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Acute Cytotoxic Effects on Morphology and Mechanical Behavior in MCF-7 Induced by TiO2NPs Exposure.

International journal of molecular sciences (2019-07-26)
Mariafrancesca Cascione, Valeria De Matteis, Giacomo Mandriota, Stefano Leporatti, Rosaria Rinaldi
ABSTRACT

The side effects induced by nanoparticle exposure at a cellular level are one of the priority research topics due to the steady increase in the use of nanoparticles (NPs). Recently, the focus on cellular morphology and mechanical behavior is gaining relevance in order to fully understand the cytotoxic mechanisms. In this regard, we have evaluated the morphomechanical alteration in human breast adenocarcinoma cell line (MCF-7) exposed to TiO2NPs at two different concentrations (25 and 50 µg/mL) and two time points (24 and 48 h). By using confocal and atomic force microscopy, we demonstrated that TiO2NP exposure induces significant alterations in cellular membrane elasticity, due to actin proteins rearrangement in cytoskeleton, as calculated in correspondence to nuclear and cytoplasmic compartments. In this work, we have emphasized the alteration in mechanical properties of the cellular membrane, induced by nanoparticle exposure.

MATERIALS
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Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture
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Triton X-100, laboratory grade
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DAPI, for nucleic acid staining
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Cell Counting Kit - 8, for quantitation of viable cell number in proliferation and cytotoxicity assays
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Glutaraldehyde solution, Grade I, 25% in H2O, specially purified for use as an electron microscopy fixative
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Dulbecco′s Phosphate Buffered Saline, 10×, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
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Phalloidin, Fluorescein Isothiocyanate Labeled, peptide from Amanita phalloides