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Abstract Recurrent caries often occurs and is a primary reason for the failure of dental composite restorations. The objectives of this study were to: (1) develop a bioactive composite containing dimethylaminohexadecyl methacrylate (DMAHDM), (2) investigate its antibacterial effects and suppression on biofilm growth, and (3) investigate its ability to modulate biofilm species composition for the first time. DMAHDM was incorporated into a composite at mass% of 0%, 0.75%, 1.5%, 2.25% and 3%. A commercial composite Heliomolar served as a comparative control. A biofilm model consisting of Streptococcus mutans (S. mutans), Streptococcus sanguinis (S. sanguinis) and Streptococcus gordonii (S. gordonii) was tested by growing biofilms for 48 h and 72 h on composites. Colony-forming units (CFUs), metabolic activity and live/dead staining were evaluated. Lactic acid and polysaccharide productions were measured to assess biofilm cariogenicity. TaqMan real-time polymerase chain reaction was used to determine the proportion of each species in the biofilm. DMAHDM-containing composite had a strong anti-biofilm function, reducing biofilm CFU by 2–3 orders of magnitude, compared to control composite. Biofilm metabolic activity, lactic acid and polysaccharides were decreased substantially, compared to control (p < 0.05). At 72 h, the cariogenic S. mutans proportion in the biofilm on the composite with 3% DMAHDM was 19.9%. In contrast, an overwhelming S. mutans proportion of 92.2% and 91.2% existed in biofilms on commercial control and 0% DMAHDM, respectively. In conclusion, incorporating DMAHDM into dental composite: (1) yielded potent anti-biofilm properties; (2) modulated the biofilm species composition toward a non-cariogenic tendency. The new DMAHDM composite is promising for applications in a wide range of tooth cavity restorations to modulate oral biofilm species and combat caries. Highlights • Developed a bioactive dental composite containing DMAHDM. • Composite with 3% DMAHDM achieved greatest reduction in multispecies biofilm. • Biofilm equilibrium shifted from a cariogenic species-dominated state to a noncariogenic species-dominated state. • The new composite is promising for cavity restorations to combat caries. [ABSTRACT FROM AUTHOR]