Engineering Highly Interconnected Neuronal Networks on Nanowire Scaffolds.

Identifying the specific role of physical guidance cues in the growth of neurons is crucial for understanding the fundamental biology of brain development and for designing scaffolds for tissue engineering. Here, we investigate the structural significance of nanoscale topographies as physical cues for neurite outgrowth and circuit formation by growing neurons on semiconductor nanowires. We monitored neurite growth using optical and scanning electron microscopy and evaluated the spontaneous neuronal network activity using functional calcium imaging. We show, for the first time, that an isotropic arrangement of indium phosphide (InP) nanowires can serve as physical cues for guiding neurite growth and aid in forming a network with neighboring neurons. Most importantly, we confirm that multiple neurons, with neurites guided by the topography of the InP nanowire scaffolds, exhibit synchronized calcium activity, implying intercellular communications via synaptic connections. Our study imparts new fundamental insights on the role of nanotopographical cues in the formation of functional neuronal circuits in the brain and will therefore advance the development of neuroprosthetic scaffolds.