The effect of osteocalcin on in vitro lipid-induced hydroxyapatite formation and seeded hydroxyapatite growth

Osteocalcin, the bone gamma-carboxy glutamic acid containing protein, is one of the major noncollagenous proteins both synthesized and localized in bony tissue. Previously, investigators have suggested, based on the in vitro and in vivo properties of this protein, that it may be involved in controlling initiation of mineralization and/or hydroxyapatite (HA) growth. In this study, the in vitro effects of osteocalcin on lipid-induced HA formation, and HA seeded growth were compared. Although osteocalcin inhibited the growth of HA, as indicated by the osteocalcin concentration-dependent decreases in the first order rate constant, kCa, osteocalcin had no effect on lipid-induced calcification, kCa remaining constant at .033 h-1. Binding studies revealed that osteocalcin did not associate with the lipid macromolecules tested (phosphatidyl serine, phosphatidyl inositol, and the Ca-acidic phospholipid-phosphate (Ca-PL-PO4) complexes prepared from these phospholipids) although the protein bound to HA with high affinity. These data suggest that a) osteocalcin is quite distinct from the gamma carboxy glutamic acid containing clotting proteins which have a high affinity for both the acidic phospholipids and for HA, and b) that osteocalcin has little effect on the initial Ca-PL-PO4-dependent formation of HA.