Abstract
This chapter provides a detailed protocol for studying the effects of carbon nanotubes (CNTs) on the proliferation, differentiation, adipocytic transdifferentiation, and mineralization of primary osteoblasts. SWNTs, DWNTs, and MWNTs with the same mean length and various diameters were shown to reduce the viability of osteoblasts and inhibit the adipocytic transdifferentiation in both time- and dose-dependent manners. The order of inhibition effect is SWNTs > DWNTs > MWNTs. CNTs were found to inhibit the formation of mineralized nodules greatly and dose-dependently during the final stage of osteoblast differentiation, causing a 50% decrease in the formation of mineralized nodules at the concentration of 50 μg/mL. The expression of important proteins such as Runx-2 and Col-I in osteoblasts was also greatly inhibited by the CNTs. TEM results revealed that the effects on cellular behavior may be exerted by the CNTs from in- and outside of the cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chen RJ, Zhan YG, Wang DW, Dai HJ (2001) Noncovalent sidewall functionalization of single-walled carbon nanotubes for protein immobilization. J Am Chem Soc 123:3838-3839
Heller DA, Baik S, Eurell TE, Strano MS (2005) Single-walled carbon nanotube spectroscopy in live cells: towards long-term labels and optical sensors. Adv Mater 17:2793-2799
Gao LZ, Nie L, Wang TH, Qin YJ, Guo ZX, Yang DL, Yan XY (2006) Carbon nanotube delivery of the GFP gene into mammalian cells. Chembiochem 7:239-242
Cui DX, Tian FR, Kong Y, Titushikin I, Gao HJ (2004) Effects of single-walled carbon nanotubes on the polymerase chain reaction. Nanotechnology 15:154-157
Murr LE, Bang JJ, Esquivel EV, Guerrero PA, Lopez A (2004) Carbon nanotubes, nanocrystal forms, and complex nanoparticle aggregates in common fuel-gas combustion sources and the ambient air. J Nanopart Res 6:241-251
Donaldson K, Li XY, Mac NW (1998) Ultrafine (nanometre) particle mediated lung injury. J Aerosol Sci 29(5/6):553-560
Nel A, Xia T, Madler L, Li N (2006) Toxic potential of materials at the nanolevel. Science 311:622-627
Wang HF, Wang J, Deng XY, Sun HF, Shi ZJ, Gu ZN, Liu YF, Zhao YL (2004) Biodistribution of carbon single-wall carbon nanotubes in mice. J Nanosci Nanotechnol 4(8):1019-1024
Shvedova AA, Castranova V, Kisin ER, Schwegler-Berry D, Murray AR, Gandelsman VZ, Maynard A, Baron P (2003) Exposure to carbon nanotube material: assessment of nanotube cytotoxicity using human keratinocyte cells. J Toxicol Environ Health 66:1909-1926
Jia G, Wang HF, Yan L, Wang X, Pei RJ, Yan T, Zhao YL, Guo XB (2005) Cytotoxicity of carbon nanomaterials: single-wall nanotube, multi-wall nanotube, and fullerene. Environ Sci Technol 39:1378-1383
Mattson MP, Haddon RC, Rao AM (2001) Molecular functionalization of carbon nanotubes and use as substrates for neuronal growth. J Mol Neurosci 14:175-182
Yi CQ, Fong CC, Chen WW, Qi SJ, Tzang CH, Lee ST, Yang MS (2007) Interactions between carbon nanotubes and DNA polymerase and restriction endonucleases. Nanotechnology 18:025102
Cui DX, Tian FR, Ozkan CS, Wang M, Gao HJ (2005) Effect of single wall carbon nanotubes on human HEK293 cells. Toxicol Lett 155:73-85
Gutwein LG, Webster TJ (2002) Osteoblast and chondrocyte proliferation in the presence of alumina and titania nanoparticles. J Nanopart Res 4:231-238
Zhang DW, Zhang JC, Chen Y, Yang MS, Yao XS (2007) Methods for anti-osteoporosis drug screening in vitro. Chin Pharm J 42(3):161-164
Zhang JC, Li XX, Xu SJ, Wang K, Yu SF, Lin Q (2004) Effects of rare earth ions on proliferation, differentiation and function expression of cultured osteoblasts in vitro. Prog Nat Sci 14(4):404-409
Ding LH, Stilwell J, Zhang TT, Elboudwarej O, Jiang HJ, Selegue JP, Cooke PA, Gray JW, Chen FF (2005) Molecular characterization of the cytotoxic mechanism of multiwall carbon nanotubes and nano-onions on human skin fibroblast. Nano Lett 5:2448-2464
Yang S, Wie D, Wang D, Phimphilai M, Krebsbach PH, Franceschi RT (2003) In vitro and in vivo synergistic interactions between the Runx2/Cbfa1 transcription factor and bone morphogenetic protein-2 in stimulating osteoblast differentiation. J Bone Miner Res 18:705-715
Park JH, Park BH, Kim HK, Park TS, Baek HS (2002) Hypoxia decreases Runx2/Cbfa1 expression in human osteoblast-like cells. Mol Cell Endocrinol 192:197-203
Zhang DW, Yi CQ, Zhang JC, Chen Y, Yao XS, Yang MS (2007) The effects of carbon nanotubes on the proliferation and differentiation of primary osteoblasts. Nanotechnology 18:475102
Acknowledgements
The authors would like to thank the National Natural Science Foundation of China (NSFC), the Key Laboratory Scheme of Science and Technology Bureau of Shenzhen Municipal Government, BTC operation fund (CityU project No. 9683001) and City University of Hong Kong (Project No.7002100) for financial support.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Zhang, D., Yi, C., Qi, S., Yao, X., Yang, M. (2010). Effects of Carbon Nanotubes on the Proliferation and Differentiation of Primary Osteoblasts. In: Balasubramanian, K., Burghard, M. (eds) Carbon Nanotubes. Methods in Molecular Biology, vol 625. Humana Press. https://doi.org/10.1007/978-1-60761-579-8_5
Download citation
DOI: https://doi.org/10.1007/978-1-60761-579-8_5
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60761-577-4
Online ISBN: 978-1-60761-579-8
eBook Packages: Springer Protocols