Chlorhexidine substantivity on salivary flora and plaque-like biofilm: an in situ model

Lucía García-Caballero, Victor Quintas, Isabel Prada-López, Juan Seoane, Nikos Donos, Inmaculada Tomás, Lucía García-Caballero, Victor Quintas, Isabel Prada-López, Juan Seoane, Nikos Donos, Inmaculada Tomás

Abstract

Objective: To evaluate the in situ antibacterial activity of a mouthrinse with 0.2% Chlorhexidine (M-0.2% CHX) on undisturbed de novo plaque-like biofilm (PL-biofilm) and on salivary flora up to 7 hours after its application.

Methods: A special acrylic appliance was designed, with 3 inserted glass disks on each buccal side, allowing for PL-biofilm growth. Fifteen healthy volunteers wore the appliance for 48 hours and then performed an M-0.2% CHX; disks were removed at 30 seconds and 1, 3, 5 and 7 hours after the mouth-rinsing. Applying a washout period, saliva samples were collected from each volunteer at 30 seconds and 1, 3, 5 and 7 hours after performing an M-0.2% CHX. The PL-biofilm and saliva samples were analysed by confocal laser scanning and epifluorescence microscopes, respectively.

Results: At 30 seconds after M-0.2% CHX, the levels of viable bacteria detected in saliva were significantly lower than those observed in PL-biofilm. The difference in the percentage of live bacteria detected in saliva was significantly higher than that observed in PL-biofilm at 5 and 7 hours after M-0.2% CHX.

Conclusion: After a single mouthrinse of the 0.2% CHX formulation tested in the present study, the 2-day PL-biofilm presented a significantly higher resistance to this antiseptic in situ than that observed in salivary flora. However, this 0.2% CHX formulation showed a higher substantivity on PL-biofilm than on salivary flora at 5 and 7 hours after mouth-rinsing, which could be related to the slower growth rate of PL-biofilm and the possible reservoir function for antimicrobial agents associated with the undisturbed de novo PL-biofilm.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Individualized splint of the lower…
Figure 1. Individualized splint of the lower arch.
A) Parts of the individualized splint: 1. internal vinyl sheet; 2. polished glass discs; 3. fenestrated external vinyl sheet. B) Clinical view of the individualized splint with the glass discs inserted (arrows).
Figure 2. Mean percentages of bacterial viability…
Figure 2. Mean percentages of bacterial viability in saliva and de novo PL-biofilm under basal conditions and at different times after a single mouthrinse of sterile water and 0.2% Chlorhexidine.
A) Intra-mouthrinse differences; B) Inter-mouthrinse differences; C) Inter-ecosystem differences.
Figure 3. Images representing the changes in…
Figure 3. Images representing the changes in bacterial viability in the saliva.
A) Sample collected under basal conditions; B), C), D) Sample collected at 30 seconds, 5 and 7 hours respectively after a single mouthrinse of 0.2% Chlorhexidine.
Figure 4. Images representing the changes in…
Figure 4. Images representing the changes in bacterial viability in de novo PL-biofilm.
A) Sample collected under basal conditions; B), C), D) Sample collected at 30 seconds, 3 and 7 hours respectively after a single mouthrinse of 0.2% Chlorhexidine.
Figure 5. Visualization of channels (A, yellow…
Figure 5. Visualization of channels (A, yellow star) and voids (B, white star) in a single cross sectional plane from X, Y and Z axis images obtained by the CLSM from a baseline sample.
The presence of both channels and voids shows a heterogeneous architecture model.

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