For the first time, in the present investigation, we have attempted the refluxing based sol-gel technique to synthesize cerium (Ce4 +) doped hydroxyapatite (Ce-HAP) with different concentrations such as 0.5, 0.75, 1 and 1.25%. The Ce-HAP powders were evaluated for in vitro biological studies such as hemocompatibility, antibacterial activity and biocompatibility. These in vitro studies have demonstrated the prominent results such as compatible nature with human blood (< 5% hemolytic ratio) and excellent pathogen inhibition efficiency towards Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus) and Bacillus subtilis (B. subtilis) respectively. Among these, E. coli shows more susceptible towards 0.5–1.25% of Ce-HAP. In vitro biocompatibility study was performed on the 1.25% of Ce-HAP using MG-63 cells at various concentrations of 200–1000 μg/ml for 24–48 h. It was found that 1.25% of Ce-HAP powder was non-toxic up to the level of 600 μg/ml concentrations towards MG-63 cells with regular cell attachment and proliferation. The Ce-HAP materials were further characterized by UV–vis DRS spectroscopy, ATR-FTIR, Powder-XRD, SEM-EDAX, TEM, ICP-OES and XPS analysis. Therefore, this study proves that the developed Ce4 + doped HAP can be widely used in orthopedic and dentistry field with improved biological properties. © 2017 Elsevier Ltd