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Medium-term culture of normal human oral mucosa: a novel three-dimensional model to study the effectiveness of drugs administration.

Abstract：

:Tissue-engineered oral mucosal equivalents have been developed for in vitro studies for a few years now. However, the usefulness of currently available models is still limited by many factors, mainly the lack of a physiological extracellular matrix (ECM) and the use of cell populations that do not reflect the properly differentiated cytotypes of the mucosa of the oral cavity. For this reason, we have developed a novel three-dimensional culture model reflecting the normal architecture of the human oral mucosa, with the main aim of creating a better in vitro model where to test cellular responses to drugs administration. This novel 3D cell culture model (3D outgrowth) was set up using an artificial extracellular matrix (Matrigel™ ), allowing the interactions required for proper differentiation of the various citotypes which form the mucosal layer. Biopsies of human oral mucosa, in fragments of about 0.5 mm3, were placed onto 6.5mm Transwells, covered with Matrigel™ and grown in a specific culture medium. A gradual formation of an architectural structure similar to that of the in vivo oral mucosa was observed. Transmission electron and confocal microscopy were employed to characterize the newly developed model: the cell components (keratinocytes and fibroblasts) differentiated properly within the outgrowth and reconstituted, in vitro, the physiological structure of the human oral mucosa, including a stratified non-keratinized squamous layer composed of four different layers, a proper basal membrane and a lamina propria where fibroblasts produce ECM. Moreover, keratinocytes expressed CK5, CK13, CK19 and E-cadherin, whereas fibroblasts expressed collagen type I and IV, laminin and fibronectin. 3D outgrowths could be considered a valid alternative to animal models, and provide useful information for researchers interested in studying the responses of the human oral mucosa to locally delivered drugs or other exogenous treatments.

journal_name

Curr Pharm Des

journal_title

Current pharmaceutical design

authors

Bucchieri F,Fucarino A,Marino Gammazza A,Pitruzzella A,Marciano V,Paderni C,De Caro V,Siragusa MG,Lo Muzio L,Holgate ST,Davies DE,Farina F,Zummo G,Kudo Y,Giannola IL,Campisi G

doi

10.2174/138161212803307482

subject

Has Abstract

pub_date

2012-01-01 00:00:00

pages

5421-30

issue

eissn

1381-6128

issn

1873-4286

pii

CPD-EPUB-20120525-7

journal_volume

pub_type

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Medium-term culture of normal human oral mucosa: a novel three-dimensional model to study the effectiveness of drugs administration.