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The growth of neurons into networks of controlled geometry is of great interest in the field of cellbased biosensors, neuroelectronic circuits, neurological implants, pharmaceutical testing as well as fundamental biological questions about neuronal interactions. The precise control of the network architecture can be achieved by defined engineering of the surface material properties: this process is called neuronal cell patterning. Different techniques can be used to produce such surface patterns. We have chosen microcontact printing (mCP), because it is a comparatively simple and universal method for patterning biomolecules.

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Functional coupling of reconstructed neuronal networks with microelectronic circuits has potential for the development of bioelectronic devices, pharmacological assays and medical engineering. Modulation of the signal processing properties of on-chip reconstructed neuronal networks is an important aspect in such applications. It may be achieved by controlling the biochemical environment, preferably with cellular resolution. In this work, we attempt to design cell-cell and cell-medium interactions in confined geometries with the aim to manipulate non-invasively the activity pattern of an individual neuron in neuronal networks for long-term modulation. Therefore, we have developed a biohybrid system in which neuronal networks are reconstructed on microstructured silicon chips and interfaced to a microfluidic system. A high degree of geometrical control over the network architecture and alignment of the network with the substrate features has been achieved by means of aligned microcontact printing. Localized non-invasive on-chip chemical stimulation of micropatterned rat cortical neurons within a network has been demonstrated with an excitatory neurotransmitter glutamate. Our system will be useful for the investigation of the influence of localized chemical gradients on network formation and long-term modulation.