The influence of antibacterial toothpastes on in vitro Streptococcus mutons biofilm formation: A continuous culture study

To evaluate the in vitro effect of five toothpastes containing antimicrobial compounds including fluoride, triclosan or hydroxyapatite nano-particles on Streptococcus mutans (S. mutans) biofilm formation. Fluoride uptake by enamel after bacterial challenge was also evaluated. Human enamel disks (n= 192) were randomly divided into six groups and brushed with five different toothpastes while the control group was brushed with distilled water. Each group was incubated for 24 and 72 hours with a S. mutans biofilm growing on a modified drip-flow reactor (MDFR). Biofilm formation was determined using a viable biomass assay based on a tetrazolium salt (MTT) and evaluated morphologically with confocal laser-scanning microscopy (CLSM) and scanning electron-microscopy (SEM). Fluoride uptake was evaluated using the enamel biopsy technique. Biofilm formation was also evaluated using 120 disks randomly divided into the same six groups. The number of viable bacteria was determined through plate count on Mitis Salivarius Bacitracin agar (MSB agar). Data from plate count showed the same overall trend of MTT assay. The latter showed that after 24 hours the effect of the tested toothpastes was significantly higher in reducing biofilm formation than after 72 hours. The toothpaste containing a high concentration of amine fluoride (AmF) had the highest performance in reducing biofilm formation. Fluoride uptake of enamel showed a positive trend related to the fluoride concentration in both incubation times.