Strawberry-Derived Exosome-Like Nanoparticles Prevent Oxidative Stress in Human Mesenchymal Stromal Cells

Abstract

Plant-derived exosome-like nanovesicles (EPDENs) have recently been isolated and evaluated as potential bioactive nutraceutical biomolecules. It has been hypothesized that EPDENs may exert their activity on mammalian cells through their specific cargo. In this study, we isolated and purified EPDENs from the strawberry juice of Fragaria x ananassa (cv. Romina), a new cultivar characterized by a high content of anthocyanins, folic acid, flavonols, and vitamin C and an elevated antioxidant capacity. Fragaria-derived EPDENs were purified by a series of centrifugation and filtration steps. EPDENs showed size and morphology similar to mammalian extracellular nanovesicles. The internalization of Fragaria-derived EPDENs by human mesenchymal stromal cells (MSCs) did not negatively affect their viability, and the pretreatment of MSCs with Fragaria-derived EPDENs prevented oxidative stress in a dose-dependent manner. This is possibly due to the presence of vitamin C inside the nanovesicle membrane. The analysis of EPDEN cargo also revealed the presence of small RNAs and miRNAs. These findings suggest that Fragaria-derived EPDENs may be considered nanoshuttles contained in food, with potential health-promoting activity.

Extracellular Vesicles from Plants: Current Knowledge and Open Questions

“…The most common method of plant vesicle isolation is differential centrifugation followed by ultracentrifugation [4][5][6][7][8][9][10][11][12]. The starting material can be fruits [4,[6][7][8][9][10], roots [5,13], stems [9,14], leaves [9,[14][15][16], seeds [16], and saps [17]; these matrices can be manually squeezed or using a mixer to obtain the juice. The juice is subjected to several differential centrifugation steps: low-speed centrifugation (about 500-3000× g for 10-15 min) to remove plant fibers and large particles; intermediate speed centrifugation (2000-10,000× g for 20-40 min) to remove large debris and subcellular organelles; and high-speed centrifugation (100,000-150,000× g for 1.5-2 h) to obtain PDEVs pellet.…”

“…The presence of these miR-NAs was then correlated with the reduction of viability of tumor cells treated with EVs [19]. Analysis of miRNAs was also performed in strawberry juice and the PDEVs, detecting only miR166g in both matrices [8].…”

“…Sugars and their derivates were also found in microvesicles, together with quinic and oxalic acid, while the nanovesicles are enriched in organic acids, such as glycolic and citric acids, and amino acids [39]. Ascorbic acid was found in strawberry-derived EVs (416 nmoles/mg EVs) [8].…”

“…It was recently found that strawberry-derived EPDENs have antioxidant properties as well. Adipose-derived mesenchymal stem cells (ADMSCs) pre-treated with strawberry EPDENs and then stimulated with H 2 O 2 showed reduced cell death and lower ROS levels than cells treated with H 2 O 2 alone, probably because EPDENs contain vitamin C [8].…”

Isolation of cabbage exosome-like nanovesicles and investigation of their biological activities in human cells

“…Blueberry-derived nanoparticles were able to modulate the expression of genes involved in inflammatory response, cytokine release, and oxidative stress [ 19 ]. Recently, exosome-like particles from strawberry were shown to prevent oxidative stress in human cells by delivering intrinsic vitamin C [ 20 ]. Lemon-derived nanoparticles have in vitro anti-neoplastic activity on a panel of different solid and hematological cancers cell lines [ 21 ].…”