Development and In Vivo Evaluation of Small-Diameter Vascular Grafts Engineered by Outgrowth Endothelial Cells and Electrospun Chitosan/Poly(ɛ-Caprolactone) Nanofibrous Scaffolds

Abstract

Successful engineering of a small-diameter vascular graft is still a challenge despite numerous attempts for decades. The present study aimed at developing a tissue-engineered vascular graft (TEVG) using autologous outgrowth endothelial cells (OECs) and a hybrid biodegradable polymer scaffold. OECs were harvested from canine peripheral blood and proliferated in vitro, as well as identified by immunofluorescent staining. Electrospun hybrid chitosan/poly(ε-caprolactone) (CS/PCL) nanofibers were fabricated and served as vascular scaffolds. TEVGs were constructed in vitro by seeding OECs onto CS/PCL scaffolds, and then implanted into carotid arteries of cell-donor dogs (n=6). After 3 months of implantation, 5 out of 6 of TEVGs remained patent as compared with 1 out of 6 of unseeded grafts kept patent. Histological and immunohistochemical analyses of the TEVGs retrieved at 3 months revealed the regeneration of endothelium, and the presence of collagen and elastin. OECs labeled with fluorescent dye before implantation were detected in the retrieved TEVGs, indicating that the OECs participated in the vascular tissue regeneration. Biomechanical testing of TEVGs showed good mechanical properties that were closer to native carotid arteries. RT-PCR and western blot analysis demonstrated that TEVGs had favorable biological functional properties resembling native arteries. Overall, this study provided a new strategy to develop small-diameter TEVGs with excellent biocompatibility and regeneration ability.