Abstract
The gingival epithelium is a physical and immunological barrier to the microbiota of the oral cavity, which interact through soluble mediators with the immune cells that patrol the tissue at the gingival epithelium. We sought to develop a three-dimensional gingivae-biofilm interface model using a commercially available gingival epithelium to study the tissue inflammatory response to oral biofilms associated with “health”, “gingivitis” and “periodontitis”. These biofilms were developed by sequential addition of microorganisms to mimic the formation of supra- and sub-gingival plaque in vivo. Secondly, to mimic the interactions between gingival epithelium and immune cells in vivo, we integrated peripheral blood mononuclear cells and CD14+ monocytes into our three-dimensional model and were able to assess the inflammatory response in the immune cells cultured with and without gingival epithelium. We describe a differential inflammatory response in immune cells cultured with epithelial tissue, and more so following incubation with epithelium stimulated by “gingivitis-associated” biofilm. These results suggest that gingival epithelium-derived soluble mediators may control the inflammatory status of immune cells in vitro, and therefore targeting of the epithelial response may offer novel therapies. This multi-cellular interface model, both of microbial and host origin, offers a robust in vitro platform to investigate host-pathogens at the epithelial surface.
Highlights
The gingival epithelium is a physical and immunological barrier to the microbiota of the oral cavity, which interact through soluble mediators with the immune cells that patrol the tissue at the gingival epithelium
The remaining bacteria were all grown on Fastidious anaerobic agar (FAA) for 2–3 days, A. naeslundii and V. dispar were sub-cultured into Brain Heart Infusion (BHI) broth and F. nucleatum, F. nucleatum spp. vicentii, P. gingivalis and P. intermedia into Schaedler anaerobic broth (Oxoid, Cambridge, UK) for a further 1–2 days, all at 37 °C in an anaerobic chamber (85% N2, 10% CO2 and 5% H2; Don Whitley Scientific Limited, Shipley, UK)
Work from the present study has shown that the multi-species biofilms containing “health-associated”, “gingivitis-associated” and “periodontitis-associated” microorganisms exhibit unique morphological features as assessed by different microscopic techniques
Summary
The gingival epithelium is a physical and immunological barrier to the microbiota of the oral cavity, which interact through soluble mediators with the immune cells that patrol the tissue at the gingival epithelium. We sought to develop a three-dimensional gingivae-biofilm interface model using a commercially available gingival epithelium to study the tissue inflammatory response to oral biofilms associated with “health”, “gingivitis” and “periodontitis”. We describe a differential inflammatory response in immune cells cultured with epithelial tissue, and more so following incubation with epithelium stimulated by “gingivitis-associated” biofilm These results suggest that gingival epithelium-derived soluble mediators may control the inflammatory status of immune cells in vitro, and targeting of the epithelial response may offer novel therapies. This multi-cellular interface model, both of microbial and host origin, offers a robust in vitro platform to investigate host-pathogens at the epithelial surface. In the current study we sought to develop an immune cell/gingival tissue/biofilm model aiming to recapture the cell interactions at the gingival margin, encapsulating the response of the local inflammatory infiltrate
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