High-throughput electrochemical sensing platform for screening nanomaterial–biomembrane interactions

Abstract

Article:Owen, J orcid.org/0000-0003-0778-0858, Kuznecovs, M, Bhamji, R et al. (7 more authors) (2020) High-throughput electrochemical sensing platform for screening nanomaterial-biomembrane interactions. Review of Scientific Instruments, 91 (2). 025002. AbstractA high-throughput, automated screening platform has been developed for the assessment of biological membrane damage caused by nanomaterials. Membrane damage is detected using the technique of analysing capacitance-current peak changes obtained through rapid cyclic voltammetry (RCV) measurements of a phospholipid self-assembled monolayer, formed on a mercury film deposited onto a microfabricated platinum electrode, after the interaction of a biomembrane-active species. To significantly improve wider usability of the screening technique, a compact, high-throughput screening platform was designed, integrating the monolayer-supporting microfabricated electrode into a microfluidic flow cell, with bespoke pumps used for precise, automated control of fluid flow. Chlorpromazine, a tricyclic antidepressant, and citrate-coated 50 nm diameter gold nanomaterial (AuNM) were screened to successfully demonstrate the platform s viability for high-throughput screening. Chlorpromazine and AuNM showed interactions with a 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) monolayer at concentrations in excess of mol dm -3 . Biological validity of the electrochemically-measured interaction of chlorpromazine with DOPC monolayers was confirmed through quantitative comparisons with HepG2 and A549 cytotoxicity assays. The platform also demonstrated desirable performance for high-throughput screening, with membrane interactions detected in <6 min per assay. Automation contributed to this significantly, by reducing * Corresponding author: Joshua Owen (Email: J.J.Owen@leeds.ac.uk) the required operating skill level when using the technique and minimising fluid consumption. KeywordsPhospholipid monolayer, gold nanomaterial, biomembrane interaction, microfluidic flow cell, mercury electrode, rapid cyclic voltammetry, high-throughput screening IntroductionAdvances in nanotechnology have resulted in widespread usage of nanomaterials, often with applications in consumer products, biomedical and sensing technologies [1-4].However, toxicity hazards associated with nanomaterials have been widely reported, with growing research in the field of nanotoxicology emphasising the need for screening techniques to characterise nanomaterial hazards [5][6][7][8][9][10][11][12]. As the applications for engineered nanomaterials continues to grow, high-throughput, in vitro screening solutions are essential to accelerate the process of evaluating the toxicity of novel engineered nanomaterials and to meet the demand for hazard identification [13,14].High-throughput in vitro toxicity sensing technologies also provide an alternative to in vivo animal toxicity studies, which have ethical implications and are not economically feasible for screening a vast range of nanomaterials [3,15,16].Understanding cytotoxicity...

Microfluidic In Vitro Platform for (Nano)Safety and (Nano)Drug Efficiency Screening

“…CFD simulations of fluid flow at a flow rate of 100 μL h −1 and species transport through the microfluidic channel, using the methodology applied to study flow through a similar microfluidic geometry by the authors, [ 96 ] confirmed no undesirable flow characteristics were observed in the microfluidic channel and cell cavity. The concentration of species within the cavity reached that of the inflow concentration after approximately 700 s, a short time compared to the overall timescale of the exposure period (24 h) during an experiment.…”

Rapid Identification of in Vitro Cell Toxicity Using an Electrochemical Membrane Screening Platform

“…All other reagents were of analytical grade and purchased from Sigma-Aldrich. The microfabricated platinum electrodes [20,21,42,43] were supplied by the Tyndall National Institute (Cork, Ireland). Hg was electrodeposited on the Pt disc of radius 0.480 mm to give a Pt/Hg electrode as described previously [20,21,42,43].…”

Advances in Microtechnology for Improved Cytotoxicity Assessment

“…Membrane-on-chip microplatforms emerge for high throughput membrane sensing, giving information of the events happening on a supported cellular membrane. The most used membrane biosensing microtechnology comprises electrochemical sensors fabricated with self-assembled monolayers supported on mercury electrodes that provide information of the interactions or damage on the membrane (Owen et al, 2020;Rashid et al, 2017). Recently, Owen et al reported an automated, rapid and HTP sensing microfluidic platform for the screening of nanomaterial-biomembrane interactions (Owen et al, 2020).…”

“…The most used membrane biosensing microtechnology comprises electrochemical sensors fabricated with self-assembled monolayers supported on mercury electrodes that provide information of the interactions or damage on the membrane (Owen et al, 2020;Rashid et al, 2017). Recently, Owen et al reported an automated, rapid and HTP sensing microfluidic platform for the screening of nanomaterial-biomembrane interactions (Owen et al, 2020). The integrated microfluidic device provided electrochemical information of the interactions of chlorpromazine and citrate coated gold nanomaterial with 1,2-dioleoyl-sn-glycero-3-phosphocoline membranes, supposing an easy-to-use interface to assess cytotoxicity based on membrane damage (less than 6 min per assay).…”