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In order to investigate the hypothesis that dynamic nanoscale stimuli can influence the functional response of the brain, in this paper we describe the development of a membrane actuator chip based on polydimethylsiloxane (PDMS) soft lithography. The chip exerts a local nanoscale mechanical load on an in vitro neuronal cell network by microfluidic pneumatic deformation of the membrane. The deformation provides a topographical change in the substrate as an input stimulus for the study of response functions of a neuronal cell network in vitro. Calcium ions (Ca2+) imaging within a neuronal cell network grown from dissociated cortical cells of the rat's brain used as a brain model indicates that a neural networks response can be provoked by means of our new method. This actuator chip provides a relatively mild and localised mechanical stimulus by means of a 2% elongation of the membrane's width during the application of a pressure pulse underneath the membrane using a microfluidic channel design. We found an average 50% increase of the intracellular Ca2+ flux activity for 2D neuronal cell networks among 4 independent samples cultured on flat membranes. Additionally, we have proven the applicability of the actuator chip for networks on nanogrooved membranes by the observation of Ca2+ traces and we also observed the Ca2+ waves response upon stimulation in a three dimensional (3D) in vivo-like neuronal cell network using Matrigel on flat membranes. Hence, the chip potentially provides a novel technology platform for the in vitro modelling of brain tissues with topographically and 3D hydrogel-defined network architectures.

Nanoscale membrane actuator for in vitro mechano-stimuli responsive studies of neuronal cell networks on chip