Recently, the PI's lab has successfully developed a whole cell based impedance system to monitor cellular behavior upon engineered nanomaterial exposure sensitive enough to measure the micro-motion of a cell and the progression of the cytotoxicity demonstrating the kinetic effects of the nanoparticles on the cells (://iopscience.iop.org/0957-4484/labtalk- article/43407labTalkTab=mostRead). In another preliminary study, ROS induced cell DNA damage was amperometrically measured at a single cell level using a microelectrochemical sensor. The objective of this proposal is to develop a cell based biosensing system with the integration of the cell based impedance sensor, the DNA damage sensor and a novel 3D in vitro BBB model to measure nanotoxicity all in real-time. The oxidative DNA damage biomarker (8-OHdG) will be introduced to monitor the genotoxic kinetics following the time course of different types of nanomaterial exposure. Furthermore, upon integrating a novel 3D cell co-culture system, the transmembrane impedance of cultured blood brain barrier (BBB) models will be measured to understand the toxic effects of nanomaterials on the structure, composition and function of the tissues. The sensing system will be capable of real-time monitoring of the behavior of cells including genomic damage, cell attachment, motility, mortality and cytotoxicity, and BBB tissue transportation ability under various nanomaterial exposures with a simpler and less labor intensive operation and more precise results compared to conventional colorimetric methods which can only provide a general sense of cytotoxicity as they show results only at a final time point The toxicity of several engineered nanomaterials such as gold, silver, graphene, etc., will be kinetically assessed for a better understanding of the toxic mechanisms and the toxic effects of the nanomaterials due to the size, shape, and particularly the surface functional groups since particles may transport across cell membranes, especially into mitochondria, causing internal damage that may affect cell behavior over time . Advances in microfabrication technology will allow up to several hundred different individual culture wells containing the detecting electrodes to be fabricated on a single chip. In this context, the methodology becomes a truly powerful and throughput analytical device. This tool will help bridge scientific gaps and elucidate potential benefits and risks related to the use of nanoengineered materials. The project will develop and promote multi-disciplinary research and training experience for graduate students at FIU. PUBLIC HEALTH RELEVANCE: We propose to develop an electrical sensing system in which the toxicity of various nanomaterials can be kinetically measured at the genomic, cellular and tissue level. This sensing system will be introduced as an indirect approach for kinetic screening of toxic properties of nanomaterials. The electrical impedance devices have important implications in solving problems in many biomedical areas such as nanotechnology based drug design and delivery, cancer diagnosis and therapy, and biomaterial development and applications.
