In Vitro Genotoxicity of Polystyrene Nanoparticles on the Human Fibroblast Hs27 Cell Line
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Polystyrene Nanoparticles
2.3. Saffron: Crocus sativus L. Stigmas Extract
2.4. Cell–Growth Curve
2.5. MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] Test
2.6. ROS (Reactive Oxygen Species) Detection
2.7. Cytokinesis-Block Micronucleus (CBMN) Assay
2.8. Analysis of Polystyrene Particles (PNPs) by Scanning Electron Microscopy SEM
2.9. Statistical Analysis
3. Results
3.1. Cell Growth Curve and MTS
3.2. Tests of Micronuclei with Block of Cytokinesis with Cytochalasin B “CBMN Assay”
3.3. ROS Detection
3.4. Analysis of PNPs by SEM Scanning Electron Microscopy
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- United Nations Environment Programme. Division of Early Warning, and Assessment. In UNEP Year Book 2011: Emerging Issues in Our Global Environment; UNEP/Earthprint: Genève, Switzerland, 2011. [Google Scholar]
- Andrady, A.L. Microplastics in the marine environment. Mar. Pollut. Bull. 2011, 62, 1596–1605. [Google Scholar] [CrossRef] [PubMed]
- Cózar, A.; Echevarría, F.; González-Gordillo, J.I.; Irigoien, X.; Úbeda, B.; Hernández-León, S.; Fernández-de-Puelles, M.L. Plastic debris in the open ocean. Proc. Natl. Acad. Sci. USA 2014, 111, 10239–10244. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ter Halle, A.; Jeanneau, L.; Martignac, M.; Jardé, E.; Pedrono, B.; Brach, L.; Gigault, J. Nanoplastic in the North Atlantic subtropical gyre. Environ. Sci. Technol. 2017, 51, 13689–13697. [Google Scholar] [CrossRef] [PubMed]
- Mattsson, K.; Jocic, S.; Doverbratt, I.; Hansson, L.A. Nanoplastics in the aquatic environment. In Microplastic Contamination in Aquatic Environments; Elsevier: Amsterdam, The Netherlands, 2018; pp. 379–399. [Google Scholar] [CrossRef]
- Browne, M.A.; Niven, S.J.; Galloway, T.S.; Rowland, S.J.; Thompson, R.C. Microplastic moves pollutants and additives to worms, reducing functions linked to health and biodiversity. Curr. Biol. 2013, 23, 2388–2392. [Google Scholar] [CrossRef] [PubMed]
- Della Torre, C.; Bergami, E.; Salvati, A.; Faleri, C.; Cirino, P.; Dawson, K.A.; Corsi, I. Accumulation and embryotoxicity of polystyrene nanoparticles at early stage of development of sea urchin embryos Paracentrotus lividus. Environ. Sci. Technol. 2014, 48, 12302–12311. [Google Scholar] [CrossRef] [PubMed]
- Phuong, N.N.; Zalouk-Vergnoux, A.; Poirier, L.; Kamari, A.; Châtel, A.; Mouneyrac, C.; Lagarde, F. Is there any consistency between the microplastics found in the field and those used in laboratory experiments? Environ. Pollut. 2016, 211, 111–123. [Google Scholar] [CrossRef] [PubMed]
- Ma, Y.; Huang, A.; Cao, S.; Sun, F.; Wang, L.; Guo, H.; Ji, R. Effects of nanoplastics and microplastics on toxicity, bioaccumulation, and environmental fate of phenanthrene in fresh water. Environ. Pollut. 2016, 219, 166–173. [Google Scholar] [CrossRef] [PubMed]
- Sussarellu, R.; Suquet, M.; Thomas, Y.; Lambert, C.; Fabioux, C.; Pernet, M.E.J.; Corporeau, C. Oyster reproduction is affected by exposure to polystyrene microplastics. Proc. Natl. Acad. Sci. USA 2016, 113, 2430–2435. [Google Scholar] [CrossRef] [Green Version]
- Li, J.; Qu, X.; Su, L.; Zhang, W.; Yang, D.; Kolandhasamy, P.; Shi, H. Microplastics in mussels along the coastal waters of China. Environ. Pollut. 2016, 214, 177–184. [Google Scholar] [CrossRef]
- Horton, A.A.; Walton, A.; Spurgeon, D.J.; Lahive, E.; Svendsen, C. Microplastics in freshwater and terrestrial environments: Evaluating the current understanding to identify the knowledge gaps and future research priorities. Sci. Total Environ. 2017, 586, 127–141. [Google Scholar] [CrossRef] [Green Version]
- Cole, M.; Galloway, T.S. Ingestion of nanoplastics and microplastics by Pacific oyster larvae. Environ. Sci. Technol. 2015, 49, 14625–14632. [Google Scholar] [CrossRef] [PubMed]
- Cole, M.; Lindeque, P.; Fileman, E.; Halsband, C.; Galloway, T.S. The impact of polystyrene microplastics on feeding, function and fecundity in the marine copepod Calanus helgolandicus. Environ. Sci. Technol. 2015, 49, 1130–1137. [Google Scholar] [CrossRef] [PubMed]
- Gigault, J.; Ter Halle, A.; Baudrimont, M.; Pascal, P.Y.; Gauffre, F.; Phi, T.L.; Reynaud, S. Current opinion: What is a nanoplastic? Environ. Pollut. 2018, 235, 1030–1034. [Google Scholar] [CrossRef] [PubMed]
- Lambert, S.; Wagner, M. Formation of microscopic particles during the degradation of different polymers. Chemosphere 2016, 161, 510–517. [Google Scholar] [CrossRef] [PubMed]
- Alimi, O.S.; Farner Budarz, J.; Hernandez, L.M.; Tufenkji, N. Microplastics and nanoplastics in aquatic environments: Aggregation, deposition, and enhanced contaminant transport. Environ. Sci. Technol. 2018, 52, 1704–1724. [Google Scholar] [CrossRef] [PubMed]
- Bouwmeester, H.; Hollman, P.C.; Peters, R.J. Potential health impact of environmentally released micro-and nanoplastics in the human food production chain: Experiences from nanotoxicology. Environ. Sci. Technol. 2015, 49, 8932–8947. [Google Scholar] [CrossRef]
- Verma, A.; Stellacci, F. Effect of surface properties on nanoparticle–cell interactions. Small 2010, 6, 12–21. [Google Scholar] [CrossRef]
- Wright, S.L.; Kelly, F.J. Plastic and human health: A micro issue? Environ. Sci. Technol. 2017, 51, 6634–6647. [Google Scholar] [CrossRef]
- Ward, J.E.; Kach, D.J. Marine aggregates facilitate ingestion of nanoparticles by suspension-feeding bivalves. Mar. Environ. Res. 2009, 68, 137–142. [Google Scholar] [CrossRef] [Green Version]
- Wegner, A.; Besseling, E.; Foekema, E.M.; Kamermans, P.; Koelmans, A.A. Effects of nanopolystyrene on the feeding behavior of the blue mussel (Mytilus edulis L.). Environ. Toxic. Chem. 2012, 31, 2490–2497. [Google Scholar] [CrossRef]
- Bhattacharya, P.; Lin, S.J.; Turner, J.P.; Ke, P.C. Physical adsorption of charged plastic nanoparticles affects algal photosynthesis. J. Phys. Chem. C 2010, 114, 16556–16561. [Google Scholar] [CrossRef]
- Brillant, M.; MacDonald, B. Postingestive selection in the sea scallop (Placopectenmagellanicus) on the basis of chemical properties of particles. Mar. Biol. 2002, 141, 457–465. [Google Scholar] [CrossRef]
- Cedervall, T.; Hansson, L.A.; Lard, M.; Frohm, B.; Linse, S. Food chain transport of nanoparticles affects behaviour and fat metabolism in fish. PLoS ONE 2012, 7, e32254. [Google Scholar] [CrossRef] [PubMed]
- Hussain, N.; Jani, P.U.; Florence, A.T. Enhanced oral uptake of tomato lectin-conjugated nanoparticles in the rat. Pharm. Res. 1997, 14, 613–618. [Google Scholar] [CrossRef] [PubMed]
- Kulkarni, S.A.; Feng, S.S. Effects of particle size and surface modification on cellular uptake and biodistribution of polymeric nanoparticles for drug delivery. Pharm. Res. 2013, 30, 2512–2522. [Google Scholar] [CrossRef] [PubMed]
- Walczak, A.P.; Kramer, E.; Hendriksen, P.J.; Helsdingen, R.; van der Zande, M.; Rietjens, I.M.; Bouwmeester, H. In vitro gastrointestinal digestion increases the translocation of polystyrene nanoparticles in an in vitro intestinal co-culture model. Nanotoxicology 2015, 9, 886–894. [Google Scholar] [CrossRef] [PubMed]
- EFSA Scientific Committee. Guidance on the risk assessment of the application of nanoscience and nanotechnologies in the food and feed chain. EFSA J. 2011, 9, 2140. [Google Scholar] [CrossRef]
- Saquib, Q.; Faisal, M.; Al-Khedhairy, A.A.; Alatar, A.A. (Eds.) Cellular and Molecular Toxicology of Nanoparticles; Springer: New York, NY, USA, 2018; Volume 1048. [Google Scholar] [CrossRef]
- Karlsson, H.L. The comet assay in nanotoxicology research. Anal. Bioanal. Chem. 2010, 398, 651–666. [Google Scholar] [CrossRef]
- Xie, H.; Mason, M.M.; Wise, J.P. Genotoxicity of metal nanoparticles. Rev. Environ. Health 2011, 26, 251–268. [Google Scholar] [CrossRef]
- Hernandez, L.M.; Yousefi, N.; Tufenkji, N. Are there nanoplastics in your personal care products? Environ. Sci. Technol. Lett. 2017, 4, 280–285. [Google Scholar] [CrossRef]
- Singh, N.; Manshian, B.; Jenkins, G.J.; Griffiths, S.M.; Williams, P.M.; Maffeis, T.G.; Doak, S.H. NanoGenotoxicology: The DNA damaging potential of engineered nanomaterials. Biomaterials 2009, 30, 3891–3914. [Google Scholar] [CrossRef] [PubMed]
- Anderson, A.G.; Grose, J.; Pahl, S.; Thompson, R.C.; Wyles, K.J. Microplastics in personal care products: Exploring perceptions of environmentalists, beauticians and students. Marine Pollut. Bullet. 2016, 113, 454–460. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Menghini, L.; Leporini, L.; Vecchiotti, G.; Locatelli, M.; Carradori, S.; Ferrante, C.; Brunetti, L. Crocus sativus L. stigmas and byproducts: Qualitative fingerprint, antioxidant potentials and enzyme inhibitory activities. Food Res. Int. 2018, 109, 91–98. [Google Scholar] [CrossRef] [PubMed]
- Lahmass, I.; Lamkami, T.; Delporte, C.; Sikdar, S.; Van Antwerpen, P.; Saalaoui, E.; Megalizzi, V. The waste of saffron crop, a cheap source of bioactive compounds. J. Funct. Foods 2017, 35, 341–351. [Google Scholar] [CrossRef]
- Cory, A.H.; Owen, T.C.; Barltrop, J.A.; Cory, J.G. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun. 1991, 3, 207–212. [Google Scholar] [CrossRef] [PubMed]
- Fenech, M. Cytokinesis-block micronucleus cytome assay. Nat. Protoc. 2007, 2, 1084–1104. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- OECD Guidelines for the Testing of Chemicals, Section 4: Health Effects. Test No. 487. Vitro Mammalian Cell Micronucleus Test. Available online: www.oecd-ilibrary.org (accessed on 4 September 2019).
- Andrady, A.L.; Neal, M.A. Applications and Societal Benefits of Plastics. Philos. Trans. R. Soc. Lond. Ser. B 2009, 364, 1977–1984. [Google Scholar] [CrossRef] [PubMed]
- Jambeck, J.R.; Geyer, R.; Wilcox, C.; Siegler, T.R.; Perryman, M.; Andrady, A.; Narayan, R.; Law, K.L. Marine pollution. Plastic waste inputs from land into the ocean. Science 2015, 347, 768–771. [Google Scholar] [CrossRef] [PubMed]
- Wilcox, C.; Van Sebille, E.; Hardesty, B.D. Threat of plastic pollution to seabirds is global, pervasive, and increasing. Proc. Natl. Acad. Sci. USA 2015, 112, 11899–11904. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gregory, M.R. Environmental implications of plastic debris in marine settings—Entanglement, ingestion, smothering, hangers-on, hitch-hiking and alien invasions. Philos. Trans. R. Soc. Lond. Ser. B Biol. Sci. 2009, 364, 2013–2025. [Google Scholar] [CrossRef] [PubMed]
- Mrosovsky, N.; Ryan, G.D.; James, M.C. Leatherback turtles: The menace of plastic. Mar. Pollut. Bull. 2009, 58, 287–289. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Laist, D.W. Impacts of marine debris: Entanglement of marine life in marine debris including a comprehensive list of species with entanglement and ingestion records. In Marine Debris; Springer: New York, NY, USA, 1997; pp. 99–139. [Google Scholar] [CrossRef]
- Mishra, P.; Vinayagam, S.; Duraisamy, K.; Patil, S.R.; Godbole, J.; Mohan, A.; Chandrasekaran, N. Distinctive impact of polystyrene nano-spherules as an emergent pollutant toward the environment. Environ. Sci. Poll. Res. 2019, 26, 1537–1547. [Google Scholar] [CrossRef] [PubMed]
- Bukhari, S.I.; Manzoor, M.; Dhar, M.K. A comprehensive review of the pharmacological potential of Crocus sativus and its bioactive apocarotenoids. Biomed. Pharmacother. 2018, 98, 733–745. [Google Scholar] [CrossRef] [PubMed]
- Frankel, E.N. Nutritional benefits of flavonoids. In Food Factors for Cancer Prevention; Springer: Tokyo, Japan, 1997; pp. 613–616. [Google Scholar] [CrossRef]
- Xia, T.; Kovochich, M.; Brant, J.; Hotze, M.; Sempf, J.; Oberley, T.; Nel, A.E. Comparison of the abilities of ambient and manufactured nanoparticles to induce cellular toxicity according to an oxidative stress paradigm. Nano Lett. 2006, 6, 1794–1807. [Google Scholar] [CrossRef] [PubMed]
- Rabinovitch, M. Professional and non-professional phagocytes: An introduction. Trends Cell Biol. 1995, 5, 85–87. [Google Scholar] [CrossRef]
- Rodriguez-Seijo, A.; Lourenço, J.; Rocha-Santos, T.A.P.; Da Costa, J.; Duarte, A.C.; Vala, H.; Pereira, R. Histopathological and molecular effects of microplastics in Eisenia andrei Bouché. Environ. Pollut. 2017, 220, 495–503. [Google Scholar] [CrossRef] [PubMed]
- Pedà, C.; Caccamo, L.; Fossi, M.C.; Gai, F.; Andaloro, F.; Genovese, L.; Maricchiolo, G. Intestinal alterations in European sea bass Dicentrarchus labrax (Linnaeus, 1758) exposed to microplastics: Preliminary results. Environ. Pollut. 2016, 212, 251–256. [Google Scholar] [CrossRef]
- Chua, E.M.; Shimeta, J.; Nugegoda, D.; Morrison, P.D.; Clarke, B.O. Assimilation of polybrominated diphenyl ethers from microplastics by the marine amphipod, Allorchestes compressa. Environ. Sci. Technol. 2014, 48, 8127–8134. [Google Scholar] [CrossRef]
- Koelmans, A.A.; Besseling, E.; Foekema, E.M. Leaching of plastic additives to marine organisms. Environ. Pollut. 2014, 187, 49–54. [Google Scholar] [CrossRef]
- Nowack, B.; Ranville, J.F.; Diamond, S.; Gallego-Urrea, J.A.; Metcalfe, C.; Rose, J.; Horne, N.; Koelmans, A.A.; Klaine, S.J. Potential scenarios for nanomaterial release and subsequent alteration in the environment. Environ. Toxicol. Chem. 2012, 31, 50–59. [Google Scholar] [CrossRef]
Time Ratio | 5 µg/mL | P Value | 25 µg/mL | P Value | ||
---|---|---|---|---|---|---|
PNPs | PNPs with Crocus s. | PNPs | PNPs with Crocus s. | |||
T15 min/T0 | 1.4 ± 0.004 | 1.01 ± 0.004 | <0.05 | 1.13 ± 0.005 | 0.99 ± 0.002 | |
T30 min/T0 | 1.60 ± 1.2 × 10−5 | 1.10 ± 0.001 | <0.0005 | 1.41 ± 0.007 | 1.16 ± 0.001 | <0.05 |
T1 h/T0 | 0.95 ± 0.0013 | 0.75 ± 0.001 | <0.005 | 0.99 ± 0.007 | 0.78 ± 0.002 | <0.05 |
© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Poma, A.; Vecchiotti, G.; Colafarina, S.; Zarivi, O.; Aloisi, M.; Arrizza, L.; Chichiriccò, G.; Di Carlo, P. In Vitro Genotoxicity of Polystyrene Nanoparticles on the Human Fibroblast Hs27 Cell Line. Nanomaterials 2019, 9, 1299. https://doi.org/10.3390/nano9091299
Poma A, Vecchiotti G, Colafarina S, Zarivi O, Aloisi M, Arrizza L, Chichiriccò G, Di Carlo P. In Vitro Genotoxicity of Polystyrene Nanoparticles on the Human Fibroblast Hs27 Cell Line. Nanomaterials. 2019; 9(9):1299. https://doi.org/10.3390/nano9091299
Chicago/Turabian StylePoma, Anna, Giulia Vecchiotti, Sabrina Colafarina, Osvaldo Zarivi, Massimo Aloisi, Lorenzo Arrizza, Giuseppe Chichiriccò, and Piero Di Carlo. 2019. "In Vitro Genotoxicity of Polystyrene Nanoparticles on the Human Fibroblast Hs27 Cell Line" Nanomaterials 9, no. 9: 1299. https://doi.org/10.3390/nano9091299