Influence of the Time-Phase Varied Magnetic Field on the Nucleic Acids Delivery Into the Cancer Cells. Summary of the Doctoral Thesis

Abstract

Gene delivery is one of the critical steps in gene therapy, including both efficient delivery of the therapeutic gene to the target cells and attachment to the cell membrane with following internalization and transfer to the nucleus. Magnetofection is one of the most effective physical gene delivery methods based on the increased delivery and concentration of nucleic acids coupling superparamagnetic iron oxide particles (SPION) onto the cell surface by the influence of an external magnetic field. In the static magnetic field generated by permanent magnets the movement of SPION in solution is axial towards the magnet. The time-phase varied magnetic field which is generated with the magnetofection device DynaFECTOR is based on the orbital rotation of the permanent magnets plate in the parallel plane of the cell culture plate. In the result of computer modelling it was shown that the orbital rotation of magnets causes the axial-lateral movement of SPION in parallel and perpendicular plane against the magnet. This led to the more uniform distribution of SPION onto the surface and could be a cause of the enhanced SPION-nucleic acids complexes internalization. In this study the influence of the time-phase varied magnetic field on the delivery efficiency of SPION-nucleic acids-liposomal component complexes into the cancer cells was experimentally investigated. Obtained results showed that in the time-phase varied magnetic field the SPION sedimentation onto the surface is more uniform and the amount of internalized SPION increases when compared to static magnetic field. Under the influence of the time-phase varied magnetic field, the delivery of SPION-nucleic acids-liposomal component complexes also increases, as evidenced by the significant increase in both the number of transfected cells and the total amount of an expressed protein in transfected cells in comparison with other gene delivery methods. While SPION-nucleic acids-liposomal component complexes delivery into the cells with less cytotoxic effect in provided by more uniform sedimentation onto the surface. In this study enhanced effective gene delivery method – the liposomal magnetofection in the time-phase varied magnetic field is reported. The method could be used for the replacement of mutated genes and for the investigation of the inhibition effect of novel therapeutic siRNAs in vitro cell lines. The method could potentially used also for the therapeutic gene delivery into the cancer cells in vivo.