Ultrasound waves and gas-filled protein bubbles will herd living cells into precise 3D patterns to build realistic models of the brain’s blood vessels. Current lab-grown models of the neurovascular unit—the interface between brain tissue and its blood supply—can mimic either the shape of vessels, the way cells talk to each other, or the barrier that keeps harmful molecules out, but never all three at once. This project uses ultrasonic standing waves to push cells into organised patterns, and exploits gas vesicles (protein bubbles from cyanobacteria) to make some cells float differently in the sound field, allowing them to be positioned separately from others. If successful, the method will produce a three-dimensional model that combines patterned endothelial cells and pericytes with unpatterned astrocytes in a hydrogel. Such a model would better replicate real tissue morphology, cell signalling, and barrier integrity. This would give pharmaceutical researchers a more reliable platform for testing drugs that need to cross the blood-brain barrier, and for modelling neurological diseases—without relying on animal experiments.
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This project will investigate the capabilities of acoustic bioassembly to create differentially patterned 3D neurovascular models. In vitro models of the neurovascular unit must accurately replicate vessel morphology, cell-cell communication, and barrier impermeability. Current approaches, including planar and bio-fabricated models, typically target one feature while compromising others. Acoustic bioassembly utilises ultrasonic (US) radiation to remotely organize cells. When applying an acoustic standing wave, cells accumulate at pressure nodes, forming regular patterns. This occurs because cell density differs from that of the media. Gas vesicles (GVs) are protein- bound, gas-filled structures found naturally in aquatic cyanobacteria. Recently, methods have been developed to engineer mammalian cells to produce GVs. GV-expressing cells exhibit lower density hence, respond differently to US compared to non-GV-expressing cells, enabling differential spatial organization. This project will establish a method to generate differentially patterned neurovascular models of patterned endothelial cells and pericytes with unpatterned astrocytes encapsulated in hydrogel, using US and GVs. The formation and function of differentially-, fully-, and un-patterned models will be compared. The differentially patterned constructs are expected to offer biomimetic morphology, cell-cell signalling, barrier integrity, and reproducibility. Ultimately, providing a complex in vitro model for neurovascular and blood-brain barrier drug testing and disease modelling.
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