OBJECTIVE To evaluate the in vitro effects of commercially available charcoal dentifrices on Streptococcus mutans biofilm development and their ability to prevent enamel demineralization. METHODS Streptococcus mutans biofilm was formed on polished bovine enamel specimens (n= 9 per treatment), and treated twice-daily for 120 seconds over the course of 5 days with: charcoal dentifrice containing fluoride (1,000 ppm F) (CF+), fluoride-free charcoal dentifrice (CF-), regular fluoride (1,100 ppm F) dentifrice (F+ ), or regular fluoride-free dentifrice (F-). Chlorhexidine (CHX, 0.12%) and deionized water (DIW) were used as positive and negative controls, respectively. Biofilms were analyzed for bacterial viability (colony-forming units, CFU). The pH of the medium was measured daily. Enamel specimens were analyzed using Vickers microhardness ( HV) and transversal microradiography (TMR). Data were analyzed using one-way ANOVA followed by post-hoc tests (α= 0.05). RESULTS F+ showed higher pH values than CF+ and CF-, and CF- presented higher pH than CF+, showing that CF+ did not have inhibitory effects on the acidogenicity of cariogenic biofilms. CFU was significantly decreased when specimens were treated with CF+, CF- and F+, compared to specimens treated with DIW (P≤ 0.035) or F- (P≤ 0.001), respectively. However, the reduction observed was minimal (approximately 1 log). CF+ and CF- were less effective than F+ in preventing enamel demineralization as determined using HV (P= 0.041 and P= 0.003, respectively) and TMR ( P≤ 0.001). Both charcoal dentifrices (CF+, CF-) did not show relevant inhibition of S. mutans biofilm growth. Additionally, neither product prevented enamel demineralization compared to a regular fluoride-containing dentifrice. CONCLUSIONS The tested charcoal dentifrices did not exhibit anticaries potential.