Biochimica et biophysica acta. General subjects

1872-8006186672022JulBiochimica et biophysica acta. General subjectsBiochim Biophys Acta Gen SubjNanomechanical characterization of exosomes and concomitant nanoparticles from blood plasma by PeakForce AFM in liquid.13013913013910.1016/j.bbagen.2022.130139S0304-4165(22)00057-5To date, EVs characterization techniques are extremely diverse. The contribution of AFM, in particular, is often confined to size distribution. While AFM provides a unique possibility to carry out measurements in situ, nanomechanical characterization of EVs is still missing.Blood plasma EVs were isolated by ultracentrifugation, analyzed by flow cytometry and NTA. Followed by cryo-EM, we applied PeakForce AFM to assess morphological and nanomechanical properties of EVs in liquid.Nanoparticles were subdivided by their size estimated for their suspended state into sub-sets of small S1-EVs (< 30 nm), S2-EVs (30-50 nm), and sub-set of large ones L-EVs (50-170 nm). Non-membranous S1-EVs were distinguished by higher Young's modulus (10.33(7.36;15.25) MPa) and were less deformed by AFM tip (3.6(2.8;4.4) nm) compared to membrane exosomes S2-EVs (6.25(4.52;8.24) MPa and 4.8(4.3;5.9) nm). L-EVs were identified as large membrane exosomes, heterogeneous by their nanomechanical properties (22.43(8.26;53.11) MPa and 3.57(2.07;7.89) nm). Nanomechanical mapping revealed a few non-deformed L-EVs, of which Young's modulus rose up to 300 MPa. Taken together with cryo-EM, these results lead us to the suggestion that two or more vesicles could be contained inside a large one being a multilayer vesicle.We identified particles similar in morphology and showed differences in nanomechanical properties that could be attributed to the features of their inner structure.Our results further elucidate the identification of EVs and concomitant nanoparticles based on their nanomechanical properties.Copyright © 2022 Elsevier B.V. All rights reserved.BairamukovViktor YuVYPetersburg Nuclear Physics Institute Named by B.P. Konstantinov of NRC "Kurchatov Institute", 1, Orlova Roshcha, 188300 Gatchina, Russia. Electronic address: bayramukov_vy@pnpi.nrcki.ru.BukatinAnton SASAlferov Saint Petersburg National Research Academic University of the Russian Academy of Sciences, 8/3, Khlopina St., 194021 Saint Petersburg, Russia; Institute for Analytical Instrumentation of the Russian Academy of Sciences, 31-33A, Ivana Chernych, 198095 Saint Petersburg, Russia.KamyshinskyRoman ARANational Research Center "Kurchatov Institute", 1, Akademika Kurchatova pl., 123182 Moscow, Russia; Moscow Institute of Physics and Technology, Institutsky lane 9, 141700, Dolgoprudny, Moscow, Russia.BurdakovVladimir SVSPetersburg Nuclear Physics Institute Named by B.P. Konstantinov of NRC "Kurchatov Institute", 1, Orlova Roshcha, 188300 Gatchina, Russia; National Research Center "Kurchatov Institute", 1, Akademika Kurchatova pl., 123182 Moscow, Russia.PichkurEvgeny BEBPetersburg Nuclear Physics Institute Named by B.P. Konstantinov of NRC "Kurchatov Institute", 1, Orlova Roshcha, 188300 Gatchina, Russia; National Research Center "Kurchatov Institute", 1, Akademika Kurchatova pl., 123182 Moscow, Russia.ShtamTatiana ATAPetersburg Nuclear Physics Institute Named by B.P. Konstantinov of NRC "Kurchatov Institute", 1, Orlova Roshcha, 188300 Gatchina, Russia; National Research Center "Kurchatov Institute", 1, Akademika Kurchatova pl., 123182 Moscow, Russia.StarodubtsevaMaria NMNDepartment of Medical and Biological Physics, Gomel State Medical University, 5, Lange St., 246000 Gomel, Belarus; Institute of Radiobiology of NAS of Belarus, 4, Fedyuninskogo St., 246007 Gomel, Belarus.engJournal ArticleResearch Support, Non-U.S. Gov't20220404