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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

查看全文 作  者:Andrea [1,2]Gärtner;Tiago [1,2]Pereira;Maria Joāo [1,2]Simōes;Paulo AS Armada-da-[3]Silva;Miguel L [1,2]França;Rosa [4]Sousa;Simone [5,6]Bompasso;Stefania [5,6]Raimondo;Yuki [7]Shirosaki;Yuri [8]Nakamura;Satoshi [7]Hayakawa;Akiyoshi [7]Osakah;Beatriz [4]Porto;Ana Lúcia [1,2]Luís;Artur SP [9]Varejāo;Ana Colette [1,2]Maurício 高影响力作者 机构地区:[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高影响力机构 出  处:《Neural Regeneration Research》索引2012年第7卷第29期,共12页高影响力期刊 基  金:supported by Technology and Science Foundation(FCT),Education and Science Ministry,Portugal,through the financed research project PTDC/DES/104036/2008;by QREN N°1372-Nucleus I&DT for the Development of Products for Regenerative Medicine and Cell Therapies-Núcleo Biomat&Cell;Andrea Grtner has a Doctoral Grantfrom Technology and Science Foundation(FCT),Education and Science Ministry,Portugal,SFRH/BD/70211/2010 摘  要: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 toform 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. 关 键 词:骨髓间充质干细胞 壳聚糖膜 神经再生 大鼠模型 果冻 学院 脐带 混合
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