Immobilization of Olive Leaf Extract with Chitosan Nanoparticles as an Adjunct to Enhance Cytotoxicity

[Image: see text] We immobilized the olive leaf extract (OLE) with chitosan nanoparticles (CNPs) by optimizing the effect of various immobilization conditions, and OLE-loaded CNPs (OLE-CNPs) were then elaborately characterized physicochemically by scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, dynamic light scattering (DLS), and atomic force microscopy (AFM). Under optimal conditions, CNPs were able to accommodate the OLE with a loading capacity of 97.5%. The resulting OLE-CNPs had a spherical morphology, and their average diameter was approximately 100 nm. The cytotoxic influence, cell cycle distribution, and apoptosis stage of OLE and OLE-CNPs were analyzed on lung carcinoma (A549) and breast adenocarcinoma (MCF-7) cell lines. In an in vitro cytotoxic assay, IC(50) values of OLE-CNPs were determined to be 540 μg/mL for A549 and 810 μg/mL for MCF-7. The treatment of both A549 and MCF-7 with OLE-CNPs caused the highest cell arrest in G0/G1 in a dose-independent manner. OLE-CNPs affected cell cycle distribution in a manner different from free OLE treatment in both cancer cells. A549 and MCF-7 cells were predominantly found in the late apoptosis and necrosis phases, respectively, upon treatment of 1000 μM OLE-CNPs. Our results suggest that CNPs enhance the utility of OLEs as nutraceuticals in cancer and that OLE-CNPs can be utilized as an adjunct to cancer therapy.