Inorganic polymeric phosphate/polyphosphate as an inducer of alkaline phosphatase and a modulator of intracellular Ca 2+ level in osteoblasts (SaOS-2 cells) in vitro

Abstract

Inorganic polymeric phosphate is a physiological polymer that accumulates in bone cells. In the present study osteoblast-like SaOS-2 cells were exposed to this polymer, complexed in a 2:1 stoichiometric ratio with Ca 2+, polyP (Ca 2+ salt). At a concentration of 100 μM, polyP (Ca 2+ salt) caused a strong increase in the activity of the alkaline phosphatase and also an induction of the steady-state expression of the gene encoding this enzyme. Comparative experiments showed that polyP (Ca 2+ salt) can efficiently replace β-glycerophosphate in the in vitro hydroxyapatite (HA) biomineralization assay. In the presence of polyP (Ca 2+ salt) the cells extensively form HA crystallites, which remain intimately associated with or covered by the plasma membrane. Only the tips of the crystallites are directly exposed to the extracellular space. Element mapping by scanning electron microscopy/energy-dispersive X-ray spectroscopy coupled to a silicon drift detector supported the finding that organic material was dispersed within the crystallites. Finally, polyP (Ca 2+ salt) was found to cause an increase in the intracellular Ca 2+ level, while polyP, as well as inorganic phosphate (P i) or Ca 2+ alone, had no effect at the concentrations used. These findings are compatible with the assumption that polyP (Ca 2+ salt) is locally, on the surface of the SaOS-2 cells, hydrolyzed to P i and Ca 2+. We conclude that the inorganic polymer polyP (Ca 2+ salt) in concert with a second inorganic, and physiologically occurring, polymer, biosilica, activates osteoblasts and impairs the maturation of osteoclasts.