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

Martin Rozman,Urban Bren,Zala Štukovnik,Ajda Sušnik,Amirhossein Pakseresht,Damjana Drobne,Matej Hočevar

doi:10.3390/bios12030160

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.

Highlights

IntroductionIn addition to liver damage caused by unhealthy lifestyles and viruses including hepatitis A, B, C, D, and E viruses, humans are frequently exposed to environmental hepatotoxins that can cause liver failure, acute cellular damage, or chronic diseases such as fibrosis and liver cancer [1]
To data presented in the neutral red test, aflatoxin B1 (AFB1) concentrations above 10 μM showed a reduction in the number of cells in the cell population and that INH concentrations above 10 mM had a significant effect on the number of cells
Neutral red test, AFB1 concentrations above 10 μM showed a reduction in the number of cells in the cell population and that INH concentrations above 10 mM had a signifi‐

Summary

Introduction

In addition to liver damage caused by unhealthy lifestyles and viruses including hepatitis A, B, C, D, and E viruses, humans are frequently exposed to environmental hepatotoxins that can cause liver failure, acute cellular damage, or chronic diseases such as fibrosis and liver cancer [1]. Hepatotoxins include mycotoxins such as aflatoxins, which are the key concern in the food industry [2,3] and are mainly produced by Aspergillus flavus and Aspergillus parasiticus [4]. Examples of hepatotoxins include monomethylhydrazine, amatoxins, orellanine [5] and amanitin, commonly present in the fungal families of Agaricaceae

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