Tissue Engineering of Neuromuscular Junctions, Development of An Alternative to The LD50 Toxicity Test

Therapeutic Botulinum neurotoxin preparations have found an increasing number of clinical uses in a large variety of neuromuscular disorders and dermatological conditions. The accurate determination of potency in the clinical application of botulinum toxins is critil;al to ensure clinical efficacy and safety, and is currently achieved using a lethal dose (LDso ) assay in mice. Operational constraints and ethical concerns associated with this assay have prompted the development of alternative assay systems that could potentially lead to its replacement. One such tissue-based alternative, the 'Intercostal Neuromuscular Junction (Nl\JJ) Assay', uses nerve:muscle tissue preparations from the rat rib cage. The individual tissue preparations and the experimental protocol were subject to a detailed analysis. The tissue sections used were found to be highly reproducible as determined through the innervation pattern and the localisation of NMJs in situ and their distribution in skeletal muscle fibre types. Furthermore, the efficacy of the assay protocol to successfully deliver the test sample to the cellular target sites ,vas critically assessed using molecular tracer molecules. Sonification during sample aplication resulted in an accelerated way of sample application.EThOS - Electronic Theses Online ServiceGBUnited Kingdo