Calcium hydroxide is often used for induction of reparative dentin formation in endodontic treatment. However, little is known about the mechanism by which calcium hydroxide works. The calcium ion (Ca2+) is an important regulator of cell functions. In this study, we examined the effect of extracellular Ca2+ on gene expression of bone-related proteins in human cultured pulp cells in serum-free conditions. A Ca2+ level elevated by 0.7 mM induced an increase in mRNA expression of osteopontin and bone morphogenetic protein (BMP)-2. However, mRNA levels of BMP-4 and alkaline phosphatase decreased under the elevated Ca2+ culture condition. The same concentration of additional magnesium ions had little effect on expressions of the examined bone-related protein mRNAs. These findings suggest that Ca2+ in Ca(OH)2 specifically modulates osteopontin and BMP-2 levels during calcification in pulp.

Little is known about the effect of aging on characteristic functions of pulp cells. When damaged pulp is recovered and mineralized tissue is formed to protect remaining pulp tissue, the general responses of pulp tissue after adequate stimuli (pulp cell proliferation and activation of alkaline phosphatase [ALPase]) are thought to be essential. In this study, we compared proliferative ability and ALPase activity between cultures of human pulp (HP) cells obtained from young and aged donors. The in vitro proliferative lifespan of HP cells from young donors was longer than HP cells from aged donors. Growth rates and ALPase activity of HP cells decreased with increasing donor age. These findings suggest that impaired repair of pulp and dentin in aged patients is partly due to a decrease in the proliferative ability and ALPase activity in aged pulp cells.