title: Cytotoxicity, oxidative stress and genotoxicity induced by glass fibers on human alveolar epithelial cell line A549.
abstract: Man-made vitreous fibers have been widely used as insulation material as asbestos substitutes; however their morphology and composition raises concerns. In 1988 the International Agency for Research on Cancer classified fiberglass, rock wool, slag wool, and ceramic fibers as Group 2B, i.e. possibly carcinogenic to humans. In 2002 it reassigned fiberglass, rock and slag wool, and continuous glass filaments to Group 3, not classifiable as carcinogenic to humans. The aim of this study was to verify the cytotoxic and genotoxic effects and oxidative stress production induced by in vitro exposure of human alveolar epithelial cells A549 to glass fibers with a predominant diameter <3 μm (97%) and length >5 μm (93%). A549 cells were incubated with 5, 50, or 100 μg/ml (2.1, 21, and 42 μg/cm(2), respectively) of glass fibers for 72 h. Cytotoxicity and DNA damage were tested by the MTT and the Comet assay, respectively. Oxidative stress was determined by measuring inducible nitric oxide synthase (iNOS) expression by Western blotting, production of nitric oxide (NO) with Griess reagent, and concentration of reactive oxygen species by fluorescent quantitative analysis with 2',7'-dichlorofluorescein-diacetate (DCFH-DA). The results showed that glass fiber exposure significantly reduced cell viability and increased DNA damage and oxidative stress production in a concentration-dependent manner, demonstrating that glass fibers exert cytotoxic and genotoxic effects related to increased oxidative stress on the human alveolar cell line A549.
authors.affiliation: Division of Occupational Medicine, University Hospital "Policlinico - Vittorio Emanuele", University of Catania, via Santa Sofia, 78 95123 Catania, Italy.
authors.forename: Venerando
authors.initials: V
authors.lastname: Rapisarda
authors.affiliation: Department of Bio-Medical Sciences, Human Anatomy and Histology Section, School of Medicine, University of Catania, via S. Sofia, 87 95123 Catania, Italy. Electronic address: carla.loreto@unict.it.
authors.forename: Carla
authors.initials: C
authors.lastname: Loreto
authors.affiliation: Hygiene and Public Health, Department of G.F. Ingrassia, University of Catania, via Santa Sofia, 87 95123 Catania, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Ledda
authors.affiliation: Department of Bio-Medical Sciences, Human Anatomy and Histology Section, School of Medicine, University of Catania, via S. Sofia, 87 95123 Catania, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Musumeci
authors.affiliation: Occupational Medicine, Department of Clinical and Molecular Sciences, Polytechnic University of Marche, via Tronto 10/a, 60020 Torrette, Ancona, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Bracci
authors.affiliation: Occupational Medicine, Department of Clinical and Molecular Sciences, Polytechnic University of Marche, via Tronto 10/a, 60020 Torrette, Ancona, Italy.
authors.forename: Lory
authors.initials: L
authors.lastname: Santarelli
authors.affiliation: Department of Drug's Sciences, Section of Biochemistry, University of Catania, Viale Andrea Doria, 6, 95125 Catania, Italy.
authors.forename: Marcella
authors.initials: M
authors.lastname: Renis
authors.affiliation: Department of Bio-Medical Sciences, Human Anatomy and Histology Section, School of Medicine, University of Catania, via S. Sofia, 87 95123 Catania, Italy.
authors.forename: Margherita
authors.initials: M
authors.lastname: Ferrante
authors.affiliation: Department of Bio-Medical Sciences, Section of Physiology, University of Catania, Viale Andrea Doria, 6, 95125 Catania, Italy.
authors.forename: Venera
authors.initials: V
authors.lastname: Cardile
mesh_terms: D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects*; D004249:DNA Damage; D005898:Glass; D006801:Humans; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010316:Particle Size; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D009153:Mutagens; D009573:Nitrites; D017382:Reactive Oxygen Species; C009753:fiberglass; D052247:Nitric Oxide Synthase Type II
keywords: DNA damage; Glass fibers; Human alveolar cells; Occupational exposure; ROS; iNOS