Attachment kinetics, proliferation rates and vinculin assembly of bovine osteoblasts cultured on different pre-coated...

Primary bovine osteoblasts were used to study in-vitro effects of attachment on vinculin assembly in cells cultured on various artificial substrates. Materials coated with fibronectin and bovine serum albumin (BSA) as well as untreated materials (tissue culture polystyrene and Aclar foils) were chosen to investigate substrate-dependent proliferation during the first 3 days of culture. Proliferation was highest on fibronectin-coated substrates, followed by BSA-coated and untreated substrates. During the first 24 h of cultivation, cell attachment kinetics revealed no significant difference between the various substrates. After 24 h detachment rates obtained by calcium depletion with ethylenediaminetetraacetic acid were highest on uncoated materials, followed by BSA- and fibronectin-coated substrates. Phase contrast microscopy revealed typical osteoblast morphology after cell adhesion for 24 h. The dynamic attachment process was concomitant with the reassembly of vinculin into streak-like focal contacts clustered on the ventral side of cells. The kinetics of vinculin reassembly were independent of the underlying coating. Thus, fibronectin coating of artificial substrates increased the attachment strength and proliferation rate of osteoblasts. While the reassembly of vinculin in focal contacts seems to be a prerequisite of osteoblast attachment in vitro, it does not seem to have profound effects on the subsequent cell behaviour on artificial substrates.