Efficacy of natural antimicrobials in toothpaste formulations against oral biofilms in vitro

Objectives: To evaluate the antimicrobial efficacies of two toothpaste formulations containing natural antimicrobials (herbal extracts and chitosan) against oral biofilms of different composition and maturational status.Methods: Bacteria from a buffer suspension or fresh saliva were adhered for 2 h to a salivary conditioning film and subsequently grown for 16 h. Dual-species biofilms were prepared from Actinomyces naeslundii T14V-J1 and Streptococcus oralis J22, whilst multi-species biofilms were grown from freshly collected human saliva. Biofilms were exposed to 25 wt% toothpaste supernatants. A chlorhexidine-containing mouthrinse and a buffer were used as positive- and negative-controls, respectively. Antibacterial efficacy was concluded from acute killing, bacterial removal, prevention of bacterial re-deposition and continued killing during re-deposition.Results: The herbal- and chitosan-based supernatants showed immediate killing of oral biofilm bacteria, comparable with chlorhexidine. Moreover, exposure of a biofilm to these supernatants or chlorhexidine, yielded ongoing killing of biofilm bacteria after exposure during re-deposition of bacteria to a matured 16 h biofilm, but not to a much thinner initial biofilm formed by 2 h adhesion only. This suggests that thicker, more matured biofilms can absorb and release oral antimicrobials.Conclusions: Supernatants based on herbal- and chitosan-based toothpastes have comparable immediate and ongoing antibacterial efficacies as chlorhexidine. Natural antimicrobials and chlorhexidine absorb in oral biofilms which contributes to their substantive action. (C) 2010 Elsevier Ltd. All rights reserved.