Design and development of novel nutritional supplements to enhance neuromuscular function using in vitro NMJ platform

Alomosh, Razan Ahmad (2025) Design and development of novel nutritional supplements to enhance neuromuscular function using in vitro NMJ platform. Doctoral thesis (PhD), Manchester Metropolitan University.

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Abstract

Normal muscle ageing in humans, known as sarcopenia, is generally associated with a gradual decline in physical and cognitive abilities, such as decreased motor skills, muscle function, and cognitive-motor coordination. These declines significantly impact daily activities and increase dependency. The neuromuscular system, particularly its neural and musculoskeletal components, is vital in the development of age-related neuromuscular disorders that contribute to reduced physical abilities. Of particular interest is the neuromuscular junction (NMJ), a specialised synapse where muscle fibres and motor neurons connect, which undergoes significant age-related deterioration affecting both neural and muscular components. This deterioration reveals insights into potential mechanisms that could be targeted with nutritional interventions, an emerging therapeutic approach. Fundamental lifestyle adjustments, including regular physical activity and balanced nutrition, are essential for maintaining health during ageing. Specific dietary nutrients have shown promise in improving neuromuscular function in older adults and individuals with neuromuscular disorders. However, despite decades of nutritional research using animal, human, and ex vivo models, effective therapeutic strategies remain limited. Therefore, this thesis aims to design and evaluate novel nutritional supplements to enhance neuromuscular function and overall muscle performance, employing a newly bioengineered NMJ model developed in collaboration with Danone Nutricia Research. The experimental design comprised three main phases. First, an NMJ co-culture platform was established using rat embryo spinal cord explants at embryonic and human immortalised myoblasts, maintained in serum-free environments without neural growth factors. Next, this model was used to screen individual nutritional supplements for their effects on myotube contractile function. Finally, the most promising nutrients identified in the second phase were combined and evaluated for their synergistic ability to enhance neuromuscular interactions and improve overall contractile performance in the co-culture system. The study successfully established a functional neuromuscular co-culture model that identified several novel compounds and prototype formulations with promising effects on muscle contraction, indicating enhanced NMJ functionality. Additionally, ELISA-based microarray results showed that tailored nutrient combinations boosted the endogenous production of specific growth and neurotrophic factors, potentially optimising NMJ formation and function. This research presents a novel approach to developing nutritional interventions for neuromuscular function. The identified nutrient combinations show significant potential for therapeutic application in mitigating sarcopenia and neuromuscular disorders, with broader implications for athletes and individuals seeking to enhance neuromuscular performance and quality of life.