Acute neuropharmacologic action of chloroquine on cortical neurons in vitro

O’Shaughnessy, Thomas J.; Zim, Bret; Ma, Wu; Shaffer, Kara M.; Stenger, David A.; Zamani, Kaveh; Gross, Guenter W.; Pancrazio, Joseph J.

doi:10.1016/s0006-8993(02)03763-0

Cited by 26 publications

(13 citation statements)

References 38 publications

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“…GFAP levels were also elevated above control in our studies implying some astrocytic activation caused by the toxin, which has also been reported previously in astrogliosis in human astrocytoma cells (Chen et al 2009). Although chloroquine is a widely used therapeutic agent for the treatment of malaria, it is also considered to be capable of exhibiting neurotoxicity within its therapeutic concentration range, although this is rare and is thought to be dependent on factors which may influence blood-brain barrier integrity and thus facilitating drug penetration into the CNS (O'Shaughnessy et al 2003). Chloroquine's elevation of GFAP levels above control in the present study suggests an activating effect on the NT.2A astrocytes, which has also been shown in other models such as terms of pro-inflammatory cytokine induction human astrocytic cells (Park et al 2003), in our own laboratory with U-251 glioma cells (Holden and Coleman 2008), as well as in primary rat astrocytes (Cookson and Pentreath 1994).…”

Section: Discussionmentioning

confidence: 99%

Single-Cell ELISA and Flow Cytometry as Methods for Highlighting Potential Neuronal and Astrocytic Toxicant Specificity

et al. 2010

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The timeline imposed by recent worldwide chemical legislation is not amenable to conventional in vivo toxicity testing, requiring the development of rapid, economical in vitro screening strategies which have acceptable predictive capacities. When acquiring regulatory neurotoxicity data, distinction on whether a toxic agent affects neurons and/or astrocytes is essential. This study evaluated neurofilament (NF) and glial fibrillary acidic protein (GFAP) directed single-cell (S-C) ELISA and flow cytometry as methods for distinguishing cell-specific cytoskeletal responses, using the established human NT2 neuronal/astrocytic (NT2.N/A) co-culture model and a range of neurotoxic (acrylamide, atropine, caffeine, chloroquine, nicotine) and non-neurotoxic (chloramphenicol, rifampicin, verapamil) test chemicals. NF and GFAP directed flow cytometry was able to identify several of the test chemicals as being specifically neurotoxic (chloroquine, nicotine) or astrocytoxic (atropine, chloramphenicol) via quantification of cell death in the NT2.N/A model at cytotoxic concentrations using the resazurin cytotoxicity assay. Those neurotoxicants with low associated cytotoxicity are the most significant in terms of potential hazard to the human nervous system. The NF and GFAP directed S-C ELISA data predominantly demonstrated the known neurotoxicants only to affect the neuronal and/or astrocytic cytoskeleton in the NT2.N/A cell model at concentrations below those affecting cell viability. This report concluded that NF and GFAP directed S-C ELISA and flow cytometric methods may prove to be valuable additions to an in vitro screening strategy for differentiating cytotoxicity from specific neuronal and/or astrocytic toxicity. Further work using the NT2.N/A model and a broader array of toxicants is appropriate in order to confirm the applicability of these methods.

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Section: Discussionmentioning

confidence: 99%

Single-Cell ELISA and Flow Cytometry as Methods for Highlighting Potential Neuronal and Astrocytic Toxicant Specificity

et al. 2010

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“…Glass, silicon substrate, and biocompatible polymer are commonly chosen as substrates because of their stiffness, fabrication convenience, and conformity with specific requirements for various applications (Heiduschka and Thanos 1998). Noble metals like platinum, gold, etc., which possess the properties of high resistance to corrosion and low chemical reactivity, are especially suited to electrical stimulation and sensing in vitro and in vivo (Yagi et al 1999;O'Shaughnessy et al 2003;Nam et al 2006;Heiduschka and Thanos 1998;Ingebrandt et al 2001). Second, surface modification techniques that are physical or chemical have been applied to perfect the cell growing environment.…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, primary cell cultures also provide direct observation of desired cells on the cultured substrate. In recent studies, various types of neuronal cells from cortex (O'Shaughnessy et al 2003), spinal cord (Chiappalone et al 2003), hippocampus (Nam et al 2006), and retina (Yagi et al 1999) have been used on MEA to characterize biocompatibility and neuronal cell growth.…”

Section: Introductionmentioning

confidence: 99%

Characterization of surface modification on microelectrode arrays for in vitro cell culture

Lin¹,

Chen²,

Liao³

et al. 2007

Biomed Microdevices

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This study aims to investigate surface-modified microelectrodes on the microelectrode arrays (MEAs) for neuronal interfaces with in vitro cell culture. The polyimide (PI) MEA was fabricated by using micro-electro-mechanical systems (MEMS) techniques. Self-assembled monolayers (SAMs) of 11-mercaptoundecanoic acid (MUA) were utilized to modify the microelectrode surface of the MEA. The SAMs' modified surface of microelectrodes offered a reliable interface to immobilize biological ligands through covalent bonding. To increase biocompatibility, the poly-D-lysine (PDL) was immobilized on the SAMs' modified microelectrodes. Several analytical techniques were used to define the physical structure and functional groups of surface-modified gold microelectrodes on the MEA. Spectra of the Fourier transform infrared reflection (FTIR) were applied to characterize the molecular structure of MUA-SAMs and PDL on the microelectrodes. The spectra, two peaks of amide I (at 1,613 cm(-1)) and amide II (at 1,548 cm(-1)), revealed that covalent amide bonding existed in PDL-MUA-SAMs modified surfaces. The thickness and formation of the MUA and PDL were also observed and quantified by using an atomic force microscope (AFM). The impedance measurement of PDL-MUA-SAMs modified MEA only increased slightly to an average of 524.6 +/- 55.8 kOmega from 352.9 +/- 34.4 kOmega of bare gold microelectrode (p < 0.05, N = 20). In addition, the time-course changes of total impedance resulting from cell sealing resistance and gap reactance were recorded for 7 days for inferring the growth of cell lines on the electrode contact of modified MEA. The experiment of 3T3 fibroblasts, PC12 cells, primary glial cells, and primary cortical neurons cultured on the modified MEAs displayed a good adhesion rate. These biocompatibility assays demonstrated that the neuronal cells are able to grow in a proximity to PDL-MUA-SAMs modified microelectrodes of the MEAs for effective electrophysiological stimulation/sensing schemes and for future implantation purposes.

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“…Therefore, in vitro models have been proposed as an appropriate paradigm to mimic in vivo-like conditions, for example, in pharmacology, toxicology, developmental biology, or basic research [Layer et al, 1992;Keefer et al, 2001;O'Shaughnessy et al, 2003;Stett et al, 2003;Martinoia et al, 2005].…”

Section: Introductionmentioning

confidence: 99%

Electromagnetic exposure of scaffold-free three-dimensional cell culture systems

Daus

Goldhammer

Layer

et al. 2011

Bioelectromagnetics

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In recent years, a number of in vitro studies have reported on the possible athermal effects of electromagnetic exposure on biological tissue. Typically, this kind of study is performed on monolayers of primary cells or cell lines. However, two-dimensional cell layer systems lack physiological relevance since cells in vivo are organized in a three-dimensional (3D) architecture. In monolayer studies, cell-cell and cell-ECM interactions obviously differ from live tissue and scale-ups of experimental results to in vivo systems should be considered carefully. To overcome this problem, we used a scaffold-free 3D cell culture system, suitable for the exploration of electrophysiological effects due to electromagnetic fields (EMF) at 900 MHz. Dissociated cardiac myocytes were reaggregated into cellular spheres by constant rotation, and non-invasive extracellular recordings of these so-called spheroids were performed with microelectrode arrays (MEA). In this study, 3D cell culture systems were exposed to pulsed EMFs in a stripline setup. We found that inhomogeneities in the EMF due to electrodes and conducting lines of the MEA chip had only a minor influence on the field distribution in the spheroid if the exposure parameters were chosen carefully.

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Acute neuropharmacologic action of chloroquine on cortical neurons in vitro

Cited by 26 publications

References 38 publications

Single-Cell ELISA and Flow Cytometry as Methods for Highlighting Potential Neuronal and Astrocytic Toxicant Specificity

Single-Cell ELISA and Flow Cytometry as Methods for Highlighting Potential Neuronal and Astrocytic Toxicant Specificity

Characterization of surface modification on microelectrode arrays for in vitro cell culture

Electromagnetic exposure of scaffold-free three-dimensional cell culture systems

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