Fibroblast growth factor-2 (FGF-2) apatite composite layers were formed on anodically oxidized titanium (Ti) rods at temperatures lower than the previously used 37 °C to reduce the risk of the inactivation of FGF-2 in calcium phosphate solution. A two-step procedure was used to coprecipitate FGF-2 apatite composite layers on Ti rods. Continuous and homogeneous carbonate-containing low-crystalline apatite layers incorporating FGF-2 were formed on the surface of the Ti. The amounts of apatite and FGF-2 coprecipitated on the Ti rod surface decreased with decreasing coprecipitation temperature. When the coprecipitation temperatures were 15 and 20 °C, the amounts of FGF-2 precipitated on the Ti rods were 0.19 ± 0.03 and 0.20 ± 0.03 μg/cm2, respectively. A cell proliferation assay for evaluating the mitogenic activity of FGF-2 immobilized in the layer showed that when FGF-2 was coprecipitated at 15 and 20 °C, the number of NIH3T3 cells cultured with FGF-2 extract was significantly larger than that at 4, 10, 25 and 30 °C. To obtain the largest amount of active FGF-2 on a Ti surface, a temperature of 15 or 20 °C should be used for coprecipitating FGF-2.
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