The effect of VEGF-immobilized nickel-free high-nitrogen stainless steel on viability and proliferation of vascular endothelial cells

Using ester bonds, vascular endothelial growth factor-A (VEGF-A) was immobilized on the surface of a novel biometal, nickel-free high-nitrogen stainless steel (HNS). The biological activity of immobilized VEGF-A was investigated after the culture of human umbilical vein endothelial cells (HUVECs) on the substrate. Immobilization of VEGF-A onto the HNS surface was performed using trisuccinimidyl citrate (TSC) as a linker. Firstly, UV irradiation was employed to amplify hydroxyl groups on the HNS surface. Next, the HNS was dipped into TSC/dimethyl sulfoxide solution at room temperature. From the results of water contact angle measurement and X-ray photoelectron spectroscopy (XPS) analysis, TSC was found to be immobilized on the HNS surface via ester bonds. Quantitative analysis demonstrated that immobilized VEGF-A remained even after immersion in culture medium for 7 days; however, it was gradually deimmobilized by hydrolysis of the ester bonds at the TSC–metal interface. As a result, VEGF-A-immobilized HNS significantly contributed to the stimulation of HUVEC growth for the initial stage of culture, even though the gradual reduction in growth stimulation of HUVECs occurred by the sequential deimmobilization of VEGF-A, which was caused by the hydrolysis of the ester groups. Therefore, VEGF-A-immobilized HNS could be applied as a basic material for coronary stents.