title: Assessment of cellular responses after short- and long-term exposure to silver nanoparticles in human neuroblastoma (SH-SY5Y) and astrocytoma (D384) cells.
abstract: Silver nanoparticle (AgNP, 20 nm) neurotoxicity was evaluated by an integrated in vitro testing protocol employing human cerebral (SH-SY5Y and D384) cell lines. Cellular response after short-term (4-48 h, 1-100 μ g/ml) and prolonged exposure (up to 10 days, 0.5-50 μ g/ml) to AgNP was assessed by MTT, calcein-AM/PI, clonogenic tests. Pulmonary A549 cells were employed for data comparison along with silver nitrate as metal ionic form. Short-term data: (i) AgNP produced dose- and time-dependent mitochondrial metabolism changes and cell membrane damage (effects starting at 25 μ g/ml after 4 h: EC50s were 40.7 ± 2.0 and 49.5 ± 2.1 μ g/ml for SH-SY5Y and D384, respectively). A549 were less vulnerable; (ii) AgNP doses of ≤ 18 μ g/ml were noncytotoxic; (iii) AgNO3 induced more pronounced effects compared to AgNP on cerebral cells. Long-term data: (i) low AgNP doses (≤ 1 μ g/ml) compromised proliferative capacity of all cell types (cell sensibility: SHSY5Y > A549 > D384). Colony number decrease in SH-SY5Y and D384 was 50% and 25%, respectively, at 1 μ g/ml, and lower dose (0.5 μ g/ml) was significantly effective towards SH-SY5Y and pulmonary cells; (ii) cell proliferation activity was more affected by AgNO3 than AgNPs. In summary, AgNP-induced cytotoxic effects after short-term and prolonged exposure (even at low doses) were evidenced regardless of cell model types.
authors.affiliation: Laboratory of Clinical Toxicology, IRCCS Salvatore Maugeri Foundation, Scientific Institute of Pavia Medical Centre, Via Maugeri 10, 27100 Pavia, Italy ; Toxicology Division, Department of Environmental Health, IRCCS Salvatore Maugeri Foundation, Scientific Institute of Pavia Medical Centre, Via Maugeri 10, 27100 Pavia, Italy.
authors.forename: Teresa
authors.identifier: 0000-0002-2039-7944
authors.initials: T
authors.lastname: Coccini
authors.affiliation: Toxicology Division, Department of Environmental Health, IRCCS Salvatore Maugeri Foundation, Scientific Institute of Pavia Medical Centre, Via Maugeri 10, 27100 Pavia, Italy ; Toxicology Unit, Department of Clinical Surgical, Diagnostic and Pediatric Sciences, Faculty of Medicine and Surgery, University of Pavia, Via Maugeri 10, 27100 Pavia, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Manzo
authors.affiliation: Department of Molecular Medicine, Institute of Biochemistry, University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Bellotti
authors.affiliation: Toxicology Unit, Department of Clinical Surgical, Diagnostic and Pediatric Sciences, Faculty of Medicine and Surgery, University of Pavia, Via Maugeri 10, 27100 Pavia, Italy.
authors.forename: Uliana
authors.identifier: 0000-0002-8423-2954
authors.initials: U
authors.lastname: De Simone
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001254:Astrocytoma / Q000473:pathology / Q000503:physiopathology*; D001681:Biological Assay / Q000379:methods; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects*; D006801:Humans; D053768:Metal Nanoparticles / Q000633:toxicity*; D009447:Neuroblastoma / Q000473:pathology / Q000503:physiopathology*; D009498:Neurotoxins / Q000633:toxicity*; D012834:Silver / Q000633:toxicity*
chemical_list: D009498:Neurotoxins; D012834:Silver