밀싹 추출물이 모발성장에 미치는 효과

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Effect of Wheat Sprout Extracts on Hair Growth

Ms. Eun-Mi Ryu
Advisor : Hyun-Jae Shin, Ph.D.
Dept. of Chemical Engineering
(Major: Cosmetic Engineering)
Graduate School of Chosun University

Wheat (wheat, Triticum spp.) the annual crops of around 1 m height has been grown around the world. Wheat sprout used in this study was named by the young shoots before generating the wheat growing areas in the development process. Traditionally, wheat sprout has been reported to be very effective for Mediterranean anemia called β-thalassemia. Also wheat sprout juice is also rich in antioxidants and has a good efficacy of vitamin E, catalase and peroxidase activity which are considered to reduce age-related cataract. There have been some reports on the health effects but little reports on the effect of wheat sprout on scalp and hair. In this study, a wheat sprout extract as a powerful natural antioxidant prepared from a hydrophobic layer has been used to examine the hair growth and scalp conditioning, and to evaluate the impact of any immune signaling pathway. Some researches have been carried out and the results are as follows.

1. Literature review
Research trends on utilization methods and physiological activities of the ingredients of wheat and wheat sprout have been summarized as follows:
Research has been confined in wheat processing characteristics and physical and chemical characteristics in case of wheat, and several works on the nutrients study have been performed using the limited domestic wheat varieties. In addition, there has been no report on the domestic wheat (Kumkang) for the domestic literature and the anticancer, antioxidant activity, and anti-ulcer activities and improved blood flow have been reported for foreign wheat. Therefore, in this study, based on the literature review some works on wheat sprout for beauty industry have been performed.

2. Establishment of wheat sprout growing conditions
Four cultivar of domestic wheats (Keumkang, Woori, Jogyung, and Baekjoong) and four cultivar of foreign wheats (NS, USA; WH, USA; ZCW, Canada; Chinese one) have been compared in terms of the leaf width and thickness of sprout. Domestic ones are superior to those of foreign ones. For sprout growth Canada one grew well but showed musty bacteria contamination during the growth process resulting in browning of blossomed wheat sprout. Thus it could not be used in the experiment. Keumkang wheat cultivar has been used throughout the study considering the growth rate and the ease of wheat supply.
Wheat sprout (Triticum aestivum) shows excellent nutritional and health effects due to the contents in amino acids, minerals, and other nutrients rich in chlorophyll and vitamins. So, the ground coffee was added to wheat sprout media as the by-product component to increase the economic feasibility. In this study, spent coffee grounds were used to cultivate the wheat sprout for 12-day. An amount of 0%, 20%, 40%, 60%, 80%, and 100% (w/w, based on commercial sterile soil media) were used under the same conditions. Total length and weight of wheat sprout, above- and below-ground length and weight, and the chloropyll contents were compared. Soil media were analyzed before and after wheat cultivation, showing that 40% and 60% (w/w) coffee media promoted wheat growth in view of length and weight. Chlorophyll contents in each group showed almost constant values, while 100% (w/w) coffee media led to a slight decrease. In summary, spent coffee grounds stimulated wheat growth, showing nearly stable contents of chlorophyll.

3. Anti-inflammatory effect of wheat sprouts' hydrophobic fraction
Wheat sprout (Triticum aestivum L., Keumkang) powder was treated with 80% ethanol at room temperature for 72 hours. They were then extracted using ethyl acetate and n-butanol. Hydrophobic and hydrophilic fractions were loaded on a silica open column (one liter for each fraction) and analyzed by thin layer chromatography (TLC). The carbohydrate fractions were detected using chromogenic reagent and concentrated by evaporator. The separated fractions from wheat sprout were evaluated for the toxicity and anti-inflammation activity by MTT/NO assay, for ingredient identification using HPLC/MS. As a result, fraction no. 2, 7, 12, 14, 17, and 23 fractions from wheat sprout showed higher anti-inflammation activity than control.

4. Hair growth effect of ethanolic extract of wheat sprout
In this study, effects of natural antioxidants from wheat sprout extract on hair growth of C57BL/6 mouse (female) were evaluated. Hydrophobic layers (80% ethanol, open column) of wheat sprout (fr. 5 and fr. 8) were used, with minoxidil (3%) as a positive control. With one week adaption, hair growth of 6-week old C57BL/6 mouse (female) was observed after 4 weeks of back hair removal. Trans-dermal examination and hematoxylin and eosin (H&E) staining and toluidin blue staining of skin tissue were used before observation with an optical microscope, which showed similar hair growth in the positive control minoxidil (3%) as wheat sprout (fr. 5), while slower in wheat sprout (fr. 8). The slaughtering cervical tissue at 4 weeks after the C57BL/6 mouse with wheat sprout (fr. 5) started the transition to growth. Hematoxylin & eosin stained histological analysis of wheat sprout (fr. 5) showed more follicles on the skin surface and smaller mast cells (shown with toluidin blue dye). In summary, wheat sprout extract (fr. 5) on C57BL/6 mouse in dermal application and histological analysis promotes the hair growth, predicting an application of wheat sprout extract in a wide range of hair remedy products.

5. Effect on immune hypersensitivity of scalp
Among 24 kinds of dermal tissue cytokine measured, C5/C5a, GM-CSF, IL-1a, IL-16, IP-10, CXCL9/MIG, MIP-2, TMP-1 have been changed after the treatment of wheat sprout (WS) extract. Among them, GM-CSF, IL-16, IP-10, CXCL9 increased with the positive control, DNCB, the result of WS was similar to that of the control group. In spleen tissue, C5/C5a, ICAM-1, IL-1b, IL-1ra, IP-10, M-CSF, CXCL9/MIG, TNF-α have been changed. Some cytokine signals were decreased (C5/C5a, ICAM-1, IL-1b, IP-10, M-CSF, CXCL9/MIG, TNF-α). And IL-1ra were very close to the normal group. It may be speculated that WS can inhibit the inflammation by a signal transduction pathway.

In conclusion, through this study, the growth conditions of domestic wheat sprout have been established and a novel media including coffee ground waste has been introduced to increase the economic feasibility. Some antioxidant and anti-inflammation activities have been verified using the hydrophobic fraction of ethanolic extracts. Several ingredients responsible for the activities were suggested and more works will be done to identify them. For some fractions, hair promoting effects were examined using a mouse model and histological and immunological results were given. Much works on the immunological cytokines should be done afterwards. Wheat sprout and its extract is very promising to cosmetic and beauty industry because the raw materials are very easy to get, the cultivation cost is very low, and several scientific findings on hair growth are available.
Alternative Title
Effect of Wheat Sprout Extracts on Hair Growth
Alternative Author(s)
Eun-Mi Ryu
조선대학교 대학원 화학공학과
일반대학원 화학공학과
Awarded Date
Table Of Contents
목 차


제 1 장 서 론
1. 연구배경 1
2. 연구동향 3
3. 연구 목적 및 내용 7

제 2 장 연구이론
1. 모발 10
1.1 모발 10
1.2 모발의 생성 11
1.3 모발의 구조 및 기능 11
1.4 모발의 주기 13
2. 두피 15
2.1 두피 15
2.2 두피의 구조 16
2.3 두피 타입별 분류 17
3. 탈모 20
3.1 탈모 20
3.2 탈모의 유형 21
4. 면역 24

제 3 장 밀싹의 문헌적 고찰
1. 밀 28
2. 국산밀 30
3. 밀싹 36
3.1 밀싹 (wheat sprout or wheat grass) 36
3.2 밀싹의 유용성분 39
3.3 밀싹의 생리활성 42

제 4 장 밀싹의 재배 조건 확립
1. 국산밀과 수입산밀의 성장 비교 48
1.1 실험재료 48
1.2 실험방법 48
1.3 실험 결과 및 고찰 52
2. 원두커피 부산물 첨가에 따른 밀싹의 성장과
엽록소 성분의 변화 55
2.1 실험재료 55
2.2 실험방법 55
2.3 실험 결과 및 고찰 57

제 5 장 RAW 246.7 대식세포에서의 밀싹 추출물의 항염증 효과
1. 실험재료 67
2. 실험방법 68
2.1 시료추출 68
2.2 세포배양 70
2.3 MTT assay 70
2.4 NO assay 70
2.5 RNA 분리 및 RT-PCR 71
2.6 Western Blot 71
3. 실험 결과 및 고찰 73
3.1 MTT assay와 NO(Nitric oxide) assay 73
3.2 RT-PCR, Western Blot 78

제 6 장 밀싹 에탄올 추출의 C57BL/6 Mouse에
대한 발모효과
1. 실험재료 80
1.1 실험시료 80
1.2 실험동물 80
2. 실험방법 81
2.1 밀싹 재배 및 추출 81
2.2 실험동물 처치 82
2.3 육안적 분석(외형 효과) 82
2.4 육안적 분석(경피 내측 효과) 82
2.5 조직학적 분석(Hematoxylin & Eosin) 84
2.6 조직학적 분석(Toluidin Blue) 84
3. 실험 결과 및 고찰 85
3.1 육안적 분석(외형 효과) 85
3.2 육안적 분석(경피 내측 효과) 89
3.3 Mouse 조직학적 분석(Hematoxylin & Eosin) 91
3.4 조직학적 분석(Toluidin Blue) 93

제 7 장 밀싹 추출물이 두피 면역 과민반응에
미치는 영향
1. 실험재료 96
1.1 실험시료 96
1.2 실험동물 96
2. 실험방법 97
2.1 밀싹 재배 및 추출 97
2.2 실험동물 처치 및 실험방법 97
2.3 Cytokine 98
3. 실험 결과 및 고찰 100
3.1 육안적 분석(외형 효과) 100
3.2 조직학적 분석(Hematoxylin & Eosin) 102
3.3 조직학적 분석(Toluidin Blue) 102
3.4 Cytokine 분석 104

제 8 장 결론 및 제언
1. 밀싹의 문헌적 고찰 110
2. 밀싹 재배조건 확립 111
3. RAW 246.7 대식세포에서의 밀싹 추출물의
항염증 효과 112
4. 밀싹 에탄올 추출의 C57BL/6 Mouse에 대한
발모효과 113
5. 밀싹 추출물이 두피 면역 과민반응에
미치는 영향 114
6. 제언 115


감사의 글
조선대학교 대학원
류은미. (2013). 밀싹 추출물이 모발성장에 미치는 효과.
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General Graduate School > 4. Theses(Ph.D)
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