Cellular uptake pathways of sepiolite nanofibers and DNA transfection improvement
| Autor: | Castro-Smirnov, Fidel Antonio, Ayache, Jeanne, Bertrand, Jean-Rémi, Dardillac, Elodie, Le Cam, Eric, Piétrement, Olivier, Aranda, Pilar, Ruiz-Hitzky, Eduardo, Lopez, Bernard S. |
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| Přispěvatelé: | Institut National du Cancer (France), Ministerio de Economía y Competitividad (España), Agence Nationale de la Recherche (France), Stabilité Génétique et Oncogenèse (UMR 8200), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Signalisation, noyaux et innovations en cancérologie (UMR8126), Vectorologie et thérapeutiques anti-cancéreuses [Villejuif] (UMR 8203), Institut Gustave Roussy (IGR), Instituto de Ciencia de Materiales de Madrid (ICMM), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC) |
| Rok vydání: | 2017 |
| Předmět: |
Drug Carriers
[SDV]Life Sciences [q-bio] Science Chloroquine CHO Cells Cell Separation DNA Flow Cytometry Transfection Time-Lapse Imaging Article Endocytosis Cell Line Cricetulus Microscopy Electron Transmission Microscopy Fluorescence Magnesium Silicates Animals Humans Medicine ComputingMilieux_MISCELLANEOUS |
| Zdroj: | Scientific Reports, Vol 7, Iss 1, Pp 1-10 (2017) Digital.CSIC. Repositorio Institucional del CSIC instname Scientific Reports Scientific Reports, Nature Publishing Group, 2017, 7 (1), ⟨10.1038/s41598-017-05839-3⟩ |
| ISSN: | 2012-3175 2045-2322 |
| DOI: | 10.1038/s41598-017-05839-3⟩ |
| Popis: | Sepiolite is a nanofibrous natural silicate that can be used as a nanocarrier because it can be naturally internalized into mammalian cells, due to its nano-size dimension. Therefore, deciphering the mechanisms of sepiolite cell internalization constitutes a question interesting biotechnology, for the use of sepiolite as nanocarrier, as well as environmental and public health concerns. Though it is low, the perfectly stable and natural intrinsic fluorescence of sepiolite nanofibers allows to follow their fate into cells by specifically sensitive technics. By combining fluorescence microscopy (including confocal analysis), time-lapse video microscopy, fluorescence activated cell sorting and transmission electron microscopy, we show that sepiolite can be spontaneously internalized into mammalian cells through both non-endocytic and endocytic pathways, macropinocytosis being one of the main pathways. Interestingly, exposure of the cells to endocytosis inhibitors, such as chloroquine, two-fold increase the efficiency of sepiolite-mediated gene transfer, in addition to the 100-fold increased resulting from sepiolite sonomechanical treatment. As sepiolite is able to bind various biological molecules, this nanoparticulate silicate could be a good candidate as a nanocarrier for simultaneous vectorization of diverse biological molecules. This work was supported by La Ligue Nationale Contre le Cancer, ANR (Agence Nationale de la Recherche, ANR-14-CE10-0010-02, ANR-16-CE18-0012-02), AFM-Téléthon and INCa (Institut National du Cancer, 2013-1-PLBIO-14), MINECO (projects MAT2012-31759 and MAT2015-71117-R), Spain. |
| Databáze: | OpenAIRE |
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