Abstract
Extracellular vesicles (EVs) are lipid bilayer nano-dimensional spherical structures and act mainly as signaling mediators between cells, in particular modulating immunity and inflammation. Milk-derived EVs (mEVs) can have immunomodulatory and anti-inflammatory effects, and milk is one of the most promising food sources of EVs. In this context, this study aimed to evaluate bovine mEVs anti-inflammatory and immunomodulating effects on an in vitro co-culture (Caco-2 and THP-1) model of intestinal inflammation through gene expression evaluation with RT-qPCR and cytokine release through ELISA. After establishing a pro-inflammatory environment due to IFN-γ and LPS stimuli, CXCL8, IL1B, TNFA, IL12A, IL23A, TGFB1, NOS2, and MMP9 were significantly up-regulated in inflamed Caco-2 compared to the basal co-culture. Moreover, IL-17, IL-1β, IL-6, TNF-α release was increased in supernatants of THP-1. The mEV administration partially restored initial conditions with an effective anti-inflammatory activity. Indeed, a decrease in gene expression and protein production of most of the tested cytokines was detected, together with a significant gene expression decrease in MMP9 and the up-regulation of MUC2 and TJP1. These results showed a fundamental capability of mEVs to modulate inflammation and their potential beneficial effect on the intestinal mucosa.
Highlights
Licensee MDPI, Basel, Switzerland.Extracellular vesicles (EVs), nano-dimensional spherical structures enclosed by a lipid bilayer membrane, include many types of vesicles that differ for the biogenesis process, dimension, tissue of origin, and function
This study aimed to evaluate the anti-inflammatory and immunomodulating effects of Milk-derived EVs (mEVs) derived from bovine milk on an in vitro co-culture model of intestinal inflammation [52]
A normalization step was performed according to the expression levels of the three reference genes (RPLP0, ribosomal protein L37a (RPL37A), and ribosomal protein S14 (RPS14)) after assessing their stability under different experimental conditions, using the Norm algorithm included in Bio-Rad CFX Maestro software
Summary
Extracellular vesicles (EVs), nano-dimensional spherical structures enclosed by a lipid bilayer membrane, include many types of vesicles that differ for the biogenesis process, dimension, tissue of origin, and function. Biomedicines 2022, 10, 570 shedding vesicles, released through the exocytosis process and ranging from 100 to 1000 nm; and apoptotic bodies greater than 1000 nm [1–3]. Vesicles associated with a specific tissue or a particular function were discerned, such as prostasomes, microparticles, tolerosomes, and oncosomes [4–9]. The confusion in nomenclature generated from the increasing number of works in the EV field, associated with the difficulties in distinguishing these different types of vesicles through the current techniques, made the International Society of Extracellular Vesicles solicit the use of the generic EV, as stated in the developed guidelines [10]
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