In vitro antiproliferative activity evaluation of functionalized nanoparticles containing phytol and methotrexate.
co-nanoencapsulation; specific targeting; HeLa cells.
Despite advances in oncological therapy, the selective delivery of drugs to tumor tissues still faces limitations due to low specificity and side effects. Methotrexate (MTX) is a chemotherapeutic agent capable of binding to folate receptors (FR), which are frequently overexpressed in cancer cells. Strategies combining chemotherapeutics with natural compounds in nanostructured systems can enhance therapeutic efficacy. In this study, co-encapsulated and functionalized nanoparticles containing MTX and phytol, a bioactive compound with anticancer potential, were developed, evaluating their antiproliferative activity in FR expressing lineages, such as HeLa cells. The nanoparticles were obtained by nanoprecipitation followed by solvent evaporation. Characterization included particle diameter, polydispersity index (PdI), zeta potential, encapsulation efficiency (EE), and morphology via scanning electron microscopy. Biological assays comprised cytocompatibility tests in L929 fibroblasts, as well as cytotoxicity and clonogenicity assays in HeLa cells. The nanoparticles presented a mean size of 250 nm, PdI below 0.2 (indicating homogeneity), physical stability, and spherical morphology, with EE exceeding 80% for both actives. The functionalized systems proved cytocompatible with L929 cells at a concentration of 5 µg/mL of the polymeric matrix and exhibited a marked antiproliferative effect on HeLa cells, with 63% cell death within 24 hours (MTT assay). After 8 days, the absence of colony formation indicated loss of clonogenic potential and prolonged cytotoxic effects. These findings highlight the potential of the MTX–phytol co encapsulated system as a promising approach for targeted antineoplastic therapies.