Clinical Oral Investigations

, Volume 12, Issue 3, pp 203–208 | Cite as

Effect of food preservatives on in situ biofilm formation

  • Nicole Birgit Arweiler
  • Ronaldo Lenz
  • Anton Sculean
  • Ali Al-Ahmad
  • Elmar Hellwig
  • Thorsten Mathias Auschill
Original Article

Abstract

The aim of this double-blind, controlled crossover study was to evaluate the influence of food preservatives on in situ dental biofilm growth. Twenty-four volunteers wore appliances with six specimens each of bovine enamel to build up intra-oral biofilms. During three test cycles, the subjects had to put one half of the appliance twice a day in one of the assigned active solutions (0.1% benzoate, BA; 0.1% sorbate, SA or 0.2% chlorhexidine, CHX) and the other into NaCl. After 5 days, the developed biofilms were stained with two fluorescent dyes to visualise vital (green) and dead bacteria (red). Biofilms were scanned by confocal laser scanning microscopy and biofilm thickness (BT) and bacterial vitality (BV%) were calculated. After a washout period of 7 days, a new test cycle was started. The use of SA, BA and CHX resulted in a significantly reduced BT and BV compared to NaCl (p < 0.001). Differences between SA and BA were not significant (p > 0.05) for both parameters, while CHX showed significantly lower values. Both preservatives showed antibacterial and plaque-inhibiting properties, but not to the extent of CHX. The biofilm model enabled the examination of undisturbed oral biofilm formation influenced by antibacterial components under clinical conditions.

Keywords

Biofilm Plaque vitality Confocal laser scanning microscopy 

Notes

Acknowledgements

This investigation was financially supported by a grant from the Deutsche Forschungsgemeinschaft (DFG; Ar 341/3-1). The authors thank Marie Follo, Ph.D., Department of Hematology and Oncology, Core facility, University of Freiburg for her help in the image analysis and preparing the manuscript.

Declaration of Interests

There are no conflicts of interest for any author.

References

  1. 1.
    Al-Ahmad A, Wunder A, Auschill TM, Follo M, Braun G, Hellwig E, Arweiler NB (2007) The in vivo dynamics of Streptococcus spp., Actinomyces naeslundii, Fusobacterium nucleatum and Veillonella spp. in dental plaque biofilm as analysed by five-colour multiplex fluorescence in situ hybridization. J Med Microbiol 56:681–687PubMedCrossRefGoogle Scholar
  2. 2.
    Arweiler NB, Hellwig E, Sculean A, Hein N, Auschill TM (2004) Individual vitality pattern of in situ dental biofilms at different locations in the oral cavity. Caries Res 38:442–447PubMedCrossRefGoogle Scholar
  3. 3.
    Auschill TM, Arweiler NB, Netuschil L, Brecx M, Reich E, Sculean A (2001) Spatial distribution of vital and dead microorganisms in dental biofilms. Arch Oral Biol 46:471–476PubMedCrossRefGoogle Scholar
  4. 4.
    Auschill TM, Hellwig E, Sculean A, Hein N, Arweiler NB (2004) Impact of the intraoral location on the rate of biofilm growth. Clin Oral Investig 8:97–101PubMedCrossRefGoogle Scholar
  5. 5.
    Auschill TM, Hein N, Hellwig E, Follo M, Sculean A, Arweiler NB (2005) Effect of two antimicrobial agents on early in situ biofilm formation. J Clin Periodontol 32:147–152PubMedCrossRefGoogle Scholar
  6. 6.
    Belli WA, Buckley DH, Marquis RE (1995) Weak acid effects and fluoride inhibition of glycolysis by Streptococcus mutans GS-5. Can J Microbiol 41:785–791PubMedCrossRefGoogle Scholar
  7. 7.
    Chichester DF, Tanner FW (1968) Antimicrobial food additives. In: Furia TE (ed) CRC handbook of food additives. Chemical Rubber Co, Cleveland, pp 142–157Google Scholar
  8. 8.
    Consensus Statement on Diet on the FDI’s Second World Conference on Oral Health Promotion (2000). Int Dent J 50:174Google Scholar
  9. 9.
    Davis BA, Raubertas RF, Pearson SK, Bowen WH (2001) The effects of benzoate and fluoride on dental caries in intact and desalivated rats. Caries Res 35:331–337PubMedCrossRefGoogle Scholar
  10. 10.
    Eklund T (1980) Inhibition of growth and uptake processes in bacteria by some chemical food preservatives. J Appl Bacteriol 48:423–432PubMedGoogle Scholar
  11. 11.
    Freese E, Sheu CW, Galliers E (1973) Function of lipophilic acids as antimicrobial food additives. Nature 241:321–325PubMedCrossRefGoogle Scholar
  12. 12.
    Jones CG (1997) Chlorhexidine: is it still the gold standard? Periodontol 15:55–62, 2000CrossRefGoogle Scholar
  13. 13.
    Koo H, Nino de Guzman P, Schobel BD, Vacca Smith AV, Bowen WH (2006) Influence of cranberry juice on glucan-mediated processes involved in Streptococcus mutans biofilm development. Caries Res 40:20–27PubMedCrossRefGoogle Scholar
  14. 14.
    Leikanger S, Bjertness E, Scheie AA (1992) Effects of food preservatives on growth and metabolism of plaque bacteria in vitro and in vivo. Scand J Dent Res 100:371–376PubMedGoogle Scholar
  15. 15.
    Main C, Geddes DA, McNee SG, Collins WJ, Smith DC, Weetman DA (1984) Instrumentation for measurement of dental plaque thickness in situ. J Biomed Eng 6:151–154PubMedCrossRefGoogle Scholar
  16. 16.
    Netuschil L, Reich E, Unteregger G, Sculean A, Brecx M (1998) A pilot study of confocal laser scanning microscopy for the assessment of undisturbed dental plaque vitality and topography. Arch Oral Biol 43:277–285PubMedCrossRefGoogle Scholar
  17. 17.
    Ostergaard E (1994) Evaluation of the antimicrobial effects of sodium benzoate and dichlorobenzyl alcohol against dental plaque microorganisms. An in vitro study. Acta Odontol Scand 52:335–345PubMedCrossRefGoogle Scholar
  18. 18.
    Prosser BL, Taylor D, Dix BA, Cleeland R (1987) Method of evaluating effects of antibiotics on bacterial biofilm. Antimicrob Agents Chemother 31:1502–1506PubMedGoogle Scholar
  19. 19.
    Roberts SK, Bass C, Brading M, Lappin-Scott H, Stoodley P (1999) Biofilm formation and structure: what’s new? In: Newman HN, Wilson M (eds) Dental plaque revisited—oral biofilms in health and disease. BioLine, Cardiff, pp 15–36Google Scholar
  20. 20.
    Robinson C, Kirkham J, Percival R, Shore RC, Bonass WA, Brookes SJ, Kusa L, Nakagaki H, Kato K, Nattress B (1997) A method for the quantitative site-specific study of the biochemistry within dental plaque biofilms formed in vivo. Caries Res 31:194–200PubMedCrossRefGoogle Scholar
  21. 21.
    Standard and Poor’s Food and Non-Alcoholic Beverages (1998) Handbook, Standard and Poor’s Corporation, New York, Sect 166, p12Google Scholar
  22. 22.
    Salmond CV, Kroll RG, Booth IR (1984) The effect of food preservatives on pH homeostasis in Escherichia coli. J Gen Microbiol 130:2845–2850PubMedGoogle Scholar
  23. 23.
    ten Cate JM, Marsh PD (1994) Procedures for establishing efficacy of antimicrobial agents for chemotherapeutic caries prevention. J Dent Res 73:695–703PubMedGoogle Scholar
  24. 24.
    Thurnheer T, Gmür R, Shapiro S, Guggenheim B (2003) Mass transport of macromolecules within an in vitro model of supragingival plaque. Appl Environ Microbiol 69:1702–1709PubMedCrossRefGoogle Scholar
  25. 25.
    Whitford GM, Schuster GS, Pashley DH, Venkateswarlu P (1977) Fluoride uptake by Streptococcus mutans 6715. Infect Immun 18:680–687PubMedGoogle Scholar
  26. 26.
    Wimpenny JW (1997) The validity of models. Adv Dent Res 11:150–159PubMedGoogle Scholar
  27. 27.
    Yeganeh S, Lynch E, Jovanovski V, Zou L (1999) Quantification of root surface plaque using a new 3-D laser scanning method. J Clin Periodontol 26:692–697PubMedCrossRefGoogle Scholar
  28. 28.
    Zaura-Arite E, ten Cate JM (2000) Effects of fluoride- and chlorhexidine-containing varnishes on plaque composition and on demineralization of dentinal grooves in situ. Eur J Oral Sci 108:154–161PubMedCrossRefGoogle Scholar
  29. 29.
    Zaura-Arite E, van Marle J, ten Cate JM (2001) Confocal microscopy study of undisturbed and chlorhexidine-treated dental biofilm. J Dent Res 80:1436–1440PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Nicole Birgit Arweiler
    • 1
    • 3
  • Ronaldo Lenz
    • 1
  • Anton Sculean
    • 2
  • Ali Al-Ahmad
    • 1
  • Elmar Hellwig
    • 1
  • Thorsten Mathias Auschill
    • 1
  1. 1.Department of Operative Dentistry and Periodontology, Dental School and HospitalUniversity Medical CenterFreiburgGermany
  2. 2.Department of PeriodontologyRadboud University Medical CenterNijmegenThe Netherlands
  3. 3.Department of Operative Dentistry and Periodontology, Dental School and HospitalAlbert-Ludwigs-UniversityFreiburgGermany

Personalised recommendations