Cellular uptake pathways of sepiolite nanofibers and DNA transfection improvement.
Fiche publication
Date publication
juillet 2017
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Dr PIETREMENT Olivier
Tous les auteurs :
Castro-Smirnov FA, Ayache J, Bertrand JR, Dardillac E, Le Cam E, Piétrement O, Aranda P, Ruiz-Hitzky E, Lopez BS
Lien Pubmed
Résumé
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.
Mots clés
Animals, CHO Cells, Cell Line, Cell Separation, Chloroquine, pharmacology, Cricetulus, DNA, chemistry, Drug Carriers, chemistry, Endocytosis, Flow Cytometry, Humans, Magnesium Silicates, chemistry, Microscopy, Electron, Transmission, Microscopy, Fluorescence, Time-Lapse Imaging, Transfection, methods
Référence
Sci Rep. 2017 07 17;7(1):5586