The aim of this study is to investigate the response of cells and bacteria on Sr, Ag, and Sr + Ag doped S53P4 sol-gel bioglass. In this study, 3 mol% of SrO and 1 mol% of Ag₂O were selected as doping for single and multiple doped S53P4 sol-gel bioglass. These doped specimens were denoted as BG, BG-3S, BG-1A, and BG-3S1A throughout the study. Cell study showed that the BG-3S1A specimen exhibited the highest proliferation of MC3T3-E1 osteoblast-like cells, ALP activity, and bone nodule formation in comparison with other specimens. Meanwhile, antibacterial study of the BG-3S1A specimen showed relatively similar antibacterial responses towards Escherichia coli and Staphylococcus aureus in comparison with the BG-1A specimen. Therefore, these findings proved that by combining appropriate amount of Sr and Ag doping ions into S53P4 bioglass composition, cellular proliferation and antibacterial response could be stimulated, which could benefit patients who suffer from bacteria-infected bone defect diseases.