title: A novel organotypic dento-epithelial culture model: effect of Fusobacterium nucleatum biofilm on B-defensin-2, -3, and LL-37 expression.
abstract: BACKGROUND
  In the present study, the expression and localization of three epithelial peptides (human β-defensin [hBD]-2 and -3, and cathelicidin [LL-37]) are studied in an organotypic dento-epithelial (OD-E) model exposed to Fusobacterium nucleatum (Fn) biofilm.
  
  
  METHODS
  Biofilm of Fn ATCC 25586 or AHN 9508 were produced by culturing each strain on semipermeable membranes. The OD-E model was constructed by seeding keratinocytes on fibroblast-containing collagen gels and by placing dentin pieces on the top. A 3-day-old biofilm was placed on the top of the OD-E and the coculture was incubated for 5 hours or 24 hours. Production of epithelial antimicrobial peptides was determined immunohistochemically.
  
  
  RESULTS
  After 5 hours of incubation, the biofilm of each Fn strain stimulated expression of hBD-2 and -3. hBD-2 was localized on superficial layers and hBD-3 on basal cell layers of the epithelium and dento-epithelial junctions, whereas LL-37 was only weakly expressed. After 24 hours, hBD-2 expression was extended toward basal cell layers of the epithelium. In contrast, hBD-3 expression extended toward superficial layers of the epithelium. In the case of Fn AHN 9508 biofilm, LL-37 was localized in the cell layers of the dento-epithelial junction.
  
  
  CONCLUSIONS
  In our OD-E model, epithelial antimicrobial peptide responses to Fn biofilms have distinct regulation and localization characteristics, resembling those known to occur in the gingival epithelium in vivo.
authors.affiliation: Department of Periodontology, Institute of Dentistry, University of Turku, Turku, Finland.
authors.forename: Ulvi K
authors.initials: UK
authors.lastname: Gursoy
authors.forename: Marja
authors.initials: M
authors.lastname: Pöllänen
authors.forename: Eija
authors.initials: E
authors.lastname: Könönen
authors.forename: Veli-Jukka
authors.initials: VJ
authors.lastname: Uitto
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis*; D018441:Biofilms / Q000254:growth & development; D018920:Coculture Techniques; D004848:Epithelium / Q000378:metabolism*; D016967:Fusobacterium nucleatum / Q000502:physiology*; D006801:Humans; D009924:Organ Culture Techniques; D014070:Tooth / Q000166:cytology; D023083:beta-Defensins / Q000096:biosynthesis*; D054804:Cathelicidins
chemical_list: D023181:Antimicrobial Cationic Peptides; C422950:DEFB103A protein, human; C106641:DEFB4A protein, human; D023083:beta-Defensins; D054804:Cathelicidins