Side view thrombosis microfluidic device with controllable wall shear rate and transthrombus pressure gradient

Hemodynamic conditions vary throughout the vasculature, creating diverse environments in which platelets must respond. To stop bleeding, a growing platelet deposit must be assembled in the presence of fluid wall shear stress (tau(w)) and a transthrombus pressure gradient (Lambda P) that drives bleeding. We designed a microfluidic device capable of pulsing a fluorescent solute through a developing thrombus forming on collagen +/- tissue factor (TF), while independently controlling Delta P and tau(w). Computer control allowed step changes in Delta P with a rapid response time of 0.26 mm Hg s(-1) at either venous (5.2 dynes cm(-2)) or arterial (33.9 dynes cm(-2)) wall shear stresses. Side view visualization of thrombosis with transthrombus permeation allowed for quantification of clot structure, height, and composition at various Delta P. Clot height was reduced 20% on collagen/TF and 28% on collagen alone when Delta P was increased from 20.8 to 23.4 mm Hg at constant arterial shear stress. When visualized with a platelet-targeting thrombin sensor, intrathrombus thrombin levels decreased by 62% as Delta P was increased from 0 to 23.4 mm Hg across the thrombus-collagen/ TF barrier, consistent with convective removal of thrombogenic solutes due to pressure-driven permeation. Independent of Delta P, the platelet deposit on collagen had a permeability of 5.45 x 10(-14) cm(2), while the platelet/fibrin thrombus on collagen/TF had a permeability of 2.71 x 10(-14) cm(2) (comparable to that of an intact endothelium). This microfluidic design allows investigation of the coupled processes of platelet deposition and thrombin/fibrin generation in the presence of controlled transthrombus permeation and wall shear stress.