The bacterial infection is one of the major problems associated with implant and reconstructive surgery of bone. Hence, the aim of this study was to develop biomaterials having antibacterial activity for bone tissue engineering. The hydroxyapatite nanoparticles (nHAp) improve the mechanical properties and incorporate nanotopographic features that mimic the nanostructure of natural bone. We report here for the first time the synthesis and characterization of nHAp and nHAp soaked with copper (nHAp-Cu) using SEM, AFM, FTIR and XRD. The antibacterial activity of nHAp and nHAp-Cu was determined using Gram-positive and Gram-negative bacterial strains. To have accelerated antibacterial activity, polyethylene glycol 400 (PEG 400), a synthetic biodegradable polymer was also added along with nHAp-Cu. The nHAp-Cu/PEG 400 had increased antibacterial activity towards Gram-positive than Gram-negative bacterial strains. The cytotoxicity of nHAp-Cu/PEG 400 was determined using MTT assay with rat primary osteoprogenitor cells and these biomaterials were found to be non-toxic. Hence, based on these results we suggest that the biomaterials containing nHAp-Cu/PEG 400 can be used as antibacterial materials in bone implant and bone regenerative medicine.