Alendronate 농도와 처리시간에 따른 MC3T3-E1와 C2C12 세포의 생존율과 Alkaline Phosphatase 활성도
ALP Activity and Percentage of Viable Cells in C2C12 and MC3T3-E1 Cells according to Alendronate Concentration and Treatment Time
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목적: 본 연구는 조골세포 분화 유도 세포주인 C2C12와 MC3T3-E1세포를 이용하여 조골세포의 증식과 분화과정 동안의 alendronate의 작용기전에 대해 조사하고자 한다. 방법: C2C12 세포와 MC3T3-E1 세포를 DMEM 배양액에 배양하면서 alendronate를 다양한 농도(10-9 M, 10-8 M, 10-7 M, 10-6 M, 10-5 M, 10-4 M)로 첨가하고 1, 2, 3일 동안 배양한 후 MTT 분석 및 alkaline phosphatase (ALP) 활성도를 측정하였다. 결과: Alendronate의 농도에 따른 세포 독성은 없었다. ALP 활성은 C2C12 세포주에서는 alendronate 처리 후 24시간까지는 모든 농도에서 대조군에 비해 2배 이상 증가하였으나 24시간 이후에는 시간이 경과함에 따라 급격히 감소되었다. MC3T3-E1에서는 ALP 활성이 10-8 M에서 48시간까지 급격하게 증가되나 10-8 M 이상의 농도에서는 시간경과에 상관없이 급격히 감소되었다. 결론: Alendronate가 농도에 의존적으로 조골세포의 분화를 촉진시키고 있음을 보여주고 있으며, 이러한 alendronate에 의한 분화 촉진 효과는 ALP의 발현 증가와 연관되는 것으로 추정된다.
Abstract
Objectives: To investigate the effect of alendronate on osteoblast proliferation and differentiation using mosue preosteoblast cell line, MC3T3-E1 and myoblastic cell line, C2C12. Methods: C2C12 and MC3T3-E1 cells were cultured in DMEM media with various concentrations(10-9 M, 10-8 M, 10-7 M, 10-6 M, 10-5 M, 10-4 M) of alendronate for one, two, or three days. After the indicated culture times, MTT assay was performed and alkaline phosphatase activity was examined. Results: Alendronate had no cytotoxicity regardless of the concentrations used. ALP activity in C2C12 cells was increased in alendronate treated group compared to the control group before 24 hours of treatment, but decreased rapidly after 24 hours of treatment. In MC3T3-E1, it was increased greatly at 10-8 M before 48 hours of treatment and decreased rapidly at more than 10-8 M concentration regardless of times of treatment. Conclusion: This study shows that alendronate has a dose-dependent effect in osteoblast differentiation. The effect may be associated with the increased expression of ALP.
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