Microscaffolds by Direct Laser Writing for Neurite Guidance Leading to Tailor-Made Neuronal Networks

While modern day integrated electronic circuits are essentially designed in a 2D fashion, the brain can be regarded as a 3D circuit. The thus enhanced connectivity enables much more complex signal processing as compared to conventional 2D circuits. Recent technological advances in the development of nano/microscale 3D structuring have led to the development of artificial neuron culturing platforms, which surpass the possibilities of classical 2D cultures. In this work, in vitro culturing of neuronal networks is demonstrated by determining predefined pathways through topological and chemical neurite guiding. Tailor-made culturing substrates of microtowers and freestanding microtubes are fabricated using direct laser writing by two-photon polymerization. The first scaffold design that allows for site-specific cell attachment and directed outgrowth of single neurites along defined paths that can be arranged freely in all dimensions, to build neuronal networks with low cell density, is presented. The neurons cultured in the scaffolds show characteristic electrophysiological properties of vital cells after 10 d in vitro. The introduced scaffold design offers a promising concept for future complex neuronal network studies on defined neuronal circuits with tailor-made design specific neurite connections beyond 2D.