MC3T3-E1 골모유사세포의 분화에 대한 산수유 추출물의 효과

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Various traditional herbal medicines have attracted interests because they have fewer side-effects and are available for safe use, and are relatively inexpensive. Cornus officinalis has been widely used as a herbal medicine exerting various pharmacological actions such as diuretic, anti-allergic, hypotensive and antibiotic activities. Thus we studied the effect of Cornus officinalis on the proliferation, differentiation and matrix mineralization of MC3T3-E1 osteoblast-like cell line.
To do the experiments, Cornus officinalis extract(COE) was prepared using ethanol at different concentrations. The cytotoxic effect and the osteogenic differentiation were examined using MTT assay, alkaline phosphatase (ALP) activity, ALP staining, Alizarin red s staining and reverse transcriptase-polymerase chain reaction for osteogenic differentiation markers such as collagent type I(Col-I), ostoecalcin(OCN).
The results indicated that there was no effect on cell proliferation on both 1st and 3rd days after treatment in MC3T-E1 cells. The level of ALP was significantly higher in COE treated group than non treated group on days 7 and 14, and the level was highest on day 14. The results from ALP staining, showed basically the same results. The level of Col-I mRNA was higher in COE treated group than non treated group on days 7, 14 and 21, and the level was highest on days 7. The level of OCN mRNA was higher in COE treated group than non treated group on day 14, and it was highest on day 21. Also, the level of mineralized nodule was higher in COE treated group than non treat group on days 21 and 28, and highest on day 28.
Our results suggest that the COE promotes the osteoblast differentiation and mineralization without observable cytotoxicity. Therefore, it may play an important role in osteoblastic bone formation, and may possibly to be an effective measure for bone healing promotion in regenerative applications.
Alternative Title
Effect of Cornus officinalis extract on the differentiation of MC3T3-E1 osteoblast-like cells
Alternative Author(s)
Ming-Zhen Zheng
일반대학원 치의생명공학과
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Table Of Contents
Ⅰ. 서 론 1
Ⅱ. 연구재료 및 방법 4
1. 실험재료 및 시약 4
1.1. 식물 재료 4
1.2. 시약 4
2. 산수유 추출물 제조 5
3. 세포 배양과 분화유도 5
4. 골모세포의 생존력 및 증식률 분석 5
5. Alkaline phosphatase(ALP) 활성도 측정 6
6. Alkaline phosphatase(ALP) 염색 6
7. Reverse transcriptase-polymerase chain reaction(RT-PCR) 분석 7
8. Alizarin Red S 염색을 통한 석회화 결절 형성 분석 9
9. 통계처리 9
Ⅲ. 실험 결과 10
Ⅳ. 고 찰 24
Ⅴ. 결 론 29
참고문헌 30
감사의 글 35
조선대학교 대학원
정명진. (2014). MC3T3-E1 골모유사세포의 분화에 대한 산수유 추출물의 효과.
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General Graduate School > 4. Theses(Ph.D)
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