Biomedical implants are the need of this era due to the increase in number of accidents and follow-up surgeries. Different types of bone diseases such as osteoarthritis, osteomalacia, bone cancer, etc. are increasing globally. Mesoporous bioactive glass nanoparticles (MBGNs) are used in biomedical devices due to their osteointegration and bioactive properties. In this study, silver (Ag) and strontium (Sr) doped mesoporous bioactive glass nanoparticles (Ag-Sr MBGNs) were prepared by a modified Stöber process. In this method, Ag+ & Sr2+ were co-substituted in pure MBGNs to harvest the antibacterial properties of Ag ions, as well as pro-osteogenic potential of Sr2 ions. The effect of the two ion concentration on morphology, surface charge, composition, antibacterial ability, and in-vitro bioactivity was studied. Scanning electron microscopy (SEM), X-Ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) confirmed the doping of Sr and Ag in MBGNs. SEM and EDX analysis confirmed the spherical morphology and typical composition of MBGNs, respectively. The Ag-Sr MBGNs showed a strong antibacterial effect against Staphylococcus carnosus and Escherichia coli bacteria determined via turbidity and disc diffusion method. Moreover, the synthesized Ag-Sr MBGNs develop apatite-like crystals upon immersion in simulated body fluid (SBF), which suggested that the addition of Sr improved in-vitro bioactivity. The Ag-Sr MBGNs synthesize in this study can be used for the preparation of scaffolds or as a filler material in the composite coatings for bone tissue engineering.