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Trauma induces overexpression of Cx43 in human fetal membrane defects
Author(s) -
Barrett David W.,
Kethees Aumie,
Thrasivoulou Christopher,
Mata Alvaro,
Virasami Alex,
Sebire Neil J.,
Engels Alex C.,
Deprest Jan A.,
Becker David L.,
David Anna L.,
Chowdhury Tina T.
Publication year - 2017
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/pd.5104
Subject(s) - fibroblast , microbiology and biotechnology , calcein , viability assay , wound healing , cell migration , immunofluorescence , cell , confocal microscopy , in vitro , mesenchymal stem cell , cell culture , pathology , andrology , chemistry , biology , immunology , medicine , membrane , biochemistry , genetics , antibody
Abstract Objective We developed an in vitro model to examine whether trauma induces connexin 43 (Cx43) expression and collagen organisation in the amniotic membrane (AM) of fetal membrane (FM) defects. Method Term human FM was traumatised in vitro . Cell morphology and Cx43 were examined in the wound edge AM by immunofluorescence (IMF) confocal microscopy and compared to control AM. Collagen microstructure was examined by second harmonic generation (SHG) imaging. Cell viability was assessed with calcein and ethidium staining. Results After trauma, the AM showed a dense region of cells, which had migrated towards the wound edge. In wound edge AM, Cx43 puncta was preferentially distributed in mesenchymal cells compared to epithelial cells with significant expression in the fibroblast layer than epithelial layer ( p < 0.001). In the fibroblast layer, the collagen fibres were highly polarised and aligned in parallel to the axis of the wound edge AM. There was an absence of cell migration across the defect with no healing after 168 h. Cell viability of the FM after trauma was maintained during culture. Conclusion Cx43 overexpression in wounded AM drives structural changes in collagen that slows down efficacy of cell migration across the FM defect. © 2017 The Authors. Prenatal Diagnosis published by John Wiley & Sons, Ltd.