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Chitosan scaffolds induce human dental pulp stem cells to neural differentiation: potential roles for spinal cord injury therapy

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Abstract

Cell-based transplantation strategies hold great potential for spinal cord injury (SCI) repair. Chitosan scaffolds have therapeutic benefits for spinal cord regeneration. Human dental pulp stem cells (DPSCs) are abundant available stem cells with low immunological incompatibility and can be considered for cell replacement therapy. The purpose of this study is to investigate the role of chitosan scaffolds in the neural differentiation of DPSCs in vitro and to assess the supportive effects of chitosan scaffolds in an animal model of SCI. DPSCs were incubated with chitosan scaffolds. Cell viability and the secretion of neurotrophic factors were analyzed. DPSCs incubated with chitosan scaffolds were treated with neural differentiation medium for 14 days and then neural genes and protein markers were analyzed by Western blot and reverse transcription plus the polymerase chain reaction. Our study revealed a higher cell viability and neural differentiation in the DPSC/chitosan-scaffold group. Compared with the control group, the levels of BDNF, GDNF, b-NGF, and NT-3 were significantly increased in the DPSC/chitosan-scaffold group. The Wnt/β-catenin signaling pathway played a key role in the neural differentiation of DPSCs combined with chitosan scaffolds. Transplantation of DPSCs together with chitosan scaffolds into an SCI rat model resulted in the marked recovery of hind limb locomotor functions. Thus, chitosan scaffolds were non-cytotoxic and provided a conducive and favorable microenvironment for the survival and neural differentiation of DPSCs. Transplantation of DPSCs might therefore be a suitable candidate for treating SCI and other neuronal degenerative diseases.

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References

  • Arthur A, Rychkov G, Shi S, Koblar SA, Gronthos S (2008) Adult human dental pulp stem cells differentiate toward functionally active neurons under appropriate environmental cues. Stem Cells 26:1787–1795

    Article  CAS  PubMed  Google Scholar 

  • Basso DM, Beattie MS, Bresnahan JC (1995) A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma 12:1–21

    Article  CAS  PubMed  Google Scholar 

  • Bozkurt G, Mothe AJ, Zahir T, Kim H, Shoichet MS, Tator CH (2010) Chitosan channels containing spinal cord-derived stem/progenitor cells for repair of subacute spinal cord injury in the rat. Neurosurgery 67:1733–1744

    Article  PubMed  Google Scholar 

  • Bray AF, Cevallos RR, Gazarian K, Lamas M (2014) Human dental pulp stem cells respond to cues from the rat retina and differentiate to express the retinal neuronal marker rhodopsin. Neuroscience 280:142–155

    Article  CAS  PubMed  Google Scholar 

  • Ceruti S, Villa G, Genovese T, Mazzon E, Longhi R, Rosa P, Bramanti P, Cuzzocrea S, Abbracchio MP (2009) The P2Y-like receptor GPR17 as a sensor of damage and a new potential target in spinal cord injury. Brain 132:2206–2218

    Article  PubMed  Google Scholar 

  • Cho SR, Kim YR, Kang HS, Yim SH, Park CI, Min YH, Lee BH, Shin JC, Lim JB (2009) Functional recovery after the transplantation of neurally differentiated mesenchymal stem cells derived from bone barrow in a rat model of spinal cord injury. Cell Transplant 18:1359–1368

    Article  PubMed  Google Scholar 

  • Clevers H (2006) Wnt/beta-catenin signaling in development and disease. Cell 127:469–480

    Article  CAS  PubMed  Google Scholar 

  • d’Aquino R, De Rosa A, Laino G, Caruso F, Guida L, Rullo R, Checchi V, Laino L, Tirino V, Papaccio G (2009) Human dental pulp stem cells: from biology to clinical applications. J Exp Zool B Mol Dev Evol 312B:408–415

    Article  PubMed  Google Scholar 

  • Espada J, Calvo MB, Diaz-Prado S, Medina V (2009) Wnt signalling and cancer stem cells. Clin Transl Oncol 11:411–427

    Article  CAS  PubMed  Google Scholar 

  • Feng X, Xing J, Feng G, Sang A, Shen B, Xu Y, Jiang J, Liu S, Tan W, Gu Z, Li L (2013) Age-dependent impaired neurogenic differentiation capacity of dental stem cell is associated with Wnt/beta-catenin signaling. Cell Mol Neurobiol 33:1023–1031

    Article  CAS  PubMed  Google Scholar 

  • Feng X, Feng G, Xing J, Shen B, Tan W, Huang D, Lu X, Tao T, Zhang J, Li L, Gu Z (2014a) Repeated lipopolysaccharide stimulation promotes cellular senescence in human dental pulp stem cells (DPSCs). Cell Tissue Res 356:369–380

    Article  CAS  PubMed  Google Scholar 

  • Feng X, Lu X, Huang D, Xing J, Feng G, Jin G, Yi X, Li L, Lu Y, Nie D, Chen X, Zhang L, Gu Z, Zhang X (2014b) 3D porous chitosan scaffolds suit survival and neural differentiation of dental pulp stem cells. Cell Mol Neurobiol 34:859–870

    Article  PubMed  Google Scholar 

  • Gaspar VM, Sousa F, Queiroz JA, Correia IJ (2011) Formulation of chitosan-TPP-pDNA nanocapsules for gene therapy applications. Nanotechnology 22:015101

    Article  CAS  PubMed  Google Scholar 

  • Giuliani A, Manescu A, Langer M, Rustichelli F, Desiderio V, Paino F, De Rosa A, Laino L, d’Aquino R, Tirino V, Papaccio G (2013) Three years after transplants in human mandibles, histological and in-line holotomography revealed that stem cells regenerated a compact rather than a spongy bone: biological and clinical implications. Stem Cells Transl Med 2:316–324

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gronthos S, Mankani M, Brahim J, Robey PG, Shi S (2000) Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A 97:13625–13630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Huang GT, Gronthos S, Shi S (2009) Mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J Dent Res 88:792–806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kas HS (1997) Chitosan: properties, preparations and application to microparticulate systems. J Microencapsul 14:689–711

    Article  CAS  PubMed  Google Scholar 

  • Khor E, Lim LY (2003) Implantable applications of chitin and chitosan. Biomaterials 24:2339–2349

    Article  CAS  PubMed  Google Scholar 

  • Kim H, Zahir T, Tator CH, Shoichet MS (2011) Effects of dibutyryl cyclic-AMP on survival and neuronal differentiation of neural stem/progenitor cells transplanted into spinal cord injured rats. PLoS One 6:e21744

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim NR, Lee DH, Chung PH, Yang HC (2009) Distinct differentiation properties of human dental pulp cells on collagen, gelatin, and chitosan scaffolds. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 108:e94–e100

    Article  PubMed  Google Scholar 

  • Koda M, Hashimoto M, Murakami M, Yoshinaga K, Ikeda O, Yamazaki M, Koshizuka S, Kamada T, Moriya H, Shirasawa H, Sakao S, Ino H (2004) Adenovirus vector-mediated in vivo gene transfer of brain-derived neurotrophic factor (BDNF) promotes rubrospinal axonal regeneration and functional recovery after complete transection of the adult rat spinal cord. J Neurotrauma 21:329–337

    Article  PubMed  Google Scholar 

  • Laino G, d’Aquino R, Graziano A, Lanza V, Carinci F, Naro F, Pirozzi G, Papaccio G (2005) A new population of human adult dental pulp stem cells: a useful source of living autologous fibrous bone tissue (LAB). J Bone Min Res 20:1394–1402

    Article  Google Scholar 

  • Lillesaar C, Arenas E, Hildebrand C, Fried K (2003) Responses of rat trigeminal neurones to dental pulp cells or fibroblasts overexpressing neurotrophic factors in vitro. Neuroscience 119:443–451

    Article  CAS  PubMed  Google Scholar 

  • Lu P, Jones LL, Snyder EY, Tuszynski MH (2003) Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury. Exp Neurol 181:115–129

    Article  CAS  PubMed  Google Scholar 

  • Miura M, Gronthos S, Zhao M, Lu B, Fisher LW, Robey PG, Shi S (2003) SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci U S A 100:5807–5812

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nosrat IV, Smith CA, Mullally P, Olson L, Nosrat CA (2004) Dental pulp cells provide neurotrophic support for dopaminergic neurons and differentiate into neurons in vitro; implications for tissue engineering and repair in the nervous system. Eur J Neurosci 19:2388–2398

    Article  PubMed  Google Scholar 

  • Pierdomenico L, Bonsi L, Calvitti M, Rondelli D, Arpinati M, Chirumbolo G, Becchetti E, Marchionni C, Alviano F, Fossati V, Staffolani N, Franchina M, Grossi A, Bagnara GP (2005) Multipotent mesenchymal stem cells with immunosuppressive activity can be easily isolated from dental pulp. Transplantation 80:836–842

    Article  PubMed  Google Scholar 

  • Sakai K, Yamamoto A, Matsubara K, Nakamura S, Naruse M, Yamagata M, Sakamoto K, Tauchi R, Wakao N, Imagama S, Hibi H, Kadomatsu K, Ishiguro N, Ueda M (2012) Human dental pulp-derived stem cells promote locomotor recovery after complete transection of the rat spinal cord by multiple neuro-regenerative mechanisms. J Clin Invest 122:80–90

    CAS  PubMed  Google Scholar 

  • Sharp J, Keirstead HS (2007) Therapeutic applications of oligodendrocyte precursors derived from human embryonic stem cells. Curr Opin Biotechnol 18:434–440

    Article  CAS  PubMed  Google Scholar 

  • Shi W, Nie D, Jin G, Chen W, Xia L, Wu X, Su X, Xu X, Ni L, Zhang X, Zhang X, Chen J (2012) BDNF blended chitosan scaffolds for human umbilical cord MSC transplants in traumatic brain injury therapy. Biomaterials 33:3119–3126

    Article  CAS  PubMed  Google Scholar 

  • Su WT, Shih YA, Ko CS (2013) Effect of chitosan conduit under a dynamic culture on the proliferation and neural differentiation of human exfoliated deciduous teeth stem cells. J Tissue Eng Regen Med (in press)

  • Sun G, Zhang XZ, Chu CC (2007) Formulation and characterization of chitosan-based hydrogel films having both temperature and pH sensitivity. J Mater Sci Mater Med 18:1563–1577

    Article  CAS  PubMed  Google Scholar 

  • Wagers AJ, Sherwood RI, Christensen JL, Weissman IL (2002) Little evidence for developmental plasticity of adult hematopoietic stem cells. Science 297:2256–2259

    Article  CAS  PubMed  Google Scholar 

  • Wei X, Zhang C, Gu Q (2010) Properties, products, and applications of chitosan. Chin J Reparative Reconstr Surg 24:1265–1270

    Google Scholar 

  • Xu Y, Gu Z, Shen B, Xu G, Zhou T, Jiang J, Xing J, Liu S, Li M, Tan W, Feng G, Sang A, Li L (2013) Roles of Wnt/beta-catenin signaling in retinal neuron-like differentiation of bone marrow mesenchymal stem cells from nonobese diabetic mice. J Mol Neurosci 49:250–261

    Article  CAS  PubMed  Google Scholar 

  • Yang Q, Du X, Fang Z, Xiong W, Li G, Liao H, Xiao J, Wang G, Li F (2014) Effect of calcitonin gene-related peptide on the neurogenesis of rat adipose-derived stem cells in vitro. PLoS One 9: e86334

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang J, Li D, Shen A, Mao H, Jin H, Huang W, Xu D, Fan J, Chen J, Yang L, Cui Z (2013) Expression of RBMX after spinal cord injury in rats. J Mol Neurosci 49:417–429

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Yang D, Nie J (2008) Chitosan/polyethylene glycol diacrylate films as potential wound dressing material. Int J Biol Macromol 43:456–462

    Article  CAS  PubMed  Google Scholar 

Download references

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    Authors and Affiliations

    1. Department of Spine Surgery, The Second Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, 226001, China

      Jinlong Zhang, Yuyu Sun, Guofeng Bao, Guanhua Xu, Jiajia Chen & Zhiming Cui

    2. Department of Stomatology, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, 226001, China

      Xiaohui Lu, Guijuan Feng, Lingfeng Xu & Xingmei Feng

    3. Department of Rheumatology, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, 226001, China

      Zhifeng Gu

    4. Department of Cardiology Medical, Tongzhou First People’s Hospital, Nantong, Jiangsu Province, 226300, China

      Yuanzhou Lu

    Authors
    1. Jinlong Zhang
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    2. Xiaohui Lu
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    3. Guijuan Feng
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    4. Zhifeng Gu
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    Correspondence to Xingmei Feng or Zhiming Cui.

    Additional information

    Jinlong Zhang, Xiaohui Lu, and Guijuan Feng contributed equally to this work.

    This work was supported by Natural Science Foundation of China Grant (nos. 81500809, 81501076), Jiangsu Natural Science Foundation (BK2011385), “Top Six Types of Talents” Financial Assistance of Jiangsu Province Grant (no. 2013-WSN-076), Graduate Student Innovation of Science and Technology Projects in Jiangsu Province and in Nantong University (nos.SJLX-0588, SJLX-0588) and Nantong Natural Science Foundation (no. BK2014038).

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    Zhang, J., Lu, X., Feng, G. et al. Chitosan scaffolds induce human dental pulp stem cells to neural differentiation: potential roles for spinal cord injury therapy. Cell Tissue Res 366, 129–142 (2016). https://doi.org/10.1007/s00441-016-2402-1

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    • DOI: https://doi.org/10.1007/s00441-016-2402-1

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