Overexpression of ASvicR combined with the antibacterial monomer DMAHDM interferes with the VicRK two-component system to attenuate the cariogenicity of Streptococcus mutans

Abstract

Background Streptococcus mutans ( S. mutans ) is a primary cariogenic pathogen responsible for acid production, exopolysaccharides (EPS) production and biofilm formation. Two-component systems (TCS) regulate EPS metabolism, especially the VicRK TCS. Overexpression of antisense vicR (AS vicR ) can reduce EPS production and thereby weaken the cariogenicity of S. mutans. Although the antimicrobial monomer dimethylaminohexadecyl methacrylate (DMAHDM) exhibits potent antibacterial properties, mature S. mutans biofilms can protect themselves by extracellular matrix. Emerging evidence suggests that genetic intervention enhances drug efficacy, yet the underlying regulatory mechanisms remain largely unexplored. Objective To investigate the chemical–genetic cooperative antibiofilm strategy inhibition and mechanisms of AS vicR overexpression combined with DMAHDM on S. mutans biofilm formation, acid and EPS metabolism, and cariogenicity through the VicRK system. Methods The minimal inhibitory concentration and minimal bactericidal concentration of DMAHDM and chlorhexidine were determined. Biofilm properties were evaluated via biomass assessment, EPS quantification, lactate production measurement, and colony-forming unit counting. Biofilm structures were examined by scanning electron microscopy. Mechanisms were investigated using RT-qPCR, zymography, and western blot. Rat caries model was employed to assess caries formation under different treatment conditions. Results The AS vicR strain exhibited an approximate 2-fold increase in susceptibility to DMAHDM and chlorhexidine. The combination treatment reduced biofilm CFU by approximately 4 log units, significantly lowered lactate and EPS levels, and resulted in a loose, porous biofilm structure. The expression levels of cariogenic virulence factors as well as the VicRK TCS genes and proteins were significantly downregulated. In vivo , the combined treatment reduced the overall caries severity score to 12.7% of the control group (p lt;0.05) without observing any systemic adverse effects. Conclusion The strategy of combining AS vicR overexpression with DMAHDM effectively modulates EPS metabolism and cariogenicity in S. mutans by interfering with the VicRK TCS, providing a potential therapeutic approach for clinical caries management.

Key findings

• ▸Overexpression of ASvicR successfully reduces EPS production by interfering with the VicRK two-component system.
• ▸The combination of genetic intervention (ASvicR) and chemical treatment (DMAHDM) overcomes the protective barrier of mature S. mutans biofilms.
• ▸The synergistic strategy significantly weakens the acid production and cariogenicity of Streptococcus mutans.

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Identifiers

DOI

10.3389/fcimb.2026.1793140

Journal

Frontiers in Cellular and Infection Microbiology

Year

2026