A HepG2 Cell-Based Biosensor That Uses Stainless Steel Electrodes for Hepatotoxin Detection

Abstract

Humans are frequently exposed to environmental hepatotoxins, which can lead to liver failure. Biosensors may be the best candidate for the detection of hepatotoxins because of their high sensitivity and specificity, convenience, time-saving, low cost, and extremely low detection limit. To investigate suitability of HepG2 cells for biosensor use, different methods of adhesion on stainless steel surfaces were investigated, with three groups of experiments performed in vitro. Cytotoxicity assays, which include the resazurin assay, the neutral red assay (NR), and the Coomassie Brilliant Blue (CBB) assay, were used to determine the viability of HepG2 cells exposed to various concentrations of aflatoxin B1 (AFB1) and isoniazid (INH) in parallel. The viability of the HepG2 cells on the stainless steel surface was quantitatively and qualitatively examined with different microscopy techniques. A simple cell-based electrochemical biosensor was developed by evaluating the viability of the HepG2 cells on the stainless steel surface when exposed to various concentrations of AFB1 and INH by using electrochemical impedance spectroscopy (EIS). The results showed that HepG2 cells can adhere to the metal surface and could be used as part of the biosensor to determine simple hepatotoxic samples.

Electrodeposited folic acid platinum electrode for rapid detection of human hepatocellular carcinoma cells

“…The CV measurement was recorded in 0.1 M PBS (pH 7.2) containing 5 mM K 4 [Fe(CN) 6 ] and 0.1 M KCl in the potential region of À 0.3 V to + 0.9 V. This potential window was chosen as the oxidation (300 mV) and reduction (200 mV) potential peak of the electrolyte solution lies in this region [39]. The use of K 4 [Fe(CN) 6 ] in an electrolyte is advantageous due to its well-defined, stable, and reversible redox potential [13,40]. Particularly the low oxidation potential of K 4 [Fe(CN) 6 ] helps avoid interference with the occurrence of electrochemical responses of desired biomolecule species in the study.…”

“…Researchers have fabricated various analytical devices to detect FA such as spectrophotometry [8], high-performance liquid chromatography (HPLC) [9], colorimetric [10], flow injection chemiluminescence [11] and fluorimetric [12]. However, these methods possess challenges like highcost instrumental setup, expertise in operating the instruments, sample pre-treatment, and time-consuming procedure [13]. To tackle these issues, electrochemical in-situ detection-based methods have gained attention to investigate the presence of biomarkers [14][15][16][17][18][19][20].…”

“…A study by Rozman et. al [24] demonstrated the capability of HepG2 cell line to adhere to a metal surface which was employed as a working electrode for sensing hepatotoxins. Hence, HepG2 cells pose as an excellent biorecognition element for cell-based biosensing applications owing to their suitability in sensing a range of biomolecules and can offer flexibility in designing the sensing strategies [25].…”

Stainless Steel Foil-Based Label-Free Modular Thin-Film Electrochemical Detector for Solvent Identification

“…Electrochemical impedance spectroscopy (EIS) was performed using the PalmSens4 instrument (PalmSens, Houten, The Netherlands). An amplitude of 10 mV against a fixed potential of 0 V was used [29,30] and scans were performed in a frequency range from 500 kHz to 1 Hz with 9.8 points per decade (57 points in total per measurement). For all measurements, the working electrode (WE) terminal was connected to one of the smaller electrodes (the electrode under study), whereas the counter electrode (CE) and the reference electrode (RE) terminals were combined and connected to the larger electrode (cf.…”

Nanomaterials and Their Recent Applications in Impedimetric Biosensing

“…Biosensors represent small and cost-effective analytical tools that convert a biological response into an electrical signal and indicate the concentration of the target analyte [5,6]. The type of analyte composition, the biologically active component, the biosensor's design, and the transducer's physical characteristics affect this information's precision [6,7].…”