DeNigris John, Yao Qingmei, Birk Erika K, Birk David E
a Department of Molecular Pharmacology & Physiology , Morsani College of Medicine, University of South Florida , Tampa , FL , USA.
Connect Tissue Res. 2016;57(1):1-9. doi: 10.3109/03008207.2015.1081901. Epub 2015 Dec 29.
Mutations in collagen V are associated with classic Ehlers-Danlos syndrome (EDS). A significant percentage of these mutations result in haploinsufficiency for collagen V. The purpose of this work was to determine if changes in collagen V expression are associated with altered dermal fibroblast behavior contributing to the poor wound healing response. A haploinsufficient Col5a1(+/-) mouse model of EDS was utilized. In vivo wound healing studies demonstrated that mutant mice healed significantly slower than Col5a1(+/+) mice. The basis for this difference was examined in vitro using dermal fibroblast strains isolated from Col5a1(+/-) and Col5a1(+/+) mice. Fibroblast proliferation was determined for each strain by counting cells at different time points after seeding as well as using the proliferation marker Ki-67. Fibroblast attachment to collagens I and III and fibronectin also was analyzed. In addition, in vitro scratch wounds were used to analyze fibroblast wound closure. Significantly decreased fibroblast proliferation was observed in Col5a1(+/-) compared to Col5a1(+/+) fibroblasts. Our data indicate that the decreased fibroblast number was not due to apoptosis. Wildtype Col5a1(+/+) fibroblasts attached significantly better to components of the wound matrix (collagens I and III and fibronectin) than Col5a1(+/-) fibroblasts. A significant difference in in vitro scratch wound closure rates also was observed. Col5a1(+/+) fibroblasts closed wounds in 22 h, while Col5a1(+/-) fibroblasts demonstrated ~80% closure. There were significant differences in closure at all time points analyzed. Our data suggest that decreased fibroblast proliferation, extracellular matrix attachment, and migration contribute to the decreased wound healing response in classic EDS.
胶原蛋白V的突变与典型的埃勒斯-当洛综合征（EDS）相关。这些突变中有很大一部分导致胶原蛋白V单倍剂量不足。这项研究的目的是确定胶原蛋白V表达的变化是否与真皮成纤维细胞行为改变有关，而这种改变会导致伤口愈合反应不佳。我们使用了EDS的单倍剂量不足的Col5a1(+/-)小鼠模型。体内伤口愈合研究表明，突变小鼠的愈合速度明显慢于Col5a1(+/+)小鼠。使用从Col5a1(+/-)和Col5a1(+/+)小鼠分离的真皮成纤维细胞系在体外研究了这种差异的原因。通过在接种后不同时间点计数细胞以及使用增殖标志物Ki-67来确定每个细胞系的成纤维细胞增殖情况。还分析了成纤维细胞与I型和III型胶原蛋白以及纤连蛋白的附着情况。此外，使用体外划痕伤口来分析成纤维细胞的伤口闭合情况。与Col5a1(+/+)成纤维细胞相比，Col5a1(+/-)成纤维细胞的增殖明显减少。我们的数据表明，成纤维细胞数量的减少不是由于细胞凋亡。野生型Col5a1(+/+)成纤维细胞比Col5a1(+/-)成纤维细胞更好地附着于伤口基质成分（I型和III型胶原蛋白以及纤连蛋白）。在体外划痕伤口闭合率方面也观察到了显著差异。Col5a1(+/+)成纤维细胞在22小时内闭合伤口，而Col5a1(+/-)成纤维细胞的伤口闭合率约为80%。在所有分析的时间点，闭合情况都存在显著差异。我们的数据表明，成纤维细胞增殖减少、细胞外基质附着和迁移减少导致了典型EDS中伤口愈合反应的降低。
