This in vitro study evaluated the influence of chlorhexidine diacetate (CDA) when blended within dentin bonding systems (DBSs) on Streptococcus mutans (S. mutans) biofilm formation. One commercially available 0.2% wt CDA-containing DBS (Peak Universal Bond) and five experimental 0.2% wt CDA-containing DBS formulations (experimental Adper Scotchbond 1XT plus experimental resins, R2, R3, R4, R5) were assessed vs their no-CDA containing counterparts. Twenty-eight DBSs disks were prepared for each group (6.4 mm×1.0 mm) and cured for 80 s at 800 mW/cm2 in a nitrogen atmosphere. A modified Drip-Flow Reactor was used to grow S. mutans biofilms on specimen surfaces for 24 h and adherent, viable biomass was evaluated using a tetrazolium salt assay (MTT). Two specimens from each of the tested materials were processed with LIVE/DEAD stain and observed using laser confocal microscopy (CLSM) while two disks from each group were examined by using scanning electron microscopy (SEM). MTT assay, CLSM and SEM observations showed that CDA addition decreased, increased or did not change S. mutans biofilm formation. The lowest biofilm formation was obtained with Peak Universal Bond and R5 (with and without CDA). It may be concluded that the chemical composition of DBSs determines their ability to promote or hamper biofilm formation. Therefore, CDA addition may be helpful in modulating biofilm formation provided that DBS formulation is tuned and optimized.
