A novel non-cytotoxic synthetic peptide, Pug-1, exhibited an antibiofilm effect on Streptococcus mutans adhesion

The aim of this study was to evaluate the effects of synthetic peptides on the growth and biofilm formation of Streptococcus mutans and their cytotoxicity on a human keratinocyte cell line. Four pomegranate-derived peptides; Pug-1, Pug-2, Pug-3 and Pug-4 were synthesized. Streptococcus mutans (ATCC 25175 and DMST 41283) were used. The antibacterial and anti-adherence activities of the peptides were evaluated. The effect of the peptides on S. mutans glucosyltransferase (Gtf) activity was measured. The peptides' cytotoxicity was evaluated on a HaCaT cell line using an MTT assay. The bactericidal penetration depth of Pug-1 into the biofilm was investigated using confocal laser scanning microscopy (CLSM). Among the four peptides, Pug-1 (200 µg ml^-1 ) exhibited antibacterial activity on S. mutans (DMST 41283). All peptides demonstrated anti-adherence activity. Streptococcus mutans Gtf activity was significantly inhibited by Pug-1. Interestingly, Pug-1 peptide was not cytotoxic to the HaCaT cell line. Pug-1 (100-200 µg ml^-1 ) exhibited bactericidal penetration at approximately 50-100% of the biofilm depth. Therefore, Pug-1 might be a candidate of anticariogenic agent. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data highlighted that pomegranate-derived antimicrobial peptides (AMPs) inhibit Streptococcus mutans adhesion, virulence-associated genes and enzymes. This in vitro study supports the hypothesis that pomegranate AMPs would be noncytotoxic to human keratinocytes, with a potent anticariogenic effect via an anti-adherence mode-of-action. The present study provides evidence for further investigation of these AMPs as therapeutic and preventive agents for dental caries.