Fabrication of stents with endothelial function in vitro

Abstract

Objective. The aim of the study is to fabricate stents with rabbit outgrowth endothelial progenitor cells (OECs) to facilitate their endothelial function in vitro. Methods. Rabbit OECs were isolated from peripheral blood and identified by immunofluorescence and flow cytometry. Cells proliferation and migration were measured by growth curve and modified Boyden chamber assay. Adhesion assay was performed by replating cells on fibronectin-coated dishes. VEGF, G-CSF and NO in supernatant were tested. OECs were poured on fibronectin-coated or uncoated stents. After six days, scanning electron microscopy (SEM) and inverted fluorescent microscopy observation were performed. Results. About three to four weeks after culture, OECs were characterized as adherent cells which were double positive for Dil-acLDL uptake and FITC-UEA-I binding, with high expression of CD34. They also showed high ability of proliferation, adhesion and migration properties. Compared with uncoated stents, more OECs migrated and adhered onto fibronectin-coated stents. OECs seeding onto the fibronectin coated stents could secret more cytokine and NO. Endothelialization of coated stents was visible both under the SEM and inverted fluorescent microscope. Conclusions. OECs can differentiate to endothelial lineage and possess high ability of proliferation, migration and adhesion. It is feasible to fabricate OECs-seeded stents in vitro, while the stents coated with fibronectin facilitate this endothelialization process.