Chitosan binding to a novel alfalfa phytoferritin nanocage loaded with baicalein: Simulated digestion and absorption evaluation

Phytoferritin was explored as an attractive nanocarrier to encapsulate bioactive compounds due to its excellent stability and biocompatibility. In the present study, a novel phytoferritin derived from alfalfa (Medicago sativa ) was successfully expressed, purified and characterized. Results confirmed that alfalfa ferritin, self-assembled by 24 subunits, formed a spherical hollow structure. Baicalein exhibits superior antioxidant properties and nutritious values, but low bioavailability and solubility limit its application. Herein, we fabricated water-soluble chitosan-ferritin-baicalein nanoparticles to overcome its drawbacks. It was calculated that one apoferritin cage could encapsulate 52 molecules of baicalein. Moreover, chitosan-ferritin-baicalein nanoparticles prolonged the release of baicalein in simulated gastrointestinal tract digestion. Caco-2 cell monolayer absorption analysis demonstrated that baicalein encapsulated within ferritin-chitosan double layers was more efficient in cellular transportation. These results indicated that alfalfa ferritin, as a novel cage-like protein, has potential application in improving the bioavailability of insoluble bioactive molecules.