Understanding the role of vascular stretch on modulation of VWF and ANGPT-2 in continuous flow left ventricular assist device (CF-VAD) patients†

Abstract

Non-surgical bleeding is a common complication in patients on continuous flow left ventricular assist device (CF-VAD) support. This study investigates how the transition from cyclic to constant stretch following CF-VAD implantation affects endothelial biosynthesis and release of Von Willebrand factor (VWF) and angiopoietin-2 (ANGPT-2), two molecules that play an essential role in the development of non-surgical bleeding. Human aortic endothelial and umbilical vein endothelial cells (HAECs and HUVECs) were cultured within a uniaxial stretch device that mimics stretch associated with both normal pulsatile and CF-VAD conditions. Following 72 hours of stretch, transcriptional regulation, intracellular accumulation, and secretion of VWF and ANGPT-2 were evaluated using molecular expression profiling and immunofluorescence microscopy. Constant stretch associated with CF-VADs upregulates transcriptional levels of VWF and ANGPT-2 in HAECs and HUVECs compared to physiological cyclic stretch (p < 0.05). Transcriptional increases in both VWF and ANGPT-2 in HAECs also resulted in increased intracellular protein levels of VWF and ANGPT-2 measured using ELISA, western blots and immunofluorescence microscopy, whereas in HUVECs, the intracellular increase was evident only with western blots and immunofluorescence microscopy. Finally, constant stretch appears to promote ANGPT-2 release and inhibit release of VWF from both HAECs and HUVECs compared to cyclic stretch. Our study found that constant stretch upregulates the production of both VWF and ANGPT-2. However, while the release of ANGPT-2 is elevated under constant stretch, the release of VWF declines, resulting in elevated extracellular levels of ANGPT-2, but not VWF.