title: Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit.
abstract: Epithelial-mesenchymal communication plays a key role in tissue homeostasis and abnormal signaling contributes to chronic airways disease such as COPD. Most in vitro models are limited in complexity and poorly represent this epithelial-mesenchymal trophic unit. We postulated that cellular outgrowth from bronchial tissue would enable development of a mucosal structure that recapitulates better in vivo tissue architecture.
  
  
  
  Bronchial tissue was embedded in Matrigel and outgrowth cultures monitored using time-lapse microscopy, electrical resistance, light and electron microscopy. Cultures were challenged repetitively with cigarette smoke extract (CSE).
  
  
  
  The outgrowths formed as a multicellular sheet with motile cilia becoming evident as the Matrigel was remodeled to provide an air interface; cultures were viable for more than one year. Immunofluorescence and electron microscopy (EM) identified an upper layer of mucociliary epithelium and a lower layer of highly organized extracellular matrix (ECM) interspersed with fibroblastic cells separated by a basement membrane. EM analysis of the mucosal construct after repetitive exposure to CSE revealed epithelial damage, loss of cilia, and ECM remodeling, as occurs in vivo.
  
  
  
  We have developed a robust bronchial mucosal model. The structural changes observed following CSE exposure suggest the model should have utility for drug discovery and preclinical testing, especially those targeting airway remodeling.
authors.affiliation: a Academic Unit of Clinical and Experimental Sciences , University of Southampton Faculty of Medicine, University Hospital Southampton , Southampton , United Kingdom.|b Dipartimento BIONEC , University of Palermo , Palermo , Italy.|c Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST) , Palermo , Italy.|d Institute of Biomedicine and Molecular Immunology (IBIM), Italian National Research Council (CNR) , Palermo , Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Bucchieri
authors.affiliation: b Dipartimento BIONEC , University of Palermo , Palermo , Italy.|c Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST) , Palermo , Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Pitruzzella
authors.affiliation: b Dipartimento BIONEC , University of Palermo , Palermo , Italy.|c Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST) , Palermo , Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Fucarino
authors.affiliation: b Dipartimento BIONEC , University of Palermo , Palermo , Italy.|c Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST) , Palermo , Italy.
authors.forename: Antonella Marino
authors.initials: AM
authors.lastname: Gammazza
authors.affiliation: b Dipartimento BIONEC , University of Palermo , Palermo , Italy.|c Istituto Euro-Mediterraneo di Scienza e Tecnologia (IEMEST) , Palermo , Italy.
authors.forename: Celeste Caruso
authors.initials: CC
authors.lastname: Bavisotto
authors.affiliation: b Dipartimento BIONEC , University of Palermo , Palermo , Italy.
authors.forename: Vito
authors.initials: V
authors.lastname: Marcianò
authors.affiliation: e Dipartimento di Discipline Chirurgiche, Oncologiche e Stomatologiche , University of Palermo , Palermo , Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Cajozzo
authors.affiliation: e Dipartimento di Discipline Chirurgiche, Oncologiche e Stomatologiche , University of Palermo , Palermo , Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Lo Iacono
authors.affiliation: f Interventional Pulmonology Unit , La Maddalena Cancer Center , Palermo , Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Marchese
authors.affiliation: b Dipartimento BIONEC , University of Palermo , Palermo , Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Zummo
authors.affiliation: a Academic Unit of Clinical and Experimental Sciences , University of Southampton Faculty of Medicine, University Hospital Southampton , Southampton , United Kingdom.|g Southampton NIHR Respiratory Biomedical Research Unit, Sir Henry Wellcome Laboratories , University of Southampton School of Medicine, University Hospital Southampton , Southampton , United Kingdom.
authors.forename: Stephen T
authors.initials: ST
authors.lastname: Holgate
authors.affiliation: a Academic Unit of Clinical and Experimental Sciences , University of Southampton Faculty of Medicine, University Hospital Southampton , Southampton , United Kingdom.|g Southampton NIHR Respiratory Biomedical Research Unit, Sir Henry Wellcome Laboratories , University of Southampton School of Medicine, University Hospital Southampton , Southampton , United Kingdom.
authors.forename: Donna E
authors.initials: DE
authors.lastname: Davies
mesh_terms: D001980:Bronchi / Q000166:cytology / Q000254:growth & development; D002478:Cells, Cultured; D003094:Collagen; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology; D006801:Humans; D007797:Laminin; D059630:Mesenchymal Stem Cells / Q000166:cytology; D008853:Microscopy; D008954:Models, Biological; D011509:Proteoglycans; D020545:Respiratory Mucosa / Q000166:cytology / Q000254:growth & development; D012906:Smoke / Q000009:adverse effects*
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D012906:Smoke; C059485:matrigel; D003094:Collagen
keywords: ECM; bronchial; cigarette smoke; electron microscopy; epithelium; fibroblast; in vitro model