VEGF analysis induced by endothelialized gas-plasma treated d,l-PLA scaffolds

Abstract

Purpose: Vascular endothelial growth factor (VEGF) isoforms play different roles in the temporal sprouting of endothelial-lined vessels in a nude mouse peritoneal model as cells respond to nontreated control and gas-plasma-treated bioresorbable poly-d,l-lactide acid 3D scaffolds with human aortic endothelial cells (HAEC). Methods and materials: Nude mice peritoneums were incubated with HAEC (CW=control; TW=gas-plasma treated) or polymer scaffolds (C_p=control; T_p=treated) for 12, 24 and 72 days. Cytoplasmic and nuclear protein fractions were isolated using NER, electrophoresized using NuPAGE–MES and analyzed by WesternBreeze Chemiluminescent. Results: Prominent VEGF bands included 28, 45 and 62 kDa; 52-kDa VEGF observed in cytoplasmic TW fractions contributed about 18.6% at 12 days, 20.0% at 24 days and 13.1% at 72 days of the total VEGF signal. Yet, it was only noted in CW at 72 days where it accounted for 6.9%. A unique 32-kDa band appeared in both C_p (24.6%) and T_p (18.3%). Significant differences between band densities occurred for cytoplasmic nuclear CW24–TW24 (P=.022), CW72–TW72 (P=.011) and, also, cytoplasmic C_p24–T_p24 (P=.038). Conclusions: The temporal and spatial organization of the TW isoforms results in more angiogenesis.