Milk-Derived Extracellular Vesicles Enhanced Sorafenib's Therapeutic Response on Hepatocellular Carcinoma Cells Publisher



Salehi M ; Negahdari B ; Hashemi M ; Seydi H ; Salarinejad S ; Piryaei A ; Moradi A ; Zarrabi A ; Shekari F ; Vosough M
Source: Journal of Drug Delivery Science and Technology Published:2026

Abstract

Sorafenib (SOR) is a commonly used medication to treat advanced hepatocellular carcinoma (HCC). However, its clinical efficacy is limited by poor water solubility, fast metabolism, and undesirable side effects. While milk and red blood cells (RBCs) have been introduced as potentially safe, scalable, and available sources of extracellular vesicles (EVs), no study has compared the feasibility of milk-derived EVs (miEVs) and RBC-derived EVs (RBCEVs) as drug delivery vehicles. Moreover, there is little information on whether loading SOR into EVs improves its therapeutic effectiveness. Herein, we isolated RBCEVs at 100,000× g (RBCEV100K) and various subpopulations of miEVs at 35,000× g (miEV35K), 70,000× g (miEV70K), and 100,000× g (miEV100K), and characterized them based on marker expression and size distribution. SOR was subsequently loaded into these EVs via incubation, and encapsulation efficiency was measured spectrophotometrically. Storage stability and cellular uptake were also assessed. Furthermore, comprehensive in vitro and in vivo assessments were performed on hepatic cell lines, Huh-7 and Hep3B. The isolated EVs were <200 nm in size and positive for EV-specific protein markers. Compared to the other EVs, miEV100K exhibited considerably higher SOR loading. Moreover, its stability during storage and cellular uptake rate in both cell lines were more efficient than RBCEV100K. Therefore, as an optimal SOR carrier, miEV100K was selected for subsequent experiments. Compared to SOR, SOR-miEV100K significantly induced apoptosis and inhibited proliferation, cell cycle progression, colony formation, and migration capacities in HCC cells by downregulating CDH2, Ki-67, CCND1 , and CD133 , while upregulating P53 expression. Moreover, SOR-miEV100K significantly attenuated the tumorigenic potential of Huh-7 cells in nude mice and was less cytotoxic to normal human fibroblasts than SOR. Therefore, miEV100K not only enhances the therapeutic efficacy of SOR but also reduces its side effects, highlighting its promise as a delivery platform. However, further investigation is required to pave the way toward clinical application. © 2026 Elsevier B.V.