Summary. Background: Plasma and other body fluids contain membranous extracellular vesicles (EVs), which are considered to derive from activated or apoptotic cells. EVs participate in physiological and pathological processes and have potential applications in diagnostics or therapeutics. Knowledge on EVs is, however, limited, mainly due to their sub-micrometer size and to intrinsic limitations in methods applied for their characterization. Objectives: Our aim was to provide a comprehensive description of EVs from plasma of healthy subjects. Methods: Cryo-transmission electron microscopy combined with receptor-specific gold labeling was used to reveal the morphology, size and phenotype of EVs. An original approach based on sedimentation on electron microscopy grids was developed for enumerating EVs. A correlation was performed between conventional flow cytometry and electron microscopy results. Results: We show that platelet-free plasma samples contain spherical EVs, 30 nm to 1 lm in diameter, tubular EVs, 1-5 lm long, and membrane fragments, 1-8 lm large. We show that only a minority of EVs expose the procoagulant lipid phosphatidylserine, in contrast to the classical theory of EV formation. In addition, the concentrations of the main EV sub-populations are determined after sedimentation on EM grids. Finally, we show that conventional flow cytometry, the main method of EV characterization, detects only about 1% of them. Conclusion: This study brings novel insights on EVs from normal plasma and provides a reference for further studies of EVs in disease situations.
International audienceHydrophobically modified maghemite (γ-Fe2O3) nanoparticles were encapsulated within the membrane of poly(trimethylene carbonate)-b-poly(L-glutamic acid) (PTMC-b-PGA) block copolymer vesicles using a nanoprecipitation process. This formation method gives a simple access to highly magnetic nanoparticles (MNPs) (loaded up to 70 wt %) together with a good control over the vesicles size (100 to 400 nm). The simultaneous loading of maghemite nanoparticles and doxorubicin was also achieved by nanoprecipitation. The deformation of the vesicle membrane under an applied magnetic field has been evidenced by small angle neutron scattering. These superparamagnetic hybrid self-assemblies display enhanced contrast properties that open potential applications for Magnetic Resonance Imaging. They can also be guided in a magnetic field gradient. The feasibility of controlled drug release by radio-frequency magnetic hyperthermia was demonstrated in the case of encapsulated doxorubicin molecules, showing the viability of the concept of magneto-chemotherapy. These magnetic polymersomes can be used as efficient multifunctional nano-carriers for combined therapy and imaging
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