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  1. Article ; Online: In Situ

    Butera, Ester / Dupont, Aurelien / Aimé, Alexis / Ducarre, Solène / Chiechio, Regina M / Even-Hernandez, Pascale / Contino, Annalinda / Maccarone, Giuseppe / Ravel, Célia / Marchi, Valérie

    ACS applied materials & interfaces

    2024  Volume 16, Issue 17, Page(s) 21643–21652

    Abstract: Extracellular vesicles (EVs) are well-known membrane-limited particles secreted by both healthy and cancerous cells. They are considered as biomarkers for early cancer diagnosis and are involved in many pathologies and physiological pathways. They could ... ...

    Abstract Extracellular vesicles (EVs) are well-known membrane-limited particles secreted by both healthy and cancerous cells. They are considered as biomarkers for early cancer diagnosis and are involved in many pathologies and physiological pathways. They could serve as diagnostic tools in liquid biopsies, as therapeutics in regenerative medicine, or as drug delivery vehicles. Our aim is here to encapsulate luminescent nanoprobes in the aqueous compartment of human EVs extracted from reproductive fluids. The analysis and labeling of the EVs content with easily detectable luminescent nanoparticles could enable a powerful tool for early diagnosis of specific diseases and also for the design of new therapeutics. In this view, gold nanoclusters (AuNCs) appear as an attractive alternative as nontoxic fluorophore probes because of their luminescence properties, large window of fluorescence lifetimes (1 ns-1 μs), ultrasmall size (<2 nm), good biocompatibility, and specific ability as X-ray photosensitizers. Here, we investigated an attractive method that uses fusogenic liposomes to deliver gold nanoclusters into EVs. This approach guarantees the preservation of the EVs membrane without any breakage, thus maintaining compartmental integrity. Different lipid compositions of liposomes preloaded with AuNCs were selected to interact electrostatically with human EVs and compared in terms of fusion efficiency. The mixture of liposomes and EVs results in membrane mixing as demonstrated by FRET experiments and fusion revealed by flux cytometry and cryo-TEM. The resulting fused EVs exhibit typical fluorescence of the AuNCs together with an increased size in agreement with fusion. Moreover, the fusion events in mixtures of EVs and AuNCs preloaded liposomes were analyzed by using cryo-electron microscopy. Finally, the ratio of released AuNCs during the fusion between the fusogenic liposomes and the EVs was estimated to be less than 20 mol % by Au titration using ICP spectroscopy.
    MeSH term(s) Gold/chemistry ; Humans ; Extracellular Vesicles/chemistry ; Extracellular Vesicles/metabolism ; Metal Nanoparticles/chemistry ; Liposomes/chemistry
    Chemical Substances Gold (7440-57-5) ; Liposomes
    Language English
    Publishing date 2024-04-16
    Publishing country United States
    Document type Journal Article
    ISSN 1944-8252
    ISSN (online) 1944-8252
    DOI 10.1021/acsami.4c02445
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Diversity of Extracellular Vesicles in Human Follicular Fluid: Morphological Analysis and Quantification.

    Neyroud, Anne-Sophie / Chiechio, Regina Maria / Moulin, Gregory / Ducarre, Solène / Heichette, Claire / Dupont, Aurélien / Budzynski, Mathieu / Even-Hernandez, Pascale / Faro, Maria Jose Lo / Yefimova, Marina / Marchi, Valérie / Ravel, Célia

    International journal of molecular sciences

    2022  Volume 23, Issue 19

    Abstract: The oocyte microenvironment constituted by the follicular fluid (FF) is a key for the optimal development of female gametes. Its composition reflects the physiological state of the ovarian follicle. The particularity of FF is to contain a huge diversity ... ...

    Abstract The oocyte microenvironment constituted by the follicular fluid (FF) is a key for the optimal development of female gametes. Its composition reflects the physiological state of the ovarian follicle. The particularity of FF is to contain a huge diversity of extracellular vesicles specific to women, in the same way as seminal plasma in men. Here, we described and compared morphological aspects of broad subcategories of human FF-related Extracellular Vesicles (EVs). EVs participate in physiological and pathological processes and have potential applications in diagnostics or therapeutics. EVs isolated from FF are involved in different biological functions related to follicular growth, oocyte maturation, and embryo development. However, knowledge on the morphology of FF-derived EVs is limited, mainly due to their sub-micrometer size and to intrinsic limitations in methods applied for their characterization. The aim of this study was to provide a comprehensive morphological description of EVs from FF of healthy subjects and quantification. EVs separation was realized by centrifugation, with comparison of the EV yield obtained from differential centrifugation and one-step ultracentrifugation. Cryo-Transmission Electron Microscopy was used to reveal the morphology, size, and phenotype of EVs. Dynamic Light Scattering (DLS) and Nanoparticle Tracking Analysis (NTA) were used to quantify and analyze the size distribution for each centrifugation step. We performed a comprehensive inventory of human follicular fluid EVs. We show that human FF contains a huge diversity of EVs. This study brings novel insights on EVs from normal FF and provides a reference for further studies of EVs in ovarian diseases.
    MeSH term(s) Extracellular Vesicles/physiology ; Female ; Follicular Fluid ; Humans ; Male ; Oocytes ; Oogenesis ; Ovarian Follicle
    Language English
    Publishing date 2022-10-02
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms231911676
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Encapsulation of Luminescent Gold Nanoclusters into Synthetic Vesicles.

    Chiechio, Regina M / Ducarre, Solène / Marets, Célia / Dupont, Aurélien / Even-Hernandez, Pascale / Pinson, Xavier / Dutertre, Stéphanie / Artzner, Franck / Musumeci, Paolo / Ravel, Célia / Faro, Maria Jose Lo / Marchi, Valérie

    Nanomaterials (Basel, Switzerland)

    2022  Volume 12, Issue 21

    Abstract: Gold nanoclusters (Au NCs) are attractive luminescent nanoprobes for biomedical applications. In vivo biosensing and bioimaging requires the delivery of the Au NCs into subcellular compartments. In this view, we explore here the possible encapsulation of ...

    Abstract Gold nanoclusters (Au NCs) are attractive luminescent nanoprobes for biomedical applications. In vivo biosensing and bioimaging requires the delivery of the Au NCs into subcellular compartments. In this view, we explore here the possible encapsulation of ultra-small-sized red and blue emitting Au NCs into liposomes of various sizes and chemical compositions. Different methods were investigated to prepare vesicles containing Au NCs in their lumen. The efficiency of the process was correlated to the structural and morphological aspect of the Au NCs' encapsulating vesicles thanks to complementary analyses by SAXS, cryo-TEM, and confocal microscopy techniques. Cell-like-sized vesicles (GUVs) encapsulating red or blue Au NCs were successfully obtained by an innovative method using emulsion phase transfer. Furthermore, exosome-like-sized vesicles (LUVs) containing Au NCs were obtained with an encapsulation yield of 40%, as estimated from ICP-MS.
    Language English
    Publishing date 2022-11-02
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2662255-5
    ISSN 2079-4991
    ISSN 2079-4991
    DOI 10.3390/nano12213875
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Luminescent Gold Nanoclusters Interacting with Synthetic and Biological Vesicles.

    Chiechio, Regina M / Ducarre, Solène / Moulin, Grégory / Dupont, Aurélien / Marets, Célia / Even-Hernandez, Pascale / Artzner, Franck / Musumeci, Paolo / Franzò, Giorgia / Ravel, Célia / LoFaro, Maria José / Marchi, Valérie

    The journal of physical chemistry letters

    2022  Volume 13, Issue 30, Page(s) 6935–6943

    Abstract: According to their high electron density and ultrasmall size, gold nanoclusters (AuNCs) have unique luminescence and photoelectrochemical properties that make them very attractive for various biomedical fields. These applications require a clear ... ...

    Abstract According to their high electron density and ultrasmall size, gold nanoclusters (AuNCs) have unique luminescence and photoelectrochemical properties that make them very attractive for various biomedical fields. These applications require a clear understanding of their interaction with biological membranes. Here we demonstrate the ability of the AuNCs as markers for lipidic bilayer structures such as synthetic liposomes and biological extracellular vesicles (EVs). The AuNCs can selectively interact with liposomes or EVs through an attractive electrostatic interaction as demonstrated by zetametry and fluorescence microscopy. According to the ratio of nanoclusters to vesicles, the lipidic membranes can be fluorescently labeled without altering their thickness until charge reversion, the AuNCs being located at the level of the phosphate headgroups. In presence of an excess of AuNCs, the vesicles tend to adhere and aggregate. The strong adsorption of AuNCs results in the formation of a lamellar phase as demonstrated by cryo-transmission electron microscopy and small-angle X-ray scattering techniques.
    MeSH term(s) Gold/chemistry ; Lipid Bilayers ; Liposomes ; Luminescence ; Metal Nanoparticles/chemistry
    Chemical Substances Lipid Bilayers ; Liposomes ; Gold (7440-57-5)
    Language English
    Publishing date 2022-07-24
    Publishing country United States
    Document type Journal Article
    ISSN 1948-7185
    ISSN (online) 1948-7185
    DOI 10.1021/acs.jpclett.2c01071
    Database MEDical Literature Analysis and Retrieval System OnLINE

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