Cell-electronic sensing of particle-induced cellular responses.

We report a new technique for the continuous and real-time measurement of microparticle-induced cellular responses using a real-time cell-electronic sensing (RT-CES) technology. The method involves the use of microelectrode-embedded microwells seeded with one of two lung cancer carcinoma cell lines (A549 and SK-MES-1), allowing for continuous measurements of impedance. The change in impedance that is automatically converted to the cell index is linearly correlated with the numbers of the seeding cells during the log phase, providing quantitative measurement of cytotoxicity. After 24 h of initial incubation in 96 microwells, the cultures are treated with microparticles, and changes in the cell index are monitored in real time. Multiple data, including dose response curves, IC(50) (a concentration inhibiting 50% cell growth), and cell-specific and particulate-specific cell responses, are obtained from a single set of experiments. SK-MES-1 cells consistently showed more severe effects and lower IC(50) values than A549 cells when they were treated with quartz particle suspensions. The different effects detected using the RT-CES technique were related to morphological change and apoptosis, supported by the scanning electronic microscopy and flow cytometry results. The method is further used to test the cytotoxicity of two PM(10) standard reference materials of urban air dust and diesel particulates, demonstrating the potential application of this new technique for biomonitoring of air particulates.

[1]  J. Cook,et al.  Viability measurements in mammalian cell systems. , 1989, Analytical biochemistry.

[2]  M. Rämet,et al.  p53Protein expression is correlated with benzo[a]pyrene-DNA adducts in carcinoma cell lines , 1995 .

[3]  D. Buchanan,et al.  Quantitative relations between exposure to respirable quartz and risk of silicosis , 2003, Occupational and environmental medicine.

[4]  Ivar Giaever,et al.  Electrical Impedance of Cultured Endothelium Under Fluid Flow , 2001, Annals of Biomedical Engineering.

[5]  S. Brimijoin,et al.  Direct evidence for an adhesive function in the noncholinergic role of acetylcholinesterase in neurite outgrowth , 2001, Journal of neuroscience research.

[6]  I. Giaever,et al.  Monitoring fibroblast behavior in tissue culture with an applied electric field. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Tanja Pless-Mulloli,et al.  Particulate Air Pollution and Fetal Health: A Systematic Review of the Epidemiologic Evidence , 2004, Epidemiology.

[8]  I Nicoletti,et al.  A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. , 1991, Journal of immunological methods.

[9]  B. Mahadevan,et al.  Long-term effects of a standardized complex mixture of urban dust particulate on the metabolic activation of carcinogenic polycyclic aromatic hydrocarbons in human cells in culture. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[10]  Xiao Xu,et al.  Real-time monitoring of morphological changes in living cells by electronic cell sensor arrays: an approach to study G protein-coupled receptors. , 2006, Analytical chemistry.

[11]  Xiao Xu,et al.  Dynamic and label-free cell-based assays using the real-time cell electronic sensing system. , 2006, Assay and drug development technologies.

[12]  I. Giaever,et al.  Effect of the pSV2-neo plasmid on NIH 3T3 cell motion detected electrically. , 1994, Experimental cell research.

[13]  R. Tepper,et al.  Cockroach extract antigen increases bronchial airway epithelial permeability. , 2002, The Journal of allergy and clinical immunology.

[14]  Li Xie,et al.  Microelectronic cell sensor assay for detection of cytotoxicity and prediction of acute toxicity. , 2006, Toxicology in vitro : an international journal published in association with BIBRA.

[15]  Miqin Zhang,et al.  Cellular impedance biosensors for drug screening and toxin detection. , 2007, The Analyst.

[16]  Stephan Gabos,et al.  Dynamic monitoring of cytotoxicity on microelectronic sensors. , 2005, Chemical research in toxicology.

[17]  V. Antony,et al.  Th2 Cytokines IL-4 and IL-13 Downregulate Paxillin Expression in Bronchial Airway Epithelial Cells , 2004, Journal of Clinical Immunology.

[18]  Wei Zheng,et al.  Application of real-time cell electronic sensing (RT-CES) technology to cell-based assays. , 2004, Assay and drug development technologies.

[19]  R. Clark,et al.  Studies in Vitro on the Role of αv and β1 Integrins in the Adhesion of Human Dermal Fibroblasts to Provisional Matrix Proteins Fibronectin, Vitronectin, and Fibrinogen , 1996 .