Oxygen consumption in human, tissue-engineered myobundles during basal and electrical stimulation conditions

During three-dimensional culture of skeletal muscle in vitro, electrical stimulation provides an important cue to enhance skeletal muscle mimicry of the in vivo structure and function. However, increased respiration can cause oxygen transport limitations in these avascular three-dimensional constructs, leading to a hypoxic, necrotic core, or nonuniform cell distributions in larger constructs. To enhance oxygen transport with convection, oxygen concentrations were measured using an optical sensor at the inlet and outlet of an 80 μl fluid volume microphysiological system (MPS) flow chamber containing three-dimensional human skeletal muscle myobundles. Finite element model simulations of convection around myobundles and oxygen metabolism by the myobundles in the 80 μl MPS flow chamber agreed well with the oxygen consumption rate (OCR) at different flow rates, suggesting that under basal conditions, mass transfer limitations were negligible for flow rates above 1.5 μl s^-1. To accommodate electrodes for electrical stimulation, a modified 450 μl chamber was constructed. Electrical stimulation for 30 min increased the measured rate of oxygen consumption by the myobundles to slightly over 2 times the basal OCR. Model simulations indicate that mass transfer limitations were significant during electrical stimulation and, in the absence of mass transfer limitations, electrical stimulation induced about a 20-fold increase in the maximum rate of oxygen consumption. The results indicate that simulated exercise conditions increase respiration of skeletal muscle and mass transfer limitations reduce the measured levels of oxygen uptake, which may affect previous studies that model exercise with engineered muscle.