NEUROTOXIC EFFECT OF TITANIUM DIOXIDE NANOPARTICLES: BIOCHEMICAL AND PATHOLOGICAL APPROACH IN MALE WISTAR RATS
Objective: Titanium dioxide nanoparticles (TiO2 NPs) are widely used in pharmaceutical, cosmeceutical, biomedical and industrial applications. The adverse effects of TiO2 NPs are also increasing alarmingly. The purpose of this study is to investigate the toxicity of TiO2 NPs on biochemical and histological changes in different regions of brain in adult male Wistar rats.
Methods: Two different doses of TiO2 NPs (50 mg/kg b. w and 100 mg/kg b. w) administered orally for 14 d along with one control group, each group consisting of six animals. Standard biochemical methods were adopted for the estimation of enzymes alkaline phosphatase, 5â€™ nucleotidase, ATPases and gamma-glutamyl transpeptidase. Trace elements calcium, sodium, potassium and magnesium as well as metals like iron, zinc and copper were also estimated.
Results: When compared with the control group, the enzymes ATPases, ALP, 5â€™-NT and GGT activities were significantly decreased in both the TiO2 NPs treated groups. Ca, Na, Fe, Cu and TiO2 contents were significantly increased in both the experimental groups, while the K, Mg and Zn contents decreased. However, the changes in the parameters studied were more in 100 mg treated group (p<0.001) when compared to the 50 mg treated group (p<0.05and p<0.01). Moreover, it is also evident that different regions responded differently due to TiO2 NPs exposure. The changes were maximum in the cerebral hemisphere (p<0.001) followed by medulla oblongata (p<0.001) and cerebrum (p<0.05).
Conclusion: The results clearly imply that TiO2 NPs could impair the electrochemical gradient, ionic homeostasis and membrane stability in different regions of the rat brain.
2. Gandamalla D, Lingabathula H, Yellu NR. Cytotoxicity evaluation of titanium and zinc oxide nanoparticles on human cell lines. Int J Pharm Pharm Sci 2017;9:240-6.
3. Robichaud CO, Uyar AL, Darby MR, Zucker LG, Wiesner MR. Estimates of upper bounds and trends in nano-TiO2 production as a basis for exposure assessment. Environ Sci Technol 2009;43:4227-33.
4. Hu R, Gong X, Duan Y, Li N, Che Y, Cui Y, et al. Neurotoxicological effects and the impairment of spatial recognition memoryin mice caused by exposure to TiO2 nanoparticles. Biomaterials 2010;31:8043-50.
5. Zhang T, You L, Zhang Y. Photocatalytic reduction of p-chloronitrobenzene on illuminated nano-titanium dioxide particles. Dyes Pigments 2006;68:95â€“100.
6. Oberdorster G, Oberdorster E, Oberdorster J. Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 2005;113:823â€“39.
7. Warheit DB, Hoke RA, Finlay C, Donner EM, Reed KL, Sayes CM. Development of a base of toxicity tests using ultrafine TiO2 particles as a component of nanoparticle risk management. Toxicol Lett 2007;171:99â€“110.
8. Grassian VH, Oâ€™Shaughnessy PT, Adamcakova-Dodd A, Pettibone JM, Thorne PS. Inhalation exposure study of titanium dioxide nanoparticles with a primary particle size of 2 to 5 nm. Environ Health Perspect 2007;115:397-402.
9. Jia X, Wang S, Zhou L, Sun L. The potential liver, brain, and embryotoxicity of titanium dioxide nanoparticles on mice. Nanoscale Res Lett 2017;12:478.
10. Oberdorster G, Sharp Z, Atudorei V, Elder A, Gelein R, Kreyling W, et al. Translocation of inhaled ultrafine particles to the brain. Inhal Toxicol 2004;16:437â€“45.
11. Chen HW, Su SF, Chien CT, Lin WH, Yu SL, Chou CC, et al. Titanium dioxide nanoparticles induce emphysema-like lung injury in mice. FASEB J 2006;20:2393â€“5.
12. Chen J, Dong X, Zhao J, Tang G. In vivo acute toxicity of titanium dioxide nanoparticles to mice after intraperitioneal injection. J App Toxicol 2009;29:330â€“7.
13. Liu H, Ma L, Zhao J, Liu J, Yan J, Ruan J. Biochemical toxicity of nano-anatase TiO2 particles in mice. Biol Trace Elem Res 2009;129:170â€“80.
14. Duan Y, Liu J, Ma L, Li N, Liu H, Wang J, et al. Toxicological characteristics of nanoparticulate anatase titanium dioxide in mice. Biomaterials 2010;31:894-9.
15. Bu Q, Yan G, Deng P, Peng R, Lin H, Xu Y, et al. NMR-based metabonomic study of the sub-acute toxicity of titanium dioxide nanoparticles in rats after oral administration. Nanotechnology 2010;21:1-12.
16. Vasantharaja D, Ramalingam V, Reddy GA. Oral toxic exposure of titanium dioxide nanoparticles on serum biochemical changes in adult male Wistar rats. Nanomed J 2015;2:46-53.
17. Lockman PR, Koziara JM, Mumper RJ, Allen DD. Nanoparticle surface charges alter blood-brain barrier integrity and permeability. J Drug Target 2004;12:635â€“41.
18. Wu J, Liu W, Xue C, Zhou S, Lan F, Bi L, et al. Toxicity and penetration of TiO2 nanoparticles in hairless mice and porcine skin after subchronic dermal exposure. Toxicol Lett 2009;191:1â€“8.
19. Gao X, Yin S, Tang M, Chen J, Yang Z, Zhang W, et al. Effects of developmental exposure to TiO2 nanoparticles on synaptic plasticity in hippocampal dentate gyrus area: an in vivo study in anesthetized rats. Biol Trace Elem Res 2011;143:1616-28.
20. Long TC, Tajuba J, Sama P, Saleh N, Swartz C, Parker J, et al. Nanosize titanium dioxide stimulates reactive oxygen species in brain microglia and damages neurons in vitro. Environ Health Perspect 2007;115:1631â€“7.
21. Oszlanczi G, Horvath E, Szabo A, Horvath E, Sapi A, Gabor K, et al. Subacute exposure of rats by metal oxide nanoparticles through the airways: general toxicity and neuro-functional effects. Acta Biol Szegediensis 2010;54:165-70.
22. Sharma HS, Sharma A. Nanoparticles aggravate heat stress-induced cognitive deficits, blood-brainbarrier disruption, edema formation and brain pathology. Prog Brain Res 2007;162:245â€“73.
23. Buzea C, Pacheco II, Robbie K. Nanomaterials and nanoparticles: sources and toxicity. Biointerphases 2007; 2:17â€“71.
24. Takeo S, Sakanashi M. Characterization of membrane-bound adenosine triphophatase activity of enriched fraction from vascular smooth muscle. Enzyme 1985;34:152â€“65.
25. Fiske CH, Subbarow Y. The colorimetric determination of phosphorous. J Biol Chem 1925;66:375-400.
26. Bessey OA, Lowery OH, Brock MJ. A method for the rapid determination of alkaline phosphatase with fire cubic millimeters of serum. J Biol Chem 1946;164:321-30.
27. Gerlach U, Hiby W. 5â€™ nucleodiase. In: Methods of enzymatic analysis, Bergmeyer HU. Eds. Vol. 2. Verlagchemie, Weinheim, Accademic Press: New York; 1974. p. 871-5.
28. Orlowski M, Meister A. Isolation of gamma-glutamyltranspeptidase from hog kidney. J Biol Chem 1965;240:338â€“47.
29. Zar JH. Biostatistical analysis. 1st edition. Prentice Hall, Eaglewood cliffs, Jersy, USA; 1974.
30. Chetty CS, Rajanna B, Rajanna S. Inhibition of rat brain microsomal Na+/K+-ATPase and brain binding by mercuric chloride. Toxicol Lett 1990;51:109-16.
31. Yallapragada PR, Vig PJ, Kodavanti PR, Desaiah D. In vivo effects of triorganotins on calmodulin activity in rat brain. J Toxicol Environ Health 1991;34:229-37.
32. Boyer PD, Chance B, Ernster L, Mitchell P, Racker E, Slater C. Oxidative phosphorylation and photophosphorylation. Ann Rev Biochem 1997;46:955â€“1026.
33. Pardini RS, Heidker JC, Baker TA, Payne B. Toxicology of various pesticides and their decomposition products on mitochondrial electron transport. Arch Environ Contam Toxicol 1980;9:87â€“97.
34. Parsons JT, Sun DA, De-Lorenzo R, Churn SB. Neuronal specific endoplasmic reticulum Mg2+/Ca2+ -ATPase Ca2+sequestration in mixed primary hippocampal culture homogenates. Anal Biochem 2004;330:130-9.
35. Mata AM, Sepulveda MR. Plasma membrane Ca-ATPases in the nervous system during development and ageing. World J Biol Chem 2010;1:229-34.
36. Clapham DE. Calcium signaling. Cell 2007;131:1047-58.
37. Abirami T, Jose AGR, Govindarajulu B, Karthikeyan J. Ecotoxicology of green synthesized silver nanoparticles on freshwater fish Mystus gulio. Int J Pharm Pharm Sci 2017; 9:192-8.
38. Kaplia AA, Morozova VS. Na+, K+ -ATPase activity in polarized cells. Ukrainskii Biokhimicheskii Zhurnal 2010;82:5â€“20.
39. Choi KL, Aldrich RW, Yellen G. Tetraethyl ammonium blockade distinguishes two inactivation mechanisms in voltage-activated K+channels. Proc Natl Acad Sci USA 1991;88:5092-5.
40. Bannister RG, Romaul CA. The localization of alkaline phosphatase activity in cerebral blood vessels. J Neurol Neurosur Psychiatry 1963;26:333-40.
41. Naido D. Alkaline phosphate at the site of cerebral injury. Acta Histochem 1963;15:182-5.
42. Goldfischer SE, Novikoff AB. The localization of phosphatase activities at the level of ultrastructure. J Histochem Cytochem 1964;12:72-95.
43. Napieralski R, Kempkes B, Gutensohn W. Evidence for coordinated induction and repression of ecto-5â€Ÿ-nucleotidase (CD73) and the A2 an adenosine receptor in a human B cell line. Biol Chem 2003;384:483-7.
44. Zatta P, Lucchini R, Rnsburg SJ, Taylor A. The role of metals in the neurodegenerative process: aluminum, manganese, and zinc. Brain Res Bull 2003;62:15-28.
45. Pal R, Nath R, Gill KD. Lipid peroxidation and antioxidant defence enzymes in various regions of adult rat brain after co-exposure to cadmium and ethanol. Basic Clin Pharmacol Toxicol 1993;73:209-14.
46. Fayed AH. Brain trace element concentration of rats treated with the plant alkaloid, vincamine. Biol Trace Elem Res 2010;136:314â€“9.
47. Madsen E, Gitlin JD. Copper and iron disorders of the brain. Ann Rev Neurosci 2007;30:317â€“7.
48. Konoha K, Sadakane Y, Kawahara M. Zinc neurotoxicity and its role in neurodegenerative diseases. J Health Sci 2006;52:1-8.
49. Thilsing Hansen T, JÃ¸rgensen RJ. Serum calcium response following oral zinc oxide administrations in dairy cows. Acta Veterinaria Scandinavica 2001;42:271â€“8.
50. Hentze MW, Mucknthaler MU, Andrews NC. Balancing acts: molecular control of mammalian iron metabolism. Cell 2004; 117:285â€“97.
51. Nappi AJ, Vass E. Iron, metalloenzymes and cytotoxic reactions. Cell Mol Biol 2002;4:637â€“47.
52. Smith MA, Zhu X, Tabaton M, Liu G, McKeel DW Jr, Cohen ML, et al. Increased iron and free radical generation in preclinical Alzheimer disease and mild cognitive impairment. J Alzheimers Dis 2010;19:363â€“72.
53. El-seweidy MM, El-Baky AE. Effect of dietary iron overload in rat brain: oxidative stress, neurotransmitter level and serum metal ion in relation to neurodegenerative disorders. Indian J Expert Biol 2008;46:855â€“8.
54. Won SM, Lee JH, Park UJ, Gwag J, Gwag BJ, Lee YB. Iron mediates endothelial cell damage and blood-brain barrier opening inthe hippocampus after transient forebrain ischemia in rats. Exp Mol Med 2011;43:121â€“8.
55. Raajshree KR, Durairaj B. Evaluation of the antityrosinase and antioxidant potential of zinc oxide nanoparticles synthesized from the brown seaweed-turbinaria conoides. Int J App Pharm 2017;9:116-20.
56. Nuttall JR, Oteiza PI. Zinc and the aging brain. Genes Nutr 2014;9:379.
57. Sandstead HH, Frederickson CJ, Penland JG. History of zinc as related to brain function. J Nutr 2000;130:496S-502S.
58. Que EL, Domaille DW, Change CJ. Metals in neurobiology: probing their chemistry and biology with molecular imaging. Chem Rev 2008;108:1517-49.
59. Wang B, Wnag Y, Feng W. Trace metal disturbance in mice brain after intranasal exposure of nano-and submicron-sized Fe2O3 particles. Chem Anal 2008;53:927-42.
60. Mohamed RH, Hussien NA. Genotoxicity studies of titanium dioxide nanoparticles (TiO2NPs) in the Brain of mice. Scientifica 2016;1-7. http://dx.doi.org/10.1155/2016/ 6710840.
61. Ma L, Liu J, Li N, Wang J, Duan Y, Yan J, et al. Oxidative stress in the brain of mice caused by translocated nanoparticulate TiO2 delivered to the abdominal cavity. Biomaterials 2010;31:99â€“105.