Changes in the structure and density of oral plaque biofilms with increasing plaque age.

In common with many biofilms in nature, oral plaque has been shown to have a heterogeneous structure as shown by confocal microscopy. In the present study we have used confocal laser scanning microscopy (CLSM) to determine changes in the structure of plaque biofilms with increasing plaque age. Natural human plaque biofilms were formed using the Leeds in situ device. Plaque biofilms were allowed to form for periods of 2 days, 1 week, 2 weeks and 4 weeks before removal of the devices and immediate imaging using the CLSM. Confocal imaging showed that the biofilms retained their heterogeneous appearance at each of the time points studied but appeared to have a tendency to become somewhat more dense with increasing time. Image analysis demonstrated that the percentage of biomass within each biofilm increased over time, particularly between 2 days and 1 week, and with increasing depth into the biofilms. In addition, using the CLSM we were able to demonstrate changes in the bacterial flora of the biofilms with time, with many more filamentous forms being present at the 2- and 4-week time points.