Versatile Microfluidic System for Creating Recirculating Unidirectional Flow for On-Chip Cultures of Barrier Tissues

The interaction of chemicals, nanoparticles, and circulating cells with the endothelium depends on the magnitude of the mechanical shear produced by the flow of blood. When simulating those interactions with microphysiological systems (MPSs), it is critical to reproduce those shear conditions faithfully. For example, unidirectional flow of a specific magnitude keeps the endothelium healthy with normal barrier tissue function, while bidirectional flow mimics disease conditions with compromised barrier function. Additionally, in MPS, recirculating fluid may be necessary to retain tissue-derived factors and metabolites. However, existing MPS designs struggle to achieve medium recirculation of small volumes of liquid with precise flow control. Here, we present an MPS design that is highly versatile and overcomes this limitation. We demonstrate how the device can produce a wide range of fluidic flow rates that can accommodate both low shear conditions suitable for tissues that typically are only exposed to interstitial flow and high shear conditions suitable for barrier tissues that experience blood flow. We demonstrate the device's functionality by culturing human umbilical vein endothelial cells (HUVEC) and confirming their flow-aligned morphology through immunostaining of the adherens junction protein (VE-cadherin) and actin filaments. Furthermore, we present a mathematical model that can be used to calculate operating parameters for culturing any tissue under optimum conditions. We also discuss how the device can be adjusted to recirculate liquid volumes ranging from 100 µL to 5 mL. This versatile system holds promise for commercial applications, including the investigation of expensive compounds that are limited to very small volume samples such as rare cells (e.g., circulating tumor cells) or engineered therapeutic cells with barrier tissues. By offering precise control over a wide range of flow conditions with medium recirculation of small liquid volumes, our device addresses a critical gap in current MPS technology.