title: A modified chemical protocol of decellularization of rat sciatic nerve and its recellularization with mesenchymal differentiated schwann-like cells: morphological and functional assessments.
abstract: The functional reconstruction of large neural defects usually requires the use of peripheral nerve autografts, though these have certain limitations. As a result, interest in new alternatives for autograft development has risen. The acellular peripheral nerve graft is an alternative for peripheral nerve injury repair, but to date there is not a standardized chemical decellularization method widely accepted. The objective of this study was to propose a modified chemical protocol of decellularization of rat sciatic nerve and its recellularization in vitro with mesenchimal differentiated Schwann-like cells. After the transplantation, an evaluation of its regeneration was performed using morphological and functional tests. The study consisted of two phases; in phase 1, different concentrations and times of exposure of rat sciatic nerves to detergents were tested, to establish a modified chemical protocol for nerve decellularization. The chemical treatment with 3% triton X-100 and 4% sodium deoxycholate for 15 days allowed a complete decellularization whilst conserving the extracellular matrix of the harvested nerve. In phase 2, the decellularized and recellularized alografts were compared against autografts. The morphological analysis showed a higher positivity to specific myelin antibodies in the recellularized group compared to the autograft. There were no differences in this parameter between the control limb and the experimental limb (recellularized group). The functional analysis showed no statistical differences at week 15 in the Sciatic Function Index in the autograft group vs the other groups. This study sets the morphological and functional bases for posterior studies about nerve defects regeneration in humans.
authors.affiliation: Plastic Surgery Service, University Hospital "Dr. José. Eleuterio González", University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: M M
authors.initials: MM
authors.lastname: García-Pérez
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: H G
authors.initials: HG
authors.lastname: Martínez-Rodríguez
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: G G
authors.initials: GG
authors.lastname: López-Guerra
authors.affiliation: Department of Histology, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: A
authors.initials: A
authors.lastname: Soto-Domínguez
authors.affiliation: Department of Biochemistry and Molecular Medicine, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: S L
authors.initials: SL
authors.lastname: Said-Fernández
authors.affiliation: Bone and Tissue Bank, Orthopedics and Traumatology Service, University Hospital "Dr. José. Eleuterio González", University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.|Department of Human Anatomy, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: R
authors.initials: R
authors.lastname: Morales-Avalos
authors.affiliation: Department of Human Anatomy, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: R E
authors.initials: RE
authors.lastname: Elizondo-Omaña
authors.affiliation: Department of Histology, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: R
authors.initials: R
authors.lastname: Montes-de-Oca-Luna
authors.affiliation: Department of Human Anatomy, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: S
authors.initials: S
authors.lastname: Guzmán-López
authors.affiliation: Department of Human Anatomy, Faculty of Medicine, University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: M L
authors.initials: ML
authors.lastname: Castillo-Galván
authors.affiliation: Bone and Tissue Bank, Orthopedics and Traumatology Service, University Hospital "Dr. José. Eleuterio González", University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México.
authors.forename: O F
authors.initials: OF
authors.lastname: Mendoza-Lemus
authors.affiliation: Bone and Tissue Bank, Orthopedics and Traumatology Service, University Hospital "Dr. José. Eleuterio González", University Autonomous of Nuevo León (U.A.N.L.), Monterrey, Nuevo León, México. vilchez.doctor@gmail.com.
authors.forename: F
authors.initials: F
authors.lastname: Vílchez-Cavazos
mesh_terms: D064591:Allografts; D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation; D017690:Cell Transplantation; D003902:Detergents / Q000737:chemistry; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009416:Nerve Regeneration; D010525:Peripheral Nerves / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D012583:Schwann Cells / Q000166:cytology*; D012584:Sciatic Nerve / Q000473:pathology*; D014182:Transplantation, Autologous
chemical_list: D003902:Detergents; D036341:Intercellular Signaling Peptides and Proteins