Well-designed two-fold crosslinked biological valve leaflets with heparin-loaded hydrogel coating for enhancing anticoagulation, endothelialization, and anticalcification

Commercial biological valve leaflets (BVLs) crosslinked with glutaraldehyde (GA) are at risk of accelerating damage and even failure, owing to the high cell toxicity of GA, acute thrombosis, and calcification in clinical applications. In this study, a novel joint strategy of double crosslinking agents (dialdehyde pectin (AP) and carbodiimide) and heparin-loaded hydrogel coating was developed, endowing BVLs with excellent mechanical properties and multiple performances. Herein, AP played two essential roles, the crosslinking agent and the main component of the hydrogel coating. Both experimental and theoretical results indicated that the mechanical properties and stability of double-crosslinked BVLs were comparable to those of GA, and heparin loaded in hydrogel coating could improve the hemocompatibility of AP + EDC/NHS-PP. Further, cytocompatibility and in vivo tests showed that compared with GA-PP, AP + EDC/NHS + CS + Hep-PP has exhibited good endothelialization ability, mild immune response and anti-calcification performance and therefore prompts it to be an extremely valuable candidate for more durable and multifunctional BVLs.