Abstract

BackgroundMicrobial biofilms are known to cause an increasing number of chronic inflammatory and infectious conditions. A classical example is chronic periodontal disease, a condition initiated by the subgingival dental plaque biofilm on gingival epithelial tissues. We describe here a new model that permits the examination of interactions between the bacterial biofilm and host cells in general. We use primary human gingival epithelial cells (HGEC) and an in vitro grown biofilm, comprising nine frequently studied and representative subgingival plaque bacteria.ResultsWe describe the growth of a mature 'subgingival' in vitro biofilm, its composition during development, its ability to adapt to aerobic conditions and how we expose in vitro a HGEC monolayer to this biofilm. Challenging the host derived HGEC with the biofilm invoked apoptosis in the epithelial cells, triggered release of pro-inflammatory cytokines and in parallel induced rapid degradation of the cytokines by biofilm-generated enzymes.ConclusionWe developed an experimental in vitro model to study processes taking place in the gingival crevice during the initiation of inflammation. The new model takes into account that the microbial challenge derives from a biofilm community and not from planktonically cultured bacterial strains. It will facilitate easily the introduction of additional host cells such as neutrophils for future biofilm:host cell challenge studies. Our methodology may generate particular interest, as it should be widely applicable to other biofilm-related chronic inflammatory diseases.

Highlights

  • Microbial biofilms are known to cause an increasing number of chronic inflammatory and infectious conditions

  • V. dispar, A. naeslundii, and S. oralis were in 1-10% range, whereas the other five organisms were all below 0.1% of the total CFU

  • confocal laser scanning microscopy (CLSM) images of a 64.5 h 9-species biofilm stained by multiplex fluorescent in situ hybridization (FISH) for (A) V. dispar, C. rectus, F. nucleatum, and P. intermedia, (B) V. dispar, A. naeslundii, S. intermedius, and S. oralis, and (C) V. dispar, T. forsythia, P. gingivalis, and C. rectus

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Summary

Introduction

Microbial biofilms are known to cause an increasing number of chronic inflammatory and infectious conditions. A classical example is chronic periodontal disease, a condition initiated by the subgingival dental plaque biofilm on gingival epithelial tissues. We use primary human gingival epithelial cells (HGEC) and an in vitro grown biofilm, comprising nine frequently studied and representative subgingival plaque bacteria. In most natural environments bacteria exist as highly structured dense surface attached aggregates designated as biofilms [1,2]. This applies to bacteria colonizing the skin and human mucosa. It induces inflammation in the adjacent host tissues and the biofilm over time extends under the gum, down the root surface, creating a niche favoring the growth of fastidious anaerobes, such as Spirochaetes and Bacteriodetes [5]

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