中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2012, Vol. 7 ›› Issue (29): 2247-2258.

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

壳聚糖杂化膜复合间充质干细胞促进轴突断伤大鼠神经再生

  

  • 收稿日期:2012-05-11 修回日期:2012-07-10 出版日期:2012-10-15 发布日期:2012-10-15

Use of hybrid chitosan membranes and human mesenchymal stem cells from the Wharton jelly of umbilical cord for promoting nerve regeneration in an axonotmesis rat model

Andrea Gärtner 1, 2, Tiago Pereira 1, 2, Maria João Simões 1, 2, Paulo AS Armada-da-Silva3, Miguel L França 1, 2, Rosa Sousa4, Simone Bompasso 5, 6, Stefania Raimondo 5, 6, Yuki Shirosaki7, Yuri Nakamura8, Satoshi Hayakawa7, Akiyoshi Osakah7, Beatriz Porto4, Ana Lúcia Luís 1, 2, Artur SP Varejão9, Ana Colette Maurício 1, 2   

  1. 1 Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal
    2 Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal
    3 Faculty of Human Kinetics, Technical University of Lisbon, Cruz Quebrada – Dafundo, 1499-002, Portugal
    4 Institute of Biomedical Sciences Abel Salazar, Cytogenetic Department, Porto University, Porto 4099-003, Portugal
    5 Neuroscience Institute of the Cavalieri Ottolenghi Foundation, Orbassano 10043, Turin, Italy
    6 Department of Clinical and Biological Sciences, University of Turin, Orbassano 10010, Turin, Italy
    7 Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan
    8 Faculty of Engineering, Okayama University, Okayama 700-8530, Japan
    9 Department of Veterinary Sciences, Research Centre in Sports, Health and Human Development, University of Trás-os-Montes and Alto Douro, Vila Real 5001-801, Portugal
  • Received:2012-05-11 Revised:2012-07-10 Online:2012-10-15 Published:2012-10-15
  • Contact: Ana Colette Maurício, Ph.D., Associate professor, Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal ana.colette@hotmail.com; acmauricio@icbas.up.pt
  • About author:Andrea G?rtner★, M.Sc, Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal Tiago Pereira, D.V.M., Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal Andrea G?rtner and Tiago Pereira contributed equally to this paper.

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Abstract:

Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to assess the effect on nerve regeneration, associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton’s jelly of umbilical cord, in peripheral nerve reconstruction after crush injury. Chromosome analysis on human mesenchymal stem cell line from Wharton’s jelly was carried out and no structural alterations were found in metaphase. Chitosan membranes were previously tested in vitro, to assess their ability in supporting human mesenchymal stem cell survival, expansion, and differentiation. For the in vivo testing, Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each: Group 1, sciatic axonotmesis injury without any other intervention (Group 1-Crush); Group 2, the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250 -1 500 human mesenchymal stem cells (total volume of 50 μL) (Group 2-CrushCell); Group 3, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane covered with a monolayer of non-differentiated human mesenchymal stem cells (Group 3-CrushChitIIICell) and Group 4, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane (Group 4-CrushChitIII). Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index, static sciatic index, extensor postural thrust, and withdrawal reflex latency. Stereological analysis was carried out on regenerated nerve fibers. Results showed that infiltration of human mesenchymal stem cells, or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration. Results obtained with chitosan type III membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising clinical tool in peripheral nerve reconstructive surgery. Yet, umbilical cord human mesenchymal stem cells, that can be expanded in culture and induced to form several different types of cells, may prove, in future experiments, to be a new source of cells for cell therapy, including targets such as peripheral nerve and muscle.

Key words: stem cells, mesenchymal stem cells, Wharton jelly, umbilical cord, biomaterials, chitosan, axonotmesis, functional analysis, rat, karyotype analysis, stereological analysis