TODENTJ-25-03-2019_TODENTJ-9-106 1553505645 Bentham Open doi@benthamopen.net Bentham Open The Open Dentistry Journal TODENTJ 1874-2106 3 31 2015 9 1 Per Rabe Svante Twetman Bertil Kinnby Gunnel Svensäter Julia R Davies 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. 3 31 2015 106 111 TODENTJ-9-106 1 10.2174/crossmark_policy opendentistryjournal.com benthamopen.com true Peer Reviewed Single blind Checked with iThenticate 2015-1-26 2015-2-21 2015-2-23 2015-3-31 http://creativecommons.org/licenses/by-nc/3.0/ 10.2174/1874210601509010106 1553505645 https://opendentistryjournal.com/VOLUME/9/PAGE/106/ https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.pdf https://opendentistryjournal.com/contents/volumes/V9/TODENTJ-9-106/TODENTJ-9-106.xml Microbiol Mol Biol Rev. Kolenbrander PE 66 486 2002 Kolenbrander PE, Andersen RN, Blehert DS, Egland PG, Foster JS, Palmer RJ. Communication among oral bacteria. Microbiol Mol Biol Rev 2002; 66 : 486-505. Scand J Dent Res. Nyvad B 95 369 1987 Nyvad B, Killian M. Microbiology of the early colonization of human enamel and root surfaces in vivo. Scand J Dent Res 1987; 95 : 369-80. Science. Costerton JW 284 1318 1999 Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms a common cause of persistent infections. Science 1999; 284 : 1318-22. Eur Arch Pediatra Dent. Marinho VC 10 183 2009 Marinho VC. Cochrane reviews of randomized trials of fluoride therapies for preventing dental caries. Eur Arch Pediatra Dent 2009; 10 : 183-91. Caries Res. Bradshaw DJ 36 81 2002 Bradshaw DJ, Marsh PD, Hodgson RJ, Visser JM. Effects of glucose and fluoride on competition and metabolism within in vitro dental bacterial communities and biofims. Caries Res 2002; 36 : 81-6. J Periodontol. Grossmann E 60 435 1989 Grossmann E, Meckel AH, Isaacs RL , et al. A clinical comparison of antibacterial mothrinses effects of chlorhexidine, phenolics and sanguinrine on dental plaque and gingivitis. J Periodontol 1989; 60 : 435-0. Caries Res. Petti S 40 514 2006 Petti S, Hausen H. Caries-preventive effect of chlorhexidine gel applications among high-risk children. Caries Res 2006; 40 : 514-21. Arch Oral Biol. Marsh PD 28 233 1983 Marsh PD, Keevil CW, McDermid AS, Williamson MI, Ellwood DC. Inhibition by the antimicrobial agent chlorhexidine of acid production and sugar transport in oral streptococcal bacteria. Arch Oral Biol 1983; 28 : 233-40. Arch Oral Biol. Netuschil L 43 277 1998 Netuschil L, Reich E, Unteregger G, Sculean A, Brecx M. A pilot study of confocal laser scanning microscopy for the assessment of undisturbed dental plaque vitality and topography. Arch Oral Biol 1998; 43 : 277-85. Arch Oral Biol. Auschill TM 46 471 2001 Auschill TM, Arweiler NB, Netuschil L, Brecx M, Reich E, Sculean A. Spatial distribution of vital and dead microorganisms in dental biofilms. Arch Oral Biol 2001; 46 : 471-6. J Clin Periodontol. Eberhard J 41 321 2014 Eberhard J, Loewen H, Kruger A , et al. Non-invasive in vivo imaging by confocal laser scanning microscopy of gingival tissues following natural plaque deposition. J Clin Periodontol 2014; 41 : 321-6. J Dent Res. Watson PS 84 451 2005 Watson PS, Pontefract HA, Devine DA , et al. Penetration of fluoride into natural plaque biofilms. J Dent Res 2005; 84 : 451-5. J Clin Periodontol. Netuschil L 16 484 1989 Netuschil L, Reich E, Brecx M. Direct measurement of the bactericidal effect of chlorhexidine on human dental plaque. J Clin Periodontol 1989; 16 : 484-8. Eur J Dent. Germano F 7 152 2013 Germano F, Bramanti E, Arcuri C, Cecchetti F, Cicciu M. Atomic force microscopy of bacteria from periodontal subgingival biofilm Preliminary study results. Eur J Dent 2013; 7 : 152-8. Eur J Oral Sci. Dige I 115 459 2007 Dige I, Nilsson H, Kilian M, Nyvad B. In situ identification of streptococci and other bacteria in initial dental biofilm by confocal laser scanning microscopy and fluorescence in situ hybridization. Eur J Oral Sci 2007; 115 : 459-67. Microbiology. Dige I 155 2116 2009 Dige I, Raarup MK, Nyengaard Jr, Kilian M, Nyvad B. Actinomyces naeslundii in initial biofilm formation. Microbiology 2009; 155 : 2116-6. Appl Environ Microbiol. Takenaka S 74 1869 2008 Takenaka S, Trivedi HM, Corbin A, Pitts B, Stewart PS. Direct visualization of spetial and temporal patterns of antimicrobial action within model oral biofilms. Appl Environ Microbiol 2008; 74 : 1869-75. Antimicrob Agents Chemother. Corbin A 55 3338 2011 Corbin A, Pitts B, Parker A, Stewart PS. Antimicrobial penetration and efficacy in an in vitro oral biofilm model. Antimicrob Agents Chemother 2011; 55 : 3338-44. Oral Microbiol Immunol. Wilson M 11 188 1996 Wilson M, Patel H, Fletcher J. Susceptibility of biofilms of Streptococcus sanguis to chlorhexidine gluconate and cetylpyridinium chloride. Oral Microbiol Immunol 1996; 11 : 188-92. J Dent Res. Zaura-Arite E 80 1436 2001 Zaura-Arite E, van Marle ten Cate JM. Confocal microscopy study of undisturbed and chlorhexidine-treated dental biofilm. J Dent Res 2001; 80 : 1436-40. Trends Microbiol. Fux CA 13 34 2005 Fux CA, Costerton JW, Stewrt PS, Stoodley P. Survical strategies of infectious biofilms. Trends Microbiol 2005; 13 : 34-40. Adv Dent Res. Koo H 20 17 2008 Koo H. Strategies to enhance the biological effects of fluoride on dental biofilms. Adv Dent Res 2008; 20 : 17-21.