title: The crocidolite fibres interaction with human mesothelial cells as investigated by combining electron microscopy, atomic force and scanning near-field optical microscopy.
abstract: In this study, we have performed a morphological analysis of crocidolite fibres interaction with mesothelial cells (MET5A) by combining conventional electron microscopy with atomic force (AFM) and scanning near-field optical microscopy (SNOM). After 6-h exposure at a crocidolite dose of 5 μg cm(-2), 90% of MET5A cells interact with fibres that under these conditions have a low cytotoxic effect. SEM images point out that fibres can be either engulfed by the cells that lose their typical morphology or they can accumulate over or partially inside the cells, which preserve their typical spread morphology. By using AFM we are able to directly visualize the entry-site of nanometric-sized fibres at the plasma membrane of the spread mesothelial cells. More importantly, the crocidolite fibres that are observed to penetrate the plasma membrane in SNOM topography can be simultaneously followed beneath the cell surface in the SNOM optical images. The analysis of SNOM data demonstrates the entrance of crocidolite fibres in proximity of nuclear compartment, as observed also in the TEM images. Our findings indicate that the combination of conventional electron microscopy with novel nanoscopic techniques can be considered a promising approach to achieve a comprehensive morphological description of the interaction between asbestos fibres and mesothelial cells that represents the early event in fibre pathogenesis.
authors.affiliation: Clinical Department of Medical, Chirurgical and Healthy Science, University of Trieste, Trieste 34127, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Andolfi
authors.forename: Elisa
authors.initials: E
authors.lastname: Trevisan
authors.forename: Marina
authors.initials: M
authors.lastname: Zweyer
authors.forename: Stefano
authors.initials: S
authors.lastname: Prato
authors.forename: Barbara
authors.initials: B
authors.lastname: Troian
authors.forename: Francesca
authors.initials: F
authors.lastname: Vita
authors.forename: Violetta
authors.initials: V
authors.lastname: Borelli
authors.forename: Maria Rosa
authors.initials: MR
authors.lastname: Soranzo
authors.forename: Mauro
authors.initials: M
authors.lastname: Melato
authors.forename: Giuliano
authors.initials: G
authors.lastname: Zabucchi
mesh_terms: D017638:Asbestos, Crocidolite / Q000378:metabolism*; D002460:Cell Line; D004848:Epithelium / Q000378:metabolism*; D006801:Humans; D008853:Microscopy
chemical_list: D017638:Asbestos, Crocidolite