title: In vitro reconstruction of human junctional and sulcular epithelium.
abstract: BACKGROUND
  The aim of this study was to develop and characterize standardized in vitro three-dimensional organotypic models of human junctional epithelium (JE) and sulcular epithelium (SE).
  
  
  METHODS
  Organotypic models were constructed by growing human normal gingival keratinocytes on top of collagen matrices populated with gingival fibroblasts (GF) or periodontal ligament fibroblasts (PLF). Tissues obtained were harvested at different time points and assessed for epithelial morphology, proliferation (Ki67), expression of JE-specific markers (ODAM and FDC-SP), cytokeratins (CK), transglutaminase, filaggrin, and basement membrane proteins (collagen IV and laminin1).
  
  
  RESULTS
  The epithelial component in 3- and 5-day organotypics showed limited differentiation and expressed Ki-67, ODAM, FDC-SP, CK 8, 13, 16, 19, and transglutaminase in a similar fashion to control JE samples. PLF supported better than GF expression of CK19 and suprabasal proliferation, although statistically significant only at day 5. Basement membrane proteins started to be deposited only from day 5. The rate of proliferating cells as well as the percentage of CK19-expressing cells decreased significantly in 7- and 9-day cultures. Day 7 organotypics presented higher number of epithelial cell layers, proliferating cells in suprabasal layers, and CK expression pattern similar to SE.
  
  
  CONCLUSIONS
  Both time in culture and fibroblast type had impact on epithelial phenotype. Five-day cultures with PLF are suggested as JE models, 7-day cultures with PLF or GF as SE models, while 9-day cultures with GF as gingival epithelium (GE) models. Such standard, reproducible models represent useful tools to study periodontal bacteria-host interactions in vitro.
authors.affiliation: The Gade Institute, Faculty of Medicine and Dentistry, University of Bergen, Bergen, Norway.
authors.forename: G
authors.initials: G
authors.lastname: Dabija-Wolter
authors.forename: V
authors.initials: V
authors.lastname: Bakken
authors.forename: M R
authors.initials: MR
authors.lastname: Cimpan
authors.forename: A C
authors.initials: AC
authors.lastname: Johannessen
authors.forename: D E
authors.initials: DE
authors.lastname: Costea
mesh_terms: D000682:Amyloid; D001485:Basement Membrane / Q000033:anatomy & histology; D015415:Biomarkers / Q000032:analysis; D002352:Carrier Proteins / Q000032:analysis; D002452:Cell Count; D049109:Cell Proliferation; D048430:Cell Shape; D018920:Coculture Techniques; D003094:Collagen; D024141:Collagen Type IV / Q000032:analysis; D004846:Epithelial Attachment / Q000033:anatomy & histology* / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology; D004848:Epithelium / Q000033:anatomy & histology; D005347:Fibroblasts / Q000502:physiology; D000091344:Filaggrin Proteins; D005881:Gingiva / Q000033:anatomy & histology* / Q000166:cytology; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis; D047908:Intracellular Signaling Peptides and Proteins; D053535:Keratin-13 / Q000032:analysis; D053536:Keratin-16 / Q000032:analysis; D053539:Keratin-19 / Q000032:analysis; D053533:Keratin-8 / Q000032:analysis; D015603:Keratinocytes / Q000502:physiology; D019394:Ki-67 Antigen / Q000032:analysis; D007797:Laminin / Q000032:analysis; D009363:Neoplasm Proteins; D010513:Periodontal Ligament / Q000166:cytology; D011506:Proteins / Q000032:analysis; D013997:Time Factors; D046509:Tissue Culture Techniques; D011503:Transglutaminases / Q000032:analysis
chemical_list: D000682:Amyloid; D015415:Biomarkers; D002352:Carrier Proteins; D024141:Collagen Type IV; C465580:FDCSP protein, human; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D047908:Intracellular Signaling Peptides and Proteins; D053535:Keratin-13; D053536:Keratin-16; D053539:Keratin-19; D053533:Keratin-8; D019394:Ki-67 Antigen; D007797:Laminin; D009363:Neoplasm Proteins; C480049:ODAM protein, human; D011506:Proteins; C121346:laminin 1; D003094:Collagen; D011503:Transglutaminases