Breaking Barriers: Proof of Concept for Selective Coronary Endothelial Decellularization in Whole Porcine Hearts for Xenotransplantation.

The critical shortage of donor hearts and the limitations of current mechanical circulatory support devices underscore the need for alternative strategies to address end-stage heart failure. Selective endothelial decellularization has emerged as a promising approach to reduce graft immunogenicity by removing donor endothelial cells while preserving myocardial architecture, thereby enabling subsequent repopulation with recipient-derived cells. However, this technique has never been applied to an intact porcine heart. This pilot study aimed to develop an ex vivo model for partial decellularization of the coronary vasculature in whole porcine hearts. Four hearts were perfused through the aortic root using a controlled circuit, applying variable concentrations of sodium dodecyl sulfate (SDS) or trypsin, followed by washout solutions including saline, distilled water, or hypothermic Buckberg cardioplegia. Histological and immunofluorescence analyses were performed on coronary arteries and ventricular myocardium to assess endothelial removal and tissue preservation. Across all experiments, SDS perfusion achieved effective qualitative and quantitative endothelial decellularization of large-caliber coronary arteries while maintaining the integrity of the internal elastic lamina, extracellular matrix, and myocardial structure. The hypothermic cardioplegic washout protocol demonstrated the highest efficacy, with marked loss of endothelial nuclei and decreased expression of CD31, without evidence of myocardial cell depletion. These findings provide preliminary evidence that selective endothelial decellularization of whole porcine hearts is feasible using SDS under controlled hypothermic conditions. This model may represent an essential step toward generating immunologically compatible xenogeneic hearts through subsequent autologous endothelial repopulation. Further studies are required to standardize the protocol, evaluate microvascular compartments, and explore effective recellularization strategies.