Abstract Monophase silver-doped hydroxyapatite (Ag x Ca 10− x (PO 4) 6(OH) 2; 0.002 ≤ x ≤ 0.04) nanoparticles were prepared using a neutralization method and investigated with respect to potential medical applications. This method consists of dissolving Ag 2O in solution of H 3PO 4, and the slow addition to suspension of Ca(OH) 2 was applied for the purpose of homogenous distribution of silver ions. Characterization studies from XRD, TEM and FTIR spectra showed that obtained crystals are monophase hydroxyapatites and that particles of all samples are of nano size, with average length of 70 nm and about 15–25 nm in diameter. Antimicrobial studies have demonstrated that all silver-doped hydroxyapatite samples exhibit excellent antimicrobial activity in vitro against the following pathogens: Staphylococcus aureus, Escherichia coli and Candida albicans. The hydroxyapatite sample with the highest content of silver has shown the highest antimicrobial activity; killed all cells of E. coli and brought to more than 99% reduction in viable counts of S. aureus and C. albicans. The atomic force microscopic studies illustrate that silver-doped hydroxyapatite sample causes considerable morphological changes of microorganism cells which might be the cause of cells’ death. Hemolysis ratios of the silver-doped hydroxyapatite samples were below 3%, indicating good blood compatibility and that are promising as biomaterials.