Abstract
This study aims to explore a sustainable and scalable approach using tomato fruit-derived sEVs (TsEVs) to deliver calcitriol for enhanced anticancer effects, addressing challenges of low yield and high costs associated with mammalian cell-derived sEVs. TsEVs were isolated by centrifugation and ultrafiltration and characterized using DLS, TEM, and biochemical assays. Calcitriol was loaded into TsEVs via loading methods, with efficiency measured by spectrophotometry and HPLC. HCT116 and HT29 colon cancer cells were treated with TsEV-calcitriol and assessed for viability, colony formation, migration, ROS levels, and apoptosis gene expression. Isolated TsEVs ranged from 30-200 nm with a protein-to-lipid ratio of ∼1. Calcitriol encapsulation efficiencies were 15.4% (passive), 34.8% (freeze-thaw), and 47.3% (sonication). TsEV-calcitriol reduced HCT116 cell viability with IC50 values of 4.05 µg/ml (24 h) and 2.07 µg/ml (48 h). Clonogenic assays showed reduced colony formation and migration. Elevated ROS levels and increased Bax/Bcl-2 ratio were observed in treated HCT116 and HT29 colon cancer cells. These findings highlight TsEVs as a promising alternative drug delivery platform to mammalian cell-derived sEV for enhancing the therapeutic efficiency of calcitriol and other anticancer agents.
Published Version
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