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      Differences in the Quantity and Composition of Extracellular Vesicles in the Aqueous Humor of Patients with Retinal Neovascular Diseases

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          Abstract

          Extracellular vesicles (EVs) are secreted by various cells in the body fluid system and have been found to influence vessel formation and inflammatory responses in a variety of diseases. However, which EVs and their subtypes are involved in vascular retinal diseases is still unclear. Therefore, the aim of this study was to explore the particle distribution of EVs in retinal neovascular diseases, including age-related macular degeneration, polypoidal choroidal vasculopathy, and central retinal vein occlusion. The aqueous humor was harvested from 20 patients with different retinal neovascular diseases and six patients with cataracts as the control group. The particle distribution was analyzed using nanoparticle tracking analysis (NTA) and transmitting electron microscopy (TEM). The results revealed that the disease groups had large amounts of EVs and their subtypes compared to the control group. After isolating exosomes, a higher expression of CD81 + exosomes was shown in the disease groups using flow cytometry. The exosomes were then further classified into three subtypes of exomeres, small exosomes, and large exosomes, and their amounts were shown to differ depending on the disease type. To the best of our knowledge, this is the first study to elucidate the dynamics of EVs in retinal neovascular diseases using clinical cases. Our findings demonstrated the possible functionality of microvesicles and exosomes, indicating the potential of exosomes in the diagnosis and therapy of retinal neovascular diseases.

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          Exosomes

          Exosomes are small, single-membrane, secreted organelles of ∼30 to ∼200 nm in diameter that have the same topology as the cell and are enriched in selected proteins, lipids, nucleic acids, and glycoconjugates. Exosomes contain an array of membrane-associated, high-order oligomeric protein complexes, display pronounced molecular heterogeneity, and are created by budding at both plasma and endosome membranes. Exosome biogenesis is a mechanism of protein quality control, and once released, exosomes have activities as diverse as remodeling the extracellular matrix and transmitting signals and molecules to other cells. This pathway of intercellular vesicle traffic plays important roles in many aspects of human health and disease, including development, immunity, tissue homeostasis, cancer, and neurodegenerative diseases. In addition, viruses co-opt exosome biogenesis pathways both for assembling infectious particles and for establishing host permissiveness. On the basis of these and other properties, exosomes are being developed as therapeutic agents in multiple disease models.
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            Biogenesis and secretion of exosomes.

            Although observed for several decades, the release of membrane-enclosed vesicles by cells into their surrounding environment has been the subject of increasing interest in the past few years, which led to the creation, in 2012, of a scientific society dedicated to the subject: the International Society for Extracellular Vesicles. Convincing evidence that vesicles allow exchange of complex information fuelled this rise in interest. But it has also become clear that different types of secreted vesicles co-exist, with different intracellular origins and modes of formation, and thus probably different compositions and functions. Exosomes are one sub-type of secreted vesicles. They form inside eukaryotic cells in multivesicular compartments, and are secreted when these compartments fuse with the plasma membrane. Interestingly, different families of molecules have been shown to allow intracellular formation of exosomes and their subsequent secretion, which suggests that even among exosomes different sub-types exist. Copyright © 2014 Elsevier Ltd. All rights reserved.
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              Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles

              Secreted membrane-enclosed vesicles, collectively called extracellular vesicles (EVs), which include exosomes, ectosomes, microvesicles, microparticles, apoptotic bodies and other EV subsets, encompass a very rapidly growing scientific field in biology and medicine. Importantly, it is currently technically challenging to obtain a totally pure EV fraction free from non-vesicular components for functional studies, and therefore there is a need to establish guidelines for analyses of these vesicles and reporting of scientific studies on EV biology. Here, the International Society for Extracellular Vesicles (ISEV) provides researchers with a minimal set of biochemical, biophysical and functional standards that should be used to attribute any specific biological cargo or functions to EVs.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Diagnostics (Basel)
                Diagnostics (Basel)
                diagnostics
                Diagnostics
                MDPI
                2075-4418
                15 July 2021
                July 2021
                : 11
                : 7
                : 1276
                Affiliations
                [1 ]Department of Ophthalmology, Chung Shan Medical University Hospital, Taichung 402306, Taiwan; amy1234575@ 123456gmail.com (Y.-P.H.); cconnie7@ 123456gmail.com (C.C.); cshy1886@ 123456csh.org.tw (C.-M.L.); rockinroll355@ 123456gmail.com (P.-Y.C.); orien1168@ 123456gmail.com (W.-H.C.); bella37245@ 123456gmail.com (B.-H.K.)
                [2 ]School of Medicine, Chung Shan Medical University, Taichung 402306, Taiwan
                [3 ]Department of Optometry, Chung Shan Medical University, Taichung 402306, Taiwan
                [4 ]Institute of Optometry, Chung Shan Medical University, Taichung 402306, Taiwan
                [5 ]Department of Radiology, Taichung Veterans General Hospital, Taichung 407219, Taiwan; drahcirxp@ 123456gmail.com
                [6 ]Department of Ophthalmology, Taichung Veterans General Hospital, Taichung 407219, Taiwan; b92401086@ 123456gmail.com
                [7 ]Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 300044, Taiwan; chaomin@ 123456mx.nthu.edu.tw
                [8 ]Institute of Nanoengineering and Microsystem, National Tsing Hua University, Hsinchu 30044, Taiwan; chihchen23@ 123456gmail.com
                [9 ]Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 300044, Taiwan
                [10 ]Biotechnology Center, National Chung Hsing University, Taichung 402202, Taiwan
                Author notes
                [* ]Correspondence: my.scott.hsu@ 123456gmail.com
                [†]

                These authors contributed equally to this article and share the first authorship.

                Author information
                https://orcid.org/0000-0002-8644-1960
                https://orcid.org/0000-0002-2195-4802
                https://orcid.org/0000-0002-5488-4257
                Article
                diagnostics-11-01276
                10.3390/diagnostics11071276
                8306174
                34359359
                c41126ba-f733-48ef-85a5-df389bdd5c84
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 27 June 2021
                : 12 July 2021
                Categories
                Article

                extracellular vesicle,exosome,aqueous humor,nanoparticle tracking analysis,retinal neovascular disease,angiogenesis

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