Antibacterial effect of chlorhexidine on bacteria adsorbed onto experimental dental plaque

The most common method used to examine the antibacterial effect of antiplaque agents is the minimal inhibitory concentration (MIC) method, which is tested on bacteria in suspensions. Examining the antibacterial effect on bacteria adsorbed onto dental surfaces models is not as popular, although it is clear that such models reflect the conditions in the oral cavity far more accurately than the conventional MIC method. The antibacterial effect of chlorhexidine (CHX) on bacteria adsorbed onto experimental dental plaque model was investigated. Hydroxyapatite beads were coated with human saliva. Next, cell-free fructosyltransferase (FTF) and glucosyltransferase (GTF) were adsorbed onto the beads, and sucrose was added to allow the synthesis of polysaccharide by the surface-bound enzymes. Following adsorption of Streptococcus sobrinus to the experimental dental plaque (EDP), the EDP was exposed to CHX at concentrations between 0.008 to 0.0002% at pH values of 5.5, 6.5, and 7.5. After 150 min incubation, growth of the adsorbed bacteria was measured by their incorporation of 3H-uridine or 3H-thymidine. Comparison of bacterial growth on the EDP with that in suspension showed that the surface-bound bacteria were less sensitive to CHX than were the bacteria in suspension. At all tested CHX concentrations, the antibacterial effect was independent of pH. In addition, under our experimental conditions, the use of 3H-uridine as an indicator of bacterial viability proved to be more sensitive than 3H-thymidine. The antibacterial effect of an agent in a model closely mimicking the tooth surface should generate results that are more relevant to the in vivo conditions than are conventional methods, thus bearing significant information concerning clinical applications of CHX.