Recently, Kaushal et al. isolated endothelial progenitor cells (EPCs) from the peripheral blood of sheep and seeded them onto decellularized porcine iliac vessels. EPC-seeded grafts remained patent for 130 days as a carotid interposition graft in sheep (non-seeded grafts occluded within 15 days), and exhibited contractile activity and nitric-oxide-mediated vascular relaxation similar to native carotid arteries. Sales et al. have demonstrated that EPCs have the potential to provide both valvular interstitial and endothelial cellular functions, demonstrating the potential for EPCs to serve as a single autologous cell source for TEPV. In addition to the identification of clinically feasible cell sources, engineered soft tissues such as the TEPV require scaffolds with anisotropic mechanical properties that undergo large deformations (not possible with current PGA/PLLA non-wovens) coupled with controllable biodegradative and cell-adhesive characteristics. As a next step in fulfilling these design criteria, the Wagner lab has recently synthesized a family of poly (ester-urethane) ureas (PEUUs), including combination with type I collagen at various ratios to enhance cell attachment and increase biodegradation rates. Electrospun PEUU scaffolds have also been produced with biaxial mechanical properties that are remarkably similar to the native pulmonary valve, including the ability to undergo large physiologic strains and pronounced mechanical anisotropy. Moreover, a novel cell micro-integration technique has been developed that allows for successful integration of the cells directly into the scaffolds at the time of fabrication, eliminating cellular penetration problems. These encouraging results suggest that ES-PEUU scaffolds micro-integrated with EPCs can serve as successful TEPV scaffolds. We hypothesize that strategic combinations of individual mechanical factors relevant to heart valves-cyclic flexure, strain, and flow-can be determined that optimize ECM synthesis, organization, and mechanical properties of EPC seeded TEPV. Moreover, we hypothesize that the use of novel elastomeric scaffolds can add a critical degree-of-freedom for TEPV designs by allowing for large strains and highly controllable mechanical anisotropy. These hypotheses will be addressed by the following specific aims:
Specific Aim 1 - Optimize ES-PEUU scaffold mechanical anisotropy, layer and pore structures, and cellular integration for EPC-seeded TEPV leaflet applications.
Specific Aim 2 - Using optimized PEUU scaffolds of specific aim 1, conduct critical in-vitro """"""""scale-up"""""""" studies in intact TEPV under simulated physiological conditions.
Specific Aim 3 - Evaluate the EPC-seeded ES-PEUU scaffold's ability to perform in-vivo using a single leaflet model. ? ? ?
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