In this study, a new artificial bone matrix was constructed with collagen and Ca(5)(PO(4))(3)OH (hydroxyapatite/HA) which are the main components of natural bone. To improve the property of the artificial bone matrix, chitosan (CS), a kind of positive charged polysaccharide, was crosslinked into the scaffolds. Solid-liquid phase separation method was used to shape 3D porous structure benefited for cells growing into. The artificial bone matrix was investigated by transmission electron microscopy, scanning electron microscopy, and electron spectroscopy for chemical analysis, etc. for structures and characteristics. And its ability of bone repair was investigated by orthotope bone defect reparation in vivo. The results showed that the artificial bone matrix was a porousscaffold with three-dimension interconnected fiber microstructure. HA particles were dispersed evenly among collagen fiber and CS was modified on the surface of collagen fiber. It was indicated that this artificial bone matrix could be used as a bone substitute with outstanding properties.