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Effect of dentin phosphoprotein on phosphate-induced apoptosis of odontoblast-like cells
Ryuichi Fujisawa1, Morimichi Mizuno, Masato Tamura
1Department of Oral Biochemistry and Molecular Biology, School of Dentistry, Hokkaido University, Sapporo, Japan. rfuji@den.hokudai.ac.jp
Abstract:
Dentin phosphoprotein, the major noncollagenous protein in dentin, has effects on differentiation of odontoblast-like cells. This study was designed to investigate the effect of the protein on apoptosis of the cells. The odontoblast-like cells were prepared from the pulp cells of rat incisors. Apoptosis was detected by measuring caspase-3 activity by using DEVD-AMC as a fluorescent substrate. The cells formed calcification nodules in the presence of 2-glycerophosphate and expressed dentin sialophosphoprotein. Apoptosis was not observed in the cells through the differentiation stages. Then, apoptosis was induced by raising inorganic phosphate concentration in the medium. Elevation of phosphate concentration to 5 mM reduced the number of viable cells and increased caspase-3 activity, indicating the induction of apoptosis. Addition of bovine dentin phosphoprotein in the medium suppressed phosphate-induced apoptosis. Phosvitin and poly(Asp) had similar antiapoptotic effects.
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