Effect of Fluoride and Chlorhexidine Digluconate Mouthrinses on Plaque Biofilms

Per Rabe 1, *, Svante Twetman 1, 2, Bertil Kinnby 3, Gunnel Svensäter 3, Julia R Davies 3
1 Maxillofacial Unit, Halland Hospital, SE-301 85, Halmstad, Sweden
2 Department of Odontology, Section of Cariology and Endodontics, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark
3 Department of Oral Biology, Faculty of Odontology, Malmö University, SE-206 05, Malmö, Sweden

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© Rabe et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Maxillofacial Unit, Halland Hospital, SE-301 85, Halmstad, Sweden; Tel: +46 35134050; E-mail:


Objective : To develop a model in which to investigate the architecture of plaque biofilms formed on enamel surfaces in vivo and to compare the effects of anti-microbial agents of relevance for caries on biofilm vitality. Materials and Methodology : Enamel discs mounted on healing abutments in the pre-molar region were worn by three subjects for 7 days. Control discs were removed before subjects rinsed with 0.1% chlorhexidine digluconate (CHX) or 0.2% sodium fluoride (NaF) for 1 minute. Biofilms were stained with Baclight Live/Dead and z-stacks of images created using confocal scanning laser micoscopy. The levels of vital and dead/damaged bacteria in the biofilms, assessed as the proportion of green and red pixels respectively, were analysed using ImageTrak® software. Results : The subjects showed individual differences in biofilm architecture. The thickness of the biofilms varied from 28-96µm although cell density was always the greatest in the middle layers. In control biofilms, the overall levels of vitality were high (71-98%) especially in the area closest to the enamel interface. Rinsing with either CHX or NaF caused a similar reduction in overall vitality. CHX exerted an effect throughout the biofilm, particularly on the surface of cell clusters whereas NaF caused cell damage/death mainly in the middle to lower biofilm layers. Conclusion : We describe a model that allows the formation of mature, undisturbed oral biofilms on human enamel surfaces in vivo and show that CHX and NaF have a similar effect on overall vitality but differ in their sites of action.

Keywords: Anti-microbial agents, confocal microscopy, dental plaque, dental implants, enamel, vitality.