Background: The aim of the present study is to assess in situ substantivity of a single mouthrinse with 0.2% chlorhexidine (CHX) on saliva and on undisturbed de novo plaque-like biofilm (PL-biofilm), differentiating between two times of application: 1) CHX mouthrinse in the morning; and 2) CHX mouthrinse at night.
Methods: The study participants were 10 healthy volunteers who wore an individualized splint with glass disks for 48 hours to boost the growth of PL-biofilm. Saliva samples were collected, and two disks were removed from each volunteer's splint at 8, 10, and 12 hours after performing a mouthrinse with 0.2% CHX at 7:00 am (M-0.2% CHX-diurnal) and 1:00 am (M-0.2% CHX-nocturnal). The saliva and plaque samples were analyzed by epifluorescence and confocal laser scanning microscopy, respectively, using a green fluorescent nucleic acid stain/propidium iodide staining.
Results: With M-0.2% CHX-diurnal, the frequency of vital bacteria in saliva was significantly higher than in the PL-biofilm at 8, 10, and 12 hours after mouthrinse. After M-0.2% CHX-nocturnal, the frequency of vital bacteria in saliva was significantly lower than in the PL-biofilm at 8 hours and higher than in the PL-biofilm at 12 hours after mouthrinse.
Conclusion: These results support the more active physiologic dynamics of the salivary flora and the possible reservoir function associated with the structure of undisturbed de novo PL-biofilm.