Despite widespread use of various concentrations of fluoride for the prevention of dental caries, the relationship between fluoride concentration and activity against cariogenic biofilms has not been much studied. Herein we investigated the relationship between fluoride concentration and activity against virulence factors and viability of Streptococcus mutans biofilms. S. mutans biofilms were formed on saliva-coated hydroxyapatite discs. The 70-hour-old biofilms were exposed to 0, 1, 3, 10, 30, 100, 300, 1,000 or 2,000 ppm F-. The changes of virulence factors and viability of the biofilms were analyzed using biochemical methods and laser scanning confocal fluorescence microscopy. At 1-2,000 ppm F-, the activity of fluoride against acid production, acid tolerance, and extracellular polysaccharide formation of S. mutans biofilms accurately followed a sigmoidal pattern of concentration dependence (R2 = 0.94-0.99), with EC50 values ranging from 3.07 to 24.7 ppm F-. Generally, the activity of fluoride against the virulence factors was concentration-dependently augmented in 10-100 ppm F- and did not increase further at concentrations higher than 100 ppm F-. However, fluoride did not alter glucosyltransferase activity and viability of S. mutans biofilm cells in all concentrations tested. These results can provide a basis for the selection of appropriate fluoride concentrations that reduce the physiological ability of cariogenic biofilms.

1.
Belli WA, Buckley HD, Marquis RE: Weak acid effects and fluoride inhibition of glycolysis by Streptococcus mutans GS-5. Can J Microbiol 1995;41:785-791.
2.
Bencini DA, Shanley MS, Wild JR, O'Donovan GA: New assay for enzymatic phosphate release: application to aspartate transcarbamylase and other enzymes. Anal Biochem 1983;132:259-264.
3.
Bowden HW: Effects of fluoride on the microbial ecology of dental plaque. J Dent Res 1990;69:653-659.
4.
Bowen WH: Do we need to be concerned about dental caries in the coming millennium? Crit Rev Oral Biol Med 2002;13:126-131.
5.
Bowen WH, Koo H: Biology of Streptococcus mutans-derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms. Caries Res 2011;45:69-86.
6.
Buzalaf MA, Pessan JP, Honório HM, ten Cate JM: Mechanisms of action of fluoride for caries control. Monogr Oral Sci 2011;22:97-114.
7.
Centers for Disease Control and Prevention: Recommendations for using fluoride to prevent and control dental caries in the United States. MMWR Morb Mortal Wkly Rep 2001;50:1-42.
8.
DeLean A, Munson PJ, Rodbard D: Simultaneous analysis of families of sigmoidal curves: application to bioassay, radioligand assay, and physiological dose-response curves. Am J Physiol 1978;235:97-102.
9.
Eisenberg AD, Oldershaw MD, Curzon ME, Handelman SL: Effects of fluoride, lithium, and strontium on growth and acid production of mutants streptococci and Actinomyces viscosus. Caries Res 1991;25:179-184.
10.
Harper DS, Loesche WJ: Growth and acid tolerance of human dental plaque bacteria. Arch Oral Biol 1984;29:843-848.
11.
Hayacibara MF, Rosa OP, Koo H, Torres SA, Costa B, Cury JA: Effects of fluoride and aluminum from ionomeric materials on S. mutans biofilm. J Dent Res 2003;82:267-271.
12.
Heydorn A, Nielsen AT, Hentzer M, Sternberg C, Givskov M, Ersbøll BK, et al: Quantification of biofilm structures by the novel computer program COMSTAT. Microbiology 2000;146:2395-2407.
13.
Jeon JG, Klein MI, Xiao J, Gregoire S, Rosalen PL, Koo H: Influences of naturally occurring agents in combination with fluoride on gene expression and structural organization of Streptococcus mutans in biofilms. BMC Microbiol 2009;9:228.
14.
Klein MI, DeBaz L, Agidi S, Lee H, Xie G, Lin AH, et al: Dynamics of Streptococcus mutans transcriptome in response to starch and sucrose during biofilm development. PLoS One 2010;5:e13478.
15.
Koo H, Xiao J, Klein MI, Jeon JG: Exopolysaccharides produced by Streptococcus mutans glucosyltransferases modulate the establishment of microcolonies within multispecies biofilms. J Bacteriol 2010;192:3024-3032.
16.
Kuramitsu HK: Virulence factors of mutans streptococci: role of molecular genetics. Crit Rev Oral Biol Med 1993;4:159-176.
17.
Lees P, Cunningham FM, Elliott J: Principles of pharmacodynamics and their applications in veterinary pharmacology. J Vet Pharmacol Ther 2004;27:397-414.
18.
Lemos JA, Abranches J, Koo H, Marquis RE, Burne RA: Protocols to study the physiology of oral biofilms. Methods Mol Biol 2010;666:87-102.
19.
Lemos JA, Burne RA: A model of efficiency: stress tolerance by Streptococcus mutans. Microbiology 2008;154:3247-3255.
20.
Loesche WJ: Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50:353-380.
21.
Marquis RE, Clock SA, Mota-Meira M: Fluoride and organic weak acids as modulators of microbial physiology. FEMS Microbiol Rev 2003;26:493-510.
22.
Marsh PD: Dental plaque as a microbial biofilm. Caries Res 2004;38:204-211.
23.
Marsh PD, Bradshaw DJ: The effect of fluoride on the stability of oral bacteria communities in vitro. J Dent Res 1990;69:668-671.
24.
Pandit S, Kim JE, Jung KH, Chang KW, Jeon JG: Effect of sodium fluoride on the virulence factors and composition of Streptococcus mutans biofilms. Arch Oral Biol 2011;56:643-649.
25.
Phan TN, Buckner T, Sheng J, Baldeck JD, Marquis RE: Physiologic actions of zinc related to inhibition of acid and alkali production by oral streptococci in suspensions and biofilms. Oral Microbiol Immunol 2004;19:31-38.
26.
Robinson C, Strafford S, Rees G, Brookes SJ, Kirkham J, Shore RC, et al: Plaque biofilms: the effect of chemical environment on natural human plaque biofilm architecture. Arch Oral Biol 2006;51:1006-1014.
27.
Song JH, Kim SK, Chang KW, Han SK, Yi HK, Jeon JG: In vitro inhibitory effects of Polygonum cuspidatum on bacterial viability and virulence factors of Streptococcus mutans and Streptococcus sobrinus. Arch Oral Biol 2006;51:1131-1140.
28.
Stewart PS, Franklin MJ: Physiological heterogeneity in biofilms. Nat Rev Microbiol 2008;6:199-210.
29.
Takahashi N, Nyvad B: Caries ecology revisited: microbial dynamics and the caries process. Caries Res 2008;42:409-418.
30.
Tten Cate JM: In vitro studies on the effect of fluoride on de- and remineralization. J Dent Res 1990;69:614-619.
31.
Ten Cate JM: Fluorides in caries prevention and control: empiricism or science. Caries Res 2004;38:254-257.
32.
Watson PS, Pontefract HA, Devine DA, Shore RC, Nattress BR, Kirkham J, et al: Penetration of fluoride into natural plaque biofilms. J Dent Res 2005;84:451-455.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.