New strategies for peripheral nerve regeneration

Nerve repair is still a major challenge in surgery, regenerative medicine and tissue engineering even if progress has been made over the last 30 years. Functional recovery after severe lesions to a nerve is often incomplete and rarely totally successful. In this thesis I present a multi-disciplinary approach to improve the regenerative potential of “nerve repair tubes” that aim to reconnect wounded nerves and refine or replace autologous nerve graft, the clinical current gold standard. The efficacy of such tubes has already been shown in the clinic especially for small gap injuries, but the outcomes are still limited, and ought to be improved by e.g. micro/nano-topography, growth factor delivery systems, supportive cells or active features such as electrical stimulation, which have individually been shown to enhance nerve regeneration. In this study organotypic cultures of dorsal root ganglions (DRG) isolated from neonatal rats were used throughout as an in vitro model of nerve regeneration. Here I tested different devices in combination with growth factors to contribute to the fundamental and technical knowledge necessary to improve the regenerative potential of such tubes. I investigated the interaction between surface features and growth factors in their joined influence on regenerating DRGs. For this polydimethylsiloxane (PDMS), a polymer with adjustable elasticity was used together with photolithography to build devices of different stiffness with different surface microgrooves, on which DRG could be grown. To optimise the use of nerve growth factor (NGF) in conjunction with these devices, and to show how NGF interacts with stiffness and topography the reaction of the DRG was tested. To ease the making of three-dimensional internally microstructured tubes I have developed up a novel, timesaving, fabrication technique for polycaprolactone (PCL) “Swiss roll” nerve repair tubes. This technique improves the reproducibility of the scaffold, and using DRG its potential for nerve regeneration is being demonstrated. The influence of time-variant, balanced, pulsed electric stimulation is a potentially useful means to influence nerve regeneration. To narrow down the parameter space the effect of various electric fields was tested in their effect on DRG regeneration using commercially available devices. In collaboration with Christopher Martin from the School of Engineering, novel custom-made devices that allowed us to quantify the directional response of the regenerating axons were developed, and the guidance effect of pulsed alternating current (AC) electrical fields on regenerating DRGs axons was investigated in vitro. This approach allows in principle to transfer the use of this nerve guiding strategy to potentially improve nerve repair tubes