@article {10552, title = {Combined use of AFM and soft X-ray microscopy to reveal fibres{\textquoteright} internalization in mesothelial cells.}, journal = {Analyst}, volume = {142}, year = {2017}, month = {2017 May 30}, pages = {1982-1992}, abstract = {

Nanotoxicology and nanomedicine investigations often require the probing of nano-objects such as fibres and particles in biological samples and cells, whilst internalization and intracellular destiny are the main issues for in vitro cellular studies. Various high resolution microscopy techniques are well suited for providing this highly sought-after information. However, sample preparation, nanomaterial composition and sectioning challenges make it often difficult to establish whether the fibres or particles have been internalized or they are simply overlaying or underlying the biological matter. In this paper we suggest a novel suitable combination of two different microscopic techniques to reveal in intact cells the uptake of asbestos fibres by mesothelial cells. After exposure to asbestos fibres and fixation, cells were first analysed under the AFM instrument and then imaged under the TwinMic soft X-ray microscope at Elettra Sincrotrone. The suggested approach combines standard soft X-ray microscopy imaging and AFM microscopy, with a common non-invasive sample preparation protocol which drastically reduces the experimental uncertainty and provides a quick and definitive answer to the nanoparticle cellular and tissue uptake.

}, keywords = {Asbestos, Cell Line, Epithelial Cells, Epithelium, Humans, Microscopy, Atomic Force, X-Rays}, issn = {1364-5528}, doi = {10.1039/c6an02661c}, author = {Gianoncelli, Alessandra and Kourousias, George and Cammisuli, Francesca and Cassese, Damiano and Rizzardi, Clara and Radillo, Oriano and Lazzarino, Marco and Pascolo, Lorella} } @article {8085, title = {Synchrotron X-ray microscopy reveals early calcium and iron interaction with crocidolite fibers in the lung of exposed mice.}, journal = {Toxicol Lett}, volume = {241}, year = {2016}, month = {2016 Jan 22}, pages = {111-20}, abstract = {

Human exposure to asbestos can cause a wide variety of lung diseases that are still a current major health concern, even if asbestos has been banned in many countries. It has been shown in many studies that asbestos fibers, ingested by alveolar macrophages, disrupt lung iron homeostasis by sequestering iron. Calcium can also be deposited on the fibers. The pathways along which iron and above all calcium interact with fibers are still unknown. Our aim was that of investigating if the iron accumulation induced by the inhaled asbestos fibers also involves calcium ions accumulation. Lung sections of asbestos-exposed mice were analyzed using an extremely sensitive procedure available at the synchrotron facilities, that provides morphological and chemical information based on X-ray fluorescence microspectroscopy (μ-XRF). In this study we show that (1) where conventional histochemical procedures revealed only weak deposits of iron and calcium, μ-XRF analysis is able to detect significant deposits of both iron and calcium on the inhaled asbestos fibers; (2) the extent of the deposition of these ions is proportionally directly related and (3) iron and calcium deposition on inhaled asbestos fibers is concomitant with the appearance of inflammatory and hyperplastic reactions.

}, issn = {1879-3169}, doi = {10.1016/j.toxlet.2015.11.016}, author = {Pascolo, Lorella and Zabucchi, Giuliano and Gianoncelli, Alessandra and Kourousias, George and Trevisan, Elisa and Pascotto, Ernesto and Casarsa, Claudia and Ryan, Chris and Lucattelli, Monica and Lungarella, Giuseppe and Cavarra, Eleonora and Bartalesi, Barbara and Zweyer, Marina and Cammisuli, Francesca and Melato, Mauro and Borelli, Violetta} }