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Fusobacterium nucleatum Biofilm Induces Epithelial Migration in an Organotypic Model of Dento‐Gingival Junction
Author(s) -
Pöllänen Marja T.,
Gursoy Ulvi K.,
Könönen Eija,
Uitto VeliJukka
Publication year - 2012
Publication title -
journal of periodontology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.036
H-Index - 156
eISSN - 1943-3670
pISSN - 0022-3492
DOI - 10.1902/jop.2012.110535
Subject(s) - fusobacterium nucleatum , epithelium , connective tissue , biofilm , basement membrane , chemistry , microbiology and biotechnology , biology , porphyromonas gingivalis , pathology , periodontitis , medicine , dentistry , bacteria , genetics
Background: Effects of Fusobacterium nucleatum (F. nucleatum) biofilm on epithelial cell proliferation, apoptotic cell death, and basement membrane constituent collagen IV production were examined in an organotypic dento‐epithelial (OD‐E) model. Methods: The OD‐E model was constructed by seeding keratinocytes on fibroblast‐containing collagen gels and by placing tooth pieces on top. A 3‐day‐old biofilm either a laboratory strain (American Type Culture Collection [ATCC] 25586) or a clinical strain (Anaerobe Helsinki Negative [AHN] 9508) of F. nucleatum was placed on the top of the model. The coculture was incubated ≤24 hours. The expression and localization of Ki‐67, caspase‐3, and collagen IV were examined by immunohistochemistry. Results: Hematoxylin and eosin staining showed epithelial migration and lateral sprouting into the connective tissue matrix in F. nucleatum OD‐E cocultures. The proliferation pattern of the in vitro dento‐epithelial junction was changed. In controls without bacterial challenge, the Ki‐67 expression was abundant in the cells attached to the tooth, whereas in F. nucleatum biofilm‐treated cultures, the Ki‐67‐expressing cells were more often in the connective tissue‐facing side of the epithelium. An apoptotic marker caspase‐3 was expressed in controls and in F. nucleatum laboratory strain ATCC cocultures throughout the epithelium, in contrast to cultures treated with F. nucleatum clinical strain AHN, in which caspase‐3 was absent. Collagen IV stainings were negative in both controls and F. nucleatum cocultures. Conclusion: F. nucleatum biofilm coculture with OD‐E model causes lateral sprouting of the epithelium with an altered epithelial proliferation pattern, resembling the histologic changes seen in vivo in the early pathogenesis of periodontal disease.