A Biomaterials Approach for an Ex-Vivo Multiple Sclerosis Model of Inflammatory Demyelination
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Multiple sclerosis (MS) is characterised by the presence of inflammatory demyelinating foci throughout the brain and spinal cord, accompanied by axonal and neuronal damage. Although inflammatory processes are thought to underlie the pathological changes, the individual mediators of this damage are unclear. In order to study the role of pro-inflammatory cytokines in demyelination in the central nervous system, in this thesis, a novel non-viral gene transfection vector was utilized to establish an inflammatory demyelinating model of MS in an ex-vivo environment. At first, a unique hyperbranched polymeric system with a linear poly 2-dimethylaminoethyl methacrylate (pDMAEMA) block and a hyperbranched polyethylene glycol methyl ether methacrylate (PEGMEMA) and ethylene dimethacrylate (EGDMA) block was designed and synthesized via deactivation enhanced atom transfer radical polymerisation (DE-ATRP) for efficient gene delivery. Using this unique structure, with a linear pDMAEMA block, which efficiently binds to plasmid DNA (pDNA) and hyperbranched polyethylene glycol (PEG) based block as a protective shell, efficient in-vitro and ex-vivo transfection was achieved with minimal cytotoxicity. Organotypic brain slices were then successfully transfected with the TNF[alpha] or IFN[gamma] genes. TNF[alpha] and IFN[gamma] expression and release in cerebellar slices via non-viral gene delivery approach resulted in inflammation mediated myelin loss, thus making it a promising ex-vivo approach for studying the underlying mechanisms of demyelination in myelin-related diseases such as MS.