title: 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.
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.
authors.affiliation: Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal ; Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Andrea
authors.initials: A
authors.lastname: Gärtner
authors.affiliation: Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal ; Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Tiago
authors.initials: T
authors.lastname: Pereira
authors.affiliation: Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal ; Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Maria João
authors.initials: MJ
authors.lastname: Simões
authors.affiliation: Faculty of Human Kinetics, Technical University of Lisbon, Cruz Quebrada - Dafundo, 1499-002, Portugal.
authors.forename: Paulo As
authors.initials: PA
authors.lastname: Armada-da-Silva
authors.affiliation: Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal ; Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Miguel L
authors.initials: ML
authors.lastname: França
authors.affiliation: Institute of Biomedical Sciences Abel Salazar, Cytogenetic Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Rosa
authors.initials: R
authors.lastname: Sousa
authors.affiliation: Neuroscience Institute of the Cavalieri Ottolenghi Foundation, Orbassano 10043, Turin, Italy ; Department of Clinical and Biological Sciences, University of Turin, Orbassano 10010, Turin, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Bompasso
authors.affiliation: Neuroscience Institute of the Cavalieri Ottolenghi Foundation, Orbassano 10043, Turin, Italy ; Department of Clinical and Biological Sciences, University of Turin, Orbassano 10010, Turin, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Raimondo
authors.affiliation: Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Shirosaki
authors.affiliation: Faculty of Engineering, Okayama University, Okayama 700-8530, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hayakawa
authors.affiliation: Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan.
authors.forename: Akiyoshi
authors.initials: A
authors.lastname: Osakah
authors.affiliation: Institute of Biomedical Sciences Abel Salazar, Cytogenetic Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Porto
authors.affiliation: Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal ; Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Ana Lúcia
authors.initials: AL
authors.lastname: Luís
authors.affiliation: 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.
authors.forename: Artur Sp
authors.initials: AS
authors.lastname: Varejão
authors.affiliation: Animal Science and Study Centre / Food and Agrarian Sciences and Technologies Institute, Porto University, 4099-003 Porto, Portugal ; Institute of Biomedical Sciences Abel Salazar, Veterinary Clinics Department, Porto University, Porto 4099-003, Portugal.
authors.forename: Ana Colette
authors.initials: AC
authors.lastname: Maurício
keywords: Wharton jelly; axonotmesis; biomaterials; chitosan; functional analysis; karyotype analysis; mesenchymal stem cells; rat; stem cells; stereological analysis; umbilical cord