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Genotoxic potential of TiO2 on bottlenose dolphin leukocytes

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Abstract

Titanium dioxide is extensively used in a variety of products, including industrial materials and cosmetics. Studies mainly performed on human cell lines and in vivo exposure on experimental animals have raised concern about the toxic effects of ultrafine titanium dioxide; however, scarce information is available about its impact on aquatic life. The aim of this article was to assess the genotoxic potential of TiO2 (anatase and rutile) on bottlenose dolphin leukocytes. Blood samples were obtained from four male and one female specimens reared at the Adriatic SeaWorld “Oltremare” (Riccione, Italy). Leukocytes were isolated by the lyses procedure and in vitro exposed to TiO2 in RPMI. Experimental solutions were sonicated immediately before dosing the cells. Three exposure times (4, 24 and 48 h) and three doses (20, 50 and 100 µg/ml) were tested. Genotoxicity was detected by the single-cell gel electrophoresis (or comet assay) at pH ≥ 13, assessing single/double-strand breaks and alkali-labile sites. Cytotoxicity was also detected by the Trypan blue exclusion method. Results showed that both the crystalline forms of TiO2 were genotoxic for bottlenose dolphin leukocytes, with a statistically significant increase of DNA fragmentation after exposure to 50 and 100 µg/ml for 24 and 48 h. Although preliminary, these are the first data regarding the genetic susceptibility of toothed cetaceans toward an “emerging” pollutant, such as TiO2 particles.

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Acknowledgments

Authors are indebted with Oltremare srl and its staff for their collaboration in providing us the bottlenose blood samples. Thanks to Dr. Paolo Lucchesi for helping us with TEM observations, and to Claudio Ghezzani with digital images processing.

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Authors and Affiliations

  1. Dipartimento di Morfologia Umana e Biologia Applicata, University of Pisa, Via Volta 4, 56126, Pisa, Italy

    Margherita Bernardeschi, Patrizia Guidi, Vittoria Scarcelli, Giada Frenzilli & Marco Nigro

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  1. Margherita Bernardeschi
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  2. Patrizia Guidi
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  3. Vittoria Scarcelli
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  4. Giada Frenzilli
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  5. Marco Nigro
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Correspondence to Marco Nigro.

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Bernardeschi, M., Guidi, P., Scarcelli, V. et al. Genotoxic potential of TiO2 on bottlenose dolphin leukocytes. Anal Bioanal Chem 396, 619–623 (2010). https://doi.org/10.1007/s00216-009-3261-3

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  • DOI: https://doi.org/10.1007/s00216-009-3261-3

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