A nano-hydroxyapatie/collagen composite was fabricated by an in situ synthesis technique. Transmission electron microscopy (TEM) and selected area electron diffraction (SAED) analyses for the composite indicated crystals on the collagen fibril matrix exhibited certain orientation. Bioactivity of the composite was investigated through in vitro tests in a sterile simulated body fluid (SBF) system. Scanning electron microscopy and energy-dispersive X-ray analysis of the composite showed that the composite had the ability to induce the formation of calcium phosphate crystals on the surface of the composite in SBF. The cytocompatibility of the composite was evaluated by in vitro cytotoxicity test. Both the results from MTT assay and scanning electron microscope (SEM) observations indicated that the composite had no adverse impact on cell proliferation and morphology. The results imply the composite is high bioactive and has good cytocompatibility