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Journal of Veterinary Science ; : 133-140, 2014.
Article in English | WPRIM | ID: wpr-56424

ABSTRACT

To investigate 1alpha,25-(OH)2D3 regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1alpha,25-(OH)2D3 during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1alpha,25-(OH)2D3 inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1alpha,25-(OH)2D3 administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.


Subject(s)
Animals , Mice , Acid Phosphatase/metabolism , Blotting, Western , Calcitriol/pharmacology , Calcium Channel Agonists/pharmacology , Cell Differentiation , Cell Line , Cell Proliferation , Gene Expression Regulation, Enzymologic/drug effects , Isoenzymes/metabolism , Matrix Metalloproteinase 9/genetics , Osteoclasts/cytology , Tetrazolium Salts , Thiazoles
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