Arthur Castleton, Brigham Young University
Engineering
More than one in three people die because of organ failures such as congestive heart failure. The major issues of heart transplants include a scarcity of donors, immunorejection and blood clot formation. Over the last decade bioartificial organs have emerged as a potential alternative to traditional transplantation because they eliminate the need for immunosuppressants, DNA testing, and the use of another human’s organs. In this study an economic, effective, and rapid decellularization process that produces minimal damage to a cardiac extracellular matrix (cECM) is described. In addition, a static blood thrombosis assay was used to verify the effect of exposed cECM on clotting. Also an aorta was recellularized and analyzed.
Pig hearts were perfused with cycles of 1X Phosphate Buffered Saline (PBS), 0.5% sodium dodecyl sulfate (SDS), and distilled water for a total of 24 hours to remove cells and nucleic acids. After decellularization, cECM samples were analyzed to verify cell removal. A static blood assay was performed on cECMs and fresh hearts to measure if exposed cECMs caused increased micro-clotting. Endothelial cells were grown at 37°C in cell medium and introduced into a decellularized aorta. Recellularized aorta samples were then tested for confluency and formation of micro-clots.
Successful decellularization of porcine hearts was achieved without significant damage to the cECM by minimalizing total detergent exposure to 6 hours. When blood was perfused, large amounts of clotting were observed in decellularized hearts while negligible amounts were found in fresh hearts. This verifies that when cECMs are exposed to blood, micro-clotting occurs; thus increased cell proliferation could be a preventative measure to heart disease. Further investigation will focus on scaling up aorta recellularization to whole hearts in order to obtain a functional organ for clinical trials.