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Induction of Calcification in MC3T3-E1 Cells by Inorganic Polyphosphate

Y. Kawazoe^1,2,3, T. Shiba^1,2,5,*, R. Nakamura⁴, A. Mizuno², K. Tsutsumi^4,, T. Uematsu⁵, M. Yamaoka⁵, M. Shindoh³ and T. Kohgo³

¹ Regenetiss Co., Ltd., 1-5-17, Akabane, Okaya, Nagano 394-0002, Japan;
² Frontier Research Division, Fujirebio Inc., Hachioji, Tokyo, Japan;
³ Graduate School of Dental Medicine, and
⁴ Graduate School of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan; and
⁵ Matsumoto Dental University, Shiojiri, Nagano, Japan;

Correspondence: ^* corresponding author, shiba{at}regenetiss.com

Relatively large amounts of inorganic polyphosphate [poly(P)] (400 µM) have been found in normal osteoblasts. The effect of poly(P) with an average chain length of 65 phosphate residues on cell calcification was therefore investigated with the use of MC3T3-E1 cells. Expression of both osteopontin and osteocalcin was induced by poly(P) (0.1 ~ 1 mM), and cells treated with poly(P) were strongly stained by alizarin red. In addition, the level of alkaline phosphatase activity induced in poly(P)-treated cells was two-fold higher than that in either orthophosphate-treated or control cells but not higher than that in cells treated with β-glycerophosphate and ascorbic acid. In contrast, however, polyphosphatase activities were activated by poly(P) treatment to levels up to six-fold greater than that in controls. MC3T3-E1 cells may utilize poly(P) as a phosphate source for calcification rather than phosphate sources that are mainly produced by ALPase. Poly(P)-dependent induction of polyphosphatase activities may therefore promote calcification in MC3T3-E1 cells.

Key Words: calcification • osteoblasts • osteopontin • osteocalcin • polyphosphate

Journal of Dental Research, Vol. 83, No. 8, 613-618 (2004)
DOI: 10.1177/154405910408300806


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