The purpose of this study was to introduce antibacterial property to pits and fissure sealant (PFS) in order to mitigate the major clinical problems associated with PFS, such as microleakage and secondary caries. We prepared a pH reliant cobalt oxide nanoparticle incorporated with minocycline (MNC@CO) and characterized to investigate its antibacterial potential against Streptococcus sobrinus. The physiochemical, morphological, and drug release kinetics at different pH (7.4, 5.0, and 3.5) from nanoparticles were investigated. The MNC@CO were added at 2.5% and 5.0% into experimental PFS and characterized for their antibiofilm capacity, biocompatibility, and mechanical properties including compressive and flexural strength. The groups 2.5% and 5.0% has shown statistically significant antimicrobial capacity against S. sobrinus compared to control (p < .05). The highest percentage of MNC release at different pH (especially at pH 5.0 and 3.5) was observed from 5.0% MNC@CO doped PFS. The PFS doped with 2.5% MNC@CO showed a highest compressive strength (110 MPa) over a period of 70 days as compared to 5.0% MNC@CO (75 MPa) and control (80 MPa). The flexural strength of both experimental groups was lower for both time points (24 h and 30 days) than control. In conclusion, the present study found that 2.5% MNC@CO doped PFS showed considerable anti-biofilm potential without compromising mechanical properties.