Effects of calcium ion implantation on human bone cell interaction with titanium

The use of calcium ion (Ca) implantation of titanium (Ti), previously reported to encourage osseointegration in vivo, has been investigated using an in vitro model in order to understand the basic mechanisms involved in the response of target cells to such surfaces. Polished Ti discs were implanted with high, medium and low (1×1017, 1×1016, 1×1015 ions cm−2) doses of Ca ions at 40 keV. The effects of different levels of Ca implantation on morphology, attachment and spreading of MG-63 cells seeded on the surface of control (non-implanted) Ti and Ca–Ti discs were assessed. Further, to understand cell–material interactions at a molecular level, the expression of β1 and α5β1 integrins and the formation of vinculin-positive focal adhesion plaques were examined. In addition, the effects of pre-immersion of the Ca (high)–Ti in tissue culture medium on cell attachment were measured and correlated with specific chemical changes at the Ti surface. Our findings suggest that Ca implantation can affect the adhesion of MG-63 cells both qualitatively and quantitatively. However, this effect appears to depend on the level at which Ca ions are implanted. Results showed that although cell adhesion on Ca (high)–Ti was initially reduced, it nevertheless was not only restored but substantially increased with progressing culture times. In addition, a significantly enhanced cell spreading, formation of focal adhesion plaques and expression of integrins were measured on this particular surface. In contrast, no marked differences were observed in cell behaviour on Ca–Ti (low and medium). Pre-immersion studies indicated that the decrease in cell attachment to Ca (high)–Ti at early time periods may be linked to the presence of Ca- and P-rich particles on the surface. The absence of these particles at 24 h was consistent with a significant increase in cell attachment.