Journal of Neuroimmune Pharmacology

, Volume 3, Issue 4, pp 286–295 | Cite as

Manufactured Aluminum Oxide Nanoparticles Decrease Expression of Tight Junction Proteins in Brain Vasculature

  • Lei Chen
  • Robert A. Yokel
  • Bernhard Hennig
  • Michal ToborekEmail author
Original Article


Manufactured nanoparticles of aluminum oxide (nano-alumina) have been widely used in the environment; however, their potential toxicity provides a growing concern for human health. The present study focuses on the hypothesis that nano-alumina can affect the blood-brain barrier and induce endothelial toxicity. In the first series of experiments, human brain microvascular endothelial cells (HBMEC) were exposed to alumina and control nanoparticles in dose- and time-responsive manners. Treatment with nano-alumina markedly reduced HBMEC viability, altered mitochondrial potential, increased cellular oxidation, and decreased tight junction protein expression as compared to control nanoparticles. Alterations of tight junction protein levels were prevented by cellular enrichment with glutathione. In the second series of experiments, rats were infused with nano-alumina at the dose of 29 mg/kg and the brains were stained for expression of tight junction proteins. Treatment with nano-alumina resulted in a marked fragmentation and disruption of integrity of claudin-5 and occludin. These results indicate that cerebral vasculature can be affected by nano-alumina. In addition, our data indicate that alterations of mitochondrial functions may be the underlying mechanism of nano-alumina toxicity.


manufactured nanoparticles nano-alumina blood-brain barrier tight junctions 



Supported in part by Kentucky Science and Engineering Foundation (KSEF-07-RDE-010), P42 ES 07380, MH63022, MH072567, and NS39254.


  1. Abbott NJ, Ronnback L, Hansson E (2006) Astrocyte-endothelial interactions at the blood-brain barrier. Nat Rev Neurosci 7:41–53 doi: 10.1038/nrn1824 PubMedCrossRefGoogle Scholar
  2. András IE, Pu H, Tian J, Deli MA, Nath A, Hennig B et al (2005) Signaling mechanisms of HIV-1 Tat-induced alterations of claudin-5 expression in brain endothelial cells. J Cereb Blood Flow Metab 25:1159–1170 doi: 10.1038/sj.jcbfm.9600115 PubMedCrossRefGoogle Scholar
  3. Banks WA, Niehoff ML, Drago D, Zatta P (2006) Aluminum complexing enhances amyloid beta protein penetration of blood-brain barrier. Brain Res 1116:215–221 doi: 10.1016/j.brainres.2006.07.112 PubMedCrossRefGoogle Scholar
  4. Baydar T, Nagymajtenyi L, Isimer A, Sahin G (2005) Effect of folic acid supplementation on aluminum accumulation in rats. Nutrition 21:406–410 doi: 10.1016/j.nut.2004.07.008 PubMedCrossRefGoogle Scholar
  5. Becaria A, Campbell A, Bondy SC (2002) Aluminum as a toxicant. Toxicol Ind Health 18:309–320 doi: 10.1191/0748233702th157oa PubMedCrossRefGoogle Scholar
  6. Bianco A, Kostarelos K, Prato M (2005) Applications of carbon nanotubes in drug delivery. Curr Opin Chem Biol 9:674–679 doi: 10.1016/j.cbpa.2005.10.005 PubMedCrossRefGoogle Scholar
  7. Braydich-Stolle L, Hussain S, Schlager JJ, Hofmann MC (2005) In vitro cytotoxicity of nanoparticles in mammalian germline stem cells. Toxicol Sci 88:412–419 doi: 10.1093/toxsci/kfi256 PubMedCrossRefGoogle Scholar
  8. Campbell A, Becaria A, Lahiri DK, Sharman K, Bondy SC (2004) Chronic exposure to aluminum in drinking water increases inflammatory parameters selectively in the brain. J Neurosci Res 75:565–572 doi: 10.1002/jnr.10877 PubMedCrossRefGoogle Scholar
  9. Colvin VL (2003) The potential environmental impact of engineered nanomaterials. Nat Biotechnol 21:1166–1170 doi: 10.1038/nbt875 PubMedCrossRefGoogle Scholar
  10. Fink B, Laude K, McCann L, Doughan A, Harrison DG, Dikalov S (2004) Detection of intracellular superoxide formation in endothelial cells and intact tissues using dihydroethidium and an HPLC-based assay. Am J Physiol Cell Physiol 287:C895–C902 doi: 10.1152/ajpcell.00028.2004 PubMedCrossRefGoogle Scholar
  11. Gault PM, Allen KR, Newton KE (2005) Plasma aluminium: a redundant test for patients on dialysis? Ann Clin Biochem 42:51–54 doi: 10.1258/0004563053026862 PubMedCrossRefGoogle Scholar
  12. Gojova A, Guo B, Kota RS, Rutledge JC, Kennedy IM, Barakat AI (2007) Induction of inflammation in vascular endothelial cells by metal oxide nanoparticles: effect of particle composition. Environ Health Perspect 115:403–409PubMedCrossRefGoogle Scholar
  13. Hawkins BT, Davis TP (2005) The blood-brain barrier/neurovascular unit in health and disease. Pharmacol Rev 57:173–185 doi: 10.1124/pr.57.2.4 PubMedCrossRefGoogle Scholar
  14. Kawahara M (2005) Effects of aluminum on the nervous system and its possible link with neurodegenerative diseases. J Alzheimers Dis 8:171–182PubMedGoogle Scholar
  15. Koziara JM, Lockman PR, Allen DD, Mumper RJ (2003) In situ blood-brain barrier transport of nanoparticles. Pharm Res 20:1772–1778 doi: 10.1023/B:PHAM.0000003374.58641.62 PubMedCrossRefGoogle Scholar
  16. Kralj M, Pavelic K (2003) Medicine on a small scale. EMBO Rep 4:1008–1012 doi: 10.1038/sj.embor.7400017 PubMedCrossRefGoogle Scholar
  17. Kreuter J (2007) Nanoparticles—a historical perspective. Int J Pharm 331:1–10 doi: 10.1016/j.ijpharm.2006.10.021 PubMedCrossRefGoogle Scholar
  18. Lockman PR, Koziara J, Roder KE, Paulson J, Abbruscato TJ, Mumper RJ et al (2003) In vivo and in vitro assessment of baseline blood-brain barrier parameters in the presence of novel nanoparticles. Pharm Res 20:705–713 doi: 10.1023/A:1023492015851 PubMedCrossRefGoogle Scholar
  19. Lockman PR, Koziara JM, Mumper RJ, Allen DD (2004) Nanoparticle surface charges alter blood-brain barrier integrity and permeability. J Drug Target 12:635–641 doi: 10.1080/10611860400015936 PubMedCrossRefGoogle Scholar
  20. Lui WY, Lee WM (2005) cAMP perturbs inter-Sertoli tight junction permeability barrier in vitro via its effect on proteasome-sensitive ubiquitination of occludin. J Cell Physiol 203:564–572 doi: 10.1002/jcp.20254 PubMedCrossRefGoogle Scholar
  21. Lukiw WJ, Percy ME, Kruck TP (2005) Nanomolar aluminum induces pro-inflammatory and pro-apoptotic gene expression in human brain cells in primary culture. J Inorg Biochem 99:1895–1898 doi: 10.1016/j.jinorgbio.2005.04.021 PubMedCrossRefGoogle Scholar
  22. Maynard AD (2007) Nanotechnology: the next big thing, or much ado about nothing? Ann Occup Hyg 51:1–12 doi: 10.1093/annhyg/mel071 PubMedCrossRefGoogle Scholar
  23. Maynard AD, Aitken RJ, Butz T, Colvin V, Donaldson K, Oberdörster G et al (2006) Safe handling of nanotechnology. Nature 444:267–269 doi: 10.1038/444267a PubMedCrossRefGoogle Scholar
  24. Moghimi SM, Hunter AC, Murray JC (2005) Nanomedicine: current status and future prospects. FASEB J 19:311–330 doi: 10.1096/fj.04-2747rev PubMedCrossRefGoogle Scholar
  25. Morten KJ, Ackrell BA, Melov S (2006) Mitochondrial reactive oxygen species in mice lacking superoxide dismutase 2: attenuation via antioxidant treatment. J Biol Chem 281:3354–3359 doi: 10.1074/jbc.M509261200 PubMedCrossRefGoogle Scholar
  26. Nel A, Xia T, Madler L, Li N (2006) Toxic potential of materials at the nanolevel. Science 311:622–627 doi: 10.1126/science.1114397 PubMedCrossRefGoogle Scholar
  27. Oberdorster G, Oberdorster E, Oberdorster J (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 113:823–839PubMedCrossRefGoogle Scholar
  28. Oesterling E, Chopra N, Gavalas V, Arzuaga X, Lim EJ, Sultana R et al (2008) Alumina nanoparticles induce expression of endothelial cell adhesion molecules. Toxicol Lett 178:160–166 doi: 10.1016/j.toxlet.2008.03.011 PubMedCrossRefGoogle Scholar
  29. Olivier JC (2005) Drug transport to brain with targeted nanoparticles. NeuroRx 2:108–119 doi: 10.1602/neurorx.2.1.108 PubMedCrossRefGoogle Scholar
  30. Persidsky Y, Heilman D, Haorah J, Zelivyanskaya M, Persidsky R, Weber GA et al (2006) Rho-mediated regulation of tight junctions during monocyte migration across the blood-brain barrier in HIV-1 encephalitis (HIVE). Blood 107:4770–4780 doi: 10.1182/blood-2005-11-4721 PubMedCrossRefGoogle Scholar
  31. Rittner MN (2002) Market analysis of nanostructured materials. Am Ceram Soc Bull 81:33–36Google Scholar
  32. Service RF (2004) Nanotoxicology. Nanotechnology grows up. Science 304:1732–1734 doi: 10.1126/science.304.5678.1732 PubMedCrossRefGoogle Scholar
  33. Silva GA (2006) Neuroscience nanotechnology: progress, opportunities and challenges. Nat Rev Neurosci 7:65–74 doi: 10.1038/nrn1827 PubMedCrossRefGoogle Scholar
  34. Uwatoku T, Shimokawa H, Abe K, Matsumoto Y, Hattori T, Oi K et al (2003) Application of nanoparticle technology for the prevention of restenosis after balloon injury in rats. Circ Res 92:e62–e69 doi: 10.1161/01.RES.0000069021.56380.E2 PubMedCrossRefGoogle Scholar
  35. Vorbrodt AW, Dobrogowska DH, Lossinsky AS (1994) Ultracytochemical studies of the effects of aluminum on the blood-brain barrier of mice. J Histochem Cytochem 42:203–212PubMedGoogle Scholar
  36. Weksler BB, Subileau EA, Perrière N, Charneau P, Holloway K, Leveque M et al (2005) Blood-brain barrier-specific properties of a human adult brain endothelial cell line. FASEB J 19:1872–1874PubMedGoogle Scholar
  37. Yang L, Watts DJ (2005) Particle surface characteristics may play an important role in phytotoxicity of alumina nanoparticles. Toxicol Lett 158:122–132 doi: 10.1016/j.toxlet.2005.03.003 PubMedCrossRefGoogle Scholar
  38. Yang Y, Estrada EY, Thompson JF, Liu W, Rosenberg GA (2007) Matrix metalloproteinase-mediated disruption of tight junction proteins in cerebral vessels is reversed by synthetic matrix metalloproteinase inhibitor in focal ischemia in rat. J Cereb Blood Flow Metab 27:697–709 doi: 10.1038/sj.jcbfm.9600440 PubMedCrossRefGoogle Scholar
  39. Yokel RA, McNamara PJ (2001) Aluminium toxicokinetics: an updated minireview. Pharmacol Toxicol 88:159–167 doi: 10.1034/j.1600-0773.2001.d01-98.x PubMedCrossRefGoogle Scholar
  40. Yokel RA, Wilson M, Harris WR, Halestrap AP (2002) Aluminum citrate uptake by immortalized brain endothelial cells: implications for its blood-brain barrier transport. Brain Res 930:101–110 doi: 10.1016/S0006-8993(02)02234-5 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Lei Chen
    • 1
  • Robert A. Yokel
    • 2
  • Bernhard Hennig
    • 3
  • Michal Toborek
    • 1
    Email author
  1. 1.Molecular Neuroscience and Vascular Biology Laboratory, Department of NeurosurgeryUniversity of Kentucky Medical CenterLexingtonUSA
  2. 2.College of PharmacyUniversity of KentuckyLexingtonUSA
  3. 3.College of AgricultureUniversity of KentuckyLexingtonUSA

Personalised recommendations