Hydroxyapatite (HA) bioceramics are widely used materials for orthopaedic applications, offering enhanced cellular adhesion and differentiation. However, the lack of antibacterial features makes the HA biomaterials prone to bacterial infections in in-vivo conditions. The osteomyelitis-like conditions are often treated with prolonged doses of antibiotics and may require surgical removal. Such problems can be tackled by inducing the antibacterial characteristics in the biomaterials which prevent such infections. Various research studies have been conducted to improve the mechanical and antibacterial characteristics of biomaterials like HA through doping. The present study focuses on the synthesis, characterization, and antibacterial study of Zn and Fe doped HA (ZFHAp) for application in the treatment of osteomyelitis. Response Surface Methodology (RSM) based Central Composite Design (CCD) approach was used to optimize the process parameters using the Design Expert software. The analyses indicated the presence of Calcium Iron Hydrogen Phosphate (CIHP) and Parascholzite (PS) as major phases. The optimized experimental conditions were estimated as 0.09 M Zn and 0.04 M Fe at 65 °C, corresponding to 71.29 % and 28.71 % of the CIHP and PS phases, respectively. The synthesized nanopowder showed mixed morphology comprising spherical and rod-shaped particles due to biphasic composition. The ZFHAp sample showed a broad-spectrum antibacterial activity, with IC50 values of 4.02 mg/ml and 4.79 mg/ml against E. coli and S. aureus, respectively. The optimized sample showed excellent biocompatibility with the osteoblast-like cells (MG-63) with improved biomineralization properties. The antibacterial and osteoinductive characteristics of the synthesized biphasic Fe and Zn doped HA suggest its potential application against osteomyelitis-related conditions.