Document Type : Original Article


1 Diabetes Research Center, Health Research Institute, Department of Physiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

2 Student Research Committee, Department of Pharmacology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

3 Student Research Committee, Department of Physiology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran

4 Student Research Committee, Department of Toxicology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran


Increasing applications of carbon nanotubes (CNTs) indicate the necessity to examine their toxicity. According to previous studies, CNTs caused oxidative stress that impaired β-cell functions and reduced insulin secretion. Our previous study indicated that single-walled carbon nanotubes (SWCNTs) could induce oxidative stress in pancreatic islets. However, there is no study on the effects of multi-walled carbon nanotubes (MWCNTs) on islets and β-cells. Therefore, the present study aims to evaluate effects of MWCNTs on the oxidative stress of islets and the protective effects of caffeic acid (CA) as an antioxidant. The effects of MWCNTs and CA on islets were investigated using MTT assay, reactive oxygen species (ROS), malondialdehyde (MDA), activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), the content of glutathione (GSH) and mitochondrial membrane potential (MMP) and insulin secretion measurements. The lower viability of islet cells was dose-dependent due to the exposure to MWCNTs according to the MTT assay. Further studies revealed that MWCNTs decreased insulin secretion and MMP, induced ROS creation, increased the MDA level, and decreased activities of SOD, GSH-Px, CAT, and content of GSH. Furthermore, the pretreatment of islets with CA returned the changes. These findings indicated that MWCNTs might induce the oxidative stress of pancreatic islets occurring diabetes and protective CA effects that were mediated by the augmentation of the antioxidant defense system of islets. Our research suggested the necessity of conducting further studies on effects of MWCNTs and CA on the diabetes.


  1. Powers KW, Brown SC, Krishna VB, et al. Research strategies for safety evaluation of nanomaterials. Part VI. Characterization of nanoscale particles for toxico-logical evaluation. Toxicol Sci 2006; 90(2): 296-303.
  2. Khalid P, Hussain MA, Suman VB, et al. Toxicology of carbon nanotubes-A review. Int J Appl Eng Res 2016; 11(1): 159-168.
  3. Jafar A, Alshatti Y, Ahmad A. Carbon nanotube toxicity: The smallest biggest debate in medical care. Cogent Med 2016; 3(1): 1217970. doi:10.1080/2331205X. 2016.1217970.
  4. Wang J, Sun P, Bao Y, et al. Cytotoxicity of single-walled carbon nanotubes on PC12 cells. Toxicol In Vitro 2011; 25(1): 242-250.
  5. Deng X, Jia G, Wang H, et al. Translocation and fate of multi-walled carbon nanotubes in vivo. Carbon 2007; 45(7): 1419-1424.
  6. Poland CA, Duffin R, Kinloch I, et al. Carbon nanotubes introduced into the abdominal cavity of mice show asbestos-like pathogenicity in a pilot study. Nat Nanotechnol 2008; 3(7): 423-428.
  7. Muller J, Huaux F, Moreau N, et al. Respiratory toxicity of multi-wall carbon nanotubes. Toxicol Appl Pharmacol 2005; 207(3): 221-231.
  8. Ryman-Rasmussen JP, Cesta MF, Brody AR, et al. Inhaled carbon nanotubes reach the subpleural tissue in mice. Nat Nanotechnol 2009; 4(11): 747-751.
  9. Shvedova AA, Castranova V, Kisin ER, et al. Exposure to carbon nanotube material: assessment of nanotube cytotoxicity using human keratinocyte cells. J Toxicol Environ Health A 2003; 66(20): 1909-1926.
  10. Wang X, Guo J, Chen T, et al. Multi-walled carbon nanotubes induce apoptosis via mitochondrial path-way and scavenger receptor. Toxicol In Vitro 2012; 26(6): 799-806.
  11. Huczko A, Lange H. Carbon nanotubes: experimental evidence for a null risk of skin irritation and allergy. Fullerene Sci Techn 2001; 9(2): 247-250.
  12. Yu LE, Lanry Yung LY, Ong CN, et al. Translocation and effects of gold nanoparticles after inhalation exposure in rats. Nanotoxicology 2007; 1(3): 235-242.
  13. Hassanpour SH, Dehghani MA, Karami SZ, et al. Role of mithochondria in diabetes and its complications. Int J Pharm Sci Res 2018; 9(6): 2185-2189.
  14. Ahangarpour A, Alboghobeish S, Oroojan AA, et al. Mice pancreatic islets protection from oxidative stress induced by single-walled carbon nanotubes through naringin. Hum Exp Toxicol 2018; 37(12): 1268-1281.
  15. Pang C, Zheng Z, Shi L, et al. Caffeic acid prevents acetaminophen-induced liver injury by activating the Keap1-Nrf2 antioxidative defense system. Free Radic Biol Med 2016; 91: 236-246.
  16. Touaibia M, Jean-François J, Doiron J. Caffeic acid, a versatile pharmacophore: an overview. Mini Rev Med Chem 2011; 11(8): 695-713.
  17. Dhungyal B, Koirala P, Sharma C, et al. Caffeic acid-A potent phytocompound against diabetes mellitus a review. SMU Med J 2014; 1(2): 152-161.
  18. Lacy PE, Kostianovsky M. Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes 1967; 16(1): 35-39.
  19. Ahangarpour A, Oroojan AA, Rezae M, et al. Effects of butyric acid and arsenic on isolated pancreatic islets and liver mitochondria of male mouse. Gastroenterol Hepatol Bed Bench 2017; 10(1): 44-53.
  20. Oboh G, Agunloye OM, Adefegha SA, et al. Caffeic and chlorogenic acids inhibit key enzymes linked to type 2 diabetes (in vitro): a comparative study. J Basic Clin Physiol Pharmacol 2015; 26(2): 165-170.
  21. Pourkhalili N, Pournourmohammadi S, Rahimi F, et al. Comparative effects of calcium channel blockers, autonomic nervous system blockers, and free radical scavengers on diazinon-induced hyposecretion of insulin from isolated islets of Langerhans in rats. Arh Hig Rada Toksikol 2009; 60(2): 157-164.
  22. Ahangarpour A, Alboghobeish S, Rezaei M, et al. Evaluation of diabetogenic mechanism of high fat diet in combination with Arsenic exposure in male mice. Iran J Pharm Res 2018; 17(1): 164-183.
  23. Pournourmohammadi S, Ostad SN, Azizi E, et al. Induction of insulin resistance by malathion: Evidence for disrupted islets cells metabolism and mitochondrial dysfunction. Pestic Biochem Phys 2007; 88(3): 346-352.
  24. Hosseini A, Baeeri M, Rahimifard M, et al. Antiapoptotic effects of cerium oxide and yttrium oxide nanoparticles in isolated rat pancreatic islets. Hum Exp Toxicol 2013; 32(5): 544-553.
  25. Xiong FL, Sun XH, Gan L, et al. Puerarin protects rat pancreatic islets from damage by hydrogen peroxide. Eur J Pharmacol 2006; 529(1-3): 1-7.
  26. Colon J, Herrera L, Smith J, et al. Protection from radiation-induced pneumonitis using cerium oxide nanoparticles. Nanomedicine 2009; 5(2): 225-231.
  27. Góth L. A simple method for determination of serum catalase activity and revision of reference range. Clina Chim Acta 1991; 196(2-3): 143-151.
  28. Thomas DW. Handbook of methods for oxygen radical research. J Pediatr Gastroenterol Nutr 1988; 7(2): 314-316.
  29. Bindokas VP, Kuznetsov A, Sreenan S, et al. Visualizing superoxide production in normal and diabetic rat islets of Langerhans. J Biol Chem 2003; 278(11): 9796-9801.
  30. Lawrence RA, Burk RF. Glutathione peroxidase activity in selenium-deficient rat liver. BiochemBiophys Res Commun 1976; 71(4): 952-928.
  31. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254.     
  32. Madani SY, Mandel A, Seifalian AM. A concise review of carbon nanotube's toxicology. Nano Rev 2013; 4: 21521. doi: 10.3402/nano.v4i0.21521.
  33. Liu Z, Davis C, Cai W, et al. Circulation and long-term fate of functionalized, biocompatible single-walled carbon nanotubes in mice probed by Raman spectroscopy. Proc Natl Acad Sci USA 2008; 105(5): 1410-1415.
  34. Wang L, Castranova V, Mishra A, et al. Dispersion of single-walled carbon nanotubes by a natural lung surfactant for pulmonary in vitro and in vivo toxicity studies. Part Fibre Toxicol 2010; 7: 31. doi: 10.1186/ 1743-8977-7-31.
  35. Haniu H, Saito N, Matsuda Y, et al. Biological responses according to the shape and size of carbon nanotubes in BEAS-2B and MESO-1 cells. Int J Nanomedicine 2014; 9: 1979-1990.    
  36. Riding MJ, Martin FL, Trevisan J, et al. Concentration-dependent effects of carbon nanoparticles in gram-negative bacteria determined by infrared spectroscopy with multivariate analysis. Environ Pollut 2012; 163: 226-234.
  37. Mercer RR, Scabilloni J, Wang L, et al. Alteration of deposition pattern and pulmonary response as a result of improved dispersion of aspirated single-walled carbon nanotubes in a mouse model. Am J Physiol Lung Cell Mol Physiol 2008; 294(1): 87-97.
  38. Cui D, Tian F, Ozkan CS, et al. Effect of single wall carbon nanotubes on human HEK293 cells. Toxicol Lett 2005; 155(1): 73-85.
  39. Davoren M, Herzog E, Casey A, et al. In vitro toxicity evaluation of single walled carbon nanotubes on human A549 lung cells. Toxicol In Vitro 2007; 21(3): 438-448.
  40. Sharma CS, Sarkar S, Periyakaruppan A, et al. Single-walled carbon nanotubes induces oxidative stress in rat lung epithelial cells. J Nanosci Nanotechnol 2007; 7(7): 2466-2472.
  41. Aschberger K, Johnston HJ, Stone V, et al. Review of carbon nanotubes toxicity and exposure--appraisal of human health risk assessment based on open literature. Crit Rev Toxicol 2010; 40(9): 759-790.
  42. Simon A, Maletz SX, Hollert H, et al. Effects of multi-walled carbon nanotubes and triclocarban on several eukaryotic cell lines: elucidating cytotoxicity, endocrine disruption, and reactive oxygen species generation. Nanoscale Res Lett 2014; 9(1): 396. doi: 10.1186/ 1556-276X-9-396.
  43. Nel A, Xia T, Mädler L, et al. Toxic potential of materials at the nanolevel. Science 2006; 311(5761): 622-627.
  44. Lièvre V, Becuwe P, Bianchi A, et al. Free radical production and changes in superoxide dismutases associated with hypoxia/reoxygenation-induced apoptosis of embryonic rat forebrain neurons in culture. Free Radic Biol Med 2000; 29(12): 1291-1301.
  45. Bi J, Wang Xb, Chen L, et al. Catalpol protects mesencephalic neurons against MPTP induced neurotoxicity via attenuation of mitochondrial dys-function and MAO-B activity. Toxicol In Vitro 2008; 22(8): 1883-1889.
  46. Grabinski C, Hussain S, Lafdi K, et al. Effect of particle dimension on biocompatibility of carbon nano-materials. Carbon 2007; 45(14): 2828-2835.
  47. Bhattacharya S, Oksbjerg N, Young JF, et al. Caffeic acid, naringenin and quercetin enhance glucose‐stimulated insulin secretion and glucose sensitivity in INS‐1E cells. Diabetes Obes Metab 2014; 16(7): 602-612.
  48. Jung UJ, Lee MK, Park YB, et al. Antihyperglycemic and antioxidant properties of caffeic acid in db/db mice. J Pharmacol Exp Ther 2006; 318(2): 476-483.
  49. Suarez-Pinzon WL, Strynadka K, Rabinovitch A. Destruction of rat pancreatic islet beta-cells by cytokines involves the production of cytotoxic aldehydes. Endocrinology 1996; 137(12): 5290-5296.