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The Scientific World Journal
Volume 2014 (2014), Article ID 259765, 13 pages
http://dx.doi.org/10.1155/2014/259765
Research Article

Assessment of Cellular Responses after Short- and Long-Term Exposure to Silver Nanoparticles in Human Neuroblastoma (SH-SY5Y) and Astrocytoma (D384) Cells

1Laboratory of Clinical Toxicology, IRCCS Salvatore Maugeri Foundation, Scientific Institute of Pavia Medical Centre, Via Maugeri 10, 27100 Pavia, Italy
2Toxicology Division, Department of Environmental Health, IRCCS Salvatore Maugeri Foundation, Scientific Institute of Pavia Medical Centre, Via Maugeri 10, 27100 Pavia, Italy
3Toxicology Unit, Department of Clinical Surgical, Diagnostic and Pediatric Sciences, Faculty of Medicine and Surgery, University of Pavia, Via Maugeri 10, 27100 Pavia, Italy
4Department of Molecular Medicine, Institute of Biochemistry, University of Pavia, Via Forlanini 6, 27100 Pavia, Italy

Received 18 November 2013; Accepted 29 December 2013; Published 13 February 2014

Academic Editors: B. Soto-Blanco, A. Takagi, and D. N. Tripathi

Copyright © 2014 Teresa Coccini et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 μg/ml) and prolonged exposure (up to 10 days, 0.5–50 μg/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 μg/ml after 4 h: EC50s were 40.7 ± 2.0 and 49.5 ± 2.1 μg/ml for SH-SY5Y and D384, respectively). A549 were less vulnerable; (ii) AgNP doses of ≤ 18 μg/ml were noncytotoxic; (iii) AgNO3 induced more pronounced effects compared to AgNP on cerebral cells. Long-term data: (i) low AgNP doses (≤1 μg/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 μg/ml, and lower dose (0.5 μg/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.