AbstractWe examined the effect of L-arginine - (i) on the growth of L. rhamnosus GG (LrG) and (ii) combined LrG synbiotic on the growth of cariogenic S. mutans. Viability of LrG was assessed using MTT/XTT assays, confocal imaging with ADS activity measurement. The effect of L-arginine (0.5%/1%/2%) (2×/24 h) with LrG on S. mutans was evaluated by measuring the colony forming units, biofilm biomass, real-time qPCR and confocal imaging. The pH of the spent media was measured immediately and 24 h post-treatment with assessment of lactic acid. The LrG viability was highest with 2% L-arginine (p < 0.001). Confocal imaging showed that 2% L-arginine increased biofilm thickness of LrG. The 2% L-arginine and LrG synbiotic significantly inhibited the growth of S. mutans (p < 0.001) reducing the viable counts (p = 0.002) and biofilm biomass (p < 0.001). The pH of spent media was the highest when treated with 2% L-arginine and LrG synbiotic (p < 0.001) with no difference between post-treatment and 24 h post-treatment (p > 0.05). Conversely, the 2% L-arginine and LrG synbiotic showed the lowest lactic acid production (p < 0.001). This study demonstrated that L-arginine enhanced the growth of LrG. The 2% L-arginine and LrG synbiotic synergistically inhibits the growth of S. mutans with significant potential to develop as an anti-caries regimen.