Osteoblast cells were separated from the neonatal rat calvaria and co-cultured on a novel mineralized hydroxyapatite/collagen/poly(lactic acid) composite scaffold. By using this static cell culture, a three-dimensional osteoblasts/composite bone-like was constructed in vitro. The culture process was observed by scanning electron microscopy, fluorescence microscopy, confocal laser scanning microscopy, and histological analysis. Cells were observed to spread and proliferate throughout the inner-pores of the scaffold material. After a 12-day culture, the cells had grown into the interior scaffold about 200–400 μm depth of the composite by histological section observation. This mobile behavior of osteoblasts appeared to be similar to the composition and hierarchical structure of bone tissue. The adherence and migration of osteoblast cells in this three-dimensional composite is clinically important for large bone defect repair based on tissue engineering.
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