Dual-Function Role of Phenolated Albumin in Hemin-Mediated Hydrogel Formation

Enzymatically crosslinked hydrogels are important in biomedical applications. However, conventional horseradish peroxidase (HRP)-based systems are expensive, unstable, and potentially immunogenic. Herein, we introduce hemin/albumin complexes as cost-effective and biocompatible catalysts for phenol-mediated hydrogel formation. Phenolated bovine serum albumins (BSA-LPh, -MPh, and-HPh) with different degrees of substitution were synthesized and complexed with hemin. Spectroscopic analysis demonstrated that phenol modification altered the hemin microenvironment, resulting in distinct shifts in the Soret band. Functional assays revealed that albumin complexation enhanced catalytic activity compared to hemin alone. Moderate phenol modification provided an optimal balance between catalytic efficiency and hydrogel integration, whereas excessive modification reduced the performance of the enzyme. Hydrogels containing hemin/BSA-Ph complexes exhibited controllable protein retention and high cytocompatibility (>90%) with mouse fibroblast 10T1/2 cells. These findings demonstrate that hemin/albumin complexes are promising, cost-effective, and cytocompatible alternatives to HRP systems for hydrogel-based biomedical and nonclinical applications.