Characterization and bioactivity of extracellular vesicles isolated from pomegranate

• Published in: Food & function Vol. 13; no. 24; pp. 1287 - 12882
• Main Authors: Sánchez-López, Christian M, Manzaneque-López, Mari Cruz, Pérez-Bermúdez, Pedro, Soler, Carla, Marcilla, Antonio
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 13.12.2022
• Subjects: Biological activity; Caco-2 Cells; Cell lines; Chromatography, Liquid; Extracellular vesicles; Extracellular Vesicles - metabolism; Humans; Inflammation; Monocytes; Nanoparticles; Plants; Proteins; Proteins - metabolism; Proteomics; Proteomics - methods; Size exclusion chromatography; Tandem Mass Spectrometry; Transmission electron microscopy; Vesicles; Wound healing
• ISSN: 2042-6496; 2042-650X
• DOI: 10.1039/d2fo01806c

In the current study, extracellular vesicles from pomegranate juice (PgEVs) were isolated for the first time using size exclusion chromatography (SEC). This method permitted us to obtain highly enriched EV samples without most of the non-EV co-isolated proteins. The characterization of PgEVs through nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) allowed the determination of vesicles' concentration/volume, size, and morphology. It was confirmed from the analytical data that PgEVs contain a homogeneous population of vesicles, with a dimension and structure comparable to plant-derived EVs. Proteomic analyses by LC-MS/MS led to the characterization of 131 proteins, and several of them were related commonly to the biogenesis and transport of EVs, and/or proposed as EV markers. PgEVs exerted anti-inflammatory, antioxidant and wound-healing effects when added to the in vitro cultures of monocytic (THP-1) and intestinal (Caco-2) cell lines, respectively. In the current study, extracellular vesicles from pomegranate juice (PgEVs) were isolated for the first time using size exclusion chromatography (SEC).