Combinatorial Glyoxal Cross-Linking and Glutamic Acid Capping for Mitigating Calcification in Bovine Pericardium

Bioprosthetic heart valves (BHVs) offer advantages over mechanical valves but are limited by long-term degeneration and calcification. This study aimed to develop a durable BHV material using decellularized bovine pericardium (BP) and alternative cross-linking strategies. BP was decellularized using a combination of sodium deoxycholate (SDC) and Triton X-100 (TX), which removed cellular components while preserving the extracellular matrix (ECM), as confirmed by histological, DNA quantification, and biochemical analyses. The impact of glutaraldehyde, glyoxal, tannic acid, and catechin cross-linking on calcification propensity and mechanical properties was evaluated. Glyoxal-cross-linked BP, further capped with 1% l-glutamic acid, demonstrated superior resistance to calcification while maintaining mechanical properties comparable to standard glutaraldehyde treatment. These results suggest that SDC-TX decellularized, glyoxal-cross-linked BP with l-glutamic acid capping presents a promising strategy for enhancing BHV durability and biocompatibility, offering a potential alternative to conventional glutaraldehyde.