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Progress in the clinical application of recording and stimulation devices for neural diseases is still limited, mainly because of suboptimal material engineering and unfavorable interactions with biological entities. Nanotechnology is providing upgraded designs of materials to better mimic the native extracellular environment and attain more intimate contacts with individual neurons, besides allowing for the miniaturization of the electrodes. However, little progress has been done to date on the understanding of the biological impact that such neural interfaces have on neural network maturation and functionality. In this work, we elucidate the effect of a gold (Au) highly ordered nanostructure on the morphological and functional interactions with neural cells and tissues. Alumina-templated Au nanostructured electrodes composed of parallel nanowires of 160 nm in diameter and 1.2 μm in length (Au-NWs), with 320 nm of pitch, are designed and characterized. Equivalent non-structured Au electrodes (Au-Flat) are used for comparison. By using diverse techniques in in vitro cell cultures including live calcium imaging, we found that Au-NWs interfaced with primary neural cortical cells for up to 14 days allow neural networks growth and increase spontaneous activity and ability of neuronal synchronization, thus indicating that nanostructured features favor neuronal network. The enhancement in the number of glial cells found is hypothesized to be behind these beneficial functional effects. The in vivo effect of the implantation of these nanostructured electrodes and its potential relevance for future clinical applicability has been explored in an experimental model of rat spinal cord injury. Subacute responses to implanted Au-NWs show no overt reactive or toxic biological reactions besides those triggered by the injury itself. These results highlight the translational potential of Au-NWs electrodes for in vivo applications as neural interfaces in contact with central nervous tissues including the injured spinal cord.

The use of nanostructured materials and nanosized-topographies has the potential to impact the performance of implantable biodevices, including neural interfaces, enhancing their sensitivity and selectivity, while reducing tissue reactivity. As a result, current trends in biosensor technology require the effective ability to improve devices with controlled nanostructures. Nanoimprint lithography to pattern surfaces with high-density and high aspect ratio nanopillars (NPs) made of polystyrene (PS-NP, insulating), or of a polystyrene/carbon-nanotube nanocomposite (PS-CNT-NP, electrically conductive) are exploited. Both substrates are challenged with cultured primary neurons. They are demonstrated to support the development of suspended synaptic networks at the NPs' interfaces characterized by a reduction in proliferating neuroglia, and a boost in neuronal emergent electrical activity when compared to flat controls. The authors successfully exploit their conductive PS-CNT-NPs to stimulate cultured cells electrically. The ability of both nanostructured surfaces to interface tissue explants isolated from the mouse spinal cord is then tested. The integration of the neuronal circuits with the NP topology, the suspended nature of the cultured networks, the reduced neuroglia formation, and the higher network activity together with the ability to deliver electrical stimuli via PS-CNT-NP reveal such platforms as promising designs to implement on neuro-prosthetic or neurostimulation devices.

Understanding neural physiopathology requires advances in nanotechnology-based interfaces, engineered to monitor the functional state of mammalian nervous cells. Such interfaces typically contain nanometer-size features for stimulation and recording as in cell-non-invasive extracellular microelectrode arrays. In such devices, it turns crucial to understand specific interactions of neural cells with physicochemical features of electrodes, which could be designed to optimize performance. Herein, versatile flexible nanostructured electrodes covered by arrays of metallic nanowires are fabricated and used to investigate the role of chemical composition and nanotopography on rat brain cells in vitro. By using Au and Ni as exemplary materials, nanostructure and chemical composition are demonstrated to play major roles in the interaction of neural cells with electrodes. Nanostructured devices are interfaced to rat embryonic cortical cells and postnatal hippocampal neurons forming synaptic circuits. It is shown that Au-based electrodes behave similarly to controls. Contrarily, Ni-based nanostructured electrodes increase cell survival, boost neuronal differentiation, and reduce glial cells with respect to flat counterparts. Nonetheless, Au-based electrodes perform superiorly compared to Ni-based ones. Under electrical stimulation, Au-based nanostructured substrates evoke intracellular calcium dynamics compatible with neural networks activation. These studies highlight the opportunity for these electrodes to excite a silent neural network by direct neuronal membranes depolarization.