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Abstract Objective (s): Recently, medicinal plants have grabbed much attention in the prevention and treatment of cancer due to their ability to increase the efficiency of chemotherapy agents. Luteolin is a flavonoid widely studied for its antitumor effects. However, luteolin has low bioavailability and poor efficacy due to its hydrophobicity. This study aimed to prepare luteolin nanoemulsion (NE) and evaluate its physicochemical and anti-tumor properties in combination with doxorubicin (DOX) in vitro. NE containing luteolin was prepared by the prob-sonicate method. The physicochemical properties of nanoparticles, including particle size, zeta potential, morphology, encapsulation efficiency, viscosity, pH, drug release profile, and thermal stability were investigated. Finally, the toxicity of free luteolin and luteolin NEs at different concentrations, with and without DOX, was assessed against normal …