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KMID : 1007020090070020111
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Korean Soceity of Osteroporosis
2009 Volume.7 No. 2 p.111 ~ p.121
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The Bone Action Mechanism of Bisphosphonate using Osteoblast Differentiation Cell Line
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Oh Young-Rim
Ock Mee-Sun
Cha Hee-Jae
Eo Wan-Kyu
Jung Min-Hyung
Jung Hyuk
Kim Heung-Yeol
Kim Tak
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Abstract
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Objectives: To investigate the effect of alendronate on osteoblast proliferation and differentiation using mosue preosteoblast cell line, MC3T3-E1 and myoblastic cell line.
Material & Methods: MC3T3-E1 cell lines 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 and ALP activity were examined. The expression of genes for ALP, osteocalcin (OCN), collagen I α1 (Col 1), inhibitor of differentiation (Id) and cathepsin K (CTSK) were examined by RT-PCR method. Transcription factors related to the increased expression of Id genes were identified with TRANSFEC program. The effect of these transcription factors on the expression of Id genes were tested by promoter assay.
Results: Alendronate had no cytotoxicity regardless of the concentrations used. In MC3T3-E1, it was increased greatly at 10-8 M before 48 hours of treatment and it was decreased rapidly at more than 10-8 M concentration regardless of times of treatment. Expressions of ALP, Col 1 and OCN genes had a tendency to increase with decreasing of alendronate concentrations before 24 hours of treatment in both cell lines. The expression of Id-1 gene began to increase from 10-6 M of alendronate compared to the control group and kept to 10-8 M. However, Id-2 gene expression was increased gradually with increasing of alendronate concentrations.
Conclusion: This study shows that alendronate has a dose-dependent effect in osteoblast differentiation. The effect may be associated with the increased expression of Id genes.
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KEYWORD
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Alendronate, Osteoblast, Proliferation, Differentiation
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