Dynamic surface tension probe for measuring the concentration of extracellular vesicles

Extracellular Vesicles

Chernyshev, Vasiliy S., Roman N. Chuprov-Netochin, Ekaterina Tsydenzhapova, Brian Van Devener, Sergey Leonov, Dmitry Gorin, and Mikhail Skliar. 2022. “Dynamic Surface Tension Probe for Measuring the Concentration of Extracellular Vesicles.” Biochemical and Biophysical Research Communications 609 (June): 189–94. https://doi.org/10.1016/j.bbrc.2022.04.017.

The concentration of extracellular vesicles (EVs) is an essential attribute of biofluids and EV preparations. EV concentration in body fluids was correlated with health status. The abundance of EV secreted by cultured cells into growth medium is vital in signaling studies, tissue and disease models, and biomanufacturing of acellular therapeutic secretome. A limited number of physical principles sensitive to EV concertation have been discovered so far. Particle-by-particle counting methods enumerate individual particles scattering light, modulating the Coulter current, or appearing in EM images. The available ensemble techniques in current use rely on the concentration-dependent signal intensity, as in the case of ELISA. In this study, we propose for the first-time the ensemble-based characterization of EV concentration by dynamic surface tension (DST) probe and demonstrate its implementation. We show that DST measurements agree with the widely used NTA measurements of EV concertation. The proposed method is low-cost and requires only basic laboratory equipment for implementation.

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Cigarette smoke (CS) represents one of the most relevant environmental risk factors for several chronic pathologies. Tissue damage caused by CS exposure is mediated, at least in part, by oxidative stress induced by its toxic and pro-oxidant components. Evidence demonstrates that extracellular vesicles (EVs) released by various cell types exposed to CS extract (CSE) are characterized by altered biochemical cargo and gained pathological properties. In the present study, we evaluated the content of oxidized proteins and phospholipid fatty acid profiles of EVs released by human bronchial epithelial BEAS-2B cells treated with CSE. This specific molecular characterization has hitherto not been performed. After confirmation that CSE reduces viability of BEAS-2B cells and elevates intracellular ROS levels, in a dose-dependent manner, we demonstrated that 24 h exposure at 1% CSE, a concentration that only slight modifies cell viability but increases ROS levels, was able to increase carbonylated protein levels in cells and released EVs. The release of oxidatively modified proteins via EVs might represent a mechanism used by cells to remove toxic proteins in order to avoid their intracellular overloading. Moreover, 1% CSE induced only few changes in the fatty acid asset in BEAS-2B cell membrane phospholipids, whereas several rearrangements were observed in EVs released by CSE-treated cells. The impact of changes in acyl chain composition of CSE-EVs accounted for the increased saturation levels of phospholipids, a membrane parameter that might influence EV stability, uptake and, at least in part, EV-mediated biological effects. The present in vitro study adds new information concerning the biochemical composition of CSE-related EVs, useful to predict their biological effects on target cells. Furthermore, the information regarding the presence of oxidized proteins and the specific membrane features of CSE-related EVs can be useful to define the utilization of circulating EVs as marker for diagnosing of CS-induced lung damage and/or CS-related diseases.

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