Background: The aims of the present study were 1) to localize human neutrophil defensins (HNP) 1-3 in gingiva and in neutrophil extract-treated epithelial cell monolayers, 2) to determine the effects of HNP-1 on the epithelial cell viability, attachment, and spreading, and 3) to analyze the effect of HNP-1 on the bacterial adherence to epithelial cells. Methods: For localization of HNP 1-3 in gingival tissue and in pre-incubated cell monolayers, immunohistochemical and immunocytochemical methods were used. Viability and proliferation of epithelial cells were determined with commercial kits after incubating the keratinocytes with different HNP-1 concentrations (low: 1-5 μg/ml, moderate: 10 μg/ml, high: 20-50 μg/ml). Attachment and spreading of keratinocytes on fibronectin-coated surfaces in the presence of HNP-1 were evaluated under microscope. Attachment of Fusobacterium nucleatum ATCC 25586 and Prevotella intermedia ATCC 25611 on keratinocytes pre-incubated with HNP-1 were determined with a standard antibiotic test. Results: HNP 1-3 localized in the junctional epithelium of clinically healthy gingiva and in the pocket epithelium of gingiva with periodontitis. When keratinocyte monolayers were incubated with neutrophil extracts, HNP 1-3 were bound to the periphery of the growing cell colonies. In low HNP-1 concentrations, the keratinocyte proliferation enhanced. Moderate and high concentrations of HNP-1 increased the cellular death significantly. HNP-1 increased the attachment and spreading of keratinocytes on fibronectin-coated surfaces, and bacterial attachment to keratinocytes, in a concentration-dependent manner. Conclusions: HNP-1 plays a role in the integrity of keratinocytes by stimulating their proliferation, attachment, and spreading, while higher doses increase the bacterial attachment and keratinocyte death.
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Gursoy, U., Könönen, E., Luukkonen, N., & Uitto, V-J. (2013). Human Neutrophil Defensins and Their Effect on Epithelial Cells. Journal of Periodontology, 84(1), 126-133. https://doi.org/10.1902/jop.2012.120017