Development of an electric stimulation system to analyze muscle damage in 3D Duchenne muscular dystrophy muscle culture

Duchenne muscular dystrophy (DMD) is a progressive muscle wasting disease that impairs the ability of myotubes to handle mechanical stress. Thus, muscle contraction leads to membrane damage in the form of sarcolemmal tears. Electrical pulse stimulation (EPS) can replace motor neuron activation in muscle tissues, allowing in vitro muscle contraction-related studies. This work aimed to provide a practical solution for the stimulation of 3D skeletal muscle tissues as an alternative to overcome the limitations of existing commercial systems. To this end, we developed an intuitive electric stimulation system tailored to the needs of the laboratory. Our system was divided into two parts designed from the ground up. The first part allowed the user to choose which samples to stimulate, while the second part delivered the stimulation signal via graphite electrodes. The system developed in this study was used to induce contraction in healthy and DMD affected samples under different stimulation regimes, and ELISA assays were used to quantify creatine kinase leakage as a muscle damage marker. Our system facilitated higher throughput experiments using multiple samples, proving its potential to serve as a valuable tool for future research in drug testing and DMD treatments.