Currently, bone tissue engineering applications comprise the development of smart materials that can induce tissue regeneration meanwhile supporting the defect site of the body. Despite of the advancements, inflammation is still a serious problem that threatens human health at the post-implantation period. To overcome potential inflammations, antibiotic therapy is commonly employed in clinical trials. However, antibiotic therapy causes some side effects such as ototoxicity and nephrotoxicity, especially when applied in high doses. Therefore, local drug delivery systems play a vital role in bone disorders due to the elimination of disadvantages introduced by conventional methods. In the presented study, it was aimed to develop chitosan-based composite nanospheres as a controlled drug delivery system against bone infections. Accordingly, chitosan and montmorillonite nanoclay was homogenized with microfluidizer and electrosprayed to obtain spherical nanoparticles. The optimum electrospraying conditions were investigated using response surface methodology. Vancomycin and Gentamicin antibiotics were incorporated in the polymeric matrix to provide controlled release at the defect region to overcome inflammations after implantation. The prepared nanospheres were characterized in terms of morphology, hydrodynamic size distribution, surface charge, drug encapsulation efficiency and release profiles. The dominant drug release mechanism was determined by empirical mathematic models. Drug loaded nanospheres have been successfully produced with a size range of 180-350 nm. High encapsulation efficiency was achieved (80-95%) with a controlled drug release up to 30 days. Fickian diffusion was found as the main mechanism in drug delivery from spherical CS/MMT nanocomposites. The in vitro release medium of nanospheres showed strong antimicrobial activity against gram-positive S. aureus and gram-negative E. coli bacteria. Furthermore, it was found that the nanospheres did not show any cytotoxic effect to 3T3 and SaOS-2 cell lines. These results demonstrated that the prepared nanospheres can be a promising option for bone infection prevention.