Flow dynamics control the effect of sphingosine-1-phosphate on endothelial permeability in a microfluidic vessel bifurcation model

Blood vessels are lined by endothelial cells that form a semipermeable barrier to restrict fluid flow across the vessel wall. The endothelial barrier is known to respond to various molecular mechanisms, but the effects of mechanical signals that arise due to blood flow remain poorly understood. Here, we report a microfluidic model that mimics the flow conditions and endothelial/extracellular matrix (ECM) architecture of a vessel bifurcation to enable systematic investigation of how flow dynamics that arise within bifurcating vessels guides the endothelial response to biochemical signals. Applying the strengths of our system, we further investigate the endothelial response to sphingosine-1-phosphate, a bioactive lipid that has demonstrated flow-dependent regulation of vascular permeability. We demonstrate that bifurcated fluid flow (BFF) that arises at the base of vessel bifurcations and laminar shear stress (LSS) that arises along downstream vessel walls induce a decrease in endothelial permeability. Furthermore, we identify that flow-dynamics and chaperone proteins regulate the endothelial response to S1P. Through pharmacological inhibition of S1P receptors 1 and 2, we report ligand-independent mechanical activation of S1P receptors 1 and 2, providing support for the role of G protein-coupled receptors as mechanosensors. These findings introduce BFF as an important regulator of vascular permeability, and establish flow dynamics as a determinant of the endothelial response to S1P.Pelotonia Fellowship ProgramBarry M. Goldwater Excellence in Education FoundationThe Ohio State University College of EngineeringA one-year embargo was granted for this item.Academic Major: Biomedical Engineerin

Competing Fluid Forces Control Endothelial Sprouting in a 3-D Microfluidic Vessel Bifurcation Model

Sprouting angiogenesis—the infiltration and extension of endothelial cells from pre-existing blood vessels—helps orchestrate vascular growth and remodeling. It is now agreed that fluid forces, such as laminar shear stress due to unidirectional flow in straight vessel segments, are important regulators of angiogenesis. However, regulation of angiogenesis by the different flow dynamics that arise due to vessel branching, such as impinging flow stagnation at the base of a bifurcating vessel, are not well understood. Here we used a recently developed 3-D microfluidic model to investigate the role of the flow conditions that occur due to vessel bifurcations on endothelial sprouting. We observed that bifurcating fluid flow located at the vessel bifurcation point suppresses the formation of angiogenic sprouts. Similarly, laminar shear stress at a magnitude of ~3 dyn/cm2 applied in the branched vessels downstream of the bifurcation point, inhibited the formation of angiogenic sprouts. In contrast, co-application of ~1 µm/s average transvascular flow across the endothelial monolayer with laminar shear stress induced the formation of angiogenic sprouts. These results suggest that transvascular flow imparts a competing effect against bifurcating fluid flow and laminar shear stress in regulating endothelial sprouting. To our knowledge, these findings are the first report on the stabilizing role of bifurcating fluid flow on endothelial sprouting. These results also demonstrate the importance of local flow dynamics due to branched vessel geometry in determining the location of sprouting angiogenesis

Modeling and optimization of parallelized immunomagnetic nanopore sorting for surface marker specific isolation of extracellular vesicles from complex media

Abstract The isolation of specific subpopulations of extracellular vesicles (EVs) based on their expression of surface markers poses a significant challenge due to their nanoscale size ( 10× relative to prior published designs without sacrificing recovery of the target EVs by selecting pore diameter, number of membranes placed in series, and flow rate. We compare TENPO-isolated EVs to those of gold-standard methods of EV isolation and demonstrate its utility for wide application and modularity by targeting subpopulations of EVs from multiple models of disease including lung cancer, pancreatic cancer, and liver cancer