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Using three-dimensional cell culture systems on Microelectrode arrays for biosensing applications
(2010)
Graphene is a promising material both as a coating for existing neural electrodes as well as for transparent electrodes made exclusively from graphene. We studied graphene‐based microelectrodes by investigating their recording and stimulation properties in order to evaluate their suitability for neuronal implants. In this work, we compare three different electrode material compositions. Microelectrode arrays (MEA) with an electrode size of about 700 μm2 were prepared of gold, graphene on gold, and plain graphene on glass substrate. In order to reduce polymer contamination during graphene transfer, we employed a polymer‐free transfer and lift‐off process. Impedance studies revealed a value of 2.3 MΩ at 1 kHz for plain, and 0.88 MΩ for graphene on gold. Neuronal recording experiments showed a sufficient SNR for both graphene‐based materials and a stable impedance, unaffected by surface degradation metal electrodes are known for. Stimulation measurements yielded a charge injection capacity of 0.15 mC/cm2 using biphasic pulses of 1 ms and 1 μA transparent graphene electrodes. Cyclic voltammetry revealed a large voltage range of −1.4 V to +1.6 V before water electrolysis occurs. Graphene‐coated gold microelectrodes show enhanced recording properties, whereas plain graphene electrodes might be better suited for stimulation applications.
Microelectrode arrays serve as an indispensable tool in electro-physiological research to study the electrical activity of neural cells, enabling measurements of single cell as well as network communication analysis. Recent experimental studies have reported that the neuronal geometry has an influence on electrical signaling and extracellular recordings. However, the corresponding mechanisms are not yet fully understood and require further investigation. Allowing systematic parameter studies, computational modeling provides the opportunity to examine the underlying effects that influence extracellular potentials. In this letter, we present an in silico single cell model to analyze the effect of geometrical variability on the extracellular electric potentials. We describe finite element models of a single neuron with varying geometric complexity in three-dimensional space. The electric potential generation of the neuron is modeled using Hodgkin-Huxley equations. The signal propagation is described with electro-quasi-static equations, and results are compared with corresponding cable equation descriptions. Our results show that both the geometric dimensions and the distribution of ion channels of a neuron are critical factors that significantly influence both the amplitude and shape of extracellular potentials.
At present, most neurotoxicological studies in the field of microelectrode array (MEA) technology are based on in vivo or in vitro animal models, particularly in mice or rats. These models contributed much to our knowledge about molecular and cellular mechanisms of neurotoxins affecting the central nervous system. Yet, animal models are not necessarily capable to forecasting the effectiveness of treatment in clinical trials. Therefore the development and characterization of suitable neuronal cell models derived from human cells is emerging in the field of toxicity testing. In this study, we present neurospheres (NS) as a new three dimensional (3D) neuronal cell model based on human embryonic stem cells (hESC), coupled onto microelectrode arrays (MEA). This cell-based assay serves as platform to investigate effects of neuroactive substances on network communication. In order to characterize the neurospheres’ reaction, the well-known GABA receptor antagonist bicuculline and the sodium channel blocker carbamazepine are applied. Preliminary results reveal anticipated and physiologically reasonable alterations in network activity. Thus, we have the first evidence that hESC derived NS are a promising 3D cell model for neurotoxicity testing. In further studies, this model will be used to investigate effects of ionizing radiation on network signaling.
Long‐term electromagnetic exposure of developing neuronal networks: A flexible experimental setup
(2016)
Neuronal networks in vitro are considered one of the most promising targets of research to assess potential electromagnetic field induced effects on neuronal functionality. A few exposure studies revealed there is currently no evidence of any adverse health effects caused by weak electromagnetic fields. Nevertheless, some published results are inconsistent. Particularly, doubts have been raised regarding possible athermal biological effects in the young brain during neuronal development. Therefore, we developed and characterized a flexible experimental setup based on a transverse electromagnetic waveguide, allowing controlled, reproducible exposure of developing neuronal networks in vitro. Measurement of S‐parameters confirmed very good performance of the Stripline in the band of 800–1000 MHz. Simulations suggested a flexible positioning of cell culture dishes throughout a large exposure area, as specific absorption rate values were quite independent of their position (361.7 ± 11.4 mW/kg) at 1 W, 900 MHz. During exposure, thermal drift inside cellular medium did not exceed 0.1 K. Embryonic rat cortical neurons were cultivated on microelectrode array chips to non‐invasively assess electrophysiological properties of electrogenic networks. Measurements were taken for several weeks, which attest to the experimental setup being a reliable system for long‐term studies on developing neuronal tissue.