Abstract
ABSTRACTPeripheral nerve injuries affect a large proportion of the global population, often causing significant morbidity and loss of function. Current treatment strategies include the use of implantable nerve guide conduits (NGC's) to direct regenerating axons between the proximal and distal ends of the nerve gap. However, NGC's are limited in their effectiveness at promoting regeneration Current NGCs are not suitable as substrates for supporting either neuronal or Schwann cell growth, as they lack an architecture similar to that of the native extracellular matrix (ECM) of the nerve. The aim of this study was to create an acellular porcine peripheral nerve using a novel decellularisation protocol, in order to eliminate the immunogenic cellular components of the tissue, while preserving the three‐dimensional histoarchitecture and ECM components. Porcine peripheral nerve (sciatic branches were decellularised using a low concentration (0.1%; w/v) sodium dodecyl sulphate in conjunction with hypotonic buffers and protease inhibitors, and then sterilised using 0.1% (v/v) peracetic acid. Quantitative and qualitative analysis revealed a ≥95% (w/w) reduction in DNA content as well as preservation of the nerve fascicles and connective tissue. Acellular nerves were shown to have retained key ECM components such as collagen, laminin and fibronectin. Slow strain rate to failure testing demonstrated the biomechanical properties of acellular nerves to be comparable to fresh controls. In conclusion, we report the production of a biocompatible, biomechanically functional acellular scaffold, which may have use in peripheral nerve repair. Biotechnol. Bioeng. 2016;113: 2041–2053. © 2016 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals, Inc.
Highlights
The nervous system is one of the most highly organised biological systems, and is subdivided into the central nervous system (CNS) and peripheral nervous system (PNS)
Excess fat and connective tissue was dissected from the nerve samples and tissues washed three times in phosphate buffered saline solution (PBS; Oxoid, Basingstoke, U.K.), containing 0.1% (w/v) ethylene diamine tetra acetic acid (EDTA; VWRi) to remove excess blood
The aim of this study was to develop a process for the decellularisation of porcine peripheral nerve, with minimal effect on the tissue properties
Summary
The nervous system is one of the most highly organised biological systems, and is subdivided into the central nervous system (CNS) and peripheral nervous system (PNS). The microenvironment surrounding an injury site in the PNS is usually permissive to axonal regeneration. While the capacity of regeneration is a possibility for PNS injuries, complete functional recovery is infrequent and often misdirected in defects greater than 30 mm (Moore et al, 2009). Current management of peripheral nerve injury includes direct end-to-end suturing of the damaged nerve ends or the use of an autologous nerve graft. For longer nerve gaps this approach is not desirable because it has been demonstrated that any tension introduced into the nerve cable inhibits nerve regeneration (Millesi, 1982). Autologous nerve grafting remains the current gold standard surgical intervention for larger nerve defects (>30 mm). A dispensable nerve, usually the sural nerve (which has sensory function), is harvested and used to bridge the defect.
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