Vascular endothelial cells (VECs), which line blood vessels and are key to understanding pathologies and treatments of various diseases, experience highly variable wall shear stress (WSS) in vivo (1-60 dyn cm(-2)), imposing numerous effects on physiological and morphological functions. Previous flow-based systems for studying these effects have been limited in range, and comprehensive information on VEC functions at the full spectrum of WSS has not been available yet. To allow rapid characterization of WSS effects, we developed the first multiple channel microfluidic platform that enables a wide range (~15×) of homogeneous WSS conditions while simultaneously allowing trans-monolayer assays, such as permeability and trans-endothelial electrical resistance (TEER) assays, as well as cell morphometry and protein expression assays. Flow velocity/WSS distributions between channels were predicted with COMSOL simulations and verified by measurement using an integrated microflow sensor array. Biomechanical responses of the brain microvascular endothelial cell line bEnd.3 to the full natural spectrum of WSS were investigated with the platform. Under increasing WSS conditions ranging from 0 to 86 dyn cm(-2), (1) permeabilities of FITC-conjugated dextran and propidium iodide decreased, respectively, at rates of 4.06 × 10(-8) and 6.04 × 10(-8) cm s(-1) per dyn cm(-2); (2) TEER increased at a rate of 0.8 Ω cm(2) per dyn cm(-2); (3) increased alignment of cells along the flow direction under increasing WSS conditions; and finally (4) increased protein expression of both the tight junction component ZO-1 (~5×) and the efflux transporter P-gp (~6×) was observed at 86 dyn cm(-2) compared to static controls via western blot. We conclude that the presented microfluidic platform is a valid approach for comprehensively assaying cell responses to fluidic WSS.

This paper presents the permeability analysis of neuroactive drugs and correlation with in vivo brain/plasma ratios in a dynamic microfluidic blood-brain barrier (BBB) model. Permeability of seven neuroactive drugs (Ethosuximide, Gabapentin, Sertraline, Sunitinib, Traxoprodil, Varenicline, PF-304014) and trans-endothelial electrical resistance (TEER) were quantified in both dynamic (microfluidic) and static (transwell) BBB models, either with brain endothelial cells (bEnd.3) in monoculture, or in co-culture with glial cells (C6). Dynamic cultures were exposed to 15 dyn/cm(2) shear stress to mimic the in vivo environment. Dynamic models resulted in significantly higher average TEER (respective 5.9-fold and 8.9-fold increase for co-culture and monoculture models) and lower drug permeabilities (average respective decrease of 0.050 and 0.052 log(cm/s) for co-culture and monoculture) than static models; and co-culture models demonstrated higher average TEER (respective 90 and 25% increase for static and dynamic models) and lower drug permeability (average respective decrease of 0.063 and 0.061 log(cm/s) for static and dynamic models) than monoculture models. Correlation of the resultant logP e values [ranging from -4.06 to -3.63 log(cm/s)] with in vivo brain/plasma ratios (ranging from 0.42 to 26.8) showed highly linear correlation (R (2) > 0.85) for all model conditions, indicating the feasibility of the dynamic microfluidic BBB model for prediction of BBB clearance of pharmaceuticals.

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