Costa E, Thrasivoulou C, Becker D L, Deprest J, David A L, Chowdhury T T
Centre for 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. 2025 Jan;45(1):102-112. doi: 10.1002/pd.6722. Epub 2024 Dec 4.
We examined the role of myofibroblasts in regulating Cx43 and collagen structure in iatrogenic preterm amniotic membrane (AM) defects subjected to mechanical stimulation.
Preterm AM specimens were collected from women undergoing planned preterm caesarean section after in utero intervention for correction of spina bifida by open fetal surgery (n = 4 patients; preterm delivery at 34 + 0 weeks to 35 + 0 weeks). Control specimens taken 5 cm away from the open fetal surgery defect site were compared with wound edge AM. In separate experiments, the effects of mechanical stimulation and co-treatment with Cx43 antisense on matrix and repair proteins were examined. Specimens were immunostained to detect αSMA and Cx43 in myofibroblasts and counterstained with DAPI to quantify nuclei shape. The direction of collagen fibrils in the wound edge region was examined by SHG imaging. Markers for matrix (collagen, elastin, GAG), inflammation (PGE) and repair (TGFβ) were examined by RT-qPCR and biochemical assays.
In iatrogenic preterm AM specimens, the diameter of the open fetal surgery defect ranged between 3.5 and 7.5 cm. At the wound edge of the open fetal surgery defect, αSMA positive myofibroblasts had deformed nuclei and showed abundant Cx43 localized in the cell bodies or formed plaques. In the fibroblast layer, collagen had degenerated in some regions or had polarity near the wound edge. In preterm AM defects, mechanical stimulation and Cx43 antisense increased the levels of collagen and elastin but not GAG or PGE release. Mechanical stimulation increased Cx43 and TGFβ gene expression.
In open fetal surgery defects, myofibroblasts were elongated with collagen fibrils that either degenerated or had polarity. Whilst cells produced substantially higher Cx43 in the fibroblast than in the epithelial layer, they formed plaques, which may prevent migration and delay healing. Mechanical stimulation of preterm AM enhanced matrix repair proteins and the mechanotransduction should be explored to understand how Cx43 contributes to membrane integrity.
我们研究了成肌纤维细胞在机械刺激下对医源性早产羊膜（AM）缺损中Cx43和胶原蛋白结构的调节作用。
收集因开放性胎儿手术进行宫内干预以纠正脊柱裂后行计划性早产剖宫产的女性的早产AM标本（n = 4例患者；34 + 0周至35 + 0周早产）。将取自距开放性胎儿手术缺损部位5 cm处的对照标本与伤口边缘AM进行比较。在单独的实验中，研究了机械刺激和Cx43反义寡核苷酸联合处理对基质和修复蛋白的影响。标本进行免疫染色以检测成肌纤维细胞中的αSMA和Cx43，并用DAPI复染以量化细胞核形状。通过二次谐波产生（SHG）成像检查伤口边缘区域胶原纤维的方向。通过逆转录定量聚合酶链反应（RT-qPCR）和生化分析检测基质（胶原蛋白、弹性蛋白、糖胺聚糖）、炎症（前列腺素E）和修复（转化生长因子β）的标志物。
在医源性早产AM标本中，开放性胎儿手术缺损的直径在3.5至7.5 cm之间。在开放性胎儿手术缺损的伤口边缘，αSMA阳性成肌纤维细胞核变形，细胞体中Cx43丰富或形成斑块。在成纤维细胞层，胶原蛋白在某些区域退化或在伤口边缘附近具有极性。在早产AM缺损中，机械刺激和Cx43反义寡核苷酸增加了胶原蛋白和弹性蛋白的水平，但未增加糖胺聚糖或前列腺素E的释放。机械刺激增加了Cx43和转化生长因子β基因的表达。
在开放性胎儿手术缺损中，成肌纤维细胞与胶原纤维一起伸长，胶原纤维要么退化要么具有极性。虽然成纤维细胞中产生的Cx43比上皮层中的高得多，但它们形成斑块，这可能会阻止迁移并延迟愈合。对早产AM的机械刺激增强了基质修复蛋白，应探索机械转导以了解Cx43如何促进膜的完整性。
