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Licensed Unlicensed Requires Authentication Published by De Gruyter April 7, 2016

Hormonal and organ-specific dysfunction induced by the interaction between titanium dioxide nanoparticles and salicylic acid in male mice

  • Nahla S. El-Shenawy EMAIL logo , Mohammad S. Al-Harbi and Fatimah F.E. Al hamayani


Background: Nanomaterials coating gained much concern in orthopedic implants and cosmetics. Drug combination may be a promising strategy for treating multi-factorial diseases. Titanium dioxide (TDN) nanoparticles are being widely used in many industries as well as in medicine and pharmacology. Therefore, increased human and environmental exposure can be expected, which has put TDN under toxicological scrutiny, and it is necessary to address the potential health and safety implications of nanomaterials used in nanomedicine. The toxicity of titanium oxide nanoparticles (TDN) and salicylic acid (SA) separately or in combination was studied for 21 days.

Methods: The liver and kidney biomarker were determined, and hormones and oxidative stress levels were detected in mice.

Results: The intraperitoneal (i.p.) injection of TDN and SA in combination had a potential toxicological effect on major organs and hormonal homeostasis of mice. TDN and SA could antagonistically interact to affect the liver and kidney functions. No synergistic damage was observed in the liver function of mice that were treated with both TDN and SA as compared to the SA group. TDN acted as a synergistic agent to SA in the case of total cholesterol and total proteins levels. SA acted as antagonistic to the effect of TDN when injected together in mice because the effect on kidney functions is less than that predicted on the basis of the additive. The effect of co-administration of SA and TDN on the following hormones; triiodothyronine, thyroxine, estradiol II and insulin various among additive, potentiation, antagonistic and no effect, respectively as compared to TDN group. The interaction of TDN and SA was also found to induce oxidative stress as indicated by the increase in lipid peroxidation (LPO) levels. The decrease in the level of the reduced glutathione in the co-treated group indicated that there were no synergistic damages. SA and TDN co-administration could induce a potential increase in LPO levels in liver, kidney, and spleen but not in heart tissue. These results have not suggested that TDN and SA have a synergistic sub-chronic toxicity in mice after i.p. administration. SA may decrease the toxicity of TDN to some degree that could be related to the potentiation chemical reaction between SA and TDN.

Conclusions: Our results suggested that the damage observed in mice treated with TDN and SA is organ-specific and associated with hormonal homeostasis and oxidative damage.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.


1. Pujalté I, Passagne I, Brouillaud B, Tréguer M, Durand E, Ohayon-Courtès C, et al. Cytotoxicity and oxidative stress induced by different metallic nanoparticles on human kidney cells. Part Fib Toxicol 2011;8:10.10.1186/1743-8977-8-10Search in Google Scholar PubMed PubMed Central

2. El-Shenawy NS, Mohsen Q, Fadl-allah SA. Oxidative stress and antioxidant responses of liver and kidney tissue after implantation of titanium or titanium oxide coated plate in rat tibiae. J Mater Sci Mater Med 2012;23:1763–74.10.1007/s10856-012-4648-9Search in Google Scholar PubMed

3. Shi H, Magaye R, Castranova V, Zhao J. Titanium dioxide nanoparticles: a review of current toxicological data. Part Fib Toxicol 2013;10:15.10.1186/1743-8977-10-15Search in Google Scholar PubMed PubMed Central

4. Gui S, Sang X, Zheng L, Ze Y, Zhao X, Sheng L, et al. Intragastric exposure to titanium dioxide nanoparticles induced nephrotoxicity in mice assessed by physiological and gene expression modifications. Part Fibre Toxicol 2013;10:5.10.1186/1743-8977-10-51Search in Google Scholar

5. Park EJ, Yoon J, Choi K, Yi J, Park K. Induction of chronic inflammation in mice treated with titanium dioxide nanoparticles by intratracheal instillation. Toxicology 2009;260:37–46.10.1016/j.tox.2009.03.005Search in Google Scholar PubMed

6. Liu H, Ma L, Liu J, Zhao J, Yan J, Hong F. Toxicity of nano anatase TiO2 to mice: liver injury, oxidative stress. Toxicol Environ Chem 2009;92:175–86.10.1080/02772240902732530Search in Google Scholar

7. 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;31:894–9.10.1016/j.biomaterials.2009.10.003Search in Google Scholar PubMed

8. Ma LL, Zhao JF, Wang J, Duan YM, Liu J, Li N. The acute liver injury in mice caused by nano-anatase TiO2. Nanoscale Res Lett 2009;4:1275–85.10.1007/s11671-009-9393-8Search in Google Scholar PubMed PubMed Central

9. Jiangxue W, Guoqiang Z, Chunying C, Hongwei Y, Tiancheng W, Yongmei M, et al. Acute toxicity and bio-distribution of different sized titanium dioxide particles in mice after oral administration. Toxicol Lett 2007;168:176–85.10.1016/j.toxlet.2006.12.001Search in Google Scholar PubMed

10. L’Azou B, Jorly J, On D, Sellier E, Moisan F, Fleury-Feith J, et al. In vitro effects of nanoparticles on renal cells. Part Fib Toxicol 2008;5:22.10.1186/1743-8977-5-22Search in Google Scholar PubMed PubMed Central

11. Zhao JF, Li N, Wang SS, Zhao XY, Wang J, Yan JY, et al. The mechanism of oxidative damage in nephrotoxicity of mice caused by nano-anatase TiO2. J Exp Nanosci 2010;5:447–62.10.1080/17458081003628931Search in Google Scholar

12. Gui SX, Zhang ZL, Zheng L, Cui YL, Liu XY, Li N, et al. Molecular mechanism of kidney injury of mice caused by exposure to titanium dioxide nanoparticles. J Hazard Mater 2011;195:365–70.10.1016/j.jhazmat.2011.08.055Search in Google Scholar PubMed

13. Guerrero A, González-Correa JA, Arrebola MM, Muñoz-Marín J, Sánchez De La Cuesta F, De La Cruz JP. Antioxidant effects of a single dose of acetylsalicylic acid and salicylic acid in rat brain slices subjected to oxygen-glucose deprivation in relation with its antiplatelet effect. Neurosci Lett 2004;358:153–6.10.1016/j.neulet.2004.01.036Search in Google Scholar PubMed

14. Dinis-Oliveira RJ, Sousa C, Remião F, Duarte JA, Sánchez Navarro A, Bastos ML, et al. Full survival of paraquat-exposed rats after treatment with sodium salicylate. Free Rad Biol Med 2007;42:1017–28.10.1016/j.freeradbiomed.2006.12.031Search in Google Scholar PubMed

15. Randjelovic P, Veljkovic S, Stojiljkovic N, Velickovic LJ, Sokolovic D, Stoiljkovic M, et al. Salicylic acid attenuates gentamicin-induced nephrotoxicity in rats. Sci World J 2012;2012:390613.10.1100/2012/390613Search in Google Scholar PubMed PubMed Central

16. Yin MJ, Yamamoto Y, Gaynor RB. The anti-inflammatory agents aspirin and salicylate inhibit the activity of IκB kinase-β. Nature 1998;396:77–80.10.1038/23948Search in Google Scholar PubMed

17. Ismagilov ZR, Tsykoza LT, Shikina NV, Zarytova VF, Zinoviev VV, Zagrebelnyi SN. Synthesis and stabilization of nano-sized titanium dioxide. Russ Chem Rev 2009;78:873–85.10.1070/RC2009v078n09ABEH004082Search in Google Scholar

18. Li Y, Li J, Yin J, Li W, Kang C, Huang Q, et al. Systematic influence induced by 3 nm titanium dioxide following intratracheal instillation of mice. J Nanosci Nanotechnol 2010;10:8544–9.10.1166/jnn.2010.2690Search in Google Scholar PubMed

19. Fadeyi O, Obafemi C, Adewunmi C, Iwalewa E. Antipyretic, analgesic, anti-inflammatory and cytotoxic effects of four derivatives of salicylic acid and anthranilic acid in mice and rats. Afr J Biotechnol 2004;3:426–31.10.5897/AJB2004.000-2081Search in Google Scholar

20. Boussarie D. Hématologie des Rongeurs et Lagomorphes de Compagnie. Bull Acad Vet 1999;72:209–16.10.4267/2042/62828Search in Google Scholar

21. Young DS, Friedman RB. Effects of disease on clinical laboratory tests, 4th ed, vol. 48. Washington DC: American Association Clinical Chemistry Press, 2001:682–3.10.1093/clinchem/48.4.682aSearch in Google Scholar

22. Reitman A, Frankel S. A colorimetric method for determination of serum glutamic oxaloacetic and glutamic pyruvic transaminase. Am J Clin Pathol 1957;28:56–63.10.1093/ajcp/28.1.56Search in Google Scholar

23. Gornal AG, Bardwil GS, David MM. Determination of serum proteins by mean of Biuret reactions. Biochem J 1949;177: 751–66.10.1016/S0021-9258(18)57021-6Search in Google Scholar

24. Lopes-Virella MF, Stone P, Ellis S, Colwell JA. Cholesterol determination in high-density lipoproteins separated by three different methods. Clin Chem 1977;23:882–4.10.1093/clinchem/23.5.882Search in Google Scholar

25. Schirmeister J. Creatinine standard and measurement of serum creatinine with picric acid. Deut Med Wochenschr 1964;89:1018–21.10.1055/s-0028-1111251Search in Google Scholar PubMed

26. Fossati P, Prencipe L, Berti G. Use of 3,5-dichloro-2- hydroxybenzene-sulfonic acid/4-aminophenazone chromogenic system in direct enzyme assay of uric acid in serum and urine. Clin Chem 1980;26:227–31.10.1093/clinchem/26.2.227Search in Google Scholar

27. Wheeler MH, Lazarus JH. Diseases of the thyroid. London, Glasgow, Weinheim, New York, Tokyo, Melbourne, Madras: Chapman and Hall Medical, 1994:108–15.Search in Google Scholar

28. Clark PM. Assays for insulin, proinsulin(s) and C-peptide. Ann Clin Biochem 1999;36:541–64.10.1177/000456329903600501Search in Google Scholar PubMed

29. Lichtenberg V, Schult-Baukloh A, Lindner CH. Discrepancies between results of serum 17β-oestradiol E2 determinations carried out using different immunoassay kits in women receiving estrogen replacement therapy. Lab Med 1992;16:412–6.10.1515/labm.1992.16.12.412Search in Google Scholar

30. Draper HH, Hadley M. Malonaldehyde determination as index of lipid peroxidation. In: Parcker L, Glazer A, editors. Methods in enzymology, vol. 186. New York: Academic Press, 1990:421–30.10.1016/0076-6879(90)86135-ISearch in Google Scholar

31. Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med 1963;61:882–90.Search in Google Scholar

32. Han C, Yan C, Xiang L, Jian-Wei C, Jin L, Ping X, et al. Antioxidation and protection against H2O2 induced oxidative stress on normal human hepatocytes cell line (LO2) by extracts and three compounds from the root of Isatis indigotica. J Med Plants Res 2012;6:1886–90.Search in Google Scholar

33. Hill AB. Principles of medical statistics, 9th ed, vol. 21. New York: Oxford University Press, 1971:687–8.Search in Google Scholar

34. Zhang R, Niu Y, Li Y, Zhao C, Song B, Li Y, et al. Acute toxicity study of the interaction between titanium dioxide nanoparticles and lead acetate in mice. Environ Toxicol Pharmacol 2010;30:52–60.10.1016/j.etap.2010.03.015Search in Google Scholar PubMed

35. Regoli F, Nigro M, Benedetti M, Gorbi S, Pretti C, Gervasi PG, et al. Interactions between metabolism of trace metals and xenobiotics agonists of the Ah receptor in the Antarctic fish Trematomus bernacchii: environmental perspectives. Environ Toxicol Chem 2005;24:1475–82.10.1897/04-514R.1Search in Google Scholar PubMed

36. Martin R, Wilson LF, Peter GB, Gary RH, Jill S, Richard JS, et al. Nanoparticle interactions with zinc and iron: implications for toxicology and inflammation. Toxicol Appl Pharmacol 2007;225:80–9.10.1016/j.taap.2007.07.012Search in Google Scholar PubMed

37. Zhao X, Ze Y, Gao G, Sang X, Li B, Gui S, et al. Nanosized TiO2-induced reproductive system dysfunction and its mechanism in female mice. PLoS One 2013;8:e59378.10.1371/journal.pone.0059378Search in Google Scholar PubMed PubMed Central

38. Crissman JW, Goodman DG, Hildebrandt PK, Maronpot RR, Prater DA, Riley JH, et al. Best practices guideline: toxicological histopathology. Toxicol Pathol 2004;32:126–31.10.1080/01926230490268756Search in Google Scholar PubMed

39. Wang JJ, Sanderson BJ, Wang H. Cyto- and genotoxicity of ultrafine TiO2 particles in cultured human lymphoblastoid cells. Mutat Res 2007;628:99–106.10.1016/j.mrgentox.2006.12.003Search in Google Scholar PubMed

40. Gao GD, Ze YG, Li B, Zhao XY, Zhang T, Sheng L, et al. Ovarian dysfunction and gene-expressed characteristics of female mice caused by long-term exposure to titanium dioxide nanoparticles. J Hazard Mater 2012;243:19–27.10.1016/j.jhazmat.2012.08.049Search in Google Scholar PubMed

41. Burns AA, Vider J, Ow H, Herz E, Penate-Medina O, Baumgart M, et al. Fluorescent silica nanoparticles with efficient urinary excretion for nanomedicine. Nano Lett 2009;9:442–8.10.1021/nl803405hSearch in Google Scholar PubMed PubMed Central

42. Kumar R, Roy I, Ohulchanskky TY, Vathy LA, Bergey EJ, Sajjad M, et al. In vivo biodistribution and clearance studies using multimodal organically modified silica nanoparticles. ACS Nano 2010;4:699–708.10.1021/nn901146ySearch in Google Scholar PubMed PubMed Central

43. WHO (World Health Organization)Global assessment of the state-of-the-science of endocrine disruptorsGeneva, SwitzerlandWHO200213Search in Google Scholar

44. Boas M, Feldt-Rasmussen U, Main KM. Thyroid effects of endocrine disrupting chemicals. Mol Cell Endocrinol 2012;355:240–8.10.1016/j.mce.2011.09.005Search in Google Scholar PubMed

45. Tassinari R, Cubadda F, Moracci G, Aureli F, D’Amato M, Valeri M, et al. Oral, short-term exposure to titanium dioxide nanoparticles in Sprague-Dawley rat: focus on reproductive and endocrine systems and spleen. Nanotoxicology 2014;8:654–62.10.3109/17435390.2013.822114Search in Google Scholar PubMed

46. Iavicoli I, Fontana L, Leso V, Bergamaschi A. The effects of nanomaterials as endocrine disruptors. Int J Mol Sci 2013;14:16732–801.10.3390/ijms140816732Search in Google Scholar PubMed PubMed Central

47. Carreau S, Lambard S, Delalande C, Denis-Galeraud I, Bilinska S, Bourguiba S. Aromatase expression and role of estrogens in male gonad: a review. Reprod Biol Endocrinol 2003;1:35.10.1186/1477-7827-1-35Search in Google Scholar PubMed PubMed Central

48. Fartkhooni FM, Noori A, Momayez M, Sadeghi L, Shirani K, Babadi VY. The effects of nano titanium dioxide (TiO2) in spermatogenesis in wistar rat. Eur J Exp Biol 2013;3:145–9.Search in Google Scholar

49. Hamza RZ, El-Shenawy NS, Ismail HA. Protective effects of blackberry and quercetin on sodium fluoride-induced oxidative stress and histological changes in the hepatic, renal, testis and brain tissue of male rat. J Basic Clin Physiol Pharmacol 2015;26:237–51.10.1515/jbcpp-2014-0065Search in Google Scholar PubMed

50. Peters K, Unger RE, Kirkpatrick CJ, Gatti AM, Monari E. Effects of nano-scaled particles on endothelial cell function in vitro: studies on viability, proliferation and inflammation. J Mater Sci Mater Med 2004;15:321–5.10.1023/B:JMSM.0000021095.36878.1bSearch in Google Scholar

51. Neenu S, Bella M, Gareth JS, Jenkins SM, Paul MW, Thierry GG, et al. Nano genotoxicology: the DNA damaging potential of engineered nanomaterials. Biomaterials 2009;30:3891–14.10.1016/j.biomaterials.2009.04.009Search in Google Scholar PubMed

52. Wang J, Li N, Zheng L, Wang S, Wang Y, Zhao X, et al. P38-Nrf-2 signaling pathway of oxidative stress in mice caused by nanoparticulate TiO2. Biol Trace Elem Res 2011;140:186–91.10.1007/s12011-010-8687-0Search in Google Scholar PubMed

53. Olmedo DG, Tasat DR, Guglielmotti MB, Cabrini RL. Effect of titanium dioxide on the oxidative metabolism of alveolar macrophages: an experimental study in rats. J Biomed Mater Res Part A 2005;73:142–9.10.1002/jbm.a.30230Search in Google Scholar PubMed

54. Limbach LK, Wick P, Manser P, Grass RN, Bruinink A, Stark WJ. Exposure of engineered nanoparticles to human lung epithelial cells: influence of chemical composition and catalytic activity on oxidative stress. Environ Sci Technol 2007;41:4158–63.10.1021/es062629tSearch in Google Scholar PubMed

55. Long TC, Tajuba J, Sama P, Saleh N, Swartz C, Parker J, et al. Nano size titanium dioxide stimulates reactive oxygen species in brain microglia and damages neurons in vitro. Environ Heal Perspect 2007;115:1631–7.10.1289/ehp.10216Search in Google Scholar PubMed PubMed Central

56. Cui Y, Gong X, Duan Y, Li NA, Hu R, Liu H, et al. Hepatocyte apoptosis and its molecular mechanisms in mice caused by titanium dioxide nanoparticles. J Hazard Mater 2010;183: 874–80.10.1016/j.jhazmat.2010.07.109Search in Google Scholar PubMed

57. Li N, Duan Y, Hong M, Zheng L, Fei M, Zhao X, et al. Spleen injury and apoptotic pathway in mice caused by titanium dioxide nanoparticles. Toxicol Lett 2010;195:161–8.10.1016/j.toxlet.2010.03.1116Search in Google Scholar PubMed

58. Liu HT, Ma LL, Zhao JF, Liu J, Yan JY, Ruan J, et al. Biochemical toxicity of nano-anatase TiO2 particles in mice. Biol Trace Elem Res 2009;129:170–80.10.1007/s12011-008-8285-6Search in Google Scholar PubMed

59. Patri A, Umbreit T, Zheng J, Nagashima K, Goering P, Francke-Carroll S, et al. Energy dispersive X-ray analysis of titanium dioxide nanoparticle distribution after intravenous and subcutaneous injection in mice. J Appl Toxicol 2009;29:662–72.10.1002/jat.1454Search in Google Scholar PubMed

Received: 2015-10-8
Accepted: 2016-1-30
Published Online: 2016-4-7
Published in Print: 2016-6-1

©2016 by De Gruyter

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