Functional characterization of a novel 3D model of the epithelial-mesenchymal trophic unit. (7th February 2017)

• Record Type: Journal Article
• Authors: Bucchieri, Fabio; Pitruzzella, Alessandro; Fucarino, Alberto; Gammazza, Antonella Marino; Bavisotto, Celeste Caruso; Marcianò, Vito; Cajozzo, Massimo; Lo Iacono, Giorgio; Marchese, Roberto; Zummo, Giovanni; Holgate, Stephen T.; Davies, Donna E.
• Is Part Of: Experimental lung research. Volume 43:Number 2(2017)
• Page Start: 82
• Page End: 92
• Publication Date: 2017-02-07
• Subjects: in vitro model -- epithelium -- fibroblast -- bronchial -- cigarette smoke -- ECM -- electron microscopy; Lungs -- Periodicals; Lungs -- Diseases -- Periodicals; Lung Diseases; Lung -- physiology; Respiratory System; 616.24
• DOI: 10.1080/01902148.2017.1303098
• Languages: English
• ISSNs: 0190-2148

Abstract

ABSTRACT: Background/Aim : 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.Materials and Methods : 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).Results : 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 .Conclusions : 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.