Dialdehyde pectin-crosslinked and hirudin-loaded decellularized porcine pericardium with improved matrix stability, enhanced anti-calcification and anticoagulant for bioprosthetic heart valves

Abstract

To conveniently and effectively cure heart valve diseases or defects, combined with transcatheter valve technology, bioprosthetic heart valves (BHVs) originated from the decellularized porcine pericardium (D-PP) have been broadly used in clinics. Unfortunately, most clinically available BHVs crosslinked with glutaraldehyde (GA) were challenged in their long-term tolerance, degenerative structural changes, and even failure, owing to the synergistic impact of multitudinous elements (cytotoxicity, calcification, immune responses, etc.). In this work, dialdehyde pectin (AP) was prepared by oxidizing the o-dihydroxy of pectin with sodium periodate. Hereafter, the AP-fixed PP model was obtained by crosslinking D-PP with AP with high aldehyde content (6.85 mmol g⁻¹), for acquiring excellent mechanical properties and outstanding biocompatibility. To further improve the hemocompatibility of the AP-fixed PP, a natural and specific inhibitor of thrombin (hirudin) was introduced to achieve surface modification of the AP-fixed PP. The feasibility of crosslinking and functionalizing AP-fixed PP, which was a potential leaflet material of BHVs, was exhaustively and systematically evaluated. In vitro studies found that hirudin-loaded and AP-fixed PP (AP + Hirudin-PP) had synchronously achieved effective fixation of collagen, highly effective anticoagulation, and good HUVECs-cytocompatibility. In vivo results revealed that the AP + Hirudin-PP specimens recruited the minimum immune cells in the implantation experiment, and also presented an excellent anti-calcification effect. Overall, AP + Hirudin-PP was endowed with competitive collagen stability (compared with GA-fixed PP), excellent hemocompatibility, good HUVECs-cytocompatibility, low immunogenicity and outstanding anti-calcification, suggesting that AP + Hirudin-PP might be a promising alternative to GA-fixed PP and exhibited a bright prospect in the clinical applications of BHVs.