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Microfluidic manufacturing of surface-functionalized graphene oxide nanoflakes for gene delivery


Graphene Oxide (GO) is a single-atomic-layered material made of a sheet of oxidized carbon atoms arranged in a honeycomb structure. Thanks to its notable physical and chemical properties of GO, the GO-based nanomaterials have application in many fields of research, including gene delivery. It has been reported that pristine GO can absorb single-stranded DNA and RNA through - stacking, which cannot be used as gene carrier because is hard to load double-stranded DNA (dsDNA). To tackle this issue, this work was aimed at developing a hybrid nanoparticle (NP) system made of GO coated by cationic lipids (hereafter referred to as GOCL) with suitable physical-chemical properties for gene delivery applications. To this end, nanosized GO flakes (nGO) were coated with the cationic lipid 1,2 -dioleoyl-3-trimethylammonium-propane (DOTAP) by microfluidic mixing. Comprehensive characterization of GOCL NPs was performed by a combination of dynamic light scattering (DLS), micro-electrophoresis and atom force microscopy (AFM). Our results show that GOCL NPs exhibit adequate size (< 150 nm) and surface charge (ξ = + 15 mV) for gene delivery purposes. Complexes made of GOCL NPs and plasmid DNA (pDNA) were used to transfect human cervical cancers cells (HeLa) and human embryonic kidney (HEK-293) cells. Pristine nGO and DOTAP cationic liposomes were used as a reference. GOCL NPs exhibited similar TE but a much higher cell viability compared with DOTAP cationic liposomes. Fluorescence confocal microscopy provided a reasonable explanation of the superior performance of GOCL/DNA complexes showing that they are much more numerous, regular in size and homogeneously distributed than DOTAP/DNA ones thus splitting their gene payload over the entire cell population. Because of the imperative demand for efficient and safe nanocarriers, this study will contribute to the development of novel surface-functionalized GO-based hybrid gene vectors.

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Publication details

The article was received on 15 Nov 2018, accepted on 06 Jan 2019 and first published on 07 Jan 2019

Article type: Paper
DOI: 10.1039/C8NR09245A
Citation: Nanoscale, 2019, Accepted Manuscript
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    Microfluidic manufacturing of surface-functionalized graphene oxide nanoflakes for gene delivery

    R. Di Santo, L. Digiacomo, S. Palchetti, V. Palmieri, G. Perini, D. Pozzi, M. Papi and G. Caracciolo, Nanoscale, 2019, Accepted Manuscript , DOI: 10.1039/C8NR09245A

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