Alendronate Enhances Osteogenic Differentiation of Bone Marrow Stromal Cells: A Preliminary Study

Chonnam National University Hwasun Hospital Center for Joint Disease 160 Ilsimri, Hwasuneup, Hwasungun 519-809 Jeonnam Korea
Clinical Orthopaedics and Related Research (Impact Factor: 2.88). 12/2009; 467(12):3121-3128. DOI: 10.1007/s11999-008-0409-y

ABSTRACT Alendronate inhibits osteoclastic activity. However, some studies suggest alendronate also has effects on osteoblast activity.
We hypothesized alendronate would enhance osteoblastic differentiation without causing cytotoxicity of the osteoblasts. We
evaluated the effect of alendronate on the osteogenic differentiation of mouse mesenchymal stem cells. D1 cells (multipotent
mouse mesenchymal stem cells) were cultured in osteogenic differentiation medium for 7days and then treated with alendronate
for 2days before being subjected to various tests using MTT assays, Alizarin Red, enzyme-linked immunosorbent assay, energy-dispersive
xray spectrophotometry, reverse transcriptase–polymerase chain reaction, confocal microscopy, and flow cytometric analysis.
D1 cells differentiated into osteoblasts in the presence of osteogenic differentiation medium as confirmed by positive Alizarin
Red S staining, increased alkaline phosphatase activity and osteocalcin mRNA expression, a calcium peak by energy-dispersive
xray spectrophotometry, and by positive immunofluorescence staining against CD44. Osteogenic differentiation was enhanced
after treatment with alendronate as confirmed by Alizarin Red S staining, elevated alkaline phosphatase activity and osteocalcin
mRNA expression, a greater calcium peak by energy-dispersive xray spectrophotometry, and by immunofluorescence staining against
CD44 by flow cytometric analysis. These data suggest alendronate enhances osteogenic differentiation when treated with mouse
mesenchymal stem cells in osteogenic differentiation medium.

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