Use of chitosan conduit combined with bone marrow mesenchymal stem cells for promoting peripheral nerve regeneration

  • Lei Zheng
  • Hui-Fei Cui


Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to develop and test chitosan conduit to use in peripheral nerve reconstruction, either alone or combined with bone marrow mesenchymal stem cells (BMSCs). In this study, the roles of the degree of deacetylation (DD) and molecular weight of chitosan on some biological properties of chitosan films, including hydrophilicity, degradation and BMSCs affinity were investigated. The molecular weight of Chitosans used are 5 × 104 Da, 2 × 105 Da, 5 × 105 Da, 1 × 106 Da, the deacetylation degrees are 85, 95%, respectively. The affinity of eight kinds of Chitosans to the BMSCs was assessed by MTT assay, the contact angle and the degradation time of the materials in vivo were also measured. Chitosans with the molecular weight of 1 × 106 Da and DD of 95% can significantly promote the survival and outgrowth of cells, which have better hydrophilicity and can remain integrity even after 8 to 16 weeks, all of above meet the requirement of nerve engineering. The BMSCs we transplanted can differentiate into neural stem cells in vivo, and the materials we selected combined with BMSCs can bridge 8-mm-long neural gap better resulting from the differentiation effects of the BMSCs.


Chitosan Contact Angle Sciatic Nerve Chitosan Film Bone Marrow Mesenchymal Stem Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Tanigawa J, Miyoshi N, Sakurai K. Characterization of chitosan/citrate and chitosan/acetate films and applications for wound healing. J Appl Polym Sci. 2008;110:608–15.CrossRefGoogle Scholar
  2. 2.
    Chalfoun CT, Wirth GA, Evans GR. Tissue engineered nerve constructs: where do we stand. J Cell Mol Med. 2006;10(2):309–17.CrossRefPubMedGoogle Scholar
  3. 3.
    Khor E, Lim LY. Implantable applications of chitin and chitosan. Biomaterials. 2003;24(13):2339–49.CrossRefPubMedGoogle Scholar
  4. 4.
    Jianga M, Zhuge X, Yang Y, Gu X, Ding F. The promotion of peripheral nerve regeneration by chitooligosaccharides in the rat nerve crush injury model. Neurosci Lett. 2009;454:239–43.CrossRefGoogle Scholar
  5. 5.
    Ciardelli G, Chiono V. Materials for peripheral nerve regeneration. Macromol Biosci. 2005;6(1):13–26.CrossRefGoogle Scholar
  6. 6.
    Amado S, Simoes MJ, Armada da Silva PA, Luís AL, Shirosaki Y, Lopes MA, et al. Use of hybrid chitosan membranes and N1E-115 cells for promoting nerve regeneration in an axonotmesis rat model. Biomaterials. 2008;29:4409–19.CrossRefPubMedGoogle Scholar
  7. 7.
    Bhatheja K, Field J. Schwann cells: origins and role in axonal maintenance and regeneration. Int J Biochem Cell Biol. 2006;38:1995–9.CrossRefPubMedGoogle Scholar
  8. 8.
    Aijun W, Qiang A, Qing H, Xiaoming G, Kai G, Yandao G, et al. Neural stem cell affinity of chitosan and feasibility of chitosan-based porous conduits as scaffolds for nerve tissue engineering. Tsinghua Sci Technol. 2006;11(4):415–20.CrossRefGoogle Scholar
  9. 9.
    Wenling C, Duohui J, Jiamou L, Yandao G, Nanming Z, Xiufang Z. Effects of the degree of deacetylation on the physicochemical properties and Schwann cell affinity of chitosan films. J Biomater Appl. 2005;20(2):157–77.CrossRefPubMedGoogle Scholar
  10. 10.
    Hsu S-h, Whu SW, Tsai C-L, Wu Y-H, Chen H-W, Hsieh K-H. Chitosan as scaffold materials: effects of molecular weight and degree of deacetylation. J Polym Res. 2004;11:141–7.CrossRefGoogle Scholar
  11. 11.
    Freier T, Koh HS, Kazazian K, Shoichet MS. Controlling cell adhesion and degradation of chitosan films by N-acetylation. Biomaterials. 2005;26(29):5872–8.CrossRefPubMedGoogle Scholar
  12. 12.
    Pountos I, Corscadden D, Emery P, Giannoudis PV. Mesenchymal stem cell tissue engineering: techniques for isolation, expansion and application Giannoudis. Injury. 2007;38:S23–33.CrossRefPubMedGoogle Scholar
  13. 13.
    Lei Z, Yongda L, Jun M, Yingyu S, Shaoju Z, Xinwen Z, et al. Culture and neural differentiation of rat bone marrow mesenchymal stem cells in vitro. Cell Biol Int. 2007;31(9):916–23.CrossRefPubMedGoogle Scholar
  14. 14.
    Wislet-Gendebien S, Leprince P, Moonen G, Rogister B. Regulation of neural markers nestin and GFAP expression by cultivated bone marrow stromal cells. J Cell Sci. 2003;116:3295–302.CrossRefPubMedGoogle Scholar
  15. 15.
    Wislet-Gendebien S, Hans G, Leprince P, Rigo JM, Moonen G, Rogister B. Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype. Stem Cells. 2005;23:392–402.CrossRefPubMedGoogle Scholar
  16. 16.
    Wang J, Ding F, Gu Y, Liu J, Gu X. Bone marrow mesenchymal stem cells promote cell proliferation and neurotrophic function of Schwann cells in vitro and in vivo. Brain Res. 2009;1262:7–15.CrossRefPubMedGoogle Scholar
  17. 17.
    Brehm M, Zeus T, Strauer BE. Stem cells-clinical application and perspectives. Herz. 2002;27:611–20.CrossRefPubMedGoogle Scholar
  18. 18.
    Jing L, Yandao G, Nanming Z, Xiufang Z. Preparation of N-butyl chitosan and study of its physical and biological properties. J Appl Polym Sci. 2005;98:1016–24.CrossRefGoogle Scholar
  19. 19.
    Evans GR, Facs MD. Challenges to nerve regeneration. Semin Surg Oncol. 2000;19:312–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Lu L, Chen X, Zhang CW, Yang WL, Wu YJ, Sun L, et al. Morphological functional characterization of predifferentiation of myelinating glia-like cells from human bone marrow stromal cells through activation of F3/Notch signaling in mouse retina. Stem Cells. 2008;26:580–90.CrossRefPubMedGoogle Scholar
  21. 21.
    Lu J, Moochhala S, Moore XL, Ng KC, Tan MH, Lee LK, et al. Adult bone marrow cells differentiate into neural phenotypes improve functional recovery in rats following traumatic brain injury. Neurosci Lett. 2006;398:12–7.CrossRefPubMedGoogle Scholar
  22. 22.
    Yang Y, Chen X, Ding F, Zhang P, Liu J, Gu X. Biocompatibility evaluation of silk fibroin with peripheral nerve tissues cells in vitro. Biomaterials. 2007;28:1643–52.CrossRefPubMedGoogle Scholar
  23. 23.
    Wang X, Hu W, Cao Y, Yao J, Wu J, Gu X. Dog sciatic nerve regeneration across a 30-mm defect bridged by a chitosan/PGA artificial nerve graft. Brain. 2005;128:1897–910.CrossRefPubMedGoogle Scholar
  24. 24.
    Bain JR, Mackinnon SE, Hunter DA. Functional evaluation of complete sciatic, peroneal, and posterior tibial nerve lesions in the rat. Plast Reconstr Surg. 1989;83:129–38.PubMedCrossRefGoogle Scholar
  25. 25.
    Chau LK, Porter MD. Surface isoelectric point of evaporated silver flms: determination by contact angle titration. J Colloid Interface Sci. 1991;145:283–6.CrossRefGoogle Scholar
  26. 26.
    Cheng M, Cao W, Gao Y, Gong Y, Zhao N, Zhang X. Studies on nerve cell affinity of biodegradable modified chitosan flms. J Biomater Sci, Polym Ed. 2003;14(10):1155–67.CrossRefGoogle Scholar
  27. 27.
    Cheng MY, Deng JG, Yang F, Gong YD, Zhao NM, Zhang XF. Study on physical properties and nerve cell affinity of composite films from chitosan and gelatin solutions. Biomaterials. 2003;24(17):2871–80.CrossRefPubMedGoogle Scholar
  28. 28.
    Drllon P, Yu X, Sridharan A, Ranieri JP, Bellamkonda RV. The influence of physical structure and charge on neurite extension in a 3D hydrogel scaffold. J Biomater Sci, Polym Ed. 1998;9(10):1049–69.CrossRefGoogle Scholar
  29. 29.
    Yuan Y, Zhang P, Yang Y, Wang X, Gu X. The interaction of Schwann cells with chitosan membranes and fibers in vitro. Biomaterials. 2004;25:4273–8.CrossRefPubMedGoogle Scholar
  30. 30.
    Kopen GC, Prockop DJ, Phinney DG. Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains. Proc Natl Acad Sci USA. 1999;96(19):10711–6.CrossRefPubMedADSGoogle Scholar
  31. 31.
    Ernst C, Christie BR. The putative neural stem cell marker, nestin, is expressed in heterogeneous cell types in the adult rat neocortex. Neuroscience. 2006;138:183–8.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Institute of Pharmacy, Shandong Traffic HospitalJinanChina
  2. 2.Institute of Biochemical and Biotechnological Drugs, School of PharmacyShandong UniversityJinanChina

Personalised recommendations