Effects of Low and High Aneurysmal Wall Shear Stress on Endothelial Cell Behavior: Differences and Similarities

Abstract

Background: Intracranial aneurysms (IAs) result from abnormal enlargement of the arterial lumen. IAs are mostly quiescent and asymptomatic, but their rupture leads to severe brain damage or death. As the evolution of IAs is hard to predict and intricates medical decision, it is essential to improve our understanding of their pathophysiology. Wall shear stress (WSS) is proposed to influence IA growth and rupture. In this study, we investigated the effects of low and supra-high aneurysmal WSS on endothelial cells (ECs).Methods: Porcine arterial ECs were exposed for 48 h to defined levels of shear stress (2, 30, or 80 dyne/cm2) using an Ibidi flow apparatus. Immunostaining for CD31 or γ-cytoplasmic actin was performed to outline cell borders or to determine cell architecture. Geometry measurements (cell orientation, area, circularity and aspect ratio) were performed on confocal microscopy images. mRNA was extracted for RNAseq analysis.Results: ECs exposed to low or supra-high aneurysmal WSS were more circular and had a lower aspect ratio than cells exposed to physiological flow. Furthermore, they lost the alignment in the direction of flow observed under physiological conditions. The effects of low WSS on differential gene expression were stronger than those of supra-high WSS. Gene set enrichment analysis highlighted that extracellular matrix proteins, cytoskeletal proteins and more particularly the actin protein family were among the protein classes the most affected by shear stress. Interestingly, most genes showed an opposite regulation under both types of aneurysmal WSS. Immunostainings for γ-cytoplasmic actin suggested a different organization of this cytoskeletal protein between ECs exposed to physiological and both types of aneurysmal WSS.Conclusion: Under both aneurysmal low and supra-high WSS the typical arterial EC morphology molds to a more spherical shape. Whereas low WSS down-regulates the expression of cytoskeletal-related proteins and up-regulates extracellular matrix proteins, supra-high WSS induces opposite changes in gene expression of these protein classes. The differential regulation in EC gene expression observed under various WSS translate into a different organization of the ECs’ architecture. This adaptation of ECs to different aneurysmal WSS conditions may affect vascular remodeling in IAs.

Effect of Blood Transfusion on Cerebral Hemodynamics and Vascular Topology Described by Computational Fluid Dynamics in Sickle Cell Disease Patients

“…The change from baseline was also not significantly different between groups ( p = 0.08 and 0.30, on the right and left respectively). It is worth noting that based on currently available data, the physiologic level of WSS is 20–30 dynes/cm 2 [ 34 ] with some studies suggesting that the lower boundary could be as low as 18 dynes/cm 2 [ 35 ]. Low (9 dynes/cm 2 ) and supra low or sub-physiologic (4.5 dynes/cm 2 ) WSS levels are associated with endothelial and consequently arterial remodeling [ 34 , 35 , 36 ] and endothelial activation [ 37 , 38 ].…”

“…It is worth noting that based on currently available data, the physiologic level of WSS is 20–30 dynes/cm 2 [ 34 ] with some studies suggesting that the lower boundary could be as low as 18 dynes/cm 2 [ 35 ]. Low (9 dynes/cm 2 ) and supra low or sub-physiologic (4.5 dynes/cm 2 ) WSS levels are associated with endothelial and consequently arterial remodeling [ 34 , 35 , 36 ] and endothelial activation [ 37 , 38 ]. Both endothelial remodeling and activation are key components of the vascular pathobiology of SCD-related cerebrovascular complications.…”

“…At study exit in Table 4 the average WSS level for the cRBC arm has increased from the sub-physiologic to the supraphysiologic levels. Unlike sub-physiologic levels of WSS, supraphysiologic levels has been reported to increase the likelihood for thrombosis [ 34 , 35 ]. The implication of this in SCD will become clear in larger sample analysis.…”

Multiplexed fluidic circuit board for controlled perfusion of 3D blood vessels-on-a-chip

“…A well-characterized response of ECs to WSS is the reorganization of the actin cytoskeleton. 61,62 A venous-like WSS of 0.3 Pa (3 dyne per cm −2 ) has been shown to alter actin morphology of primary venous-ECs significantly. 29,63,64 By forming actin stress fibres, the ECs can react to haemodynamic forces.…”

Effect of Blood Transfusion on Cerebral Hemodynamics and Vascular Topology Described by Computational Fluid Dynamics in Sickle Cell Disease Patients.

“…The change from baseline was also not significantly different between group (p = 0.08 and 0.30, on the right and left respectively). It is worth noting that based on currently available data, physiologic level of WSS is 20 -30 dynes/cm 2 [35] with some studies suggesting that the lower boundary could be as low as 18 dynes/cm 2 [36]. Low (9 dynes/cm 2 ) and supra low or sub-physiologic (4.5 dynes/cm 2 ) WSS levels have are associated with endothelial and consequently arterial remodeling [35; 36; 37] and endothelial activation [38; 39].…”