The effect of fluoride on the patterns of adherence of osteoclasts cultured on and resorbing dentine: a 3-D assessment of vinculin-labelled cells using confocal optical microscopy.

Osteoclasts were isolated mechanically from chick long bones and cultured on dentine for three days in MEM/10% FCS with or without 1 mg/l NaF, (near to therapeutic ranges for serum fluoride levels) or 15 mg/l NaF (which has been found to reduce the volume:plan-area ratios of resorption pits in vitro). The distribution of close contacts with the substratum was determined by immunolabelling of vinculin and confocal fluorescence microscopy. Four characteristic patterns of contacts were observed: infilled discs, rings, crescents and patches: these reflect the progress of the cell in the process of pit formation. Cell-substratum adherence was more extensive than previously reported with contacts often extending down the sides of pits and sometimes across the floor. This distribution explains the curved shape of unilocular pits and the kidney-shaped extensions of multilocular pits. The peripheral creeping of the contact region was more marked in the osteoclasts of the 15 mg/l NaF cultures, and these cultures showed a decrease in pit depth and inhibition of osteoclastic movement in the presence of fluoride.