The theranostic ability of a new fluorescently labeled 17 cationic cyclodextrin−graphene nanoplatform (GCD@Ada-Rhod) was 18 investigated by studying its intracellular trafficking and its ability to 19 deliver plasmid DNA and microRNA. The nanoplatform was 20 synthesized by both covalent and supramolecular approaches, and its 21 chemical structure, morphology, and colloidal behavior were 22 investigated by TGA, TEM, spectroscopic analysis such as UV−vis, 23 fluorescence emission, DLS, and ζ-potential measurements. The 24 cellular internalization of GCD@Ada-Rhod and its perinuclear 25 localization were assessed by FLIM, Raman imaging, and fluorescence 26 microscopy. Biological experiments with pCMS-EGFP and miRNA-15a 27 evidenced the excellent capability of GCD@Ada-Rhod to deliver both pDNA and microRNA without significant cytotoxicity. 28 The biological results evidenced an unforeseen caveolae-mediated endocytosis internalization pathway (generally expected for 29 particles <200 nm), despite the fact that the GCD@Ada-Rhod size is about 400 nm (by DLS and TEM data). We supposed that 30 the internalization pathway was driven by physical−chemical features of GCD@Ada-Rhod, and the caveolae-mediated uptake 31 enhanced the transfection efficiency, avoiding the lysosomal acid degradation. The cellular effects of internalized miRNA-15a on 32 the oncogene protein BCL-2 were investigated at two different concentrations (N/P = 10 and 5), and a reduction of the BCL-2 33 level was detected at a low concentration (i.e., N/P = 10). miRNA-15a is considered an ideal cancer therapy molecule due to its 34 activity on multiple transcription factors, and the elucidation of the correlation between the concentration of delivered miRNA35 15a and the down-/up-regulation of the BCL-2 level, documented for the first time in this work, could be an important 36 contribution to guide its clinical application.
|Titolo:||Casting Light on Intracellular Tracking of a New Functional Graphene-Based MicroRNA Delivery System by FLIM and Raman Imaging|
|Data di pubblicazione:||2019|
|Appare nelle tipologie:||1.1 Articolo in rivista|