Decellularisation and histological characterisation of porcine peripheral nerves

Abstract

Peripheral 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.

Investigation of cell adhesion in chitosan membranes for peripheral nerve regeneration

“…Related to fresh healthy nerves, the values of Young Modulus in mice and rats are very low [63] when compared to pigs [64] creating a range that goes from 0.5 MPa (in mice) to 7.75 MPa (in pigs). In this respect, comparing our data with the values of mechanical properties to fresh healthy in vivo nerves in pigs [64,65] the values were similar and in the same range of the ones reported in the literature for porcine peroneal and tibial nerves (≈ 7.43-7.75 MPa).…”

Comparison of systematically combined detergent and nuclease‐based decellularization methods for acellular nerve graft: An ex vivo characterization and in vivo evaluation

“…The basic problem with allogenic nerve development is that a standard evaluation protocol for the decellularized nerve has not been established (Philips, Cornelissen, & Carriel, ). A comprehensive approach including various evaluation tools was recently emphasized for verification of the decellularized tissue or organs (Zilic, Wilshaw, & Haycock, ). Crapo et al () suggested a criteria for evaluation of cell removal composed of histological analysis with H&E or DAPI staining and DNA quantification.…”

Combination of BMSCs‐laden acellular nerve xenografts transplantation and G‐CSF administration promotes sciatic nerve regeneration

“…It has been proven that the nerve defects can be repaired by acellular nerve allografts to some degree (Saheb‐Al‐Zamani et al, ; Tajdaran, Gordon, Wood, Shoichet, & Borschel, ). The acellular nerve grafts of different mammalian species that are chemically extracted possess favorable biocompatibility, thus the utilization of heterogenetic acellular nerves seems to be capable of augmenting the insufficient source of nerve grafts in clinical practice (Zhang, Tong et al, ; Zilic, Wilshaw, & Haycock, ). However, there are few reports about nerve repairs by ANX.…”