National Engineering Laboratory for Modern Silk, College of Textile and Clothing Engineering, Soochow University, Suzhou, China.
Cell and Molecular Biology Institute, College of Medicine, Soochow University, Suzhou, China.
J Tissue Eng Regen Med. 2019 May;13(5):715-728. doi: 10.1002/term.2819. Epub 2019 Apr 8.
Vascularization remains a critical challenge in dermal tissue regeneration. In this study, a vascular endothelial growth factor (VEGF165) and angiopoietin-1 (Ang-1) dual gene coexpression vector that encoded green fluorescent protein (GFP) was constructed from an arginine-glycine-aspartic acid-modified adenovirus. Silk fibroin (SF) scaffolds loaded with adenovirus vectors were fabricated by freeze-drying method. In vitro, the human endothelial-derived cell line EA.hy926 was infected with adenovirus vectors and then expressed GFP, secreted VEGF165 and Ang-1, and promoted cell proliferation effectively. The VEGF165 and Ang-1 genes loaded in the SF scaffolds significantly promoted the formation of abundant microvascular networks in the chick embryo chorioallantoic membrane. In vivo, angiogenic genes loaded in the scaffolds promoted vascularization and collagen deposition in scaffolds, thus effectively accelerating dermal tissue regeneration in a dorsal full-thickness skin defect wound model in Sprague-Dawley rats. In conclusion, SF scaffolds loaded with arginine-glycine-aspartic acid-modified adenovirus vectors encoding VEGF165 and Ang-1 could stimulate the formation of vascular networks through the effective expression of target genes in vascular endothelial cells, thereby accelerating the regeneration of dermal tissue.
血管化仍然是皮肤组织再生的一个关键挑战。在这项研究中,构建了一种血管内皮生长因子(VEGF165)和血管生成素-1(Ang-1)双基因共表达载体,该载体由精氨酸-甘氨酸-天冬氨酸修饰的腺病毒编码,并表达绿色荧光蛋白(GFP)。通过冷冻干燥法制备了负载腺病毒载体的丝素蛋白(SF)支架。体外,人血管内皮细胞系 EA.hy926 被腺病毒载体感染,然后有效表达 GFP,分泌 VEGF165 和 Ang-1,并促进细胞增殖。SF 支架中负载的 VEGF165 和 Ang-1 基因显著促进了鸡胚绒毛尿囊膜中丰富的微血管网络的形成。体内,支架中负载的血管生成基因促进了支架中的血管化和胶原沉积,从而有效加速了 Sprague-Dawley 大鼠背部全层皮肤缺损模型中皮肤组织的再生。总之,负载精氨酸-甘氨酸-天冬氨酸修饰的腺病毒载体编码 VEGF165 和 Ang-1 的 SF 支架可以通过血管内皮细胞中靶基因的有效表达刺激血管网络的形成,从而加速皮肤组织的再生。