Qualitative and Quantitative Evaluation of a Novel Detergent-Based Method for Decellularization of Peripheral Nerves
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Tissue engineering is an emerging strategy for the development of nerve substitutes for peripheral nerve repair. Especially decellularized peripheral nerve allografts are interesting alternatives to replace the gold standard autografts. In this study, a novel decellularization protocol was qualitatively and quantitatively evaluated by histological, biochemical, ultrastructural and mechanical methods and compared to the protocol described by Sondell et al. and a modified version of the protocol described by Hudson et al. Decellularization by the method described by Sondell et al. resulted in a reduction of the cell content, but was accompanied by a loss of essential extracellular matrix (ECM) molecules such as laminin and glycosaminoglycans. This decellularization also caused disruption of the endoneurial tubes and an increased stiffness of the nerves. Decellularization by the adapted method of Hudson et al. did not alter the ECM composition of the nerves, but an efficient cell removal could not be obtained. Finally, decellularization by the method developed in our lab by Roosens et al. led to a successful removal of nuclear material, while maintaining the nerve ultrastructure and ECM composition. In addition, the resulting ECM scaffold was found to be cytocompatible, allowing attachment and proliferation of adipose-derived stem cells. These results show that our decellularization combining Triton X-100, DNase, RNase and trypsin created a promising scaffold for peripheral nerve regeneration.
KeywordsPeripheral nerve regeneration Decellularized nerve allografts Tissue engineering Detergents Extracellular matrix Biomechanics
This study was supported by the Grant FIS PI14/1343 and FIS PI17/0393 of the Spanish Plan Nacional de Investigación Científica, Desarrollo e Innovación Tecnológica from the Ministerio de Economía y Competitividad, Instituto de Salud Carlos III (co-financed by FEDER funds, European Union) and by the Special Research Fund (BOF 14/IOP/045) from Ghent University, Belgium. The authors would like to thank Dr. Víctor Domingo Roa, Amalia de la Rosa and Concepción Villegas (Experimental Unit of the University Hospital Virgen de las Nieves, Granada) for their assistance with the laboratory animals, Leen Pieters (Ghent University) for the technical assistance with the TEM and Lisa Van Vlaenderen (Ghent University) for the assistance with the ex vivo cytocompatibility.
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