In this study, a novel biocompatible hydroxyapatite (HA) was synthesized by using chitosan oligosaccharide (COS) as a template. These HA samples were studied by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and transmission electron microscopy (TEM). The biocompatibility of HA samples was evaluated via cell viability, cell morphology and alkaline phosphatase staining of MG-63 cell lines. The results show that HA synthesized in the presence of COS was favorable to proliferation and osteogenic differentiation of MG-63 cells. These hydroxyapatites are potentially attractive biomaterials for bone tissue engineering applications.