Assessment of whole blood thrombosis in a microfluidic device lined by fixed human endothelium

Abstract

The vascular endothelium and shear stress are critical determinants of physiological hemostasis and platelet function in vivo, yet current diagnostic and monitoring devices do not fully incorporate endothelial function under flow in their assessment and, therefore, they can be unreliable and inaccurate. It is challenging to include the endothelium in assays for clinical laboratories or point-of-care settings because living cell cultures are not sufficiently robust. Here, we describe a microfluidic device that is lined by a human endothelium that is chemically fixed, but still retains its ability to modulate hemostasis under continuous flow in vitro even after few days of storage. This device lined with a fixed endothelium supports formation of platelet-rich thrombi in the presence of physiological shear, similar to a living arterial vessel. We demonstrate the potential clinical value of this device by showing that thrombus formation and platelet function can be measured within minutes using a small volume (0.5 mL) of whole blood taken from subjects receiving antiplatelet medications. The inclusion of a fixed endothelial microvessel will lead to biomimetic analytical devices that can potentially be used for diagnostics and point-of-care applications.Electronic supplementary materialThe online version of this article (doi:10.1007/s10544-016-0095-6) contains supplementary material, which is available to authorized users.

Mimicking arterial thrombosis in a 3D-printed microfluidic in vitro vascular model based on computed tomography angiography data

“…With this approach thrombosis can be recapitulated in 3D vessel geometries in a way that is not possible in microfluidic chips fabricated with typical 2D wafer-based soft lithography, or even in microfluidic chips produced with acrylic fibres or by advanced biofabrication methods. 14,15,49 Furthermore, because it is based on 3D models, this methodology allows for easy correlation and comparison of in vitro studies with in silico fluid flow simulations. In the future, the devices and method described herein could be used to employ medical CTA imaging data to produce patient-specific microfluidic chips, and as such, the method and proof-ofprinciple can be considered to be an important stepping stone towards patient-specific arterial thrombosis studies.…”

Point-of-Care Diagnostics: Recent Developments in a Connected Age

“…The detection method was Raman spectroscopy, performed on a fluorescent microscope for imaging and subsequent Raman spectroscopic fingerprinting 104 . In another advance, a microfluidic chip was developed for thrombosis detection from whole blood 105 . This device used endothelial cells to support formation of platelet-rich thrombi, a design choice made to increase longevity and storage of the device 105 .…”

“…In another advance, a microfluidic chip was developed for thrombosis detection from whole blood 105 . This device used endothelial cells to support formation of platelet-rich thrombi, a design choice made to increase longevity and storage of the device 105 . Detection was based on fluorescence microscopy and computer algorithms 105 .…”

Understanding the hemotoxicity of graphene nanomaterials through their interactions with blood proteins and cells

“…This is because microfluidic devices allow experiments to be conducted in a controlled threedimensional in vitro setting, mimicking the dynamic in vivo physiological environment. [60][61][62][63][64][65] Consequently, in addition to the biological effects and toxicity of graphene nanomaterials on plasma proteins and individual blood cells under static in vitro condition, the hemocompatibility of graphene nanomaterials has been assessed on whole blood under active in vivo-mimicking conditions.…”