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bioRxiv · 10.1101/2025.10.30.685564

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

Montesi, L.·Lattanzi, D.·Picchi, M.·Sartini, S.·Mekler, T.·Korin, N.·Rauti, R.
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Abstract

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.

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Montesi, L., Lattanzi, D., Picchi, M., Sartini, S., Mekler, T., Korin, N., Rauti, R.. 2025-10-31. Engeneering the neurovascular unit: a novel sensorized microfluidic platform to study barrier function and maturation. https://doi.org/10.1101/2025.10.30.685564

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