Mosahebi-Mohammadi, Afshin; (2001) Genetic labelling and transplantation of Schwann cells to enhance peripheral nerve regeneration. Doctoral thesis (Ph.D), UCL (University College London).

Text Genetic_labelling_and_transpla.pdf Download (21MB)

Abstract

There is a clinical need to improve functional outcome after peripheral nerve reconstruction, and also to find a replacement for autologous nerve grafts currently used for this purpose. Schwann cells (SC) are essential for adequate axonal regeneration and the aim of this work was to develop a methodology to enable investigation and characterisation of the effects of SC transplantation and genetic manipulation on peripheral nerve regeneration. Initially an identifiable and pure population of cultured SC was obtained. In vitro evaluation showed that chemical labelling adversely affected SC properties. Transduction of lacZ genetic label was carried out and a stable population of genetically modified SC was obtained. Transduced SC properties and lacZ expression were preserved in vitro for 6 months of continuous culture. Suspension matrix is required for SC transplantation and the suitability of alginate hydrogel was confirmed by in vitro tests to support SC proliferation and neurite sprouting in a neuron-glial co-culture. Defects in the rat sciatic nerve were bridged using resorbable poly-3-hydroxybutyrate (PHB) conduits containing SC. The results showed that the optimal number of SC required to enhance axonal regeneration was 80×l0⁶/ml. Following transplantation of transduced SC, syngeneic SC could be identified for up to 6 weeks and allogeneic SC for up to 3 weeks, as identified by X-gal staining. Immunohistochemistry was used to characterise SC, immune response, and axonal regeneration and the staining quantified by image analysis. Transplantation of both syngeneic and allogeneic SC improved axonal regeneration distance, the quantity of regeneration was better and more sustained with syngeneic SC. Furthermore, addition of liquid fibronectin to alginate improved regeneration which was further enhanced when SC were present. Finally a new technique for nonviral gene delivery of insulin-like growth factor I and mechano growth factor gene with alginate hydrogel matrix showed promising results in improving peripheral nerve regeneration.

Download activity - last month

Export as JSONXMLCSV

Download activity - last 12 months

Export as CSVXMLJSON

Downloads by country - last 12 months

Export as JSONCSVXML

Archive Staff Only

View Item