An approach to differentiate between antibacterial and antiadhesive effects of mouthrinses in vivo

Abstract

An experimental set-up allowing differentiation in vivo between antibacterial and antiadhesive properties of mouthrinses is described. The percentage of vital bacteria (=microbial vitality) and the bacterial counts were microscopically evaluated in saliva and in supragingival dental plaque both collected simultaneously at various times during de novo plaque formation. In a cross-over design, 12 healthy participants refrained from all oral hygiene for four separate periods of 2×4 h and 2×72 h after having rinsed with either an amine fluoride/stannous fluoride solution (Meridol®) or 0.9% NaCl (placebo). Stimulated whole saliva was collected before and after the rinse. Together with whole-saliva samples, representative 4, 24 and 72-h-old plaque samples were separately taken from defined vestibular tooth surfaces that had been either exposed to the mouthrinse (unprotected sites) or temporarily covered with inert plastic films (protected sites) during rinsing. The pooled plaque and saliva were stained with fluorescent dyes to differentiate vital from dead micro-organisms which permitted the estimation of the percentages of vital bacteria. The total bacterial counts were quantified under the darkfield microscope. The Wilcoxon test was used for selected pairwise comparisons ( α=0.05). The percentage of vital bacteria in saliva fell significantly from 80–95% to about 50–60% as a result of the antibacterial activity of the test solution. These baseline values and those found in the presence of 4 and 24-h-old plaque were frequently lower than those recorded after the placebo rinse. In comparison to the placebo, microbial vitality was significantly reduced in early supragingival plaque formed on unprotected sites after applying the test solution. The similar total bacterial counts in 4-h-old plaque recorded after the use of the test solution on the unprotected and the protected areas did not point to an antiadhesive effect of the agent. It is concluded that this new experimental set-up allows decoding of the mode of action of a mouthrinse.