Investigation into the effects of varying frequency of mechanical stimulation in a cycle-by-cycle manner on engineered cardiac construct function.

Mechanical stimulation has been used extensively to improve the function of cardiac engineered tissue, as it mimics the physical environment in which the tissue is situated during normal development. However, previous mechanical stimulation has been carried out under a constant frequency that more closely resembles a diseased heart. The goal of this study was to create a bioreactor system that would allow us to control the mechanical stimulation of engineered cardiac tissue on a cycle-by-cycle basis. This unique system allows us to determine the effects on cardiac construct function of introducing variability to the mechanical stretch. To test our bioreactor system, constructs created from neonatal rat cardiomyocytes entrapped in fibrin hydrogels were stimulated under various regimes for 2 weeks and then assessed for functional outcomes. No differences were observed in the final cell number in each condition, indicating that variability in frequency did not have a negative effect on viability. The forces were higher for all mechanical stimulation groups compared to static controls, although no differences were observed between the mechanically stimulated conditions, indicating that variable frequency on a cycle-by-cycle basis has limited effects on the resulting force. Although differences in the observed twitch force were not observed, differences in the protein expression indicate that variable-frequency mechanical stimulation had an effect on cell-cell coupling and growth pathway activation in the constructs. Thus, this bioreactor system provides a valuable tool for further development and optimization of engineered myocardial tissue as a repair or replacement strategy for patients undergoing heart failure.