Comparison of Impedance-based Sensors for Cell Adhesion Monitoring andIn VitroMethods for Detecting Cytotoxicity Induced by Chemicals

Abstract

The recent European Commission REACH (Registration, Evaluation and Authorisation of Chemicals) policy outlines a plan for toxicological testing by using alternative non-animal in vitro methods. In this context, there is a need to develop and standardise high-throughput screening (HTS) methods for studying the cytotoxicity induced by chemicals. Electrochemical impedance spectroscopy (EIS) can be considered as a complementary technique to alternative in vitro testing for studying cell adhesion to the substrate, and can give real-time and kinetic information on cell responses to a toxicant. This paper describes the development of a home-made chip based on impedance spectroscopy, and its application in studying the kinetics of BALB/3T3 cell adhesion and the cellular responses to a toxic product as a function of time. Concentrations of sodium arsenite, ranging from 10μM up to 1000μM, were tested in the system, and the results were compared with those obtained with standard protocols used to study basal cytotoxicity induced by chemicals in the BALB/3T3 cell line. The results show that the sensitivity of the developed chip was better than that with the MTT test, with the additional advantages of online monitoring.

Amorphous silica nanoparticles do not induce cytotoxicity, cell transformation or genotoxicity in Balb/3T3 mouse fibroblasts

“…It has been reported that cytotoxicity not only depends on the physicochemical properties of particles, but also varies with the cellular type investigated [22]. Therefore, even if our results are in contrast to other studies performed using different cell lines and different silica NPs, we are confident of the reliability and reproducibility of the CFE assay for testing NPs [49,54].…”

A panel ofin vitrotests to evaluate genotoxic and morphological neoplastic transformation potential onBalb/3T3cells by pristine and remediated titania and zirconia nanoparticles

“…In fact the cytome assay measures the frequency of cell division and the evaluation occurs directly after the end of the NP exposure. On the contrary, the CFE assay does not refer to a specific mechanism of cytotoxicity, and colonies can origin from any surviving cell following 7 days of culture and after 4 days by the end of the exposure period (20). Taken together these data suggest that ZrO 2 NP are able to induce cell death, however cells not too heavily damaged remain viable, continue to replicate themselves and are able to form colonies.…”

High throughput toxicity screening and intracellular detection of nanomaterials

“…The results were consistent with findings from the traditional MTT assay. 105 In summary, impedance-based methods for cytotoxicity testing are simple, cost-effective, labelfree, lend themselves to HTS, and importantly allow in situ monitoring of not just cytotoxicity but also other aspects of cell physiology such as proliferation, morphology, attachment, and intercellular adhesion. These methods are widely used in pharmaceutical research, oncology, cardiology, and other fields of biomedicine and clinical research, to investigate responses of cells to exposure to bioactive chemical agents and toxicants.…”