Materials and methods
A three-species biofilm (Porphyromonas gingivalis, Fusobacterium nucleatum and Streptococcus sanguinis) was formed in vitro on protein-coated titanium disks using a flow chamber system combined with a static biofilm growth model. The oscillation frequencies of three commercial side-to-side toothbrushes were evaluated by means of a dose response. The frequency was decreased in steps (100, 85, 70, 55, and 40%). Subsequently, the biofilm-coated substrates were exposed to the side-to-side toothbrushes. The biofilm volumes were measured using volumetric analyses (Imaris 8.1.2) with confocal laser scanning microscope images (Zeiss LSM700).
Compared to maximum oscillation frequency (100%), lower oscillation frequencies (up to 40%) resulted in reduced median percentages of biofilm reduction (median biofilm reduction up to 53% for maximum oscillation frequency, and up to 13% for 40% oscillation frequency) (p ≥ 0.03). In addition, decreasing the oscillation frequencies of the side-to-side toothbrushes showed an enhanced variety in the results of repeated experiments.