Engeneering the neurovascular unit: a novel sensorized microfluidic platform to study barrier function and maturation

Central nervous system diseases pose a significant challenge for the development of effective drugs and therapies. A major limiting factor is the neurovascular unit (NVU), which is both anatomically complex and characterized by a highly selective barrier. Conventional 2D in-vitro models and in-vivo animal models do not adequately replicate its pathophysiology.

Organ-on-a-Chip technology provides a powerful platform to model the NVU, enabling replication of its anatomical and functional features within a dynamic microenvironment that closely mimics the human brain. However, the requirement for specialized facilities and technical expertise limits accessibility, reducing broader translational applications. Additionally, conventional endpoint analyses constrain real-time monitoring of cellular behavior.

Here, we present and validate a novel bi-modular microfluidic chip that offers an easy-to-use and scalable solution for studying cellular cross-talk, while enabling live imaging and real-time measurements. The model incorporates human endothelial cells and primary neurons that were investigated through immunofluorescence and live imaging. The design overcomes key fabrication challenges and integrates a simplified method for Trans-Epithelial/Endothelial Electrical Resistance (TEER) monitoring, allowing in situ real-time assessment of barrier integrity. Overall, this platform represents a robust and versatile tool for in-vitro studies of the NVU, facilitating comprehensive evaluation of its structural and functional dynamics. Our microfluidic NVU-on-chip represents a significant advancement in NVU modelling, providing a versatile platform for CNS drug screening, disease modelling, and personalized medicine applications.