Monitoring the efficacy of magnetofection with Au-Fe3O4 nanoparticles

• Main Author: Sim, Stanley Siong Wei
• Other Authors: Xu Chenjie
• Format: Final Year Project
• Language: English
• Published: 2015
• Subjects: DRNTU::Engineering::Bioengineering
• Online Access: http://hdl.handle.net/10356/65070
• Institution: Nanyang Technological University

One of the key biomedical applications in today’s world is genetic therapy, which is a promising technology that can be used to treat congenital and acquired diseases. However, there are not many efficient gene or drug delivery vectors around in the industry for such application. In this paper, the uniquely structured Au-Fe3O4 dumbbell nanoparticles were modified and used to test its efficacy of magnetofection, which may prove to have the potential for future applications of genetic therapy. While proving its efficacy, the Au-Fe3O4 nanoparticles are made up of both magnetic and optically active units, which have been widely used as functional probes for diagnostic and therapeutic purposes. In the experiment, the Au-Fe3O4 nanoparticles were modified by coating them with polyethylenimine (PEI), the as-synthesized polyplex were then conjugated with plasmid DNA. The binding and coating of the PEI and plasmid DNA was verified through the measurements of hydrodynamic diameter and zeta-potential using the dynamic light scattering (DLS) machine. The Au-Fe3O4/PEI/pDNA complexes then undergoes magnetofection with HEK293T cells. The in vitro results have proven that the expression of GFP gene can be boosted to up to 70% after 2 hours of magnetofection and 16 hours of transfection. The GFP gene expression was detected using a fluorescence microscope. The data collected also have shown that the Au-Fe3O4/PEI/pDNA complexes have negligible cytotoxic effect on HEK293T cells, and is a viable technology for the application of a non-viral delivery vector.