Sequence of ultrastructural changes of enamel crystals and Streptococcus mutans biofilm in early enamel caries in vitro.

Undecalcified mature enamel sections were used to observe the sequence of ultrastructural changes of enamel crystals and Streptococcus mutans biofilm in the early stages of caries. Human enamel blocks were incubated from 1 to 7 days with S. mutans suspension, and the pH of biofilm was measured. They were processed for light microscopic and transmission electron microscopy observations, and the number of bacteria located in the area adjacent to enamel surface counted. It was observed that the pH of the biofilm dropped to 4, after 1-day of incubation and the S. mutans number increased until 4-day. Round shaped enamel crystals were observed in the 2-day specimens and from the 4-day, images of crystals showing defects and perforations were visualized, becoming more defective along the incubation days. The length of time that the enamel was exposed to biofilm was the main factor for enamel crystals demineralization. Current in vitro caries induction system could standardize time-related changes of the property of Streptococcus mutans biofilm and its relation to enamel crystals demineralization at the ultrastructural level, and thus provide a useful model for the evaluation of the effects of various anti-cariogenic agents.