Alexandria University, Egypt
Noha Attia has completed her MD in 2014 from Alexandria University, Egypt and Postdoctoral studies at University of the Basque Country, Spain. She is a Lecturer at Histology & Cell Biology Department, Alexandria Medical School and the Manager of Tissue Culture Lab of Alexandria Center of Excellence in Regenerative Medicine. She is the PI of research project entitled “Gene transfection of mesenchymal stem cells in 3D culture using non-viral vectors” funded by Science & Technology Development Fund, 2015.
Niosomes represent a recent promising approach for gene delivery purposes. Our group formulated a novel cationic niosome based on N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA) cationic lipid combined with polysorbate 60 and lycopene to evaluate the transfection efficiency in retinal pigment cells. Cationic niosomes prepared by emulsification-evaporation technique were mixed with the anionic pCMS-EGFP plasmid to form by electrostatic interaction nioplexes which were characterized in terms of morphology, size, surface charge and ability for DNA condensation, release and protection. In vitro studies were conducted to evaluate cell uptake, cytoplasmic distribution, transfection efficiency and viability in ARPE-19 cells. Nioplexes at 18/1 w/w ratio (cationic lipid/plasmid ratio) were 94 nm in size, +27 mV in zeta potential and exhibited imperfectly spherical morphology. At such mass ratio, niosomes condensed, released and protected the DNA from enzymatic digestion. When compared with commercial Lipofectamine 2000 lipoplexes, our nioplexes efficiently transfected ARPE-19 cells without hampering their viability. Nioplexes were quickly internalized and homogeneously distributed in the cytoplasm of cells. Furthermore, our results show that macropinocytosis, caveola-mediated endocytosis and to lesser extent clathrin-dependent endocytosis, all contribute to nioplexes uptake and transport into lysosomes. This novel niosome formulation represents a promising approach to deliver genetic material into the retina to treat inherited retinal diseases.