Phytoferritin has a natural cagelike architecture for carrying bioactive molecules, and it is uniquely suited to function as a carrier due to its multiple interfaces and channels. In this study, a novel approach was proposed to prepare ferritin-salvianolic acid B-epigallocatechin gallate (EGCG) three-layer nanoparticles (FSE) through the steric hindrance of ferritin channels. Urea (30 mM) could expand the ferritin channel size evidenced by the improved iron release rate vo and promote the EGCG penetration into the ferritin cavity without disassembly of the ferritin cage. The encapsulation ratio of EGCG was 16.0 ± 0.14% (w/w). Salvianolic acid B attached to the outer interface of ferritin through weak bonds with a binding constant of (2.91 ± 0.04) × 105 M-1. The FSE maintained a spherical structure with a diameter of 12 nm. Moreover, when subjected to heat (40-70 °C) there was a significant increase in the stability of EGCG in the FSE due to the binding of salvianolic acid B. Through this interesting approach, two molecules are simultaneously attached and encapsulated in ferritin in a multilayer form under moderate conditions, which is conducive to the protection of unstable molecules for potential encapsulation and delivery utilization.
Keywords: bioactive molecules; ferritin channel; multilayer; steric hindrance; urea.