Harnessing tomato-derived small extracellular vesicles as drug delivery system for cancer therapy
Creators: name: Sarwareddy, Kartik Kumar
name: Singh, Anula Divyash
name: Patnam, Sreekanth
name: Sesuraj, Babiola Annes
name: Ponamgi, spd
name: Thakur, Basant Kumar
name: Manda, Venkata Sasidhar
Publisher: name: Taylor & Francis
Year: 2025
Subjects: Biochemistry
Space Science
Cell Biology
Molecular Biology
Pharmacology
Biotechnology
Chemical Sciences not elsewhere classified
Biological Sciences not elsewhere classified
Cancer

description: 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. This study explores using tiny particles from tomatoes, known as small extracellular vesicles (TsEVs), to deliver calcitriol, a type of vitamin D, as a treatment for colon cancer. These tomato-derived particles offer a cost-effective and sustainable alternative to drug carriers traditionally made from animal cells. TsEVs were successfully loaded with calcitriol, which effectively reduced the growth, spread, and survival of colon cancer cells in lab tests. Additionally, treated cancer cells showed increased levels of stress markers and signals leading to cell death. This approach demonstrates a promising, natural way to improve cancer treatment while addressing cost and supply challenges. This study developed an environmentally friendly, scalable, and cost-effective drug delivery system using tomato-derived extracellular vesicles (TsEVs).TsEVs were efficiently isolated using differential centrifugation, ultrafiltration, and PEG precipitation, resulting in 30–200 nm vesicles suitable for drug delivery.Calcitriol was loaded into the TsEVs using sonication, achieving a high loading efficiency of 47%.TsEVs-calcitriol showed enhanced drug release at acidic pH (4.5) and higher cytotoxicity in colon cancer cells than free calcitriol.TsEVs-calcitriol more effectively reduced colony formation, inhibited cell migration, and elevated ROS levels than free calcitriol.TsEV-calcitriol treatment activated the cell death switch involving survivin and Bax/Bcl-2, indicating superior apoptosis induction compared to free calcitriol. This study developed an environmentally friendly, scalable, and cost-effective drug delivery system using tomato-derived extracellular vesicles (TsEVs). TsEVs were efficiently isolated using differential centrifugation, ultrafiltration, and PEG precipitation, resulting in 30–200 nm vesicles suitable for drug delivery. Calcitriol was loaded into the TsEVs using sonication, achieving a high loading efficiency of 47%. TsEVs-calcitriol showed enhanced drug release at acidic pH (4.5) and higher cytotoxicity in colon cancer cells than free calcitriol. TsEVs-calcitriol more effectively reduced colony formation, inhibited cell migration, and elevated ROS levels than free calcitriol. TsEV-calcitriol treatment activated the cell death switch involving survivin and Bax/Bcl-2, indicating superior apoptosis induction compared to free calcitriol.