KR20240127216A - Method for enhancing the fucntion of dermal papilla cell, and the use of dermal papilla cell therefrom - Google Patents
Method for enhancing the fucntion of dermal papilla cell, and the use of dermal papilla cell therefrom Download PDFInfo
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Abstract
본 발명은 모유두세포의 배양 중에 초음파 자극을 가함으로써 상기 모유두세포의 기능을 유지 또는 강화하는 배양 방법에 관한 것으로, 본 발명에 따르면 초음파 자극에 의해 모유두세포의 증식이 증가하고 모유두세포의 Wnt/β-catenin 신호전달을 촉진하여 탈모 방지 또는 발모 촉진의 세포 기능 강화 효과를 가질 수 있다.The present invention relates to a culturing method for maintaining or strengthening the function of breast papilla cells by applying ultrasonic stimulation during the culturing of the breast papilla cells. According to the present invention, ultrasonic stimulation increases the proliferation of breast papilla cells and promotes Wnt/β-catenin signaling of the breast papilla cells, thereby having the effect of strengthening cell function for preventing hair loss or promoting hair growth.
Description
본 발명은 모유두세포의 배양 중에 초음파 자극을 가함으로써 상기 모유두세포의 기능을 유지 또는 강화하는 배양 방법에 관한 것이다.The present invention relates to a culturing method for maintaining or strengthening the function of breast papilla cells by applying ultrasonic stimulation during the culturing of the breast papilla cells.
모발은 일정한 모주기(hair cycle)를 거치게 되며, 모유두(dermal papilla)의 자극을 받은 모모세포(keratinocyte)가 활발히 분열 및 증식하여 모발이 자라는 성장기(anagen), 모구(hair bulb)에 혈액공급이 끊기고 모유두가 모낭으로부터 분리되는 퇴행기(catagen) 및 세포 증식이 멈춰 모발이 성장하지 않는 휴지기(telogen)를 거쳐 다시 성장기로 들어가거나 모발이 두피로부터 빠지는 탈락기(exogen)로 가게 된다. Hair goes through a regular hair cycle, with the growth phase (anagen) in which keratinocytes stimulated by the dermal papilla actively divide and proliferate, the regression phase (catagen) in which the blood supply to the hair bulb is cut off and the dermal papilla separates from the hair follicle, and the resting phase (telogen) in which cell proliferation stops and hair does not grow, before either entering the growth phase again or going into the shedding phase (exogen) in which the hair falls out from the scalp.
사람의 모발은 각각 독립된 성장주기를 가지며, 일부는 탈락기로, 일부는 성장기로 들어가며 전체적으로 동일한 수준의 모발수를 유지하게 되는데, 이러한 균형이 탈락기로 기울어 정상적으로 모발이 존재해야 할 부위에 모발이 없어지는 상태를 탈모증(alopecia)이라 한다.Each human hair has an independent growth cycle, with some entering the shedding phase and some entering the growth phase, while the overall number of hairs is maintained at the same level. When this balance tilts toward the shedding phase, hair disappears in areas where hair should normally be present, this condition is called alopecia.
탈모(alopecia)는 유전적 요인과 더불어 환경적 요인 스트레스 등의 원인으로 오늘날 사회 진출이 활발한 젊은 층에서도 발명율이 증가하는 추세이다. 기존의 탈모 치료 방법은 일반적으로 모발이식, 수술, 약물 치료, 세포치료제 및 레이저 치료 기술 등으로 구분된다. 하지만, 기존 기술은 고비용, 낮은 치료 효율, 극심한 통증 및 부작용, 기술 수준 미비와 같은 문제점을 가지고 있어 새로운 치료 방법의 연구가 요구되고 있다,Hair loss (alopecia) is on the rise among young people who are actively participating in society today due to environmental factors such as stress as well as genetic factors. Existing hair loss treatment methods are generally divided into hair transplantation, surgery, drug treatment, cell therapy, and laser treatment. However, existing technologies have problems such as high cost, low treatment efficiency, severe pain and side effects, and insufficient technology level, so research on new treatment methods is required.
생물체 유래 단백질 유전자 세포 등을 원료로 한 의약품인 생물의약품(biologics)은 합성의약품처럼 체내에서 발생되는 대사 산물이 없어 독성이 낮으며 부작용이 적다는 장점을 가진다. 최근 이러한 장점이 주목받으면서 줄기세포를 이용하여 탈모를 치료하는 방법이 개발되고 있다. Biologics, which are medicines made from proteins, genes, cells, etc. derived from living organisms, have the advantage of being less toxic and having fewer side effects because they do not produce metabolic products in the body like synthetic medicines. Recently, as these advantages have been gaining attention, methods for treating hair loss using stem cells are being developed.
초음파는 기존 기술과 비교하여, 비침습성, 무고통, 무독성, 지속성 및 적용 다양성 등의 특징을 가지므로 최근 다양한 질병 치료 분야에 적용되고 있다.Compared to existing technologies, ultrasound has the characteristics of non-invasiveness, painlessness, non-toxicity, sustainability, and application versatility, and is therefore being applied to various disease treatment fields.
종래기술로서 특허문헌 1은 엑소좀을 얻기 위해 HDF세포에 초음파 처리하는 방법만을 개시하고 있다. HDF세포에 초음파을 처리하여 효능이 있다는 것이 다른 세포도 가능하다는 것을 암시하지는 않으므로 향후 많은 연구가 요구되고 있다.As a prior art, Patent Document 1 only discloses a method of treating HDF cells with ultrasound to obtain exosomes. The fact that treating HDF cells with ultrasound is effective does not imply that it is also effective for other cells, so much research is required in the future.
본 발명은 모유두세포의 배양 중에 초음파 자극을 가함으로써 상기 모유두세포의 기능을 유지 또는 강화하는 배양 방법을 제공하는 것이다.The present invention provides a culturing method for maintaining or strengthening the function of breast papilla cells by applying ultrasonic stimulation during the culturing of the breast papilla cells.
또한, 본 발명은 상기 방법으로 얻어지며, 모낭형성 및 모낭주기를 조절한다고 알려진 Wnt5a, Wnt10b, β-catenin 및 VEGF 중 1종 이상의 발현을 증가시키는 탈모 방지 및 발모 촉진용 조성물을 제공하는 것이다.In addition, the present invention provides a composition for preventing hair loss and promoting hair growth, which is obtained by the above method and increases the expression of at least one of Wnt5a, Wnt10b, β-catenin and VEGF, which are known to regulate hair follicle formation and hair follicle cycle.
또한, 본 발명은 상기 조성물을 포함하는 약학적 조성물 또는 화장료 조성물 중 어느 하나를 제공하는 것이다.In addition, the present invention provides either a pharmaceutical composition or a cosmetic composition comprising the composition.
본 발명에 따른 모유두세포의 기능 향상 방법은,The method for improving the function of mammary papilla cells according to the present invention is as follows:
(A) 모유두세포를 배지에 접종하는 단계;(A) A step of inoculating mammary papilla cells into a medium;
(B) 상기 모유두세포 및 배지에 초음파 처리하는 단계; 및(B) a step of sonicating the above-mentioned mammary gland cells and medium; and
(C) 초음파 처리된 세포를 배양시키는 단계;를 포함하는 것이 바람직하다.(C) It is preferable to include a step of culturing ultrasonically treated cells.
상기 초음파는 20~60 kHz 및 0.25~0.5 W/cm2의 음파인 것이 바람직하다.It is preferable that the above ultrasound be a sound wave of 20 to 60 kHz and 0.25 to 0.5 W/cm 2 .
상기 초음파 처리는 10~70초 동안 지속하는 것이 바람직하다.It is preferable that the above ultrasonic treatment last for 10 to 70 seconds.
상기 방법에 따르면 모유두세포의 증식을 증가시키는 것이 바람직하다.According to the above method, it is desirable to increase the proliferation of mammary papilla cells.
본 발명의 다른 측면으로서, 본 발명은 상기 방법에 따라 제조된 모유두세포를 포함하는 발모 촉진 또는 탈모 방지용 조성물을 제공하는 것이 바람직하다.As another aspect of the present invention, it is preferable to provide a composition for promoting hair growth or preventing hair loss comprising mammary papilla cells prepared according to the above method.
상기 조성물은 모낭형성 및 모낭주기의 조절인자인 Wnt5a, Wnt10b, β-catenin 및 VEGF 중 하나 이상의 발현을 증가시키는 것이 바람직하다.It is preferable that the composition increases the expression of one or more of Wnt5a, Wnt10b, β-catenin and VEGF, which are regulators of hair follicle formation and hair follicle cycle.
상기 탈모는 안드로겐성 탈모, 휴지기 탈모, 약제성 탈모, 기계적 탈모, 외상성 탈모, 압박성 탈모, 생장기 탈모, 비강성 탈모, 매독성 탈모, 지루 탈모, 증후성 탈모, 반흔성 탈모, 선천성 탈모, 원형 탈모, 머리 백선, 전두 탈모, 감모증, 유전성 단순 감모증 및 전신 탈모로 이루어진 군에서 선택되는 어느 하나 이상인 것이 바람직하다.The above hair loss is preferably at least one selected from the group consisting of androgenetic alopecia, telogen effluvium, drug-induced alopecia, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, nasal alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, congenital alopecia, alopecia areata, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex and alopecia universalis.
상기 조성물은 약학적 조성물 및 화장료 조성물 중 하나 이상에 포함되는 것이 바람직하다.It is preferable that the above composition be included in at least one of a pharmaceutical composition and a cosmetic composition.
상기 조성물은 팩, 밴드 또는 패치 중 어느 하나에 포함되는 것이 바람직하다.It is preferred that the composition be included in any one of a pack, a band or a patch.
상기 팩은 국소형 패치, 시트타입, 거즈타입, 크림타입 및 젤타입 중 어느 하나인 것이 바람직하다.It is preferable that the above pack be one of the following types: local patch, sheet type, gauze type, cream type, and gel type.
상기 조성물은 에어로졸, 액체, 용액, 젤, 페이스트, 크림, 연고, 폼 및 분말 중 어느 하나의 제형으로 되는 것이 바람직하다.The composition is preferably in the form of any one of an aerosol, liquid, solution, gel, paste, cream, ointment, foam, and powder.
상기 조성물은 헤어토닉, 헤어컨디셔너, 헤어에센스, 헤어로션, 헤어영양로션, 헤어샴푸, 헤어린스, 헤어트리트먼트, 헤어크림, 헤어영양크림, 헤어모이스처크림, 헤어맛사지크림, 헤어왁스, 헤어에어로졸, 헤어팩, 헤어영양팩, 헤어비누, 헤어클렌징폼, 머릿기름, 모발건조제, 모발보존처리제, 모발염색제, 모발용 웨이브제, 모발탈색제, 헤어겔, 헤어글레이즈, 헤어드레싱어, 헤어래커, 헤어모이스처라이저, 헤어무스 및 헤어스프레이를 포함하는 군에서 선택되는 제형에 포함되는 것이 바람직하다.The above composition is preferably included in a formulation selected from the group including hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair drying agent, hair preservation treatment agent, hair dye, hair waving agent, hair bleaching agent, hair gel, hair glaze, hair dressing agent, hair lacquer, hair moisturizer, hair mousse and hair spray.
본 발명은 모유두세포의 배양 중에 초음파 자극을 가함으로써 상기 모유두세포의 기능을 유지 또는 강화하는 배양 방법에 관한 것으로, 본 발명에 따르면 초음파 자극에 의해 모유두세포의 증식이 증가하고 모유두세포의 Wnt/β-catenin 신호전달을 촉진하여 탈모 방지 또는 발모 촉진의 세포 기능 강화 효과를 가질 수 있다.The present invention relates to a culturing method for maintaining or strengthening the function of breast papilla cells by applying ultrasonic stimulation during the culturing of the breast papilla cells. According to the present invention, ultrasonic stimulation increases the proliferation of breast papilla cells and promotes Wnt/β-catenin signaling of the breast papilla cells, thereby having the effect of strengthening cell function for preventing hair loss or promoting hair growth.
도 1은 실험예 1에 따라 모유두세포 증식능을 확인한 결과를 개략적으로 도시한 것이다.
도 2는 실험예 2에 따라 모낭형성 및 모발생장에 관여하는 인자(Wnt5a, Wnt10b, β-catenin 및 VEGF)들의 발현을 확인한 결과를 개략적으로 도시한 것이다.Figure 1 schematically illustrates the results of confirming the proliferation ability of mammary papilla cells according to Experimental Example 1.
Figure 2 schematically illustrates the results of confirming the expression of factors (Wnt5a, Wnt10b, β-catenin, and VEGF) involved in hair follicle formation and hair growth according to Experimental Example 2.
이하, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본 발명의 실시형태를 들어 상세히 설명한다. 본 발명의 실시형태는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 더욱 완전하게 설명하기 위해서 제공되는 것이다. 따라서, 본 발명의 실시형태는 여러 가지 다른 형태로 변형될 수 있으며, 본 발명의 범위가 이하 설명하는 실시형태로 한정되는 것은 아니다.Hereinafter, embodiments of the present invention will be described in detail so that those with ordinary knowledge in the technical field to which the present invention pertains can easily practice it. Embodiments of the present invention are provided so that those with average knowledge in the art can more completely explain the present invention. Therefore, embodiments of the present invention can be modified in various different forms, and the scope of the present invention is not limited to the embodiments described below.
본 발명의 명세서 전체에서, 어떤 부분이 어떤 구성요소를 “포함”한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외하는 것이 아니라 다른 구성요소를 더 포함하는 것을 의미한다.Throughout the specification of the present invention, when a part is said to “include” a certain component, this means that it further includes other components, rather than excluding other components, unless specifically stated otherwise.
본 발명의 명세서 전체에서, 어떤 단계가 다른 단계와 “상에” 또는 “전에” 위치하고 있다고 할 때, 이는 어떤 단계가 다른 단계와 직접적 시계열적인 관계에 있는 경우뿐만 아니라, 각 단계 후의 혼합하는 단계와 같이 두 단계의 순서에 시계열적 순서가 바뀔 수 있는 간접적 시계열적 관계에 있는 경우와 동일한 권리를 포함할 수 있다.Throughout the specification of the present invention, when a step is said to be located “on” or “before” another step, this may include the same rights as when a step has a direct time-series relationship with the other step, as well as when the two steps have an indirect time-series relationship where the time-series order may be changed, such as a mixing step after each step.
본 발명의 명세서 전체에서 사용되는 정도의 용어 “약”, “실질적으로” 등은 언급된 의미에 고유한 제조 및 물질 허용 오차가 제시될 때 그 수치에서 또는 그 수치에 근접한 의미로 사용되고, 본 발명의 이해를 돕기 위해 정확하거나 절대적인 수치가 언급된 개시 내용을 비양심적인 침해자가 부당하게 이용하는 것을 방지하기 위해 사용된다. 본원 명세서 전체에서 사용되는 용어 “~(하는) 단계” 또는 “~의 단계”는 “~를 위한 단계”를 의미하지 않는다.The terms “about,” “substantially,” etc., used throughout the specification of the present invention are used in a meaning close to or at the numerical value when manufacturing and material tolerances inherent in the stated meaning are presented, and are used to prevent unscrupulous infringers from unfairly using the disclosure contents in which exact or absolute values are mentioned to help understanding of the present invention. The terms “~ (doing) step” or “~ step” used throughout the specification of the present invention do not mean “~ step.”
본 발명에 따른 모유두세포의 기능 향상 방법은,The method for improving the function of mammary papilla cells according to the present invention is as follows:
(A) 모유두세포를 배지에 접종하는 단계;(A) A step of inoculating mammary papilla cells into a medium;
(B) 상기 모유두세포 및 배지에 초음파 처리하는 단계; 및(B) a step of sonicating the above-mentioned mammary gland cells and medium; and
(C) 초음파 처리된 세포를 추가로 배양시키는 단계;를 포함하는 것이 바람직하다.(C) It is preferable to include a step of additionally culturing the sonicated cells.
초음파는 주파수가 20 kHz 이상 10 MHz 미만인 기계적 음파로, 다양한 산업분야에서 사용되고 있다. 특히 식품 분야에서는 식물 등에서 생리 활성 물질들을 추출하는데 이용하기도 하고, 식품의 유통기한 연장을 위해 미생물을 파괴하는데 이용하기도 한다. 본 발명에서 초음파는 20~60 kHz 및 0.25~0.5 W/cm2의 음파인 것이 바람직하다. 구체적으로는, 40 kHz 주파수로 처리하였다.Ultrasound is a mechanical sound wave with a frequency of 20 kHz or more and less than 10 MHz, and is used in various industrial fields. In particular, in the food field, it is used to extract physiologically active substances from plants, etc., and is also used to destroy microorganisms to extend the shelf life of food. In the present invention, it is preferable that the ultrasound is a sound wave of 20 to 60 kHz and 0.25 to 0.5 W/cm 2 . Specifically, it was processed at a frequency of 40 kHz.
상기 초음파 처리는 10~70초 동안 지속하는 것이 바람직하다. 구체적으로는, 15~60초 동안 처리하였다.It is preferable that the above ultrasonic treatment last for 10 to 70 seconds. Specifically, the treatment was performed for 15 to 60 seconds.
상기 방법에 따르면 모유두세포의 증식이 증가되어 생존 및 지속가능성이 높아진다.According to the above method, the proliferation of mammary papilla cells increases, thereby enhancing their survival and sustainability.
본 발명의 다른 측면으로서, 본 발명은 상기 방법에 따라 제조된 모유두세포를 포함하는 발모 촉진 또는 탈모 방지용 조성물을 제공하는 것이 바람직하다.As another aspect of the present invention, it is preferable to provide a composition for promoting hair growth or preventing hair loss comprising mammary papilla cells prepared according to the above method.
상기 조성물은 모낭형성 및 모낭주기의 조절인자인 Wnt5a, Wnt10b, β-catenin 및 VEGF 중 하나 이상의 발현을 증가시키는 것이 바람직하다.It is preferable that the composition increases the expression of one or more of Wnt5a, Wnt10b, β-catenin and VEGF, which are regulators of hair follicle formation and hair follicle cycle.
본 발명의 조성물이 적용되는 탈모는 안드로겐성 탈모, 휴지기 탈모, 약제성 탈모, 기계적 탈모, 외상성 탈모, 압박성 탈모, 생장기 탈모, 비강성 탈모, 매독성 탈모, 지루 탈모, 증후성 탈모, 반흔성 탈모, 선천성 탈모, 원형 탈모, 머리 백선, 전두 탈모, 감모증, 유전성 단순 감모증 및 전신 탈모를 포함할 수 있으나, 이에 제한되는 것은 아니다.Hair loss to which the composition of the present invention is applied may include, but is not limited to, androgenetic alopecia, telogen effluvium, drug-induced alopecia, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, nasal alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, congenital alopecia, alopecia areata, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex and alopecia universalis.
상기 조성물은 약학적 조성물 및 화장료 조성물 중 하나 이상에 포함되는 것이 바람직하다.It is preferable that the above composition be included in at least one of a pharmaceutical composition and a cosmetic composition.
본 발명에서의 약학 조성물은 정제, 캅셀제, 주사제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제 등을 포함하는 군에서 선택되는 제형일 수 있으나 이에 제한되지는 않는다.The pharmaceutical composition in the present invention may be a dosage form selected from the group including, but not limited to, tablets, capsules, injections, creams, gels, patches, sprays, ointments, pastes, lotions, liniments, pastes, and cataplasmas.
본 발명의 약학적 조성물은 제제시에 통상적으로 사용되는 약제학적으로 허용되는 담체, 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutical composition of the present invention may additionally contain pharmaceutically acceptable carriers, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. commonly used in formulation.
본 발명의 약학적 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여인 경우에는 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여, 점막 투여 및 점안 투여, 국소투여 등의 방식으로 투여할 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, mucosal administration, eye drop administration, topical administration, etc., but is not limited thereto.
본 발명의 약학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 섭취 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 바람직하게는, 본 발명의 약학적 조성물의 투여량은 성인 기준으로 0.0001-100 ㎎/kg(체중)일 수 있으나 이에 제한되는 것은 아니다.The appropriate dosage of the pharmaceutical composition of the present invention can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food intake, administration time, administration route, excretion rate, and reaction sensitivity. Preferably, the dosage of the pharmaceutical composition of the present invention can be 0.0001-100 mg/kg (body weight) for adults, but is not limited thereto.
본 발명에 추가될 수 있는 적합한 성분의 선택은, 당업자에게 자명한 범위에서, 본 발명의 피부 외용제 조성물이 용액, 로션, 림, 연고, 겔, 폼 또는 경피 패치로 제형화 되는지 여부에 어느 정도 의존할 것이다.The selection of suitable ingredients that may be added to the present invention will depend, to a degree, on whether the topical skin composition of the present invention is formulated as a solution, lotion, rim, ointment, gel, foam or transdermal patch, to the extent that will be apparent to those skilled in the art.
예를 들어, 착색제(색소), 착향제(향료), 현탁화제, 유화제, 용해보조제, 안정제, 등장제, pH조절제, 점도조절제, 용제 등을 포함할 수 있다. 화장료 조성물에 통상적으로 부가되는 성분이나 함량 등은 당업자에게 자명할 것이다.For example, it may include a coloring agent (pigment), a flavoring agent (fragrance), a suspending agent, an emulsifier, a solubilizing agent, a stabilizer, an isotonic agent, a pH adjusting agent, a viscosity adjusting agent, a solvent, etc. Components or contents commonly added to cosmetic compositions will be apparent to those skilled in the art.
본 발명에 따른 화장료 조성물은 에어로졸, 액체, 용액, 젤, 페이스트, 크림, 연고, 폼 및 분말 중 어느 하나의 제형으로 가능하며, 이에 제한되지 않는다.The cosmetic composition according to the present invention may be in any one of the formulations of an aerosol, liquid, solution, gel, paste, cream, ointment, foam, and powder, but is not limited thereto.
예를 들어, 로션은 용액과 유사하지만 용액보다 더 진하고 본질적으로 더 완화시키는 경향이 있다. 로션에는 일반적으로 물과 오일이 혼합되어 있으며 용액보다 알코올이 적은 경우가 많다.For example, lotions are similar to solutions, but they tend to be thicker and more emollient in nature. Lotions are usually a mixture of water and oil, and often contain less alcohol than solutions.
예를 들어, 크림은 거의 같은 비율의 오일과 물의 에멀젼을 가지고 있다. 피부의 각질층 외층까지 잘 침투한다. 크림은 로션보다 두껍고 용기에서 꺼낼 때 모양이 유지된다.For example, creams have an emulsion of oil and water in roughly equal proportions. They penetrate well into the outer layer of the skin's stratum corneum. Creams are thicker than lotions and retain their shape when taken out of the container.
또한, 본 발명의 조성물은 팩, 밴드 또는 패치 중 어느 하나에 포함될 수 있으며, 이에 제한되지 않는다.Additionally, the composition of the present invention may be included in any one of a pack, a band or a patch, but is not limited thereto.
상기 팩은 국소형 패치, 시트타입, 거즈타입, 크림타입 및 젤타입 중 어느 하나의 형태일 수 있다.The above pack may be in the form of a local patch, sheet type, gauze type, cream type, or gel type.
본 발명의 조성물은 헤어토닉, 헤어컨디셔너, 헤어에센스, 헤어로션, 헤어영양로션, 헤어샴푸, 헤어린스, 헤어트리트먼트, 헤어크림, 헤어영양크림, 헤어모이스처크림, 헤어맛사지크림, 헤어왁스, 헤어에어로졸, 헤어팩, 헤어영양팩, 헤어비누, 헤어클렌징폼, 머릿기름, 모발건조제, 모발보존처리제, 모발염색제, 모발용 웨이브제, 모발탈색제, 헤어겔, 헤어글레이즈, 헤어드레싱어, 헤어래커, 헤어모이스처라이저, 헤어무스 및 헤어스프레이 등을 포함하는 군에서 선택되는 제형일 수 있으나, 이에 제한되는 것은 아니다.The composition of the present invention may be a formulation selected from the group including, but not limited to, hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair drying agent, hair preservation treatment agent, hair dye, hair waving agent, hair bleaching agent, hair gel, hair glaze, hair dressing agent, hair lacquer, hair moisturizer, hair mousse, and hair spray.
이하, 본 발명의 내용에 대해 하기 실시예 또는 실험예를 통해 예시적으로 보다 상세히 설명하고자 한다.Hereinafter, the contents of the present invention will be explained in more detail through the following examples or experimental examples.
(실험예 1) 초음파에 의한 모유두세포 증식 효과 확인(Experimental Example 1) Confirmation of the effect of ultrasound on the proliferation of mammary papilla cells
모유두세포(dermal papilla cell)는 양모 주기를 조절하는 중요한 세포로 모낭 줄기 세포(hair follicle stem cell)와 상호작용하고 지시하는 역할을 하여 발모 주기에서 중요한 세포이다. 따라서, 모유두세포의 증식이 증가하는지 여부를 분석하면, 후보 물질의 모발 생성 촉진 효능을 평가할 수 있다.Dermal papilla cells are important cells that control the hair cycle and play a role in interacting with and directing hair follicle stem cells, making them important cells in the hair growth cycle. Therefore, by analyzing whether the proliferation of dermal papilla cells increases, the hair growth promotion efficacy of a candidate substance can be evaluated.
초음파가 처리된 모유두세포를 제조하기 위해, 먼저 플레이트에 모유두세포를 접종하고 초음파기기를 이용하여 40 KHz, 0.25~0.5 W/cm2의 초음파 자극을 15~60초간 직접 가한 뒤, 새로운 배지로 교체하였다. To prepare ultrasound-treated mammary papilla cells, mammary papilla cells were first seeded on a plate, and ultrasound stimulation of 40 KHz, 0.25–0.5 W/cm 2 was directly applied for 15–60 seconds using an ultrasound device, and then the medium was replaced with new medium.
배지 교체 후 37℃에서 24~72시간 동안 배양시킨 후, CCK-8 용액을 2시간 동안 반응시킨 뒤, 450 nm에서 흡광도를 측정하였다.After culture at 37°C for 24 to 72 hours after replacing the medium, the CCK-8 solution was reacted for 2 hours, and then the absorbance was measured at 450 nm.
도 1에서 보는 바와 같이, 초음파 처리에 의해 모유두세포 증식이 증가함을 확인하였다.As shown in Figure 1, it was confirmed that the proliferation of mammary papilla cells increased by ultrasonic treatment.
(실험예 2) 모낭형성능 및 모낭주기능(모발생장) 확인(Experimental Example 2) Confirmation of hair follicle formation performance and hair follicle main function (hair growth)
초음파 처리에 의한 모유두세포의 기능 강화 효과를 확인하기 위하여 모낭형성 및 모낭주기를 조절한다고 알려진 Wnt5a, W nt10b, β-catenin 및 VEGF의 총 4종의 유전자 발현을 확인하였다.To confirm the effect of enhancing the function of hair papilla cells by ultrasonic treatment, the expression of four genes, Wnt5a, Wnt10b, β-catenin, and VEGF, which are known to regulate hair follicle formation and hair follicle cycle, was confirmed.
구체적으로, 적정 농도로 접종된 모유두세포에 초음파기기를 이용하여 40 KHz, 0.25~0.5 W/cm2의 초음파 자극을 15~60 초간 직접 가한 뒤, 새로운 배지로 교체하였다.Specifically, ultrasound stimulation of 40 KHz, 0.25 to 0.5 W/ cm2 was directly applied for 15 to 60 seconds to mammary papilla cells inoculated at an appropriate concentration using an ultrasound device, and then the medium was replaced with a new medium.
배지 교체 후, 37℃에서 48시간 동안 배양시킨 후, 확보된 세포에 클로로포름 및 이소프로판올을 이용하여 총 RNA를 분리하였다. 상기 RNA를 주형으로 Maxima First Strand cDNA Synthesis Kit (Thermo Fisher)를 이용해 cDNA를 합성하였다.After replacing the medium, the cells were cultured at 37°C for 48 hours, and total RNA was isolated from the obtained cells using chloroform and isopropanol. cDNA was synthesized using the RNA as a template using the Maxima First Strand cDNA Synthesis Kit (Thermo Fisher).
이후, PowerUp?? SYBR?? Green Master Mix (Thermo Fisher)를 이용해 실시간 역전사 중합 효소 연쇄 반응 분석을 실시하였다.Afterwards, real-time reverse transcription polymerase chain reaction analysis was performed using PowerUp?? SYBR?? Green Master Mix (Thermo Fisher).
도 2에서 보는 바와 같이 모유두세포에 초음파 처리시 Wnt5a, Wnt10b, β-catenin 및 VEGF의 총 4종의 유전자 발현이 증가함을 확인하였다.As shown in Figure 2, it was confirmed that when ultrasonic treatment was applied to mammary papilla cells, the expression of four genes, Wnt5a, Wnt10b, β-catenin, and VEGF, increased.
이상에서 본 발명의 실시예에 대하여 상세하게 설명하였지만 본 발명의 권리범위는 이에 한정되는 것은 아니고, 청구범위에 기재된 본 발명의 기술적 사상을 벗어나지 않는 범위 내에서 다양한 수정 및 변형이 가능하다는 것은 당 기술분야의 통상의 지식을 가진 자에게는 자명할 것이다.Although the embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and it will be apparent to those skilled in the art that various modifications and variations are possible within a scope that does not depart from the technical spirit of the present invention described in the claims.
Claims (12)
(B) 상기 모유두세포 및 배지에 초음파 처리하는 단계; 및
(C) 초음파 처리된 세포를 배양시키는 단계;를 포함하는, 모유두세포의 기능 향상 방법.(A) A step of inoculating mammary papilla cells into a medium;
(B) a step of sonicating the above-mentioned mammary gland cells and medium; and
(C) A method for improving the function of mammary papilla cells, comprising the step of culturing ultrasonically treated cells.
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