title: Acute effects of cigarette smoke on three-dimensional cultures of normal human oral mucosa.
abstract: BACKGROUND
  Human oral mucosa is the combustion chamber of cigarette, but scanty evidence is available about the early smoke effects.
  
  
  OBJECTIVE
  The present work aimed at evaluating from a morphological point of view whole smoke early effects on epithelial intercellular adhesion and keratinocyte terminal differentiation in a three-dimensional model of human oral mucosa.
  
  
  METHODS
  Biopsies of keratinized oral mucosa of healthy nonsmoking women (n = 5) were collected. After culturing in a Transwell system, one fragment of each biopsy was exposed to the smoke of one single cigarette; the remnant represented the internal control. The distribution of epithelial differentiation markers (keratin-10, K10, and keratin-14, K14, for suprabasal and basal cells respectively), desmosomes (desmoglein-1, desmoglein-3), tight junctions (occludin), adherens junctions (E-cadherin, β-catenin), and apoptotic cells (p53, caspase 3) were evaluated by immunofluorescence.
  
  
  RESULTS
  Quantitative analysis of K14 immunolabeling revealed an overexpression in the suprabasal layers as early as 3 h after smoke exposure, without impairment of the epithelial junctional apparatus and apoptosis induction.
  
  
  CONCLUSIONS
  These results suggested that the first significant response to cigarette smoke came from the basal and suprabasal layers of the human oral epithelium. The considered model maintained the three-dimensional arrangement of the human mucosa in the oral cavity and mimicked the inhalation/exhalation cycle during the exposure to cigarette smoke, offering a good possibility to extrapolate the reported observations to humans.
authors.affiliation: Dipartimento di Morfologia Umana e Scienze Biomediche - Città Studi, Italy.
authors.forename: Alice
authors.initials: A
authors.lastname: Gualerzi
authors.forename: Michele
authors.initials: M
authors.lastname: Sciarabba
authors.forename: Gianluca
authors.initials: G
authors.lastname: Tartaglia
authors.forename: Chiarella
authors.initials: C
authors.lastname: Sforza
authors.forename: Elena
authors.initials: E
authors.lastname: Donetti
mesh_terms: D000328:Adult; D015820:Cadherins / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D051183:Desmoglein 1 / Q000378:metabolism; D051184:Desmoglein 3 / Q000378:metabolism; D005260:Female; D006801:Humans; D053550:Keratin-10 / Q000378:metabolism; D053547:Keratin-14 / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D009061:Mouth Mucosa / Q000187:drug effects* / Q000378:metabolism; D062793:Occludin; D012906:Smoke / Q000009:adverse effects*; D012907:Smoking / Q000009:adverse effects*; D046509:Tissue Culture Techniques; D014026:Nicotiana; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
chemical_list: D015820:Cadherins; C495297:DSG1 protein, human; C071894:DSG3 protein, human; D051183:Desmoglein 1; D051184:Desmoglein 3; C507493:KRT10 protein, human; C507540:KRT14 protein, human; D053547:Keratin-14; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D012906:Smoke; D016159:Tumor Suppressor Protein p53; D051176:beta Catenin; D053550:Keratin-10; D053148:Caspase 3