Biomechanical determinants of endothelial permeability assessed in standard and modified hollow-fibre bioreactors.

Effects of mechanical stress on the permeability of vascular endothelium are important to normal physiology and in the development of atherosclerosis. Here we elucidate novel effects using commercially available and modified hollow-fibre bioreactors, in which endothelial cells form confluent monolayers lining plastic capillaries with porous walls, contained in a cartridge. The capillaries were perfused with a near-aortic waveform, and permeability was assessed by the movement of rhodamine-labelled albumin from the intracapillary to the extracapillary space. Permeability was increased by acute application of shear stress and decreased by chronic shear stress compared with a static control: this has previously been shown only for multidirectional flows. Increasing viscosity reduced permeability under both acute and chronic shear; since shear rate remained unchanged, these effects resulted from altered shear stress. Reducing pulsatility increased permeability, contrary to the widely held assumption that flow which is highly oscillatory causes endothelial dysfunction. Chronic convection across the monolayer increased effective permeability more than could be explained by the addition of advective transport, contrary to results from previous acute experiments. The off-the-shelf and modified bioreactors provide an excellent tool for investigating the biomechanics of endothelial permeability and have revealed novel effects of flow duration, viscosity, pulsatility and transmural flow.

Visualization of three pathways for macromolecule transport across cultured endothelium and their modification by flow.

Transport of macromolecules across vascular endothelium and its modification by fluid mechanical forces are important for normal tissue function and in the development of atherosclerosis. However, the routes by which macromolecules cross endothelium, the hemodynamic stresses that maintain endothelial physiology or trigger disease, and the dependence of transendothelial transport on hemodynamic stresses are controversial. We visualized pathways for macromolecule transport and determined the effect on these pathways of different types of flow. Endothelial monolayers were cultured under static conditions or on an orbital shaker producing different flow profiles in different parts of the wells. Fluorescent tracers that bound to the substrate after crossing the endothelium were used to identify transport pathways. Maps of tracer distribution were compared with numerical simulations of flow to determine effects of different shear stress metrics on permeability. Albumin-sized tracers dominantly crossed the cultured endothelium via junctions between neighboring cells, high-density lipoprotein-sized tracers crossed at tricellular junctions, and low-density lipoprotein-sized tracers crossed through cells. Cells aligned close to the angle that minimized shear stresses across their long axis. The rate of paracellular transport under flow correlated with the magnitude of these minimized transverse stresses, whereas transport across cells was uniformly reduced by all types of flow. These results contradict the long-standing two-pore theory of solute transport across microvessel walls and the consensus view that endothelial cells align with the mean shear vector. They suggest that endothelial cells minimize transverse shear, supporting its postulated proatherogenic role. Preliminary data show that similar tracer techniques are practicable in vivo.NEW & NOTEWORTHY Solutes of increasing size crossed cultured endothelium through intercellular junctions, through tricellular junctions, or transcellularly. Cells aligned to minimize the shear stress acting across their long axis. Paracellular transport correlated with the level of this minimized shear, but transcellular transport was reduced uniformly by flow regardless of the shear profile.