Institute of Bioengineering, School of Engineering and Materials Science, Queen Mary University of London, London, UK.
Department of Cell and Developmental Biology, University College London, London, UK.
Prenat Diagn. 2017 Sep;37(9):899-906. doi: 10.1002/pd.5104. Epub 2017 Aug 1.
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.
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.
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.
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.
我们建立了一个体外模型，以研究创伤是否会引起胎儿膜（FM）缺陷羊膜（AM）中的连接蛋白 43（Cx43）表达和胶原组织。
体外创伤足月人类 FM。通过免疫荧光共聚焦显微镜（IMF）检查伤口边缘 AM 中的细胞形态和 Cx43，并与对照 AM 进行比较。通过二次谐波产生（SHG）成像检查胶原微观结构。用钙黄绿素和 ethidium 染色评估细胞活力。
创伤后，AM 显示出细胞密集区，这些细胞已向伤口边缘迁移。在伤口边缘 AM 中，与上皮细胞相比，Cx43 斑点优先分布在间充质细胞中，而在成纤维细胞层中的表达明显高于上皮细胞层（p<0.001）。在成纤维细胞层中，胶原纤维高度极化并与伤口边缘 AM 的轴平行排列。在没有愈合的情况下，细胞在 168 小时后不能穿过缺陷迁移。创伤后 FM 的细胞活力在培养过程中得以维持。
受伤 AM 中的 Cx43 过表达会导致胶原结构发生变化，从而减缓细胞穿过 FM 缺陷的迁移效率。©2017 作者。产前诊断由 John Wiley & Sons，Ltd. 出版。
