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Synthesis and Characterization of Dextran Based Magnetic Nanogels as a Gene Delivery Vector and Investigating its Gene Therapy Efficiency

Azadpour, Behnam | 2021

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 54639 (07)
  4. University: Sharif University of Technology
  5. Department: Materials Science and Engineering
  6. Advisor(s): Madaah Hosseini, Hamid Reza; Arefian, Ehsan
  7. Abstract:
  8. The use of magnetic nanoparticles modified with proper surface agent and the ability of controlling by magnetic field, that can induce colloidal stability, is considered as a vector to transfer DNA plasmid, or pDNA in short, in the field of gene therapy. In thiss research, pH-responsive dextran-based magnetic nanogels (dextMNGs) were synthesized via inverse mini-emulsion method. Fourier transformation infrared spectroscopy (FTIR) showed that magnetite nanoparticles (MNPs) were successfully modified with arginine and had amine terminals. FTIR, also, proved that aldehyded dextran was crosslinked by arginine modified magnetite nanoparticles (RMNPs) via pH sensitive imine bonds. X-ray diffraction (XRD) showed that synthesized nanoparticles were magnetite with inverse spinel crystalline structure, and with 4 nanometers (nm) crystallite size. Hydrodynamic mean particle size of 20 nm and 112 nm were measured for RMNPs and dextMNGs via dynamic light scattering (DLS) respectively. Functionalization of magnetite with arginine and crosslinking of dextran via RMNPs were also proved with thermal gravimetry (TGA), which showed more weight loss for dextMNGs, related to dextran, compared to RMNPs that has the weight loss around 220˚C to 600˚C relates to arginine. Field emission scanning electron micrsocopy (FESEM) showed that particles have spherical morphology and mean particle size of 28 nm. FESEM also proved the pH-sensitivity of dextMNGs via investigating them in different pH. Saturation magnetization for RMNPs and dextMNGs were measured 32 emu/g and 26 emu/g respectively via vibrating sample magnetometer (VSM). Superparamagnesim property of RMNPs and dextMNGs was also proved by VSM. Gel retardation essay showed the limited ability of dextMNGs to uptake and capture of pDNA and magnetofection complex formation. Inverted fluorescence microscopy reealed that dextMNGs could transfect HEK293T cells in high dextMNGs:pDNA ratios; although the number of transfected cells was limited. It is clear that the investigated platform has the ability of transfecting HEK293T cells and innovative; however it is necessary to investigate its surface properties such as electrical charge, functional groups, and surface ligands to optimize the magnetofection
  9. Keywords:
  10. Superparamagnetic Iron Oxide Nanoparticle ; Magnetite Nanoparticle ; Magnetofection Method ; Arginine ; Dextrances ; Inverse Mini-Emulsion ; Magnetic Nanogels ; Gene Therapy

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