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

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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).


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).


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.

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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.

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The authors declare that they have no conflicts of interest.

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Correspondence to Nicole Birgit Arweiler.

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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

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