Bioactive materials in combination with antibiotics are widely developed for the treatment of osteomyelitis which play a dual role: as bone substitutes at the affected site and as local drug delivery systems for antibiotic delivery. In the present study, a series of ciprofloxacin loaded hydroxyapatite (HA) nanoparticles with sustained and prolonged release behavior has been synthesized by an in situ precipitation method. The amount of ciprofloxacin loaded on HA nanoparticles can be easily adjusted by changing the initial concentration during the synthesis. It was observed that as the loaded concentration of ciprofloxacin increases the release is sustained and prolonged. The in situ loading of ciprofloxacin onto HA nanoparticles does not significantly affect the bioactivity and cytocompatibility of HA whereas it provides antibacterial activity to HA against S. aureus and E. coli that causes osteomyelitis. Consequently synthesized ciprofloxacin loaded HA nanoparticles can be potential candidates for the treatment of osteomyelitis.