Abstract
With the increasing applications of carbon nanotubes (CNTs) in fields of biomedical engineering and medical chemistry, it is important to understand the response of mammalian cells to the CNTs exposure and treatment. In this study, the influences of multiwalled carbon nanotubes (MWCNTs) on cellular behavior of human dermal fibroblasts and NIH 3T3 murine fibroblasts were investigated. Results showed that the MWCNTs treatment induced dose-dependent cytotoxicity and arrested the cell cycle in the G1 phase, indicating inhibition of DNA synthesis. The presence of MWCNTs also down regulated the expression level of adhesion-related genes, and simultaneously caused cytoskeleton damage and disturbance of actin stress fibers, thereby inducing dramatically adverse effects on the cell physiological functions such as cell spreading, adhesion, migration, and wound healing ability.
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Baughman, R. H., A. A. Zakhidov, and W. A. de Heer. Carbon nanotubes—the route toward applications. Science 297(5582):787–792, 2002.
Bianco, A., and M. Prato. Can carbon nanotubes be considered useful tools for biological applications? Adv. Mater. 15(20):1765–1766, 2003.
Bottini, M., S. Bruckner, K. Nika, N. Bottini, S. Bellucci, A. Magrini, A. Bergamaschi, and T. Mustelin. Multi-walled carbon nanotubes induce T lymphocyte apoptosis. Toxicol. Lett. 160(2):121–126, 2006.
Chang, J. S., K. L. Chang, D. F. Hwang, and Z. L. Kong. In vitro cytotoxicitiy of silica nanoparticles at high concentrations strongly depends on the metabolic activity type of the cell line. Environ. Sci. Technol. 41(6):2064–2068, 2007.
Cheng, C., K. H. Muller, K. K. Koziol, J. N. Skepper, P. A. Midgley, M. E. Welland, and A. E. Porter. Toxicity and imaging of multi-walled carbon nanotubes in human macrophage cells. Biomaterials 30(25):4152–4160, 2009.
Colognato, H., and P. D. Yurchenco. Form and function: the laminin family of heterotrimers. Dev. Dyn. 218(2):213–234, 2000.
Cui, D. X., F. R. Tian, C. S. Ozkan, M. Wang, and H. J. Gao. Effect of single wall carbon nanotubes on human HEK293 cells. Toxicol. Lett. 155(1):73–85, 2005.
Ding, L., J. Stilwell, T. Zhang, O. Elboudwarej, H. Jiang, J. P. Selegue, P. A. Cooke, J. W. Gray, and F. F. Chen. Molecular characterization of the cytotoxic mechanism of multiwall carbon nanotubes and nano-onions on human skin fibroblast. Nano Lett. 5(12):2448–2464, 2005.
Even-Ram, S., and K. M. Yamada. Cell migration in 3D matrix. Curr. Opin. Cell. Biol. 17(5):524–532, 2005.
Fujimoto, L. M., R. Roth, J. E. Heuser, and S. L. Schmid. Actin assembly plays a variable, but not obligatory role in receptor-mediated endocytosis in mammalian cells. Traffic 1(2):161–171, 2000.
Gao, H. J., Y. Kong, and D. X. Cui. Spontaneous insertion of DNA oligonucleotides into carbon nanotubes. Nano Lett. 3(4):471–473, 2003.
Gotlieb, A. I. The endothelial cytoskeleton: organization in normal and regenerating endothelium. Toxicol. Pathol. 18(4 Pt 1):603–617, 1990.
Gupta, A. K., M. Gupta, S. J. Yarwood, and A. S. Curtis. Effect of cellular uptake of gelatin nanoparticles on adhesion, morphology and cytoskeleton organisation of human fibroblasts. J. Control Rel. 95(2):197–207, 2004.
Hafner, J. H., C. L. Cheung, A. T. Woolley, and C. M. Lieber. Structural and functional imaging with carbon nanotube AFM probes. Prog. Biophys. Mol. Biol. 77(1):73–110, 2001.
Hu, H., Y. Ni, S. K. Mandal, V. Montana, B. Zhao, R. C. Haddon, and V. Parpura. Polyethyleneimine functionalized single-walled carbon nanotubes as a substrate for neuronal growth. J. Phys. Chem. B 109(10):4285–4289, 2005.
Iijima, S. Helical microtubules of graphitic carbon. Nature 354:56–58, 1991.
Jia, G., H. F. Wang, L. Yan, X. Wang, R. J. Pei, T. Yan, Y. L. Zhao, and X. B. Guo. Cytotoxicity of carbon nanomaterials: single-wall nanotube, multi-wall nanotube, and fullerene. Environ. Sci. Technol. 39(5):1378–1383, 2005.
Kagan, V. E., Y. Y. Tyurina, V. A. Tyurin, N. V. Konduru, A. I. Potapovich, A. N. Osipov, E. R. Kisin, D. Schwegler-Berry, R. Mercer, V. Castranova, and A. A. Shvedova. Direct and indirect effects of single walled carbon nanotubes on RAW 264.7 macrophages: role of iron. Toxicol. Lett. 165(1):88–100, 2006.
Kaiser, J. P., P. Wick, P. Manser, P. Spohn, and A. Bruinink. Single walled carbon nanotubes (SWCNT) affect cell physiology and cell architecture. J. Mater. Sci. 19(4):1523–1527, 2008.
Kaiser, J. P., H. F. Krug, and P. Wick. Nanomaterial cell interactions: how do carbon nanotubes affect cell physiology? Nanomedicine 4(1):57–63, 2009.
Kam, N. W., and H. Dai. Carbon nanotubes as intracellular protein transporters: generality and biological functionality. J. Am. Chem. Soc. 127(16):6021–6026, 2005.
Kisin, E. R., A. R. Murray, M. J. Keane, X. C. Shi, D. Schwegler-Berry, O. Gorelik, S. Arepalli, V. Castranova, W. E. Wallace, V. E. Kagan, and A. A. Shvedova. Single-walled carbon nanotubes: geno- and cytotoxic effects in lung fibroblast V79 cells. J. Toxicol. Environ. Health A 70(24):2071–2079, 2007.
Kostarelos, K., L. Lacerda, G. Pastorin, W. Wu, S. Wieckowski, J. Luangsivilay, S. Godefroy, D. Pantarotto, J. P. Briand, S. Muller, M. Prato, and A. Bianco. Cellular uptake of functionalized carbon nanotubes is independent of functional group and cell type. Nat. Nanotechnol. 2(2):108–113, 2007.
Laaksonen, T., H. Santos, H. Vihola, J. Salonen, J. Riikonen, T. Heikkila, L. Peltonen, N. Kurnar, D. Y. Murzin, V. P. Lehto, and J. Hirvonent. Failure of MTT as a toxicity testing agent for mesoporous silicon microparticles. Chem. Res. Toxicol. 20(12):1913–1918, 2007.
Li, Y., X. B. Zhang, X. Y. Tao, J. M. Xu, W. Z. Huang, J. H. Luo, Z. Q. Luo, T. Li, F. Liu, Y. Bao, and H. J. Geise. Mass production of high-quality multi-walled carbon nanotube bundles on a Ni/Mo/MgO catalyst. Carbon 43(2):295–301, 2005.
Manna, S. K., S. Sarkar, J. Barr, K. Wise, E. V. Barrera, O. Jejelowo, A. C. Rice-Ficht, and G. T. Ramesh. Single-walled carbon nanotube induces oxidative stress and activates nuclear transcription factor-kappaB in human keratinocytes. Nano Lett. 5(9):1676–1684, 2005.
Mao, Z. W., B. Wang, L. Ma, C. Y. Gao, and J. C. Shen. The influence of polycaprolactone coating on the internalization and cytotoxicity of gold nanoparticles. Nanomedicine 3(3):215–223, 2007.
Monteiro-Riviere, N. A., and A. O. Inman. Challenges for assessing carbon nanomaterial toxicity to the skin. Carbon 44(6):1070–1078, 2006.
Monteiro-Riviere, N. A., A. O. Inman, Y. Y. Wang, and R. J. Nemanich. Surfactant effects on carbon nanotube interactions with human keratinocytes. Nanomedicine 1(4):293–299, 2005.
Oberlin, A., M. Endo, and T. Koyama. Filamentous growth of carbon through benzene decomposition. J. Cryst. Growth 32(3):335–349, 1976.
Oh, J. M., S. J. Choi, S. T. Kim, and J. H. Choy. Cellular uptake mechanism of an inorganic nanovehicle and its drug conjugates: enhanced efficacy due to clathrin-mediated endocytosis. Bioconjug. Chem. 17(6):1411–1417, 2006.
Pan, Z., W. Lee, L. Slutsky, R. A. Clark, N. Pernodet, and M. H. Rafailovich. Adverse effects of titanium dioxide nanoparticles on human dermal fibroblasts and how to protect cells. Small 5(4):511–520, 2009.
Pankov, R., and K. M. Yamada. Fibronectin at a glance. J. Cell. Sci. 115(20):3861–3863, 2002.
Pantarotto, D., R. Singh, D. McCarthy, M. Erhardt, J. P. Briand, M. Prato, K. Kostarelos, and A. Bianco. Functionalized carbon nanotubes for plasmid DNA gene delivery. Angew. Chem. Int. Ed. 43(39):5242–5246, 2004.
Patlolla, A., B. Patlolla, and P. Tchounwou. Evaluation of cell viability, DNA damage, and cell death in normal human dermal fibroblast cells induced by functionalized multiwalled carbon nanotube. Mol. Cell Biochem. 331:207–214, 2009.
Pernodet, N., X. Fang, Y. Sun, A. Bakhtina, A. Ramakrishnan, J. Sokolov, A. Ulman, and M. Rafailovich. Adverse effects of citrate/gold nanoparticles on human dermal fibroblasts. Small 2(6):766–773, 2006.
Raja, P. M., J. Connolley, G. P. Ganesan, L. Ci, P. M. Ajayan, O. Nalamasu, and D. M. Thompson. Impact of carbon nanotube exposure, dosage and aggregation on smooth muscle cells. Toxicol. Lett. 169(1):51–63, 2007.
Reddy, A. R., Y. N. Reddy, D. R. Krishna, and V. Himabindu. Multi wall carbon nanotubes induce oxidative stress and cytotoxicity in human embryonic kidney (HEK293) cells. Toxicology 272(1–3):11–16, 2010.
Ridley, A. J., M. A. Schwartz, K. Burridge, R. A. Firtel, M. H. Ginsberg, G. Borisy, J. T. Parsons, and A. R. Horwitz. Cell migration: integrating signals from front to back. Science 302(5651):1704–1709, 2003.
Sayes, C. M., F. Liang, J. L. Hudson, J. Mendez, W. Guo, J. M. Beach, V. C. Moore, C. D. Doyle, J. L. West, W. E. Billups, K. D. Ausman, and V. L. Colvin. Functionalization density dependence of single-walled carbon nanotubes cytotoxicity in vitro. Toxicol. Lett. 161(2):135–142, 2006.
Schneider, M., F. Stracke, S. Hansen, and U. F. Schaefer. Nanoparticles and their interactions with the dermal barrier. Dermatoendocrinology 1(4):197–206, 2009.
Shi, H., Y. Huang, H. Zhou, X. Song, S. Yuan, Y. Fu, and Y. Luo. Nucleolin is a receptor that mediates antiangiogenic and antitumor activity of endostatin. Blood 110(8):2899–2906, 2007.
Small, J. V., T. Stradal, E. Vignal, and K. Rottner. The lamellipodium: where motility begins. Trends Cell Biol. 12(3):112–120, 2002.
Stadelmann, W. K., A. G. Digenis, and G. R. Tobin. Physiology and healing dynamics of chronic cutaneous wounds. Am. J. Surg. 176(2A Suppl):26S–38S, 1998.
Tabet, L., C. Bussy, N. Amara, A. Setyan, A. Grodet, M. J. Rossi, J. C. Pairon, J. Boczkowski, and S. Lanone. Adverse effects of industrial multiwalled carbon nanotubes on human pulmonary cells. J. Toxicol. Environ. Health A 72(2):60–73, 2009.
Tian, F. R., D. X. Cui, H. Schwarz, G. G. Estrada, and H. Kobayashi. Cytotoxicity of single-wall carbon nanotubes on human fibroblasts. Toxicol. In Vitro 20(7):1202–1212, 2006.
Varedi, M., A. Ghahary, P. G. Scott, and E. E. Tredget. Cytoskeleton regulates expression of genes for transforming growth factor-beta 1 and extracellular matrix proteins in dermal fibroblasts. J. Cell Physiol. 172(2):192–199, 1997.
Wehrle-Haller, B., and B. A. Imhof. Actin, microtubules and focal adhesion dynamics during cell migration. Int. J. Biochem. Cell Biol. 35(1):39–50, 2003.
Xu, L. H., X. Yang, R. J. Craven, and W. G. Cance. The COOH-terminal domain of the focal adhesion kinase induces loss of adhesion and cell death in human tumor cells. Cell Growth Differ. 9(12):999–1005, 1998.
Zhao, B., H. Hu, K. M. Swadhin, and R. C. Haddon. A bone mimic based on the self-assembly of hydroxyapatite on chemically functionalized single-walled carbon nanotubes. Chem. Mater. 17(12):3235–3241, 2005.
Acknowledgments
This work is financially supported by the Natural Science Foundation of China (50903069, 50873087), Zhejiang Provincial Natural Science Foundation of China (Z4090177) and Frame Work Program 7 of European Commission (FP7-NMP-SMALL-2).
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Associate Editor Mona Kamal Marei oversaw the review of this article.
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Zhang, Y., Wang, B., Meng, X. et al. Influences of Acid-Treated Multiwalled Carbon Nanotubes on Fibroblasts: Proliferation, Adhesion, Migration, and Wound Healing. Ann Biomed Eng 39, 414–426 (2011). https://doi.org/10.1007/s10439-010-0151-y
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DOI: https://doi.org/10.1007/s10439-010-0151-y