Spatial distribution of dead and vital bacteria in the native dental biofilm

Objective To examine the spatial distribution of dead and vital bacteria in the early formation of native dental biofilm. Methods An experimental dental biofilm model in the oral cavity was established by enamel slabs and the spatial distribution of dead and vital bacteria in the early colonization of native dental biofilm on the enamel surface was observed by in situ real-time and dynamic observations and optical sections utilizing confocal laser scanning microscope (CLSM) and live and dead bacterial fluorescence staining technique. Results At the initial stage of dental biofilm formation, the structure was sparse and the percentage of dead cells reached 70% - 80% at the inner layers. In the middle layers the structure became denser than in the inner layers, which was mainly composed of vital cells (40% - 70%), and void-like structures were surrounded by vital bacteria. In the outer layers, the structure was sparse and vital cells occupied 20% - 40%. Conclusions Native dental biofilms showed an uneven spatial distribution of vital and dead microorganisms. The percentage of vital microorganisms was lower adjacent to the enamel surface, increased in the z-axis towards the central parts, and decreased again towards the outer layers. The dead bacteria is an integral component in the early formation of native dental biofilm. Bacteria in the biofilms increased with time forming abundant channels.