Stability of Blueberry Extracellular Vesicles and Their Gene Regulation Effects in Intestinal Caco-2 Cells

Abstract

Plant extracellular vesicles (P-EVs) are considered promising functional food ingredients due to their various health benefits. In this study, blueberry extracellular vesicles (B-EVs) were collected and purified by size exclusion chromatography (SEC). The chemical compounds in B-EV extracts were analyzed by LC-MS/MS. In addition, the stability of B-EVs was evaluated during short- and long-term storage, heating, and in vitro digestion. The results showed that the B-EVs had a desirable particle size (88.2 ± 7.7 nm). Protein and total RNA concentrations were 582 ± 11.2 μg/mL and 15.4 μg/mL, respectively. The optimal storage temperatures for B-EVs were 4 °C and −80 °C for short- and long-term storage, respectively. Fluorescent labeling and qRT-PCR tests showed that B-EVs could be specifically internalized by Caco-2 cells, whereas virtually no cytotoxic or growth-inhibitory effects were observed. B-EVs down-regulated the expression levels of IL-1β and IL-8 and up-regulated the expression levels of NF-κβ and TLR5 in Caco-2 cells. Overall, the results proved that the intact structure of B-EVs could be preserved during food storage and processing conditions. B-EVs had the ability to reach the human intestine through oral delivery. Moreover, they could be absorbed by intestinal cells and affect human intestinal function.

Exploring new avenues of health protection: plant-derived nanovesicles reshape microbial communities

“…Existing preservation of PDNVs is mainly done by simply lowering the temperature, with most of the PDNVs being stored at −80 °C, some of the PDNVs undergo liquid nitrogen flash freezing and then stored at −80 °C, and there are also studies of storing them at −20 °C or place them at 4 °C for short-term preservation. There have been some studies evaluating the storage of PDNVs at different temperatures for short periods of time, and they found that 1 month's storage at some temperatures caused these PDNVs to undergo morphological changes, mainly in the form of aggregation or fusion [ 112 ]. Of course, this is related to the origin of the plant, some PDNVs are relatively more stable, and different PDNVs may be suitable for storage at different temperatures [ 112 ].…”

“…There have been some studies evaluating the storage of PDNVs at different temperatures for short periods of time, and they found that 1 month's storage at some temperatures caused these PDNVs to undergo morphological changes, mainly in the form of aggregation or fusion [ 112 ]. Of course, this is related to the origin of the plant, some PDNVs are relatively more stable, and different PDNVs may be suitable for storage at different temperatures [ 112 ]. Some studies have shown that the effect of transdermal treatment of melanoma with aloe vera-derived nanovesicles as a delivery vehicle for indocyanine green (ICG) remains after storage for some time [ 113 ].…”