It has been suggested that enamel would resist higher frequencies of sucrose exposure if fluoride from water or dentifrice is being used. However, the effect of increasing frequencies of sugar on dental biofilm composition is not well known. Ten volunteers living in a fluoridated area wore palatal appliances bearing human enamel slabs during 14 days. The slabs were exposed to 20% sucrose solution 0 (control), 2, 4, 6, 8 or 10 times/day and the volunteers used fluoride dentifrice 3 times/day. Enamel demineralization was significantly greater than control for sucrose frequencies higher than 6 times/day. However, biofilm mass, total microbiota, total streptococci, lactobacilli counts and insoluble extracellular polysaccharide concentration increased, while Ca, P i and F concentration in whole biofilm decreased significantly, with frequencies of sucrose exposure lower than 6 times/day. The findings confirm that fluoride can reduce enamel demineralization if sucrose consumption is not higher than 6 times/day, but changes in the biochemical and microbiological composition of the biofilm are observed with lower frequencies of sucrose use. Copyright © 2007 S. Karger AG.