Evaluation of epigenetic changes of liver tissue induced by oral administration of Titanium dioxide nanoparticles and possible protective role of Nigella Sativa oil, in adult male albino rats

Document Type : Research Paper


1 Associate Professor of Forensic Medicine & Clinical Toxicology, Alexandria University, Egypt

2 Lecturer of Forensic Medicine & Clinical Toxicology, Alexandria University, Egypt

3 Lecturer of Medical Biochemistry, Alexandria University, Egypt


Objective (s): Titanium dioxide nanoparticles (TiO2 NPs) have been recognized as biologically inert material and have been used in a multitude of applications. Nevertheless, the negative impact on the human health is not yet well understood. Aim of the work: The study attempted to evaluate the epigenetic changes of the genome, in the form of DNA methylation in liver tissue samples, resulting from oral administration of TiO2 NPs (mixed rutile and anatase) in adult male albino rats. Furthermore, whether the Nigella sativa oil (NSO) can prevent the toxic effects of TiO2 NPs.
Materials and Methods: Thirty-two adult male albino rats were divided into four groups. (I) control, (II) nigella sativa oil, (III) TiO2 NPs and (IV) TiO2 NPs + nigella sativa oil. The impact of TiO2 NPs on the global DNA methylation and the oxidative status were assessed.
Results: Among the study groups, TiO2 NPs exposure provoked oxidative stress; increased blood levels of MDA and decreased reduced glutathione (GSH) level. The global DNA methylation levels decreased after exposure to titanium nanoparticles. Significant differences were recorded between the control group and the group receiving TiO2 NPs. Marked improvement was noticed after supplementation of nigella sativa oil in terms of DNA methylation and oxidative stress markers.
Conclusion: Oral administration of TiO2 NPs caused global DNA hypo-methylation in liver tissue samples. The epigenetic damage raises the concern about the safety associated with applications of the TiO2 NPs. The maintenance of DNA methylation patterns by Nigella sativa oil has a role in protection against genomic instability.


1.Elsaesser A, Howard CV. Toxicology of nanoparticles. Adv Drug Deliv Rev. 2012; 64: 129-137.
2.Xu A, Chai YF, Nohmi T, Hei TK. Genotoxic responses to titanium dioxide nanoparticles and fullerene in gpt delta transgenic MEF cells. Part Fibre Toxicol. 2009; 6: 3.
3. Trouiller B, Reliene R, Westbrook A, Solaimani P, Schiestl RH. Titanium dioxide nanoparticles induce DNA damage and genetic instability in vivo in mice. Cancer Res. 2009; 69: 8784-8789.
4.Song B1, Zhou T2, Yang W3, Liu J4, Shao L5. Contribution of oxidative stress to TiO2 nanoparticle-induced toxicity. Environ Toxicol Pharmacol. 2016; 48: 130-140.
5.Jaenisch R, Bird A. Epigenetic regulation of gene expression: how the genome integrates intrinsic and environmental signals. Nat Genet. 2003; 33: 245–254.
6.Ebru U1, Burak U, Yusuf S, Reyhan B, Arif K, Faruk TH, Emin M, Aydin K, Atilla II, Semsettin S, Kemal E. Cardioprotective effects of Nigella sativa oil on cyclosporine A-induced cardiotoxicity in rats. Basic Clin Pharmacol Toxicol. 2008; 103(6): 574-580.
7.Yaman I, Balikci E. Protective effects of nigella sativa against gentamicin-induced nephrotoxicity in rats. Exp Toxicol Pathol. 2010; 62(2): 183-190.
8.Farooqui Z, Shahid F, Khan AA, Khan F. Oral administration of Nigella sativa oil and thymoquinone attenuates long term cisplatin treatment induced toxicity and oxidative damage in rat kidney. Biomed Pharmacother. 2017; 96: 912-923.
9.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(1); 46-53.
10.Ohkawa H, Ohishi N, Yagi K: Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979; 95:351.
11.Shen GX. Oxidative stress and diabetes cardiovascular disorders: Roles of mitochondria and NADPH oxidase. Can J Physiol Pharmacol. 2010; 88: 241-248.
12.REF: Kriaucionis S, and Heintz N. The nuclear DNA base, 5-hydroxymethylcytosine is present in brain and enriched in Purkinje neurons.Science. 2009; 324(5929): 929–930.
13.Jovanovic B. Critical review of public health regulations of titanium dioxide, a human food additive. Integr Environ Assess Manag. 2015; 11(1): 10–20.
14.Jones K, Morton J, Smith I, Jurkschat K, Harding AH, Evans G. Human in vivo and in vitro studies on gastrointestinal absorption of titanium dioxide nanoparticles. Toxicol Lett. 2015; 233(2): 95–101.
15.Powell JJ, Faria N, Thomas-McKay E, Pele LC. Origin and fate of dietary nanoparticles and microparticles in the gastrointestinal tract. J Autoimmun 2010; 34(3): 226–330.
16.Smolkova B, Dusinska M, Gabelova A. Nanomedicine and epigenome. Possible health risks. Food Chem Toxicol 2017; 109 (1): 780-796.
17.Wang J, Zhou G, Chen C, Yu H, Wang T, Ma Y, Jia G, Gao Y, Li B, Sun J, Li Y, Jiao F, Zhao Y, Chai Z. Acute toxicity and biodistribution of different sized titanium dioxide particles in mice after oral administration. Toxicol Lett. 2007; 168(2): 176-185.
18.Patil NA, Gade WN, Deobagkar DD. Epigenetic modulation upon exposure of lung fibroblasts to TiO2 and ZnO nanoparticles: alterations in DNA methylation. Int J Nanomedicine 2016; 11: 4509-4519.
19.Song, Bin; Zhou, Ting; Liu, Jia; Shao, Long Quan. Measuring Global DNA Methylation to Assess Neurotoxicity of Titanium Dioxide Nanoparticles. Science of Advanced Materials. 2017; 1051-1056(6).
20.Yue Ma, Yinsheng Guo, Shuang Wu, Ziquan Lv, Qian Zhang and Yuebin Ke. Titanium dioxide nanoparticles induce size-dependent cytotoxicity and genomic DNA hypomethylation in human respiratory cells. RSC Adv. 2017; 7, 23560–23572.
21.Cheung HH, Yang Y, Lee TL, Rennert O, Chan WY. Hypermethylation of genes in testicular embryonal carcinomas. Br J Cancer. 2016; 114: 230–236.
22.Feinberg AP, Ohlsson R, and Henikoff S. The epigenetic progenitor origin of human cancer. Nat Rev Genet 2006; 7, 21–33.
23. Wang Y, Yu Q, Cho AH, Rondeau G, Welsh J, Adamson E, Mercola D, and McClelland M. Survey of differentially methylated promoters in prostate cancer cell lines. Neoplasia. 2005; 7, 748–760.
24.Halušková J. Epigenetic studies in human diseases. Folia Biol (Praha). 2010; 56(3): 83–96.
25.Deobagkar D. DNA methylation and toxicogenomics. In: Sahu SC, editor. Toxicology and Epigenetics. Chichester, UK: John Wiley & Sons, Ltd; 2012.
26.Saquib Q, Al-Khedhairy AA, Siddiqui MA, Abou-Tarboush FM, Azam A, Musarrat J, Titanium Dioxide Nanoparticles Induced Cytotoxicity, Oxidative Stress and DNA Damage in Human Amnion Epithelial (WISH) Cells. Toxicology in Vitro. 2012; 26 (2): 351-361.
27.Solarska K, Gajewska A, Kaczorowski W, Bartosza G, Mitura K, Effect of Nanodiamond Powders on the Viability and Production of Reactive Oxygen and Nitro- gen Species by Human Endothelial Cells, Diamond and Related Materials. 2012; 21: 107-113.
28.de Planque MR, Aghdaei S, Roose T, Morgan H, “Electrophysiological Characterization of Membrane Dis- ruption by Nanoparticles,” ACS Nano. 2011; 5(5): 3599-3606.
29.Sayes CM, Gobin AM, Ausman KD, Mendez J, West JL, Colvin VL. “Nano-C60 Cytotoxicity Is Due to Lipid Peroxidation,” Biomaterials. 2005; 26(36): 7587-7595.
30.Simkó M, Gazsó A, Fiedeler U, Nentwich M. Nanoparticles, Free Radicals and Oxidative Stress,” NanoTrust-Dossie. 2011; 012.
31.Fratelli M, Goodwin LO, Orom UA, Lombardi S, Tonelli R, Mengozzi M, Ghezzi P. Gene Expression Profiling Reveals a Signaling Role of Glutathione in Re- dox Regulation, Proceedings of the National Academy of Sciences of USA. 2005; 102(39): 13998- 14003.
32.Puppel K, Kapusta A, KuczyƄska B. The etiology of oxidative stress in the various species of Animals, a review. J. Sci Food Agric. 2015; 95: 2179-2184.
33.Al-Okbi SY, Mohamed DA, Hamed TE, Edris AE, Fouda K. Hepatic Regeneration and Reno-Protection by Fish oil, Nigella sativa Oil and Combined Fish Oil/Nigella sativa Volatiles in CCl4 treated Rats. J Oleo Sci. 2018; 67(3): 345-353.
34.Majdalawieh AF, Fayyad MW. Immunomodulatory and anti-inflammatory action of Nigella sativa and thymoquinone: A comprehensive review. Int Immunopharmacol. 2015; 28(1): 295-304.