Deconstructing the Tissue Engineered Vascular Graft: Evaluating Scaffold Pre-Wetting, Conditioned Media Incubation, and Determining the Optimal Mononuclear Cell Source.

Stenosis limits widespread use of tissue-engineered vascular grafts (TEVGs), and bone marrow mononuclear cell (BM-MNC) seeding attenuates this complication. Yet seeding is a multistep process, and the singular effects of each component are unknown. We investigated which components of the clinical seeding protocol confer graft patency and sought to identify the optimal MNC source. Scaffolds composed of polyglycolic acid and ε-caprolactone/ι-lactic acid underwent conditioned media (CM) incubation (n = 25) and syngeneic BM-MNC (n = 9) or peripheral blood (PB)-MNC (n = 20) seeding. TEVGs were implanted for 2 weeks in the mouse IVC. CM incubation and PB-MNC seeding did not increase graft patency compared to control scaffolds prewet with PBS (n = 10), while BM-MNC seeding reduced stenosis by suppressing inflammation and smooth muscle cell, myofibroblast, and pericyte proliferation. IL-1β, IL-6, and TNFα were elevated in the seeded BM-MNC supernatant. Further, BM-MNC seeding reduced platelet activation in a dose-dependent manner, possibly contributing to TEVG patency.