Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Action of food preservatives on 14-days dental biofilm formation, biofilm vitality and biofilm-derived enamel demineralisation in situ

  • 482 Accesses

  • 5 Citations

Abstract

Aims

The aims of this double-blind, controlled, crossover study were to assess the influence of food preservatives on in situ dental biofilm growth and vitality, and to evaluate their influence on the ability of dental biofilm to demineralize underlying enamel over a period of 14 days.

Materials and methods

Twenty volunteers wore appliances with six specimens each of bovine enamel to build up intra-oral biofilms. During four test cycles of 14 days, the subjects had to place the appliance in one of the assigned controls or active solutions twice a day for a minute: negative control 0.9 % saline, 0.1 % benzoate (BA), 0.1 % sorbate (SA) and 0.2 % chlorhexidine (CHX positive control). After 14 days, the biofilms on two of the slabs were stained to visualize vital and dead bacteria to assess biofilm thickness (BT) and bacterial vitality (BV). Further, slabs were taken to determine mineral loss (ML), by quantitative light-induced laser fluorescence (QLF) and transversal microradiography (TMR), moreover the lesion depths (LD).

Results

Nineteen subjects completed all test cycles. Use of SA, BA and CHX resulted in a significantly reduced BV compared to NaCl (p < 0.001). Only CHX exerted a statistically significant retardation in BT as compared to saline. Differences between SA and BA were not significant (p > 0.05) for both parameters. TMR analysis revealed the highest LD values in the NaCl group (43.6 ± 44.2 μm) and the lowest with CHX (11.7 ± 39.4 μm), while SA (22.9 ± 45.2 μm) and BA (21.4 ± 38.5 μm) lay in between. Similarly for ML, the highest mean values of 128.1 ± 207.3 vol% μm were assessed for NaCl, the lowest for CHX (−16.8 ± 284.2 vol% μm), while SA and BA led to values of 83.2 ± 150.9 and 98.4 ± 191.2 vol% μm, respectively. With QLF for both controls, NaCl (−33.8 ± 101.3 mm2 %) and CHX (−16.9 ± 69.9 mm2 %), negative values were recorded reflecting a diminution of fluorescence, while positive values were found with SA (33.9 ± 158.2 mm2 %) and BA (24.8 ± 118.0 mm2 %) depicting a fluorescence gain. These differences were non-significant (p > 0.05).

Conclusion

The biofilm model permited the assessment of undisturbed oral biofilm formation influenced by antibacterial components under clinical conditions for a period of 14 days. An effect of BA and SA on the demineralization of enamel could be demonstrated by TMR and QLF, but these new findings have to be seen as a trend. As part of our daily diet, these preservatives exert an impact on the metabolism of the dental biofilm, and therefore may even influence demineralization processes of the underlying dental enamel in situ.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3

References

  1. 1.

    Wimpenny J, Manz W, Szewzyk U (2000) Heterogeneity in biofilms. FEMS Microbiol Rev 24:661–671

  2. 2.

    Costerton JW, Cook G, Lamont R (1999) The community architecture of biofilms. In: Newman HN, Wilson M (eds) Dental plaque revisited. BioLine, Antony Rowe Ltd., Chippenham, pp 5–14

  3. 3.

    ten Cate JM (2006) Biofilms, a new approach to the microbiology of dental plaque. Odontology 94:1–9

  4. 4.

    Paster BJ, Olsen I, Aas JA (2000) Dewhirst FE (2006) The breadth of bacterial diversity in the human periodontal pocket and other oral sites. Periodontol 42:80–87

  5. 5.

    Zaura E, Keijser BJ, Huse SM, Crielaard W (2009) Defining the healthy “core microbiome” of oral microbial communities. BMC Microbiol 9:259

  6. 6.

    Donlan RM, Costerton JW (2002) Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15:167–193

  7. 7.

    Obst U, Schwartz T, Volkmann H (2006) Antibiotic resistant pathogenic bacteria and their resistance genes in bacterial biofilms. Int J Artif Organs 29:387–394

  8. 8.

    del Pozo JL, Patel R (2007) The challenge of treating biofilm-associated bacterial infections. Clin Pharmacol Ther 82:204–209

  9. 9.

    Brecx M, Netuschil L, Hoffmann T (2003) How to select the right mouthrinses in periodontal prevention and therapy. Int J Dent Hygiene 1:188–194

  10. 10.

    Arweiler NB, Pergola G, Kuenz J, Hellwig E, Sculean A, Auschill TM (2011) Clinical and antibacterial effect of an anti-inflammatory toothpaste formulation with Scutellaria baicalensis extract on experimental gingivitis. Clin Oral Invest 15:909–913

  11. 11.

    De Liefde B (1998) The decline of caries in New Zealand over the past 40 years. N Z Dent J 94:109–113

  12. 12.

    Leikanger S, Bjertness E, Scheie AA, Eriksen HM, Von Der Fehr FR (1990) Preservatives and dental caries. Can preservatives in food and beverages influence oral health? Tidsskr Nor Laegeforen 110:3378–3382

  13. 13.

    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 100:371–376

  14. 14.

    Eklund T (1980) Inhibition of growth and uptake processes in bacteria by some chemical food preservatives. J Appl Bacteriol 48:423–432

  15. 15.

    Phan TN, Nguyen PT, Abranches J, Marquis RE (2002) Fluoride and organic weak acids as respiration inhibitors for oral streptococci in acidified environments. Oral Microbiol Immunol 17:119–124

  16. 16.

    Suhr KI, Nielsen PV (2004) Effect of weak acids preservatives on growth of bakery product spoilage fungi at different water activities and pH values. Int J Food Microbiol 95:67–78

  17. 17.

    Piper P, Calderon CO, Hatzixanthis K, Mollapour M (2001) Weak acid adaptation: the stress response that convers yeasts with resistance to organic acid food preservatives. Microbiology 147:2635–2442

  18. 18.

    Krebs HA, Wiggins D, Stubbs M, Sols A, Bedoya F (1983) Studies on the mechanism of the antifungal action of benzoate. Biochem J 214:657–663

  19. 19.

    Arweiler NB, Lenz R, Sculean A, Al-Ahmad A, Hellwig E, Auschill TM (2008) Effect of food preservatives on in situ biofilm formation. Clin Oral Invest 12:203–208

  20. 20.

    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 52:335–345

  21. 21.

    Lerouge S, Wertheimer MR, Yahia LH (2001) Plasma sterilisation: a review of parameters, mechanisms and limitations. Plasmas Polymers 6:175–188

  22. 22.

    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 56:681–687

  23. 23.

    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 43:277–285

  24. 24.

    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 46:471–476

  25. 25.

    Kühnisch J, Heinrich-Weltzien R (2004) Quantitative light-induced fluorescence (QLF)—a literature review. Quantitative licht-induzierte Fluoreszenz (QLF)—ein Literatur-überblick. Internat J Comput Dent 7:325–338

  26. 26.

    Gmür R, Giertsen E, van der Veen MH, Josselin D, de Jong E, Ten Cate JM, Guggenheim B (2006) In vitro quantitative light-induced fluorescence to measure changes in enamel mineralisation. Clin Oral Invest 10:187–195

  27. 27.

    Buchalla W, Attin T, Schulte-Mönting J, Hellwig E (2002) Fluoride uptake, retention, and remineralization efficacy of a hihgly concentrated fluoride solution on enamel lesions in situ. J Dent Res 81:329–333

  28. 28.

    Hellwig E, Altenburger M, Attin T, Lussi A, Buchalla W (2010) Remineralization of initial carious lesions in deciduous enamel after application of dentifrices of different fluoride concentrations. Clin Oral Invest 14:265–269

  29. 29.

    Lennon AM, Wiegand A, Buchalla W, Attin T (2007) Approximal caries development in surfaces in contact with fluoride-releasing and non-fluoride-releasing restorative matrerials: an in situ study. Eur J Oral Sci 115:497–501

  30. 30.

    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 31:194–200

  31. 31.

    Zaura-Arite E, van Marle J, ten Cate JM (2001) Confocal microscopy study of undisturbed and chlorhexidine-treated dental biofilm. J Dent Res 80:1436–1440

  32. 32.

    Thurnheer T, Gmür R, Shapiro S, Guggenheim B (2003) Mass transport of macromolecules within an in vitro model of supragingival plaque. Appl Environ Microbiol 69:1702–1709

  33. 33.

    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 38:442–447

  34. 34.

    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 8:97–101

  35. 35.

    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 32:147–152

  36. 36.

    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 108:154–161

  37. 37.

    Lamont RJ, Jenkinson HF (1998) Life below gum line: pathogenic mechanisms of Porphyromonas gingivalis. Microbiol Mol Biol Rev 62:1244–1263

  38. 38.

    Weiger R, Netuschil L, Brecx M (1992) Relationship between bacterial counts, microbial vitality and the accumulation of supragingival dental plaque in humans. J Periodontal Res 27:575–580

  39. 39.

    Mikkelsen L (1993) Influence of sucrose intake on saliva and number of microorganisms and acidogenic potential in early dental plaque. Microb Ecol Health Dis 6:253–264

  40. 40.

    Davies A (1973) The mode of action of chlorhexidine. J Periodont Res 8(Suppl 12):68–75

  41. 41.

    Brecx M, Theilade J (1984) Effect of chlorhexidine rinses on the morphology of early dental plaque formed on plastic film. J Clin Periodontol 11:553–564

  42. 42.

    Altenburger MJ, Klasser M, Schirrmeister JF, Hellwig E (2006) Remineralisation of carious enamel lesions after application of a CHX/F-mouthrinse compared with sole CHX- and placebo-application. Oral Health Prev Dent 4:255–263

  43. 43.

    Marquis RE, Clock SA, Mota-Meira M (2003) Fluoride and organic weak acids as modulators of microbial physiology. FEMS Microbiol Rev 26:493–510

  44. 44.

    Phan TN, Nguyen PT, Abranches J, Marquis RE (2002) Fluoride and organic weak acids as inhibitors for oral streptococci in acidified environments. Oral Microbiol Immunol 17:119–124

  45. 45.

    Belli WA, Buckley DH, Marquis RE (1995) Weak acid effects and fluoride inhibition of glycolysis by Streptococcus mutans GS-5. Can J Microbiol 41:785–791

  46. 46.

    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 35:331–337

  47. 47.

    Hafström-Björkman U, Sundström F, de Josselin de Jong E, Oliveby A, Angmar-Mansson B (1992) Comparison of laser fluorescence and longitudinal microradiography for quantitative assessment of in vitro enamel caries. Caries Res 26:241–247

  48. 48.

    Van der Veen MH, de Josselin de Jong E (2000) Application of quantitative light-induced fluorescence for assessing early caries lesions. Monogr Oral Sci 17:144–162

  49. 49.

    Brecx M, Netuschil L, Reichert B, Schreil G (1990) Efficacy of Listerine, Meridol and Chlorhexidine mouthrinses on plaque, gingivitis and plaque bacteria vitality. J Clin Periodontol 17:292–297

  50. 50.

    Emilson CG (1994) Potential efficacy of chlorhexidine against mutans streptococci and human dental caries. J Dent Res 73:682–691

  51. 51.

    Taylor AM, Ellwood RP, Pretty IA, Mohan N (2009) Quantitative stain detection in vivo using fluorescent imaging. J Dent 37:397–405

  52. 52.

    Sheen SA, Owens J, Addy M (2001) The effect of toothpastes on the propensity of chlorhexidine and cetyl pyridinium chloride to produce staining in vitro: a possible predictor of inactivation. J Clin Periodontol 28:46–51

  53. 53.

    Brecx M, Netuschil L, Hoffmann T (2003) How to select the right mouthrinses in periodontal prevention and therapy. Int J Dent Hyg 1:188–194

  54. 54.

    Buchalla W, Lennon AM, Attin T (2004) Fluorescence spectroscopy of dental calculus. J Periodontal Res 39:327–332

  55. 55.

    Buchalla W (2005) Comparative fluorescence spectroscopy shows differences in noncavitated enamel lesions. Caries Res 39:150–156

  56. 56.

    Netuschil L, Weiger R, Preisler R, Brecx M (1995) Plaque bacteria counts and vitality during chlorhexidine, Meridol and Listerine mouthrinses. Eur J Oral Sci 103:355–361

  57. 57.

    Arweiler NB, Reich E, Brecx M, Dörner M, Netuschil L (2001) Influence of ethanol on the antibacterial and anti-plaque efficacy of an amine fluoride/stannous fluoride mouthrinse solution. J de Parodontologie d’Implantologie orale 20:331–340

Download references

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, Albert Ludwigs University, Freiburg, Germany for her help in the image analysis. Special thanks also to Kristina Schmidt, MPH, RDH, praxisHochschule für Gesundheit und Soziales, Cologne, Germany, for helping to prepare the final version of the manuscript.

Conflicts of interests

The authors declare that they have no conflicts of interest.

Author information

Correspondence to Nicole Birgit Arweiler.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Arweiler, N.B., Netuschil, L., Beier, D. et al. Action of food preservatives on 14-days dental biofilm formation, biofilm vitality and biofilm-derived enamel demineralisation in situ. Clin Oral Invest 18, 829–838 (2014). https://doi.org/10.1007/s00784-013-1053-9

Download citation

Keywords

  • Dental biofilm
  • Biofilm in situ model
  • Food preservatives
  • Plaque vitality
  • Confocal laser scanning microscopy
  • Enamel lesions
  • TMR
  • Mineral content
  • QLF
  • Lesion depth