturing method

Publications (1)

Publication Number Publication Date
KR100903283B1 true KR100903283B1 (en) 2009-06-17

Family

ID=40982862

Family Applications (1)

Application Number Title Priority Date Filing Date
KR1020080088645A KR100903283B1 (en) 2008-09-09 2008-09-09 Hair growing material which contains a deer antler ingredient and its manufacturing method

Country Status (2)

Country Link
KR (1) KR100903283B1 (en)
WO (1) WO2010030092A2 (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180047952A (en) * 2016-11-02 2018-05-10 (주)프로스테믹스 Functional composition comprising deer antlers derived stem cell culture medium
KR20180048431A (en) * 2017-11-16 2018-05-10 (주)프로스테믹스 Functional composition comprising deer antlers derived stem cell culture medium
KR102170417B1 (en) * 2019-05-28 2020-10-27 주식회사 미오내츄럴 Gel-type hairdye using natural materials and its manufacturing method
KR102192317B1 (en) * 2020-04-29 2020-12-21 주식회사 메디컬오 Composition for Skin Soothing Comprising Exosomes Derived from Natural Extracts

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007001011A (en) * 2006-10-10 2007-01-11 Mitsubishi Materials Corp Drilling tool

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1018801B (en) * 1989-03-09 1992-10-28 陈峰林 Releasing powder for metallic cast
CN1242057C (en) * 2003-09-15 2006-02-15 梁国坚 Simple process for cultivating organization cell of antler
KR100806125B1 (en) * 2005-12-29 2008-02-22 현천근 Manufacture method shampoo for stopping the hair from falling out and shampoo
KR100747651B1 (en) * 2006-02-01 2007-08-08 조점화 Material compositions for hair health care and hair growth acceleration
KR20070091056A (en) * 2006-03-04 2007-09-07 허문표 Hair restorer and method of producing the same

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007001011A (en) * 2006-10-10 2007-01-11 Mitsubishi Materials Corp Drilling tool

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
"줄기세포 통한 ‘녹용’ 인공배양 성공", 2007.10.11.자 한의신문*

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180047952A (en) * 2016-11-02 2018-05-10 (주)프로스테믹스 Functional composition comprising deer antlers derived stem cell culture medium
KR101894229B1 (en) * 2016-11-02 2018-09-04 (주)프로스테믹스 Functional composition comprising deer antlers derived stem cell culture medium
KR20180048431A (en) * 2017-11-16 2018-05-10 (주)프로스테믹스 Functional composition comprising deer antlers derived stem cell culture medium
KR101885501B1 (en) 2017-11-16 2018-08-06 (주)프로스테믹스 Functional composition comprising deer antlers derived stem cell culture medium
KR102170417B1 (en) * 2019-05-28 2020-10-27 주식회사 미오내츄럴 Gel-type hairdye using natural materials and its manufacturing method
KR102192317B1 (en) * 2020-04-29 2020-12-21 주식회사 메디컬오 Composition for Skin Soothing Comprising Exosomes Derived from Natural Extracts

Also Published As

Publication number Publication date
WO2010030092A2 (en) 2010-03-18
WO2010030092A3 (en) 2010-07-15

Similar Documents

Publication Publication Date Title
US20210220256A1 (en) Hair growth promoting capacity of conditioned media of stimulated stem cells and use thereof
CN105106028B (en) A kind of peptide composition for hair growth
JP4974532B2 (en) Use of at least one cypress plant extract as an anti-glycation agent
JPH10265347A (en) Agent for extending hair growth period
EP3146973B1 (en) Hair growth-promoting function of small-sized stem cells and use thereof
JPH07330554A (en) Hair tonic
JP2017500882A (en) Method for inducing pluripotent stem cells and pluripotent stem cells produced by the method
KR20130088997A (en) Cosmetic composition for hair growth and restoration from caviar extracts and its fermentation
KR100903283B1 (en) Hair growing material which contains a deer antler ingredient and its manufacturing method
KR20000038214A (en) Hair growth facilitation composition
KR20100096447A (en) Cosmetic composition comprising matrials cultured adult stem cells derived from swine placenta tissue and proteins extracted therefrom
EP0867169B1 (en) Hair growth phase extender composition
KR101114307B1 (en) Hair growth agent composition
US9161900B2 (en) Cosmetic composition for inhibiting hair loss and enhanced hair growth
KR101066797B1 (en) Hair growth agent composition
KR101953257B1 (en) Composition for preventing hair loss and growing hair comprising tulip, saw palmetto, nettle extracts
JP2009091325A (en) Hair growth agent
CN113876611A (en) Application of sialic acid in preparation of preparation for promoting collagen production
JPH10226628A (en) Hair papilla-activating agent
KR101570808B1 (en) Hair Care Composition Containing Human Adipocyte Conditioned Media Extracts and Pleuropterus Multiflorus Stem Cell Culture Extracts for Hair Growth
JP2016065019A (en) Integrin expression promoter
CN115554380B (en) Composition for preventing and treating alopecia by acting on hair follicle tissue and application thereof
CN114948932B (en) Use of a composition comprising mulberenone for the preparation of a product for the prevention and treatment of hair loss by acting on the hair follicle tissue
JP7178087B2 (en) Stem cell undifferentiated state maintenance agent and growth promoter
JP7202610B2 (en) Stem cell undifferentiated state maintenance agent and growth promoter

Legal Events

Date Code Title Description
A201 Request for examination
A302 Request for accelerated examination
E902 Notification of reason for refusal
E90F Notification of reason for final refusal
E701 Decision to grant or registration of patent right
GRNT Written decision to grant
FPAY Annual fee payment

Payment date: 20120910

Year of fee payment: 4

FPAY Annual fee payment

Payment date: 20131209

Year of fee payment: 5

FPAY Annual fee payment

Payment date: 20141209

Year of fee payment: 6

FPAY Annual fee payment

Payment date: 20151208

Year of fee payment: 7

FPAY Annual fee payment

Payment date: 20161208

Year of fee payment: 8

FPAY Annual fee payment

Payment date: 20171207

Year of fee payment: 9

FPAY Annual fee payment

Payment date: 20180809

Year of fee payment: 10

FPAY Annual fee payment

Payment date: 20190515

Year of fee payment: 11