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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

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Neural Regeneration Research 2012年第29期7卷 2247-2258页

作者： Andrea Gärtner Tiago Pereira Maria Joāo Simōes Paulo AS Armada-da-Silva Miguel L França Rosa Sousa Simone Bompasso Stefania Raimondo Yuki Shirosaki Yuri Nakamura Satoshi Hayakawa Akiyoshi Osakah Beatriz Porto Ana Lúcia Luís Artur SP Varejāo Ana Colette Maurício Animal Science and Study Centre/Food and Agrarian Sciences and Technologies Institute Porto University4099-003 PortoPortugal Institute of Biomedical Sciences Abel Salazar Veterinary Clinics DepartmentPorto UniversityPorto 4099-003Portugal Faculty of Human Kinetics Technical University of LisbonCruz Quebrada-Dafundo1499-002Portugal Institute of Biomedical Sciences Abel Salazar Cytogenetic DepartmentPorto UniversityPorto 4099-003Portugal Neuroscience Institute of the Cavalieri Ottolenghi Foundation Orbassano 10043TurinItaly Department of Clinical and Biological Sciences University of TurinOrbassano 10010TurinItaly Graduate School of Natural Science and Technology Okayama UniversityOkayama 700-8530Japan Faculty of Engineering Okayama UniversityOkayama 700-8530Japan Department of Veterinary Sciences Research Centre in SportsHealth and Human DevelopmentUniversity of Trás-os-Montes and Alto DouroVila Real 5001-801Portugal

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 pL）（Group 2-CrushCell）; Group 3, axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ill membrane covered with a monolayer of non-differentiated human mesenchymal stem cells （Group 3-CrushChitlllCell） and Group 4, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane （Group 4-CrushChiUll）. 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 carded 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 ner

关键词： stem cells mesenchymal stem cells Wharton jelly umbilical cord biomaterials chitosan axonotmesis functional analysis rat karyotype analysis stereological analysis

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Decellularized peripheral nerve grafts by a modified protocol for repair of rat sciatic nerve injury

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Neural Regeneration Research 2021年第6期16卷 1086-1092页

作者： Arash Zaminy Sara Sayad-Fathi Farshad Moharrami Kasmaie Zohreh Jahromi Adib Zendedel Neuroscience Research Center School of MedicineGuilan University of Medical SciencesRashtIran Student Research Committee School of MedicineGuilan University of Medical SciencesRashtIran Institute of Neuroanatomy Faculty of MedicineRheinisch-Westfälische Technische Hochschule(RWTH)Aachen UniversityAachenGermany

Studies have shown that acellular nerve xenografts do not require immunosuppression and use of acellular nerve xenografts for repair of peripheral nerve injury is safe and effective.However,there is currently no widely accepted standard chemical decellularization method.The purpose of this study is to investigate the efficiency of bovine-derived nerves decellularized by the modified Hudson’s protocol in the repair of rat sciatic nerve injury.In the modified Hudson’s protocol,Triton X-200 was replaced by Triton X-100,and DNase and RNase were used to prepare accelular nerve xenografts.The efficiency of bovine-derived nerves decellularized by the modified Hudson’s protocol was tested in vitro by hematoxylin&eosin,Alcian blue,Masson’s trichrome,and Luxol fast blue staining,immunohistochemistry,and biochemical assays.The decellularization approach excluded cells,myelin,and axons of nerve xenografts,without affecting the organization of nerve xenografts.The decellularized nerve xenograft was used to bridge a 7 mm-long sciatic nerve defect to evaluate its efficiency in the repair of peripheral nerve injury.At 8 weeks after transplantation,sciatic function index in rats subjected to transplantation of acellular nerve xenograft was similar to that in rats undergoing transplantation of nerve allograft.Morphological analysis revealed that there were a large amount of regenerated myelinated axons in acellular nerve xenograft;the number of Schwann cells in the acellular nerve xenograft was similar to that in the nerve allograft.These findings suggest that acellular nerve xenografts prepared by the modified Hudson’s protocol can be used for repair of peripheral nerve injury.This study was approved by the Research Ethics Committee,Research and Technology Chancellor of Guilan University of Medical Sciences,Iran(approval No.IR.GUMS.REC.1395.332)on February 11,2017.

关键词： acellular scaffold axonotmesis bovine crush injury detergent rat sciatic nerve xenograft

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