| Summary: | Epithelial-mesenchymal transition is reportedly important in loss of epithelial integrity and cell migration in inflammatory/infectious diseases and cancer. Since Gram negative anaerobic periodontal pathogens are well-recognized to induce intense inflammatory responses; the present study investigated their ability to induce EMT in vitro. A 2D chronic inflammatory model was developed using either the H400 oral keratinocyte cell-line or primary rat oral keratinocytes which were exposed to heat-killed Fusobacterium nucleatum, Porphyromonas gingivalis and Escherichia coli LPS for up to 8-days. EMT-associated changes were determined using semi-quantitative-RT-PCR, PCR-arrays, ELISA, scratch/transwell migration assays, immunocytochemistry/immunofluorescence, and transepithelial electrical resistance. Chronically stimulated cultures increased extracellular levels of the EMT regulatory cytokines, TGF-β1, TNF-α and EGF, whilst subsequent EMT-induction was indicated by up-regulation of mesenchymal markers, including vimentin and N-cadherin, and concomitant down-regulation of epithelial markers including E-cadherin and β-catenin. In addition, intracellular signaling activity of key EMT regulatory transcription factors, Snail-1 and NF-ĸB, increased following chronic bacterial exposure and was associated with enhanced cellular migratory activity and reduced epithelial barrier integrity. These results indicated for the first time that EMT may be involved in the compromised epithelial barrier function observed during periodontitis pathogenesis which may occur in response to prolonged local bacterial exposure.
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