Silk fibroin scaffolds loaded with angiogenic genes in adenovirus vectors for tissue regeneration.

Abstract

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.