Advertisement

Cellular and Molecular Neurobiology

, Volume 34, Issue 1, pp 43–50 | Cite as

Effects of Carbon Nanotubes in a Chitosan/Collagen-Based Composite on Mouse Fibroblast Cell Proliferation

  • Wen Zhao
  • Wenwen Yu
  • Jiawei Zheng
  • Ying Wang
  • Zhiyuan ZhangEmail author
  • Dongsheng ZhangEmail author
Original Research

Abstract

This study investigated the in vitro cytocompatibility of carbon nanotubes (CNTs) in a chitosan/collagen-based composite. Mouse fibroblasts were cultured on the surface of a novel material consisting of CNTs in a chitosan/collagen-based composite (chitosan/collagen+CNTs group). Chitosan/collagen composites without CNTs served as the control material (chitosan/collagen group) and cells cultured normally in tissue culture plates served as blank controls (blank control group). Cell adhesion and proliferation were observed, and cell apoptosis was measured. The doubling time (DT1) of cells was significantly shorter in the chitosan/collagen+CNTs group than in the chitosan/collagen group, and that in the chitosan/collagen group was shorter than in the blank control group. The CNTs in the chitosan/collagen-based composites promoted mouse fibroblast adhesion, producing a distinct cytoskeletal structure. At 24 h after culture, the cytoskeleton of the cells in the chitosan/collagen+CNTs group displayed typical fibroblastic morphology, with clear microfilaments. Cells in the chitosan/collagen group were typically round, with an unclear cytoskeleton. The blank control group even had a few unattached cells. At 4 days after incubation, no early apoptosis of cells was detected in the blank control group, whereas early apoptosis of cells was observed in the chitosan/collagen+CNTs and chitosan/collagen groups. No significant difference in the proportion of living cells was detected among the three groups. After entering the plateau stage, the average cell number in the chitosan/collagen+CNTs group was similar to that in the chitosan/collagen group and significantly smaller than that in the blank control group. Early apoptosis of cells in the blank control group was not detectable. There were significant differences in early apoptosis among the three groups. These results suggest that CNTs in a chitosan/collagen-based composite did not cause significant cytotoxic effects on mouse fibroblasts. Compared with chitosan/collagen composites, early adhesion and proliferation of fibroblasts were increased on chitosan/collagen+CNTs. However, at relatively high cell densities, the CNTs in the chitosan/collagen-based composite might exert an inhibitory effect on mouse fibroblast proliferation by inducing apoptosis.

Keywords

Carbon nanotubes Biodegradable material Cytocompatibility Bioactivity Nerve repair 

Notes

Acknowledgments

The project was supported by the Research Award Fund for outstanding young scientists of Shandong Province (No. BS2011SW037) and the National Natural Science Foundation of China (No. 81270290).

Conflict of interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work; there are no conflicts of interest regarding this manuscript.

References

  1. Bonelli G, Sacchi MC, Barbiero G, Duranti F, Goglio G, Verdun di Cantogno L, Amenta JS, Piacentini M, Tacchetti C, Baccino FM (1996) Apoptois of L929 cells by etoposide:a quantitative and kinetic approach. Exp Cell Res 228(2):292–305PubMedCrossRefGoogle Scholar
  2. Cellot G, Cilia E, Cipollone S, Rancic V, Sucapane A, Giordani S, Gambazzi L, Markram H, Grandolfo M, Scaini D, Gelain F, Casal L (2009) Carbon nanotubes might improveneuronal performance by favouring electrical shortcuts. Nat Nano-technol 4(2):126–133CrossRefGoogle Scholar
  3. Cellot G, Toma FM, Varley ZK, Laishram J, Villari A, Quintana M, Cipollone S, Prato M, Ballerini L (2011) Carbon nanotube scaffolds tune synaptic strength in cultured neural circuits: novel frontiers in nanomaterial-tissue interactions. J Neurosci 31(36):12945–12953PubMedCrossRefGoogle Scholar
  4. Chang CJ, Hsu SH, Lin FT, Chang H, Chang CS (2005) Low-intensity-ultrasound-accelerated nerve regeneration using cell-seeded poly (D, L-lactic acid-co-glycolic acid) conduits: an in vivo and in vitro study. J Biomed Mater Res B 75(1):99–107CrossRefGoogle Scholar
  5. Chen CS, Soni S, Le C, Biasca M, Farr E, Chen EY, Chin WC (2012) Human stem cell neuronal differentiation on silk-carbon nanotube composite. Nanoscale Res Lett 7(1):126PubMedCrossRefGoogle Scholar
  6. Cregg JM, Wiseman SL, Pietrzak-Goetze NM, Smith MR, Jaroch DB, Clupper DC, Gilbert RJ (2010) A rapid, quantitative method for assessing axonal extension on biomaterial platforms. Tissue Eng Part C 16(2):167–172CrossRefGoogle Scholar
  7. Dienstknecht T, Klein S, Vykoukal J, Gehmert S, Koller M, Gosau M, Prantl L (2013) Type I collagen nerve conduits for median nerve repairs in the forearm. J Hand Surg 38(6):1119–1124CrossRefGoogle Scholar
  8. Ding F, Wu J, Yang Y, Hu W, Zhu Q, Tang X, Liu J, Gu X (2010) Use of tissue-engineered nerve grafts consisting of a chitosan/poly(lactic-co-glycolic acid)-based scaffold included with bone marrow mesenchymal cells for bridging 50-mm dog sciatic nerve gaps. Tissue Eng Part A 16(12):3779–3790PubMedCrossRefGoogle Scholar
  9. Galvan-Garcia P, Keefer EW, Yang F, Zhang M, Fang S, Zakhidov AA, Baughman RH, Romero MI (2007) Robust cell migration andneuronal growth on pristine carbon nanotube sheets and yarns. J Bio-mater Sci Polym Ed 18(10):1245–1261CrossRefGoogle Scholar
  10. Lee DY, Choi BH, Park JH, Zhu SJ, Kim BY, Huh JY, Lee SH, Jung JH, Kim SH (2006) Nerve regeneration with the use of a poly(l-lactide-co-glycolic acid)-coated collagen tube filled with collagen gel. J Craniomaxillofac Surg 34(1):50–56PubMedCrossRefGoogle Scholar
  11. Malarkey EB, Fisher KA, Bekyarova E, Liu W, Haddon RC, Parpura V (2009) Conductive single-walled carbon nanotube substrates modulate neuronal growth. Nano Lett 9(1):264–268PubMedCentralPubMedCrossRefGoogle Scholar
  12. Nie X, Zhang YJ, Tian WD, Jiang M, Dong R, Chen JW, Jin Y (2007) Improvement of peripheral nerve regeneration by a tissue-engineered nerve filled with ectomesenchymal stem cells. Int J Oral Maxillofac Surg 36(1):32–38PubMedCrossRefGoogle Scholar
  13. Paluch D, Szosland L, Staniszewska-Kuś J, Solski L, Szymonowicz M, Gebarowska E (2000) The biological assessment of the chitin fibres. Polim Med 30(3):3–6PubMedGoogle Scholar
  14. Runge MB, Dadsetan M, Baltrusaitis J, Ruesink T, Lu L, Windebank AJ, Yaszemski MJ (2010) Development of electrically con-ductive oligo (polyethylene glycol) fumarate-polypyrrole hydrogels fornerve regeneration. Biomacromolecules 11(11):2845–2853CrossRefGoogle Scholar
  15. Sager T, Wolfarth MG, Friend S, Hubbs AF, Hamilton RF, Wu N, Yang F, Porter DW, Holian A (2013) Effect of multi-walled carbon nanotube surface modification on bioactivity in the C57Bl/6 mouse model. Nanotoxicology 39(1):48–57Google Scholar
  16. Sorkin R, Gabay T, Blinder P, Baranes D, Ben-Jacob E, Hanein Y (2006) Compact self-wiring in cultured neural networks. J Neural Eng 3(2):95–101PubMedCrossRefGoogle Scholar
  17. Wahlström O, Linder C, Kalén A, Magnusson P (2007) Variation of pH in lysed platelet concentrates influence proliferation and alkaline phosphatase activity in human osteoblast-like cells. Platelets 18(2):113–118PubMedCrossRefGoogle Scholar
  18. Yu W, Zhao W, Zhu C, Zhang X, Ye D, Zhang W, Zhou Y, Jiang X, Zhang Z (2011) Sciatic nerve regeneration in rats by a promising electrospun collagen/poly(ε-caprolactone) nerve conduit with tailored degradation rate. BMC Neurosci 12:68PubMedCentralPubMedCrossRefGoogle Scholar
  19. Zhao W, Zhang ZY, Sun J, Zheng JW, Jiang XQ, Zhu YQ, Wang Y, Jiang LX (2009) Peripheral nerve regeneration using carbon nanotubes enhanced chitosan/collagen composite nerve conduit. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 13(47):9236–9240Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Department of StomatologyProvincial Hospital Affiliated to Shandong UniversityJinanChina
  2. 2.Department of Oral and Maxillo-facial Surgery, The Ninth People’s Hospital of Shanghai, School of StomatologyShanghai Jiaotong UniversityShanghaiChina
  3. 3.Key Laboratory for Thin Film and Microfabrication Technology, Ministry of Education, Research Institute of Micro/Nanometer Science and TechnologyShanghai Jiao Tong UniversityShanghaiChina

Personalised recommendations