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Vishal R. Chauhan, Suber S. Huang; The Use of Magnetic Nanoparticles to Enhance Ocular Gene Delivery. Invest. Ophthalmol. Vis. Sci. 2011;52(14):1419.
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© ARVO (1962-2015); The Authors (2016-present)
To study the efficiency of gene transfer using magnetic nanoparticles in ARPE-19 cells.
Green-fluorescence protein (GFP) plasmids were associated with paramagnetic nanoparticles (purchased from Chemicell). Gene delivery was then targeted to cultured human diploid retinal pigment epithelial (ARPE-19) cells by application of a magnetic field. Cells were imaged using fluorescence microscopy to assay for GFP expression. Magnetic nanoparticles were then tagged with red fluorescence and delivered to cultured ARPE-19 cells with application of a magnetic field. Uptake of particles into cells was detected using fluorescence microscopy. This procedure was repeated using Polyethylenamine (PEI)-GFP nanoparticles complexes as a standard control. Transfection efficiencies were compared using fluorescence microscopy.
Micrographs quantitating transfection efficiency of ARPE-19 cells using GFP-magnetic nanoparticle complexes will be presented and demonstrate significantly higher integration than that of PEI-RPE nanoparticles. In addition, fluorescently tagged magnetic nanoparticles showed nearly diffuse uptake by ARPE-19 cells. Finally, transfection of ARPE-19 cells using GFP-magnetic nanoparticle complexes required only 15 minutes of incubation time while exposed to magnetic field in order to achieve optimal gene delivery. In contrast, PEI-GFP nanoparticle complexes required 4 hours (as per standard protocol) of incubation time to achieve optimal delivery.
The use of magnetic nanoparticles as vehicles for gene delivery in ARPE19 cells leads to high levels of nanoparticle uptake, increased transfection efficiencies and short incubation times. All of these characteristics may make magnetic nanoparticles particularly suitable to ocular gene therapy.
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