OBJECTIVE: Composite restorations are increasingly popular, but recurrent caries is a main reason for composite restoration failures. The objectives of this study were to investigate a dual strategy of combining rnc gene-deletion for Streptococcus mutans (S. mutans) with antibacterial dimethylaminohexadecyl methacrylate (DMAHDM) composite, and determine the effects of rnc gene-deletion alone, DMAHDM composite alone, and rnc-deletion plus DMAHDM composite, on biofilm growth and lactic acid production. METHODS: Parent S. mutans (UA159, ATCC 700610) and rnc-deleted S. mutans were used. DMAHDM was incorporated into a composite at mass fractions of 0%, 1.5%, and 3%. Gene expressions for biofilm formation and drug resistance were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). Biofilms were grown on composite surfaces for 2 days. Live/dead, biomass, polysaccharide, metabolic activity (MTT), colony-forming units (CFU) and lactic acid production of biofilms were evaluated. RESULTS: Compared to the parent S. mutans, the rnc-deletion technique yielded significantly less biofilm biomass, polysaccharides, metabolic activity, CFU, and lactic acid for biofilms grown on control composite (p < 0.05). With no gene modification, the biofilm CFU was decreased by 5-6 logs at 3% DMAHDM, when compared to control composite group. The dual strategy of combining rnc-deletion with 3% DMAHDM composite achieved the strongest biofilm-inhibition, with the greatest reduction in CFU by 8 logs. The combination of rnc-deletion with 3% DMAHDM composite decreased the biofilm lactic acid production by 95% (p < 0.05). CONCLUSIONS: The dual strategy of rnc-deletion plus DMAHDM composite produced synergistic effects and achieved the strongest biofilm-inhibition. This method has great potential to inhibit dental caries and is promising to reduce secondary caries and protect tooth structures.
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