In this study, a good combination consisting of electrospun titanium dioxide (TiO2) nanofibers incorporated with high purity hydroxyapatite (HAp) nanoparticles (NPs) and antimicrobial silver NPs is introduced for hard tissue engineering applications. The synthesized nanofibers were characterized by various state of art techniques like; SEM, XRD, TEM, TEM EDS and XPS analyses. SEM results confirmed well oriented nanofibers and good dispersion of HAp and silver NPs, respectively. XRD results demonstrated well crystalline feature of three components used for electrospinning. Silver NPs were having a diameter in range of 5-8 nm indicated by TEM analysis. Moreover, TEM EDS analysis demonstrated the presence of each component with good dispersion over TiO2 nanofiber. The surface analyses of nanofibers were investigated by XPS which indicated the presence of silver NPs on the surfaces of nanofibers. The obtained nanofibers were checked for antimicrobial activity by using two model organisms E. coli and S. aureus. Subsequently, antimicrobial tests have indicated that the prepared nanofibers do posses high bactericidal effect. Accordingly, these results strongly recommend the use of obtained nanofiber mats as future implant materials.