Impedance-Based Cell Culture Platform To Assess Light-Induced Stress Changes with Antagonist Drugs Using Retinal Cells

Abstract

This Article describes an unprecedented, simple, and real-time in vitro analytical tool to measure the luminous effect on the time responses function of retinal ganglion cells (RGC-5) by electric cell substrate impedance sensing (ECIS) system. The ECIS system was used for the continuous measurement of different color light-induced effects on the response of cells that exposed to protective drugs. The measurement suggests that the association of photo-oxidative stress was mediated by reactive oxygen species (ROS), which plays a critical role that leads to cell stress, damages, and retinopathy, resulting in eye degenerative diseases. Continuous light radiation caused time-dependent decline of RGC-5 response and resulted in photodamage within 10 h due to adenosine 5'-triphosphate depletion and increased ROS level, which is similar to in vivo photodamage. The ECIS results were correlated with standard cell viability assay. ECIS is very helpful to determine the protective effects of analyzed drugs such as β-carotene, quercetin, agmatine, and glutathione in RGC-5 cells, and the maximum drug activity of nontoxic safer drug concentrations was found to be 0.25, 0.25, 0.25, and 1.0 mM, respectively. All drugs show protection against light radiation toxicity in a dose-dependent manner; the most effective drug was found to be glutathione. The proposed system identifies the phototoxic effects in RGC-5 and provides high throughput drug screening for photo-oxidative stress during early stages of drug discovery. This study is convenient and potential enough for the direct measurements of the photoprotective effect in vitro and would be of broad interest in the field of therapeutics.

Electric Cell-Substrate Impedance Sensing To Monitor Viral Growth and Study Cellular Responses to Infection with Alphaherpesviruses in Real Time

“…The absorbance at 570 nm was then measured spectrophotometrically and used to construct a dose-response curve, and the CC 50 was determined. Cidofovir toxicity was evaluated by ECIS in a manner similar to that previous described (43, 44). Briefly, 20,000 CRFK cells were plated and monitored for 24 h as described above.…”

Photoprotective effects of apple peel nanoparticles

“…Photoprotective effects in real-time ECIS was used to measure the changes in the photoprotective activity in vitro, as previously reported by Bennet et al 17 Precise amounts of HDFn cells were seeded to each electrode well, after which volume was adjusted to 400 µL using culture medium. Cells adhered to the surface of the electrode after 10 hours of incubation, resulting in confluence.…”

“…Bennet and Kim have developed the real-time light setup with electric cell-substrate impedance sensing (ECIS), a model for continuous monitoring of lightinduced changes in cells. 17 The proposed model is an ideal system for real-time continuous monitoring of HDFn cell behavior study with various natural drug concentrations, and this system imitates the in vivo environment.…”

In vitro hyperthermia studied in a continuous manner using electric impedance sensing

“…7 Previous studies reveal that the cell apoptosis temperature threshold is 43 C for many cell lines, such as HeLa cells. 22 With the aid of an automatic sensing system, detection and analyses could be automatic. Thus further research on mechanisms is needed to fully utilize and optimize hyperthermia treatments on a particular cancer.…”

“…22 With the aid of an automatic sensing system, detection and analyses could be automatic. 22 Although having been used to study drug-cell, 32 compound-cell, 19 electric pulses-cell, 33,34 or light-cell 22 interactions, the ECIS technique has never been published to research heat-cell interactions or hyperthermia. [23][24][25] Further, the high speed sensing system allows us to shorten the detection duration, which makes it possible to detect rapid changes in cellular impedance, for example, the rapid Ca 2+ concentration changes during hyperthermia can affect impedance.…”