A novel technique for decellularization of allogenic nerves and in vivo study of their use for peripheral nerve reconstruction

Autografts represent the gold standard for peripheral nerve reconstruction but their limited availability, the discrepancy of nerve caliber, and long surgical times are drawbacks. Allografts have therefore become a valid alternative option. In particular, acellular nerve allografts (ANAs) rather tha...

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Bibliographic Details
Published in:Journal of biomedical materials research. Part A 2017-08, Vol.105 (8), p.2228-2240
Main Authors: Boriani, F., Fazio, N., Fotia, C., Savarino, L., Nicoli Aldini, N., Martini, L., Zini, N., Bernardini, M., Baldini, N.
Format: Article
Language:English
Subjects:
Allografts
Animals
Architecture
Asepsis
Autografts
Axons
Bundles
Bundling
decellularization
Defects
Detergents - chemistry
Detritus
Fibrosis
Grafting
Grafts
Guided Tissue Regeneration - methods
Immunohistochemistry
Immunosuppression
In vitro methods and tests
In vivo methods and tests
Leukocytes
Lysis
Macrophages
Male
Microscopy
Myelin
nerve defect
Nerve Regeneration
Nerves
Peripheral nerves
Peripheral Nerves - cytology
Peripheral Nerves - physiology
Peripheral Nerves - surgery
Peripheral Nerves - transplantation
Rabbits
Reconstruction
Sonication
Sonication - methods
Stirring
Surgery
Surgical implants
Tibial nerve
Tissue Scaffolds - chemistry
Transplantation, Homologous - methods
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Summary:Autografts represent the gold standard for peripheral nerve reconstruction but their limited availability, the discrepancy of nerve caliber, and long surgical times are drawbacks. Allografts have therefore become a valid alternative option. In particular, acellular nerve allografts (ANAs) rather than fresh allografts do not need immunosuppression and appear to be safe and effective based on recent studies. An innovative method was conceived to obtain ANAs, so as to speed up nerve decellularization, without compromising nerve architecture, and without breaking the asepsis chain. Several detergent‐based techniques, integrated with sonication and mechanical stirring, were tested in vitro on rabbit nerves, to identify, by microscopy and immunohistochemistry, the most effective protocol in terms of cell lysis and cellular debris clearance, while maintaining nerve architecture. Furthermore, a pilot in vivo study was performed: ANAs were implanted into tibial nerve defects of three rabbits, and autografts, representing the gold standard, in other three animals. Twelve weeks postoperatively, rabbits were clinically evaluated and euthanasized; grafts were harvested and microscopically and histomorphometrically analyzed. The method proved to be effective in vitro: the treatment removed axons, myelin and cells, without altering nerve architecture. The in vivo study did not reveal any adverse effect: animals maintained normal weight and function of posterior limb during the entire experimental time. A mild fibrotic reaction was observed, macrophages and leukocytes were rare or absent; ANAs regenerated fascicles and bundles were comparable versus autografts. Based on these results, this decellularization protocol is encouraging and deserves deeper investigations with further preclinical and clinical studies. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2228–2240, 2017.
ISSN:1549-3296
1552-4965
DOI:10.1002/jbm.a.36090