Shear Preconditioning Restores Endothelial Function of HLHS Patient-Derived iPSC-ECs for Successful Conduit Implantation

Thrombosis, stenosis, and unsatisfied remodeling remain major challenges limiting the success of tissue engineered vascular grafts (TEVGs) in clinical application requiring vascular reconstruction. A key contributing factor is the Patients with single ventricle congenital heart defects (SVCHDs) urgently require TEVGs for cardiopulmonary reconstruction during Fontan procedure. However, endothelial dysfunction has been reported in both primary and hiPSC-derived endothelial cells (ECs) from this patient population, especially in the subgroup with hypoplastic left heart syndrome (HLHS), one severe form of SVCHD. Building on our prior work demonstrating that shear preconditioning enhances the antithrombotic and remodeling capacity of endothelialized conduits, we extended this approach to HLHS patient-derived iPSC-ECs. Compared with healthy controls, HLHS iPSC-ECs seeded on the luminal surface of decellularized human umbilical artery (dHUA) scaffolds and subjected to physiological shear preconditioning maintained patency and supported remodeling following implantation into the rat inferior vena cava (IVC), modeling Fontan circulation. In vitro, shear-preconditioned HLHS iPSC-ECs exhibited enhanced resistance to hemodynamic stress, improved anti-inflammatory and antithrombotic function, and partial restoration of mechanosensitive NOTCH1 signaling. These findings demonstrate that shear preconditioning can restore critical endothelial functions in HLHS iPSC-ECs and support the translational potential of autologous, patient-derived endothelialized TEVGs as future therapeutic solutions for SVCHDs. absence of a functional endothelium. We aim to establish a “ready-to-use” endothelial layer on TEVGs suitable for diverse clinical settings. The primary barriers to clinical translation include source fo endothelia and achieving a mature, stable endothelial layer. Human induced pluripotent stem cells (hiPSCs) provide an renewable source capable of generating patient-specific endothelial cells (iPSC-ECs) for TEVG endothelialization.