Proteomics-guided characterization of Hericium erinaceus protein-capped silver nanoparticles for cytotoxicity against cancer cells and improved hemocompatibility

Biological synthesis offers an alternative approach for producing silver nanoparticles (AgNPs) using naturally derived biomolecules. In this study, a protein-rich extract of Hericium erinaceus (PHEx) was used for the biosynthesis of protein-capped AgNPs, followed by physicochemical, proteomic, cytotoxicity, and hemocompatibility analyses. The AgNPs exhibited a surface plasmon resonance band at approximately 410 nm, a hydrodynamic diameter of 70-90 nm, and a moderately negative surface charge. FTIR and proteomic analyses indicated the association of extract-derived biomolecules, particularly proteins, with the nanoparticle surface, supporting the formation of a protein-associated capping layer. Comparative LC-MS/MS identified 2,374 proteins, with 1,203 shared between PHEx and the AgNPs-associated fraction, further supporting the involvement of PHEx-derived proteins in AgNPs surface capping and stabilization. The AgNPs showed concentration-dependent cytotoxicity against SW480, NCI-N87, and HepG2 cells, with IC 50 values of 30.63, 23.21, and 2.01 µg/mL, respectively, compared with 223.19 µg/mL in CCD 841 CoN cells. AgNPs treatment was associated with increased total oxidative status and changes in Bax and Bcl-2 mRNA expression. Hemolysis remained below 1% up to 250 µg/mL. Collectively, PHEx-derived protein-capped AgNPs showed defined physicochemical characteristics, in vitro cytotoxic activity, and low hemolytic activity under the conditions examined.