In the present work, the novel calcium phosphate cement (CPC) composite was prepared and the microstructure characteristics and bioactivity behaviour was evaluated after soaking in simulated body fluid (SBF). The cement powder phase was prepared by mixing natural hydroxyapatite extracted from fish bone (HAp) and dicalcium phosphate dehydrate (DCPD). As hardening, the liquid cement phase containing chitosan (2 w/v %) in acetic acid solution was used. The microstructure properties of natural hydroxyapatite prepared by heat treatment of fish bone and bioactivity evaluation of obtained cements were investigated. Fourier Transform Infrared (FTIR) Spectroscopy and X-ray Diffraction (XRD) results indicated that formation of crystal structure of HAp phase in the bone matrix particularly when annealed above 600 ⁰C. After preparation and hardening of calcium phosphate/natural hydroxyapatite/chitosan composite, the bioactivity of cements were investigated with incubation in SBF. The results of 28 days incubation in SBF revealed that, the biomimetic apatite layer were formed. The average crystallite sizes of samples after 28 days incubation in SBF estimated by Scherrer’s equation were found to be less than 100 nm. The results were obtained from this research indicate that new nanoscale biological composite cement can be a good candidate for biomedical applications.