Passive and active contributions to generated force and retraction in heart valve tissue engineering

In tissue engineered heart valves, cell-mediated stress development during culture results in leaflet retraction at time of implantation. This tissue retraction is partly active due to traction forces exerted by the cells and partly passive due to release of residual stress in the extracellular matrix and the cells. Within this study, we unraveled the passive and active contributions of cells and matrix to generated force and retraction in engineered heart valve tissues. Tissue engineered rectangular strips, fabricated from PGA/P4HB scaffolds and seeded with human myofibroblasts, were cultured for 4 weeks, after which the cellular contribution was changed at different levels. Elimination of the active cellular traction forces was achieved with Cytochalasin D and inhibition of the Rho-associated kinase pathway. Both active and passive cellular contributions were eliminated by lysation and/or decellularization of the tissue. Maximum cell activity was reached by increasing the fetal bovine serum concentration to 50%. The generated force decreased similar to 20% after elimination of the active cellular component, similar to 25% when the passive cellular component was removed as well and remained unaffected by increased serum concentrations. Passive retraction accounted for similar to 60% of total retraction, of which similar to 15% was residual stress in the matrix and similar to 45% was passive cell retraction. Cell traction forces accounted for the remainder similar to 40% of the retraction. Full activation of the cells increased retraction by similar to 45%. These results illustrate the importance of the cells in the process of tissue retraction, not only actively retracting the tissue, but also in a passive manner to a large extent.