Journal
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
Volume 1861, Issue 11, Pages 2802-2810Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbagen.2017.07.025
Keywords
Magnetic nanoparticles; Transfection; DNA chemisorption; Mesenchymal stem cell; Green fluorescent protein (GFP); Fluorescent nanoparticles
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Funding
- Italian Institutional Ministry [60A06-7411, 60A06-8055]
- University of Padua (Italy), grant PRAT (progetti di Ateneo) [CPDA159850, CPDR148959]
- CARIPARO Foundation
- Ministry of Education, Youth and Sports of the Czech Republic [LO1305, LM2015073]
- Internal Student Grant of Palacky University in Olomouc, Czech Republic [IGA_PrF_2015_017]
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Background: Conversely to common coated iron oxide nanoparticles, novel naked surface active maghemite nanoparticles (SAMNs) can covalently bind DNA. Plasmid (pDNA) harboring the coding gene for GFP was directly chemisorbed onto SAMNs, leading to a novel DNA nanovector (SAMN@pDNA). The spontaneous internalization of SAMN@pDNA into cells was compared with an extensively studied fluorescent SAMN derivative (SAMN@RITC). Moreover, the transfection efficiency of SAMN@pDNA was evaluated and explained by computational model. Methods: SANIN@pDNA was prepared and characterized by spectroscopic and computational methods, and molecular dynamic simulation. The size and hydrodynamic properties of SAMN@pDNA and SAMN@RITC were studied by electron transmission microscopy, light scattering and zeta-potential. The two nanomaterials were tested by confocal scanning microscopy on equine peripheral blood-derived mesenchymal stem cells (ePB-MSCs) and GFP expression by SAMN@pDNA was determined. Results: Nanomaterials characterized by similar hydrodynamic properties were successfully internalized and stored into mesenchymal stem cells. Transfection by SAMN@pDNA occurred and GFP expression was higher than lipofectamine procedure, even in the absence of an external magnetic field. A computational model clarified that transfection efficiency can be ascribed to DNA availability inside cells. Conclusions: Direct covalent binding of DNA on naked magnetic nanoparticles led to an extremely robust gene delivery tool. Hydrodynamic and chemical-physical properties of SAMN@pDNA were responsible of the successful uptake by cells and of the efficiency of GFP gene transfection.
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