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 endothelia...

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Bibliographic Details
Published in:American journal of physiology. Heart and circulatory physiology 2017-11, Vol.313 (5), p.H959-H973
Main Authors: Ghim, Mean, Alpresa, Paola, Yang, Sung-Wook, Braakman, Sietse T, Gray, Stephen G, Sherwin, Spencer J, van Reeuwijk, Maarten, Weinberg, Peter D
Format: Article
Language:English
Subjects:
Algorithms
Animals
Aorta - cytology
Aorta - metabolism
Arteriosclerosis
Atherosclerosis
Biological Transport, Active
Cell Nucleus - metabolism
Cell Nucleus - ultrastructure
Cells
Cells, Cultured
Computer simulation
Endothelial cells
Endothelial Cells - metabolism
Endothelium
Endothelium, Vascular - metabolism
Endothelium, Vascular - ultrastructure
Fluids
Fluorescence
Fluorescent indicators
In vivo methods and tests
Intercellular Junctions - metabolism
Lipoproteins, LDL - metabolism
Macromolecular Substances - metabolism
Macromolecules
Permeability
Shear stress
Solute transport
Stress, Mechanical
Substrates
Swine
Tracer techniques
Tracers
Transport
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Summary: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. 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.
ISSN:0363-6135
1522-1539
DOI:10.1152/ajpheart.00218.2017