Investigation of endothelial cell seeding and attachment on decellularized porcine carotid artery using a rotating bioreactor

Abstract

The prevalence of cardiovascular diseases around the world had highlighted the need for more small-diameter vascular grafts (<6mm). Porcine carotid artery (PCA) after a treatment process called decellularization could provide a potential off the shelf solution to the shortage. The goal of this project was to decellularize PCA with three different treatment solutions while examining the effects of decellularization on extracellular matrix and cytotoxicity of the decellularized artery. Furthermore, human umbilical vein endothelial cells were seeded onto the decellularized artery in a rotating bioreactor to determine whether the decellularized PCA is conducive for endothelial cell attachment. Qualitative analysis of the nucleus and F-actin staining showed that 0.05M NaOH + 50% human serum was the most effective treatment to achieve decellularization, while immunofluorescence indicated the preservation of collagen type I, IV, fibronectin but not von willebrand factor. Decellularized PCA was also shown to be non-cytotoxic and seeding of cells at 10rpm resulted in the most even cell distribution within the lumen. Finally, attached cells were shown to be viable and immunofluorescence indicated the expression of integrin receptors α5β1 and α6β1. To conclude, decellularized PCA retained bioactive ECM proteins and supported cell adhesion, demonstrating its potential use as a small-diameter vascular graft.