title: Human neutrophil defensins and their effect on epithelial cells.
abstract: BACKGROUND
  The aims of the present study include: 1) to localize human neutrophil defensin-1 (HNP-1) through HNP-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 through HNP-3 in gingival tissue and in preincubated 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 to 5 μg/mL; moderate, 10 μg/mL; high, 20 to 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 ATCC25586 and Prevotella intermedia ATCC25611 on keratinocytes preincubated with HNP-1 were determined with a standard antibiotic test.
  
  
  RESULTS
  HNP-1 through HNP-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 through HNP-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, whereas higher doses increase the bacterial attachment and keratinocyte death.
authors.affiliation: Department of Periodontology, Institute of Dentistry, University of Turku, Turku, Finland. ulvi.gursoy@utu.fi
authors.forename: Ulvi K
authors.initials: UK
authors.lastname: Gursoy
authors.forename: Eija
authors.initials: E
authors.lastname: Könönen
authors.forename: Noora
authors.initials: N
authors.lastname: Luukkonen
authors.forename: Veli-Jukka
authors.initials: VJ
authors.lastname: Uitto
mesh_terms: D000328:Adult; D000890:Anti-Infective Agents / Q000032:analysis / Q000494:pharmacology*; D001422:Bacterial Adhesion / Q000187:drug effects; D001485:Basement Membrane / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D055113:Chronic Periodontitis / Q000473:pathology; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004846:Epithelial Attachment / Q000187:drug effects / Q000473:pathology; D004847:Epithelial Cells / Q000187:drug effects / Q000473:pathology; D005260:Female; D005353:Fibronectins / Q000737:chemistry; D016967:Fusobacterium nucleatum / Q000187:drug effects; D005881:Gingiva / Q000187:drug effects* / Q000473:pathology; D005888:Gingival Pocket / Q000473:pathology; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000502:physiology*; D018713:Prevotella intermedia / Q000187:drug effects; D023084:alpha-Defensins / Q000032:analysis / Q000494:pharmacology*
chemical_list: D000890:Anti-Infective Agents; D020099:Coated Materials, Biocompatible; D005353:Fibronectins; D023084:alpha-Defensins; C406413:human neutrophil peptide 1; C406416:human neutrophil peptide 2; C406427:human neutrophil peptide 3