Substance P regulates the function of rabbit cultured osteoclast; increase of intracellular free calcium concentration and enhancement of bone resorption.

The effects of substance P on cultured rabbit osteoclasts were investigated. The intracellular Ca(2+) concentration (Ca(2+)](i)) which was monitored by the microfluorometric technique using fura-2, in osteoclasts elevated by the addition of substance P (0.3-5 microM). The EC(50) value of substance P was about 200 nM. This substance P-evoked Ca(2+)](i) elevation was not observed in the Ca(2+)-free medium. Simultaneous application of spantide, a substance P receptor antagonist, blocked the Ca(2+) response. The addition of substance P (0.1-10 microM) to cultured osteoclasts enhanced their bone resorption activity which was evaluated by the pit formation assay. Maximum enhancement of the pit formation by substance P (5 microM) peaked at about 170% of the control level. The addition of substance P receptor antagonists also inhibited the enhancement of the bone resorption by substance P addition. Substance P possibly stimulates the bone remodeling by osteoclasts via the Ca(2+)](i) elevation.