Alterations in the mechanical properties of erythrocytes occurring in inflammatory and haematological disorders such as sickle-cell disease (SCD) and malaria often lead to increased endothelial permeability, haemolysis and microvascular obstruction. However, the associations among these pathological phenomena remain unknown. Here, we show that a perfusable, endothelialized microvasculature-on-a-chip featuring an interpenetrating-polymer-network hydrogel that recapitulates the stiffness of blood vessel intima, basement membrane self-deposition and self-healing endothelial barrier function for longer than one month enables the real-time visualization, with high spatiotemporal resolution, of microvascular obstruction and endothelial permeability under physiological flow conditions. We found that extracellular haem—a haemolytic by-product—induces delayed yet reversible endothelial permeability in a dose-dependent manner, and demonstrate that endothelial interactions with SCD or malaria-infected erythrocytes cause reversible microchannel occlusion and increased in situ endothelial permeability. The microvasculature-on-a-chip enables mechanistic insight into the endothelial barrier dysfunction associated with SCD, malaria and other inflammatory and haematological diseases.
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This work was performed in part at the Georgia Tech Institute for Electronics and Nanotechnology (a member of the National Nanotechnology Coordinated Infrastructure, which is supported by the National Science Foundation (grant ECCS-1542174)). We acknowledge the clinical research personnel at Emory University and the Children’s Healthcare of Atlanta who helped to obtain samples, and the patients for donating blood. We acknowledge D. Archer and L. A. Brown for valuable discussions. We acknowledge the rest of the Lam Lab for technical support and suggestions. Financial support was provided by National Science Foundation CAREER Award 1150235 (to W.A.L.), National Institutes of Health grants U01HL117721 (to S.F.O.-A., C.H.J. and W.A.L.), U54HL112309 (to W.A.L.) and R01HL121264 (to W.A.L.), and the National Institute for Neurological Disorders and Strokes grant R21NS085382 (to T.J.L.).
The authors declare no competing interests.
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Supplementary figures, tables and video captions.
3D rendering of confocal microscopy immunostaining images of the adherens-junction protein VE-cadherin in the IPN-hydrogel-based endothelialized microfluidic system.
Diffusion of BSA-AF594 from acellular (non-endothelialized) microchannels can be used as a positive control to measure permeability.
Perfused BSA-AF594 was maintained in the ‘vascular’ space of the endothelialized microchannels during the permeability assay.
Perfusion of RBCs isolated from the sickle-cell disease patients with lower percentages of ISCs (~2.5%) into the engineered microvasculature (4-hour perfusion).
Perfusion of RBCs isolated from the sickle-cell disease patients with higher percentages of ISCs into the engineered microvasculature (4-hour perfusion).
Perfusion of malaria-infected RBCs into the engineered microvasculature (4-hour perfusion).
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Qiu, Y., Ahn, B., Sakurai, Y. et al. Microvasculature-on-a-chip for the long-term study of endothelial barrier dysfunction and microvascular obstruction in disease. Nat Biomed Eng 2, 453–463 (2018). https://doi.org/10.1038/s41551-018-0224-z
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