Neurotoxicity Research

, Volume 32, Issue 2, pp 187–203 | Cite as

Neurobehavioural Toxicity of Iron Oxide Nanoparticles in Mice

  • Vasanth Dhakshinamoorthy
  • Vijayprakash Manickam
  • Ekambaram Perumal
ORIGINAL ARTICLE

Abstract

Iron oxide nanoparticles (Fe2O3-NPs) are widely used in various biomedical applications, extremely in neurotheranostics. Simultaneously, Fe2O3-NP usage is of alarming concern, as its exposure to living systems causes deleterious effects due to its redox potential. However, study on the neurobehavioural impacts of Fe2O3-NPs is very limited. In this regard, adult male mice were intraperitoneally administered with Fe2O3-NPs (25 and 50 mg/kg body weight) once a week for 4 weeks. A significant change in locomotor behaviour and spatial memory was observed in Fe2O3-NP-treated animals. Damages to blood–brain barrier permeability by Fe2O3-NPs and their accumulation in brain regions were evidenced by Evan’s blue staining, iron estimation and Prussian blue staining. Elevated nitric oxide, acetylcholinesterase, lactate dehydrogenase leakage and demyelination were observed in the Fe2O3-NP-exposed brain tissues. Imbalanced levels of ROS generation and antioxidant defence mechanism (superoxide dismutase and catalase) cause damages to lipids, proteins and DNA. PARP and cleaved caspase 3 expression levels were found to be increased in the Fe2O3-NP-exposed brain regions which confirms DNA damage and apoptosis. Thus, repeated Fe2O3-NP exposure causes neurobehavioural impairments by nanoparticle accumulation, oxidative stress and apoptosis in the mouse brain.

Keywords

Apoptosis Fe2O3-NPs Locomotor behaviour Oxidative stress Spatial memory 

Notes

Acknowledgements

This work was supported by the University Grants Commission—Special Assistance Programme (UGC-SAP-II:F-3-20/2013) and Department of Science and Technology, Fund for Improvement of S&T infrastructure in universities and higher educational institutions (DST-FIST:SR/FST/LSI-618/2014), New Delhi, India. Vijayprakash Manickam acknowledges the UGC-BSR fellowship (UGC-BSR-No.F.7-25/2007) funded by UGC-BSR, New Delhi, India.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Supplementary material

12640_2017_9721_MOESM1_ESM.docx (522 kb)
ESM 1 (DOCX 522 kb)
12640_2017_9721_MOESM2_ESM.docx (331 kb)
ESM 2 (DOCX 330 kb)
12640_2017_9721_MOESM3_ESM.docx (196 kb)
ESM 3 (DOCX 196 kb)

References

  1. Acker CI, Souza AC, Pinton S, Rocha JT, Friggi CA, Zanella R, Nogueira CW (2011) Repeated malathion exposure induces behavioral impairment and AChE activity inhibition in brains of rat pups. Ecotoxicol Environ Safety 74:2310–2315CrossRefPubMedGoogle Scholar
  2. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefPubMedGoogle Scholar
  3. Ali YO, Escala W, Ruan K, Zhai RG (2011) Assaying locomotor, learning, and memory deficits in drosophila models of neurodegeneration. J Vis Exp 49:2504–2509Google Scholar
  4. Anderson GJ (2007) Mechanisms of iron loading and toxicity. Am J Hematol 82:1128–1131CrossRefPubMedGoogle Scholar
  5. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287CrossRefPubMedGoogle Scholar
  6. Budni P, de Lima MN, Polydoro M, Moreira JC, Schroder N, Dal-Pizzol F (2007) Antioxidant effects of selegiline in oxidative stress induced by iron neonatal treatment in rats. Neurochem Res 32:965–972CrossRefPubMedGoogle Scholar
  7. Charlotte SJ, Lone G, Erik B (1997) Acetylcholinestrase inhibition and altered locomotr behaviour in the carabid beetle pterostichus cupres. A linkage between biomarkers at two levels of biological complexity. Environ Toxicol Chem 16:1727–1732CrossRefGoogle Scholar
  8. Chen Z, Meng H, Xing G, Chen C, Zhao Y, Jia G, Wang T, Yuan H, Ye C, Zhao F, Chai Z, Zhu C, Fang X, Ma B, Wan L (2006) Acute toxicological effects of copper nanoparticles in vivo. Toxicol Lett 163:109–120CrossRefPubMedGoogle Scholar
  9. Comănescu MV, Mocanu MA, Anghelache L, Marinescu B, Dumitrache F, Bădoi AD, Manda G (2015) Toxicity of L-dopa coated iron oxide nanoparticles in intraperitoneal delivery setting—preliminary preclinical study. Romanian J Morphol Embryol 56:691–696Google Scholar
  10. De Lima MN, Polydoro M, Laranja DC, Bonatto F, Bromberg E, Moreira JC, Dal-Pizzol F, Schröder N (2005) Recognition memory impairment and brain oxidative stress induced by postnatal iron administration. Eur J Neurosci 21:2521–2528CrossRefPubMedGoogle Scholar
  11. Deacon RM (2013) Measuring motor coordination in mice. J Vis Exp 75:2609Google Scholar
  12. Devasagayam TP, Tarachand U (1987) Decreased lipid peroxidation in rat kidney during gestation. Biochem Biophys Res Commun 145:134–138CrossRefPubMedGoogle Scholar
  13. Dharmalingam P, Kulasekaran G, Ganapasam S (2013) Fisetin enhances behavioral performances and attenuates reactive gliosis and inflammation during aluminum chloride-induced neurotoxicity. NeuroMolecular Med 15:192–208CrossRefGoogle Scholar
  14. Drechsel DA, Est’evez AG, Barbeito L, Beckman JS (2012) Nitric oxide-mediated oxidative damage and the progressive demise of motor neurons in ALS. Neurotox Res 22:251–264CrossRefPubMedPubMedCentralGoogle Scholar
  15. Dunham NW, Miya TS (1957) A note on a simple apparatus for detecting neurological defects in rats and mice. J Am Pharmaceutical Assoc; Scientific Edition XIVI:208–209Google Scholar
  16. Ellman GL, Courtney D, Jr Andres V, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95CrossRefPubMedGoogle Scholar
  17. Fan CH, Ting CY, Lin HJ, Wang CH, Liu HL, Yen TC, Yeh CK (2013) SPIO-conjugated, doxorubicin-loaded microbubbles for concurrent MRI and focused-ultrasound enhanced brain-tumor drug delivery. Biomaterials 34:3706–3715CrossRefPubMedGoogle Scholar
  18. Federica Ma. Antonella P, Anna C, Franca C, Giuseppina DG, Sara C, Stefania R, Dario F, Letterio SP, Francesca C, Ottavio C, Fabio G, Sonia L (2015) A novel neuroferritinopathy mouse model (FTL 498InsTC) shows progressive brain iron dysregulation, morphological signs of early neurodegeneration and motor coordination deficits. Neurobiol Dis 81:119–133CrossRefGoogle Scholar
  19. Fiset C, Rioux FM, Surette ME, Fiset S (2015) Prenatal iron deficiency in guinea pigs increases locomotor activity but does not influence learning and memory. PLoS One 10:0133168Google Scholar
  20. Frechou M, Beray-Berthat V, Raynaud JS, Meriaux S, Gombert F, Lancelot E, Plotkine M, Marchand-Leroux C, Ballet S, Robert P, Louin G, Margaill I (2013) Detection of vascular cell adhesion molecule-1 expression with USPIO enhanced molecular MRI in a mouse model of cerebral ischemia. Contrast Media Mol Imaging 8:157–164CrossRefPubMedGoogle Scholar
  21. Fredriksson A, Schroder N, Eriksson P, Izquierdo I, Archer T (1999) Neonatal iron exposure induces neurobehavioural dysfunctions in adult mice. Toxicol Appl Pharmacol 159:25–30CrossRefPubMedGoogle Scholar
  22. Fredriksson A, Schroder N, Eriksson P, Izquierdo I, Archer T (2000) Maze learning and motor activity deficits in adult mice induced by iron exposure during a critical postnatal period. Brain Res Dev 119:65–74CrossRefGoogle Scholar
  23. Fretham SJ, Carlson ES, Georgieff MK (2011) The role of iron in learning and memory. Adv Nutr 2:112–121CrossRefPubMedPubMedCentralGoogle Scholar
  24. Frings M, Boenisch R, Gerwig M, Diener HC, Timmann D (2004) Learning of sensory sequences in cerebellar patients. Learn Mem 11:347–355CrossRefPubMedPubMedCentralGoogle Scholar
  25. Fukui K, Omoi NO, Hayasaka T, Shinnkai T, Suzuki S, Abe K, Urano S (2002) Cognitive impairment of rats caused by oxidative stress and aging, and its prevention by vitamin E. Ann N Y Acad Sci 959:275–284CrossRefPubMedGoogle Scholar
  26. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR (1982) Analysis of nitrate, nitrite and [15N] nitrate in biological fluids. Anal Biochem 126:131–138CrossRefPubMedGoogle Scholar
  27. Halliwell B (1992) Oxygen radicals as key mediators in neurological disease: fact or fiction. Ann Neurol 32:10–15CrossRefGoogle Scholar
  28. Haseeb AK, Abdullah SA, Samia HS, Syed SH, Zohair AA, Adnan AK, Abdulrahman AM (2013) Serum markers of tissue damage and oxidative stress in patients with acute myocardial infarction. Biomed Res 24:15–20Google Scholar
  29. Hautot D, Pankhurst QA, Morris CM, Curtis A, Burn J, Dobson J (2007) Preliminary observation of elevated levels of nanocrystalline iron oxide in the basal ganglia of neuroferritinopathy patients. Biochim Biophys Acta 1:21–25CrossRefGoogle Scholar
  30. Jain KK (2009) Cell therapy for CNS trauma. Mol Biotechnol 42:367–376CrossRefPubMedGoogle Scholar
  31. Joseph R, Prohaska AAG (2005) Rat brain iron concentration is lower following perinatal copper deficiency. J Neurochem 93:698–705CrossRefGoogle Scholar
  32. Jung-Jin H, So-Young C, Jae-Young K (2002) The role of NADPH oxidase, neuronal nitric oxide synthase and poly (ADP ribose) polymerase in oxidative neuronal death induced in cortical cultures by brain-derived neurotrophic factor and neurotrophin-4/5. J Neurochem 82:894–902CrossRefGoogle Scholar
  33. Kircher MF, Mahmood U, King RS, Weissleder R, Josephson L (2003) A multimodal nanoparticle for preoperative magnetic resonance imaging and intraoperative optical brain tumor delineation. Cancer Res 63:8122–8125PubMedGoogle Scholar
  34. Korfias S, Papadimitriou A, Stranjalis G, Bakoula C, Daskalakis G, Antsaklis A, Sakas DE (2009) Serum biochemical markers of brain injury. Mini Rev Med Chem 9:227–234CrossRefPubMedGoogle Scholar
  35. Kovacic P, Somanathan R (2013) Nanoparticles: toxicity, radicals, electron transfer, and antioxidants. Methods Mol Biol 1028:15–35CrossRefPubMedGoogle Scholar
  36. Lekawanvijit S, Chattipakorn N (2009) Iron overload thalassemic cardiomyopathy: iron status assessment and mechanisms of mechanical and electrical disturbance due to iron toxicity. Can J Cardiol 25:213–218CrossRefPubMedPubMedCentralGoogle Scholar
  37. Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464–478CrossRefPubMedGoogle Scholar
  38. Linas R, Lang EJ, Welsh JP (1997) The cerebellum, LTD and memory: alternative views. Learn Mem 3:444–445Google Scholar
  39. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  40. Manaenko A, Chen H, Kammer J, Zhang JH, Tang J (2011) Comparison Evans blue injection routes: intravenous vs. intraperitoneal, for measurement of blood-brain barrier in a mice hemorrhage model. J Neurosci Methods 195:206–210CrossRefPubMedGoogle Scholar
  41. Ma P, Luo Q, Chen J, Gan Y, Du J, Ding S, Xi Z, Yang X (2012) Intraperitoneal injection of magnetic Fe3O4-nanoparticle induces hepatic and renal tissue injury via oxidative stress in mice. Int J Nanomedicine 7:4809–4818Google Scholar
  42. Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autooxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474CrossRefPubMedGoogle Scholar
  43. McCullough B, Kolokythas O, Maki J, Green D (2012) Ferumoxytol in clinical practice: implications for MRI. J Magn Reson Imaging 36:1476–1479Google Scholar
  44. Megraw RE (1971) Reaction of reduced nicotinamide adenine dinucleotide with 2,4-dinitrophenylhydrazine in serum lactate dehydrogenase assays. Am J Clin Pathol 56:225–226CrossRefPubMedGoogle Scholar
  45. Micaela G, Hadas S, Noa MC, Shlomo M, Edward AS (2013) Age-dependent effects of microglial inhibition in vivo on Alzheimer’s disease neuropathology using bioactive conjugated iron oxide nanoparticles. J Nanobiotech 11:32–44CrossRefGoogle Scholar
  46. Miwa CP, de Lima MN, Scalco F, Vedana G, Mattos R, Fernandez LL, Hilbig A, Schröder N, Vianna MR (2011) Neonatal iron treatment increases apoptotic markers in hippocampal and cortical areas of adult rats. Neurotox Res 19:527–535CrossRefPubMedGoogle Scholar
  47. Movasaghi Z, Rehman S, Rehman I (2008) Fourier transform infrared (FTIR) spectroscopy of biological tissues. Appl Spectrosc Rev 43:134–179CrossRefGoogle Scholar
  48. Nel A, Xia T, Madler L, Li N (2006) Toxic potential of materials at the nanolevel. Science 311:622–627CrossRefPubMedGoogle Scholar
  49. Papandreoua MA, Tsachaki M, Efthimiopoulos S, Cordopatisc P, Lamaric FN, Margarity M (2011) Memory enhancing effects of saffron in aged mice are correlated with antioxidant protection. Behav Brain Rese 219:197–204CrossRefGoogle Scholar
  50. Paul V, Ekambaram P, Jayakumar AR (1998) Effects of sodium fluoride on locomotor behaviour and a few biochemical parameters in rats. Env Toxicol Pharmacol 6:187–191Google Scholar
  51. Perez VP, De Lima MN, Da Silva RS, Dornelles AS, Vedana G, Bogo MR, Bonan CD, Schröder N (2010) Iron leads to memory impairment that is associated with a decrease in acetylcholinesterase pathways. Curr Neurovasc Res 7:15–22CrossRefPubMedGoogle Scholar
  52. Prina-Mello A, Crosbie-Staunton K, Salas G, Morales MP, Volkov Y (2013) Multiparametric toxicity evaluation of SPIONs by high content screening technique: identification of biocompatible multifunctional nanoparticles for nanomedicine. IEEE Trans Magn 49:377–382CrossRefGoogle Scholar
  53. Rahmat AK, Muhammad RK, Sumaira S (2012) Brain antioxidant markers, cognitive performance and acetylcholinesterase activity of rats: efficiency of Sonchus asper. Behav Brain Funct 8:8–21CrossRefGoogle Scholar
  54. Rinne JO, Kaasinen V, Järvenpää T, Någren K, Roivainen A, Yu M, Oikonen V, Kurki T (2003) Brain acetylcholinesterase activity in mild cognitive impairment and early Alzheimer’s disease. J Neurol Neurosurg Psychiatry 74:113–115CrossRefPubMedPubMedCentralGoogle Scholar
  55. Rudi DH, Peter PDD (2001) Applications of the Morris water maze in the study of learning and memory. Brain Res Rev 36:60–90CrossRefGoogle Scholar
  56. Schipper HM (2012) Neurodegeneration with brain iron accumulation-clinical syndromes and neuroimaging. Biochim Biophys Acta 1822:350–360CrossRefPubMedGoogle Scholar
  57. Schroder N, Fredriksson A, Vianna MR, Roesler R, Izquierdo I, Archer T (2011) Memory deficits in adult rats following postnatal iron administration. Behav Brain Res 124:77–85CrossRefGoogle Scholar
  58. Shahidi S, Komaki A, Mahmoodi M, Atrvash N, Ghodrati M (2008) Ascorbic acid supplementation could affect passive avoidance learning and memory in rat. Brain Res Bull 76:109–113CrossRefPubMedGoogle Scholar
  59. Sripetchwandee J, Pipatpiboon N, Chattipakorn N, Chattipakorn S (2014) Combined therapy of iron chelator and antioxidant tcompletely restores brain dysfunction induced by iron toxicity. PLoS One 9:85115CrossRefGoogle Scholar
  60. Stephane LB, Umar I, James NR, Michael AA, Kanji N (2008) Perinatal iron deficiency affects locomotor behavior and water maze performance in adult male and female rats. J Nutr 138:931–937Google Scholar
  61. Sundarraj K, Manickam V, Raghunath A, Periyasamy M, Viswanathan MP, Perumal E (2017a) Repeated exposure to iron oxide nanoparticles causes testicular toxicity in mice. Environ Toxicol 32(2):594–608CrossRefPubMedGoogle Scholar
  62. Sundarraj K, Raghunath A, Panneerselvam L, Perumal E (2017b) Iron oxide nanoparticles modulate heat shock proteins and organ specific markers in mice male accessory organs. Toxicol Appl Pharmacol 317:12–24CrossRefPubMedGoogle Scholar
  63. Svitlana GD, Maria COR, Diana GH, Naoki T, Aric FD, Sean MB, Jerry VA, Mibel P, Andrew W, Hiroto I, Kazutaka S, Edward H, Dunham RS, Yuji K, Cesario VB (2013) Blood-brain barrier alterations provide evidence of subacute diaschisis in an ischemic stroke rat model. PLoS One 8:53–68Google Scholar
  64. Thach WT (1996) On the specific role of cerebellum in motor learning and cognition: clues from PET activation and lesion studies in man. Behav Brain Sci 19:411–431CrossRefGoogle Scholar
  65. Thompson KJ, Shoham S, Connor JR (2011) Iron and neurodegenerative disorders. Brain Res Bull 55:155–164CrossRefGoogle Scholar
  66. Toiber D, Berson A, Greenberg D, Melamed-Book N, Diamant S, Soreq H (2008) N-acetylcholinesterase-induced apoptosis in Alzheimer’s disease. PLoS One 3:108CrossRefGoogle Scholar
  67. Varallyay CG, Nesbit E, Fu R, Gahramanov S, Moloney B, Earl E, Muldoon LL, Li X, Rooney WD, Neuwelt EA (2013) High-resolution steady-state cerebral blood volume maps in patients with central nervous system neoplasms using ferumoxytol, a superparamagnetic iron oxide nanoparticle. J Cereb Blood Flow Metab 33:780–786CrossRefPubMedPubMedCentralGoogle Scholar
  68. Wadghiri Y, Li J, Wang J, Hoang D, Sun Y, Xu H, Tsui W, Li Y, Boutajangout A, Wang A, de Leon M, Wisniewski T (2013) Detection of amyloid plaques targeted by bifunctional USPIO in Alzheimer’s disease transgenic mice using magnetic resonance microimaging. PLoS One 8:57097CrossRefGoogle Scholar
  69. Wang B, Feng W, Zhu M, Wang Y, Wang M, Gu Y, Ouyang H, Wang H, Li M, Zhao Y, Chai Z, Wang H (2009) Neurotoxicity of low-dose repeatedly intranasal instillation of nano- and submicron-sized ferric oxide particles in mice. J Nanopart Res 11:41–53CrossRefGoogle Scholar
  70. Wang B, Feng WY, Wang M, Shi JW, Zhang F, Ouyang H, Zhao YL, Chai ZF, Huang YY, Xie YN, Wang HF, Wang J (2007) Transport of intranasally instilled fine Fe2O3 particles into the brain: micro-distribution, chemical states, and histopathological observation. Biol Trace Elem Res 118:233–243CrossRefPubMedGoogle Scholar
  71. Wenk GL (2004) Assessment of spatial memory using the radial arm maze and Morris water maze. Current Protocols in Neuroscience 26:8.5A:8.5A.1–8.5A8.5A.12Google Scholar
  72. Williamson SM, Moffat C, Gomersall MA, Saranzewa N, Connolly CN, Wright GA (2013) Exposure to acetylcholinesterase inhibitors alters the physiology and motor function of honeybees. Front Physiol 4:1–10CrossRefGoogle Scholar
  73. Winer J, Kim P, Law M, Liu C, Apuzzo M (2011) Visualizing the future: enhancing neuroimaging with nanotechnology. World Neurosurg 75:626–637CrossRefPubMedGoogle Scholar
  74. Winer JL, Liu CY, Apuzzo ML (2012) The use of nanoparticles as contrast media in neuroimaging: a statement on toxicity. World Neurosurg 78:709–711CrossRefPubMedGoogle Scholar
  75. Won SM, Lee JH, Park UJ, Gwag J, Gwag BJ (2011) Iron mediates endothelial cell damage and blood-brain barrier opening in the hippocampus after transient forebrain ischemia in rats. Exp Mol Med 43:121–128CrossRefPubMedPubMedCentralGoogle Scholar
  76. Xin F, Jiang L, Liu X, Geng C, Wang W, Zhong L, Yang G, Chen M (2014) Bisphenol A induces oxidative stress-associated DNA damage in INS-1 cells. Mutat Res Genet Toxicol Environ Mutagen 769:29–33CrossRefPubMedGoogle Scholar
  77. Yehuda S, Youdim ME, Mostofsky DI (1986) Brain iron-deficiency causes reduced learning capacity in rats. Pharmacol Biochem Behav 25:141–144CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Vasanth Dhakshinamoorthy
    • 1
  • Vijayprakash Manickam
    • 1
  • Ekambaram Perumal
    • 1
  1. 1.Molecular Toxicology Laboratory, Department of BiotechnologyBharathiar UniversityCoimbatoreIndia

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