Guiding the Patterned Growth of Neuronal Axons and Dendrites Using Anisotropic Micropillar Scaffolds

The patterning of axonal and dendritic growth is an important topic in neural tissue engineering and critical for generating directed neuronal networks in vitro. Evidence shows that artificial micro/nanotopography can better mimic the environment for neuronal growth in vivo. However, the potential mechanisms by which neurons interact with true three dimensional (3D) topographical cues and form directional networks are unclear. Herein, 3D micropillar scaffolds are designed to guide the growth of neural stem cells and hippocampal neurons in vitro. Discontinuous and anisotropic micropillars are fabricated by femtosecond direct laser writing to form patterned scaffolds with various spacings and heights, which are found to affect the branching and orientation of axons and dendrites. Interestingly, axons and dendrites tend to grow on an array of 3D micropillar scaffolds of the same height and form functionally connected neuronal networks, as reflected by synchronous neuronal activity visualized by calcium imaging. This method may represent a promising tool for studying neuron behavior and directed neuronal networks in a 3D environment.