Defranchi Enrico, Novellino Antonio, Whelan Maurice, Vogel Sandra, Ramirez Tzutzuy, van Ravenzwaay Ben, Landsiedel Robert
ETT s.r.l. Genova, Italy.
Front Neuroeng. 2011 Apr 28;4:6. doi: 10.3389/fneng.2011.00006. eCollection 2011.
Detection and characterization of chemically induced toxic effects in the nervous system represent a challenge for the hazard assessment of chemicals. In vivo, neurotoxicological assessments exploit the fact that the activity of neurons in the central and peripheral nervous system has functional consequences. And so far, no in vitro method for evaluating the neurotoxic hazard has yet been validated and accepted for regulatory purpose. The micro-electrode array (MEA) assay consists of a culture chamber into which an integrated array of micro-electrodes is capable of measuring extracellular electrophysiology (spikes and bursts) from electro-active tissues. A wide variety of electrically excitable biological tissues may be placed onto the chips including primary cultures of nervous system tissue. Recordings from this type of in vitro cultured system are non-invasive, give label free evaluations and provide a higher throughput than conventional electrophysiological techniques. In this paper, 20 substances were tested in a blinded study for their toxicity and dose-response curves were obtained from fetal rat cortical neuronal networks coupled to MEAs. The experimental procedure consisted of evaluating the firing activity (spiking rate) and modification/reduction in response to chemical administration. Native/reference activity, 30 min of activity recording per dilution, plus the recovery points (after 24 h) were recorded. The preliminary data, using a set of chemicals with different mode-of-actions (13 known to be neurotoxic, 2 non-neuroactive and not toxic, and 5 non-neuroactive but toxic) show good predictivity (sensitivity: 0.77; specificity: 0.86; accuracy: 0.85). Thus, the MEA with a neuronal network has the potency to become an effective tool to evaluate the neurotoxicity of substances in vitro.
检测和表征化学物质对神经系统的毒性作用是化学物质危害评估面临的一项挑战。在体内，神经毒理学评估利用了中枢和外周神经系统中神经元活动具有功能后果这一事实。到目前为止，尚未有用于评估神经毒性危害的体外方法得到监管目的的验证和认可。微电极阵列（MEA）检测由一个培养室组成，其中集成的微电极阵列能够测量电活性组织的细胞外电生理（尖峰和爆发）。多种电兴奋性生物组织可以放置在芯片上，包括神经系统组织的原代培养物。这种体外培养系统的记录是非侵入性的，提供无标记评估，并且比传统电生理技术具有更高的通量。在本文中，在一项盲法研究中测试了20种物质的毒性，并从与MEA耦合的胎鼠皮质神经元网络中获得了剂量反应曲线。实验程序包括评估给药后放电活动（放电率）和变化/降低情况。记录天然/参考活动、每次稀释30分钟的活动记录以及恢复点（24小时后）。使用一组具有不同作用模式的化学物质（13种已知具有神经毒性、2种无神经活性且无毒、5种无神经活性但有毒）的初步数据显示出良好的预测性（敏感性：0.77；特异性：0.86；准确性：0.85）。因此，带有神经元网络的MEA有潜力成为体外评估物质神经毒性的有效工具。
