Project Summary/Abstract The goals of the proposed research are to develop functional autologous trilayered heart valve leaflets with collagen fibril orientations of a native leaflet using trilayered nanofibrous substrates and to extend this approach in developing fully autologous heart valves with native heart valve functionality. The proposed work will develop a technology to fabricate trilayered nanofibrous substrates from a FDA approved polymer mimicking trilayered structure and orientations of collagen fibrils of native heart valve leaflets. The proposed work will then apply leaflet-shaped trilayered nanofibrous substrates to develop non-contractile autologous valve leaflets mimicking the structure of native leaflets by in-body tissue engineering. The leaflet constructs will be tested in-vitro to verify their morphological, structural, and functional properties and contractility. The proposed work will then develop heart valve-shaped nanofibrous substrates containing leaflet-shaped trilayered nanofibrous substrates and circumferentially oriented tubular nanofibrous substrates to engineer autologous non-contractile heart valves with comparable properties of native heart valves through in-body tissue engineering. The engineered valves will be tested for their morphological, structural, mechanical and functional properties in-vitro. The engineered autologous valves will also be tested for clinically-relevant outcomes including function, thrombus formation, and calcification in an ovine implantation model. These valves are expected to be an important step in the development toward clinical translation. The proposed research focuses the candidate's research in a novel direction to provide training on new skills required to begin the transition to independence. The candidate holds a Ph.D. in Materials Science and Engineering from the University of Washington and is currently a research associate at Mayo Clinic. His Ph.D. thesis work involved development of biomaterials for tissue engineering and regenerative medicine. This led to his postdoctoral work that involves design and development of nanofibrous biomaterials for biological cardiac valve development. His postdoctoral work also includes development of decellularized heart valve, pericardium tissue-based heart valve and stent graft, and their functionality testing in an ovine/porcine implantation model. The candidate's immediate career goal is to transition from mentored to independent research by completing his postdoctoral training and beginning a tenure track faculty position at a major research university. This will require focusing his current projects into a novel research direction while also receiving additional training needed to successfully complete the current and future projects in cardiovascular tissue engineering as an independent investigator. The K99/R00 mechanism is the ideal means of achieving this goal. The candidate's long-term career objective is to establish an indepe...