Vol. 44, No. 2, 2010
Issue release date: May 2010
Free Access
Caries Res 2010;44:116–126
(DOI:10.1159/000296306)
Original Paper
Add to my selection

Influence of Cranberry Proanthocyanidins on Formation of Biofilms by Streptococcus mutans on Saliva-Coated Apatitic Surface and on Dental Caries Development in vivo

Koo H.a–c · Duarte S.a · Murata R.M.a · Scott-Anne K.b · Gregoire S.b · Watson G.E.a · Singh A.P.d · Vorsa N.d, e
aEastman Department of Dentistry, bCenter for Oral Biology, and cDepartment of Microbiology and Immunology, University of Rochester Medical Center, Rochester, N.Y., dDepartment of Plant Biology and Plant Pathology, Rutgers University, New Brunswick, N.J., and ePhilip E. Marucci Center for Blueberry and Cranberry Research and Extension, Rutgers University, Chatsworth, N.J., USA
email Corresponding Author


 goto top of outline Key Words

  • Biofilm
  • Cranberry
  • Dental caries
  • Extracellular matrix
  • Glucans
  • Glucosyltransferases
  • Proanthocyanidins

 goto top of outline Abstract

Cranberry crude extracts, in various vehicles, have shown inhibitory effects on the formation of oral biofilms in vitro. The presence of proanthocyanidins (PAC) in cranberry extracts has been linked to biological activities against specific virulence attributes of Streptococcus mutans, e.g. the inhibition of glucosyltransferase (Gtf) activity. The aim of the present study was to determine the influence of a highly purified and chemically defined cranberry PAC fraction on S. mutans biofilm formation on saliva-coated hydroxyapatite surface, and on dental caries development in Sprague-Dawley rats. In addition, we examined the ability of specific PAC (ranging from low-molecular-weight monomers and dimers to high-molecular-weight oligomers/polymers) to inhibit GtfB activity and glycolytic pH drop by S. mutans cells, in an attempt to identify specific bioactive compounds. Topical applications (60-second exposure, twice daily) with PAC (1.5 mg/ml) during biofilm formation resulted in less biomass and fewer insoluble polysaccharides than the biofilms treated with vehicle control had (10% ethanol, v/v; p < 0.05). The incidence of smooth-surface caries in rats was significantly reduced by PAC treatment (twice daily), and resulted in less severe carious lesions compared to the vehicle control group (p < 0.05); the animals treated with PAC also showed significantly less caries severity on sulcal surfaces (p < 0.05). Furthermore, specific A-type PAC oligomers (dimers to dodecamers; 0.1 mg/ml) effectively diminished the synthesis of insoluble glucans by GtfB adsorbed on a saliva-coated hydroxyapatite surface, and also affected bacterial glycolysis. Our data show that cranberry PAC reduced the formation of biofilms by S. mutans in vitro and dental caries development in vivo, which may be attributed to the presence of specific bioactive A-type dimers and oligomers.

Copyright © 2010 S. Karger AG, Basel


 goto top of outline References
  1. Ajdić D, McShan WM, McLaughlin RE, Savić G, Chang J, Carson MB, Primeaux C, Tian R, Kenton S, Jia H, Lin S, Qian Y, Li S, Zhu H, Najar F, Lai H, White J, Roe BA, Ferretti JJ: Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc Natl Acad Sci USA 2002;99:14434–14439.
  2. Belli WA, Buckley DH, Marquis RE: Weak acid effects and fluoride inhibition of glycolysis by Streptococcus mutans GS-5. Can J Microbiol 1995;41:785–791.
  3. Bennick A: Interaction of plant polyphenols with salivary proteins. Crit Rev Oral Biol Med 2002;13:184–196.
  4. Bowen WH: Do we need to be concerned about dental caries in the coming millennium? Crit Rev Oral Biol Med2002;13:126–131.
  5. Bowen WH, Madison KM, Pearson SK: Influence of desalivation in rats on incidence of caries in intact cagemates. J Dent Res 1988;67:1316–1318.
  6. Brecx M: Strategies and agents in supragingival chemical plaque control. Periodontol2000 1997;15:100–108.
  7. Cegelski L, Marshall GR, Eldridge GR, Hultgren SJ: The biology and future prospects of antivirulence therapies. Nat Rev Microbiol 2008;6:17–27.
  8. Cunningham DG, Vannozzi SA, Turk R, O’Shea E, Brilliant K: Cranberry phytochemicals and their health benefits; in Shahidi F, Weerasinghe DK (eds): Nutraceutical Beverages. Chemistry, Nutrition, and Health Effects. ACS Symposium Series 871. Washington, American Chemical Society, 2004, pp 35–50.
  9. Delehanty JB, Johnson BJ, Hickey TE, Pons T, Ligler FS: Binding and neutralization of lipopolysaccharides by plant proanthocyanidins. J Nat Prod 2007;70:1718–1724.
  10. DiPersio JR, Mattingly SJ, Higgins ML, Shockman GD: Measurement of intracellular iodophilic polysaccharide in two cariogenic strains of Streptococcus mutans by cytochemical and chemical methods. Infect Immun1974;10:597–604.
  11. Duarte S, Gregoire S, Singh AP, Vorsa N, Schaich K, Bowen WH, Koo H: Inhibitory effects of cranberry polyphenols on formation and acidogenicity of Streptococcus mutans biofilms. FEMS Microbiol Lett 2006;257:50–56.
  12. Duarte S, Klein MI, Aires CP, Cury JA, Bowen WH, Koo H: Influences of starch and sucrose on Streptococcus mutans biofilms. Oral Microbiol Immunol 2008;23:206–212.
  13. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F: Colorimetric method for determination of sugars and related substances. Anal Chem 1956;28:350–356.
  14. Emilson CG, Nilsson B, Bowen WH: Carbohydrate composition of dental plaque from primates with irradiation caries. J Oral Pathol1984;13:213–220.
  15. Flemming HC, Neu TR, Wozniak DJ: The EPS matrix: the ‘house of biofilm cells’. J Bacteriol2007;189:7945–7947.
  16. Foo LY, Lu Y, Howell AB, Vorsa N: The structure of cranberry proanthocyanidins which inhibit adherence of uropathogenic P-fimbriated Escherichia coli in vitro. Phytochemistry 2000;54:173–181.
  17. Gibbons RJ: Adherent interactions which may affect microbial ecology in the mouth. J Dent Res 1984;63:378–385.
  18. Gregoire S, Singh AP, Vorsa N, Koo H: Influence of cranberry phenolics on glucan synthesis by glucosyltransferases and Streptococcus mutans acidogenicity. J Appl Microbiol 2007;103:1960–1968.
  19. Guggenheim B: Extracellular polysaccharides and microbial plaque. Int Dent J 1970;20:657–678.
  20. Hamada S, Slade HD: Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev 1980;44:331–384.
  21. Haslam E: Natural polyphenols (vegetable tannins) as drugs: possible modes of action. J Nat Prod 1996;59:205–215.
  22. Hayacibara MF, Koo H, Vacca Smith AM, Kopec LK, Scott-Anne K, Cury JA, Bowen WH: The influence of mutanase and dextranase on the production and structure of glucans synthesized by streptococcal glucosyltransferases. Carbohydr Res 2004;12:2127–2137.

    External Resources

  23. Hotz P, Guggenheim B, Schmid R: Carbohydrates in pooled dental plaque. Caries Res1972;6:103–121.
  24. Koo H: Strategies to enhance the biological effects of fluoride on dental biofilms. Adv Dent Res 2008;20:17–21.
  25. Koo H, Hayacibara MF, Schobel BD, Cury JA, Rosalen PL, Park YK, Vacca Smith AM, Bowen WH: Inhibition of Streptococcus mutans biofilm accumulation and polysaccharide production by apigenin and tt-farnesol. J Antimicrob Chemother 2003;52:782–789.
  26. Koo H, Nino de Guzman P, Schobel BD, Vacca Smith AV, Bowen WH: Influence of cranberry juice on glucan-mediated processes involved in Streptococcus mutans biofilm development. Caries Res 2006;40:20–27.
  27. Koo H, Rosalen PL, Cury JA, Park YK, Bowen WH: Effects of compounds found in propolis on Streptococcus mutans growth and on glucosyltransferase activity. Antimicrob Agents Chemother 2002;46:1302–1309.
  28. Koo H, Schobel B, Scott-Anne K, Watson G, Bowen WH, Cury JA, Rosalen PL, Park YK: Apigenin and tt-farnesol with fluoride effects on S mutans biofilms and dental caries. J Dent Res 2005;84:1016–1020.
  29. Koo H, Vacca Smith AM, Bowen WH, Rosalen PL, Cury JA, Park YK: Effects of Apis mellifera propolis on the activities of streptococcal glucosyltransferases in solution and adsorbed onto saliva-coated hydroxyapatite. Caries Res 2000;34:418–426.
  30. Kopec LK, Vacca Smith AM, Bowen WH: Structural aspects of glucans formed in solution and on the surface of hydroxyapatite. Glycobiology 1997;7:929–934.
  31. La VD, Howell AB, Grenier D: Cranberry proanthocyanidins inhibit MMP production and activity. J Dent Res 2009;88:627–632.
  32. Larson RM: Merits and modifications of scoring rat dental caries by Keyes’ method; in Tanzer JM (ed): Animal Models in Cariology. Microbiology Abstracts (special suppl.). Washington, IRL, 1981, pp 195–203.
  33. Marquis RE, Clock SA, Mota-Meira M: Fluoride and organic weak acids as modulators of microbial physiology. FEMS Microbiol Rev2003;760:1–18.
  34. Marsh PD: Are dental diseases examples of ecological catastrophes? Microbiology 2003;149:279–294.
  35. Moore S, Stein WH: A modified ninhydrin reagent for the photometric determination of amino acids and related compounds. J Biol Chem 1954;211:907–913.
  36. Murata RM, Branco de Almeida LS, Yatsuda R, Dos Santos MH, Nagem TJ, Rosalen PL, Koo H: Inhibitory effects of 7-epiclusianone on glucan synthesis, acidogenicity and biofilm formation by Streptococcus mutans. FEMS Microbiol Lett 2008;282:174–181.
  37. Osawa K, Miyazaki K, Shimura S, Okuda J, Matsumoto M, Ooshima T: Identification of cariostatic substances in the cacao bean husk: their anti-glucosyltransferase and antibacterial activities. J Dent Res 2001;80:2000–2004.
  38. Paes Leme AF, Koo H, Bellato CM, Bedi G, Cury JA: The role of sucrose in cariogenic dental biofilm formation: new insight. J Dent Res 2006;85:878–887.
  39. Quivey RG Jr, Kuhnert WL, Hahn K: Adaptation of oral streptococci to low pH. Adv Microb Physiol 2000;42:239–274.
  40. Raubertas RF, Davis BA, Bowen WH, Pearson SK, Watson GE: Litter effects on caries in rats and implications for experimental design. Caries Res 1999;33:164–169.
  41. Reese S, Guggenheim B: A novel TEM contrasting technique for extracellular polysaccharides in in vitro biofilms. Microsc Res Tech 2007;70:816–822.
  42. Rölla G, Ciardi JE, Eggen K, Bowen WH, Afseth J: Free glucosyl- and fructosyltransferase in human saliva and adsorption of these enzymes to teeth in vivo; in Doyle RJ, Ciardi JE (eds): Glucosyltransferases, Glucans, Sucrose, and Dental Caries. Washington, Chemical Senses, 1983, pp 21–30.
  43. Schilling KM, Bowen WH: The activity of glucosyltransferase adsorbed onto saliva-coated hydroxyapatite. J Dent Res 1988;67:2–8.
  44. Schilling KM, Bowen WH: Glucans synthesized in situ in experimental salivary pellicle function as specific binding sites for Streptococcus mutans. Infect Immun 1992;60:284–295.
  45. Schmidt BM, Ribnicky DM, Lipsky P, Raskin I: Revisiting the ancient concept of botanical therapeutics. Nat Chem Biol 2007;3:360–366.
  46. Singh AP, Singh RK, Kim KK, Satyan KS, Nussbaum R, Torres M, Brard L, Vorsa N: Cranberry proanthocyanidins are cytotoxic to human cancer cells and sensitize platinum-resistant ovarian cancer cells to paraplatin. Phytother Res 2009;23:1066–1074.
  47. Steinberg D, Rozen R, Bromshteym M, Zaks B, Gedalia I, Bachrach G: Regulation of fructosyltransferase activity by carbohydrates, in solution and immobilized on hydroxyapatite surfaces. Carbohydr Res 2002;337:701–710.
  48. Tanzer JM, Freedman ML, Fitzgerald RJ: Virulence of mutants defective in glucosyltransferase, dextran-mediated aggregation, or dextranase activity; in Mergenhagen SE, Rosan B (eds): Molecular Basis of Oral Microbial Adhesion. Washington, American Society for Microbiology, 1985, pp 204–211.
  49. Vacca Smith AM, Bowen WH: Binding properties of streptococcal glucosyltransferases for hydroxyapatite, saliva-coated hydroxyapatite, and bacterial surfaces. Arch Oral Biol 1998;43:103–110.
  50. Weiss EL, Lev-Dor R, Sharon N, Ofek I: Inhibitory effect of a high-molecular-weight constituent of cranberry on adhesion of oral bacteria. Crit Rev Food Sci Nutr 2002;42:285–292.
  51. Wilson T, Singh AP, Vorsa N, Goettl CD, Kittleson KM, Roe CM, Kastello GM, Ragsdale FR: Human glycemic response and phenolic content of unsweetened cranberry juice. J Med Food 2008;11:46–54.
  52. Xiao J, Koo H: Structural organization and dynamics of exopolysaccharide matrix and microcolonies formation by Streptococcus mutans in biofilms. J Appl Microbiol 2009, E-pub ahead of print.
  53. Yamanaka A, Kimizuka R, Kato T, Okuda K: Inhibitory effects of cranberry juice on attachment of oral streptococci and biofilm formation. Oral Microbiol Immunol 2004;19:150–154.
  54. Yamashita Y, Bowen WH, Burne RA, Kuramitsu HK: Role of the Streptococcus mutans gtf genes in caries induction in the specific-pathogen-free rat model. Infect Immun 1993;61:3811–3817.
  55. Yanagida A, Kanda T, Tanabe M, Matsudaira F, Oliveira Cordeiro JG: Inhibitory effects of apple polyphenols and related compounds on cariogenic factors of mutans streptococci. J Agric Food Chem 2000;48:5666–5671.
  56. Zero DT: Dentifrices, mouthwashes, and remineralization/caries arrestment strategies. BMC Oral Health 2006;15:S9.

    External Resources


 goto top of outline Author Contacts

Hyun Koo
University of Rochester Medical Center
Center for Oral Biology, 601 Elmwood Avenue
Box 611, Rochester, NY 14620 (USA)
Tel. +1 585 273 4216, Fax +1 585 276 0190, E-Mail hyun_koo@urmc.rochester.edu


 goto top of outline Article Information

Received: September 11, 2009
Accepted after revision: January 27, 2010
Published online: March 16, 2010
Number of Print Pages : 11
Number of Figures : 2, Number of Tables : 4, Number of References : 56


 goto top of outline Publication Details

Caries Research

Vol. 44, No. 2, Year 2010 (Cover Date: May 2010)

Journal Editor: Beighton D. (London)
ISSN: 0008-6568 (Print), eISSN: 1421-976X (Online)

For additional information: http://www.karger.com/CRE


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 or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
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 goverment 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.