Abstract In this paper, the nano-hydroxyapatite/chitosan-chondroitin sulfate (Hap/Cs-Chs) composite materials with different mass ratios were prepared by liquid co-precipitation method. On the basis of the microstructure, phase and compressive strength of the composite materials were investigated respectively. The safety of composite materials was evaluated by MMT method. The results showed that Hap/Cs-Chs composite materials exhibited excellent mechanical performance and good biocompatibility. The nano-Hap particles were dispersed uniformly in the organic phase Cs and Chs with relatively weak crystallinity. It is combined with relatively closely between Hap particles and Cs-Chs. The particle size is about 40nm-100 nm with spherical shape. The crystallization, particle size, and dispersion of Hap are influenced directly by the mass ratio of Hap and Cs-Chs. The uniformity, compactness, and thermal stability can reach the optimum when mass ratio is 50/50. EDS analysis indicated that Ca/P ratio is around 1.80±0.03, belonging to the calcium-rich type of Hap. The Hela cell toxicity experiment confirmed that the composite material has good biocompatibility and safety, and the physical and chemical performance can meet the requirements of bone tissue engineering materials.