Abstract
Purpose: We investigated the functional potential of poly-4-hydroxybutyrate (P4HB) scaffolds by determining the most suitable thickness and ratio of co-cultured progenitor cells for construction of pulmonary leaflets and annular sewing ring in tissue engineered (TE) stentless PV. Methods: Melt-blown fibronectin (FN)-coated P4HB scaffolds of varying thickness were seeded with different percentages of characterized ovine bone marrow-derived mesenchymal stem cells (MSC) and peripheral blood-derived endothelial progenitor cells (EPC) for 21 days in a laminar flow system. Results: HE α-SMA+ cells were found both on the surface and in the interstitium evidenced by IHC. Immunoblotting revealed increased expression of α-SMA+. Hence, autologous TE stentless PV using 220µM for PV leaflet covered with 100µM for annular sewing ring was successfully placed in PV position. The neo-tissue integrity allowed secure anastomosis with adequate tensile strength. Echocardiography demonstrated normal biventricular function, and adequate leaflet coaptation with trivial central regurgitation. Conclusion: This demonstrates the successful creation of an anatomically functional, autologous TE stentless PV using sequentially seeded progenitor cell sources. This suggest the importance of scaffold thickness and ratio of seeded cells in the suturability of TE stentless PV.
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