Effect of ZnO Nanoparticles on Human Bone Marrow Mesenchymal Stem Cells: Viability, Morphology, Particles Uptake, Cell Cycle and Metabolites

Abstract

Despite the growing interest in nanoparticles (NPs), the evaluation of their safety use has to be deeply considered, but standardized procedures for the evaluation of their toxicity have not been defined. In vitro methods are ideal in toxicology research because they can rapidly provide reproducible results while preventing the use of animals. Primary cells are considered a better option as model systems for predicting toxicological behavior, although several cell types do not survive enough in culture and isolated cells can have substantial variability when obtained from different donors. Recently, a new test for acute toxicity based on the use of human bone marrow mesenchymal stem cells (hBMMSCs) has been developed and successfully tested in our laboratory following the ICCVAM (Interagency Coordinating Committee on the Validation of Alternative Methods) guidelines [1]. Along these lines, the aim of this study is to evaluate the acute cytotoxicity of ZnO nanoparticles using the new toxicity test based on hBMMSCs, while comparing their behavior with respect to the toxicity of ZnO micrometer ones. For this reason, we assessed the citotoxicity by performing Neutral Red assay, the cellular uptake by transmission electron microscopy and the effects on hBMMSCs cycle by FACS analysis. Furthermore, we also analyzed by means of GC-MS the polar metabolite profile of hBMMSCs samples treated with ZnO micro- and nanoparticles. Our results show that despite the slight differences in terms of cytotoxicity, nano and microparticles show a very different behavior with respect to their effects on hBMMSCs cycle, metabolite profile and cellular uptake.