Stability of Plant Leaf-Derived Extracellular Vesicles According to Preservative and Storage Temperature

Abstract

Plant-derived extracellular vesicles (EVs) are capable of efficiency delivering mRNAs, miRNAs, bioactive lipids, and proteins to mammalian cells. Plant-derived EVs critically contribute to the ability of plants to defend against pathogen attacks at the plant cell surface. They also represent a novel candidate natural substance that shows potential to be developed for food, cosmetic, and pharmaceutical products. However, although plant-derived EVs are acknowledged as having potential for various industrial applications, little is known about how their stability is affected by storage conditions. In this study, we evaluated the stability of Dendropanax morbifera leaf-derived extracellular vesicles (LEVs) alone or combined with the preservatives, 1,3-butylene glycol (to yield LEVs-1,3-BG) or TMO (LEVs-TMO). We stored these formulations at −20, 4, 25, and 45 °C for up to 4 weeks, and compared the stability of fresh and stored LEVs. We also assessed the effect of freeze-thawing cycles on the quantity and morphology of the LEVs. We found that different storage temperatures and number of freeze-thawing cycles altered the stability, size distribution, protein content, surface charge, and cellular uptake of LEVs compared to those of freshly isolated LEVs. LEVs-TMO showed higher stability when stored at 4 °C, compared to LEVs and LEVs-1,3-BG. Our study provides comprehensive information on how storage conditions affect LEVs and suggests that the potential industrial applications of plant-derived EVs may be broadened by the use of preservatives.

Plant-derived nanovesicles: Current understanding and applications for cancer therapy

“…Kim et al investigated Dendropanax morbifera leaf PELN stability at various temperatures: -20 °C, 4 °C, 25 °C, and 45 °C [ 95 ]. After a purification process, these PELNs were diluted in distilled water, stored at different temperatures, and assessed in terms of their pH, size distribution, and total protein.…”

Isolation of high-purity and high-stability exosomes from ginseng

“…Colloidal stability is an important factor influencing the functional maintenance and storage stability of isolated exosomes. Flocculation of exosomes negatively affects their colloidal stability ( Mu et al., 2014 ; Wang et al., 2015 ; Helwa et al., 2017 ; Umar et al., 2018 ; Kim et al., 2022 ). Zeta potential is key to estimating the colloidal stability of isolated exosomes ( Clogston and Patri, 2011 ; Soares Martins et al., 2018 ; Samimi et al., 2019 ; Midekessa et al., 2020 ).…”

Emergence of Edible Plant-Derived Nanovesicles as Functional Food Components and Nanocarriers for Therapeutics Delivery: Potentials in Human Health and Disease

“…In addition, studies on storage conditions for PDNVs are limited. Kim et al tested the efficacy of preservatives on the stability of nanovesicles derived from Dendropanax morbifera leaf with varying temperatures at −20, 4, 25, and 45 °C by comparing physico-chemical properties (pH, size, protein content, and surface charge) and cellular uptake ability, but did not test whether the post-storage PDNVs preserve biological activity in recipient cells [ 111 ]. Similarly, another study showed that ginseng-derived nanovesicles retained stability after 60 days of storage post freeze-dry, but their bioactivity after the storage was not measured [ 112 ].…”