Reducing retraction in engineered tissues through design of sequential growth factor treatment

Abstract Heart valve disease is associated with high morbidity and mortality worldwide, resulting in hundreds of thousands of heart valve replacements each year. Tissue engineered heart valves (TEHVs) have the potential to overcome the major limitations of traditional replacement valves; however, leaflet retraction has led to the failure of TEHVs in preclinical studies. Sequentially varying growth factors over time has been utilized to promote maturation of engineered tissues and may be effective in reducing tissue retraction, yet it is difficult to predict the effects of such treatments due to complex interactions between the cells and the extracellular matrix (ECM), biochemical environment, and mechanical stimuli. We hypothesize that sequential treatments of fibroblast growth factor 2 (FGF-2) and transforming growth factor beta 1 (TGF-β1) can be used to minimize cell-generated tissue retraction by decreasing active cell contractile forces exerted on the ECM and by inducing the cells to increase the ECM stiffness. Using a custom culturing and monitoring system for 3D tissue constructs, we designed and tested various TGF-β1 and FGF-2 based growth factor treatments, and successfully reduced tissue retraction by 85% and increased the ECM elastic modulus by 260% compared to non-growth factor treated controls, without significantly increasing the contractile force. We also developed and verified a mathematical model to predict the effects of various temporal variations in growth factor treatments and analyzed relationships between tissue properties, the contractile forces, and retraction. These findings improve our understanding of growth factor-induced cell-ECM biomechanical interactions, which can inform the design of next generation TEHVs with reduced retraction. The mathematical models could also potentially be applied toward fast screening and optimizing growth factors for use in the treatment of diseases including fibrosis.