Since pits and fissures are the areas most commonly affected by caries due to their structural irregularity, bioactive resin-based sealant (RBS) may contribute to the prevention of secondary caries. This study aims to investigate the mechanical, physical, ion-release, enamel remineralisation, and antibacterial capabilities of the novel RBS with bioactive glass (BAG) and 2-methacryloyloxyethyl phosphorylcholine (MPC). For the synthesis, 12.5 wt% BAG and 3 wt% MPC were incorporated into RBS. The contact angle, flexural strength, water sorption, solubility, and viscosity were investigated. The release of multiple ions relating to enamel remineralisation was investigated. Further, the attachments of bovine serum albumin, brain heart infusion broth, and Streptococcus mutans on RBS were studied. Finally, the thickness and biomass of a human saliva-derived microsm biofilm model were analysed before aging, with static immersion aging and with thermocycling aging. In comparison to commercial RBS, BAG+MPC increased the wettability, water sorption, solubility, viscosity, and release of multiple ions, while the flexural strength did not significantly differ. Furthermore, RBS with MPC and BAG+MPC significantly reduced protein and bacteria adhesion and suppressed multi-species biofilm attachment regardless of the existence of aging and its type. The novel RBS has great potential to facilitate enamel remineralisation and suppress biofilm adhesion, which could prevent secondary dental caries.