We are first to develop 3D numerical models for optimization of gene electrotransfer in vivo. It is the first study that presents parametrization and optimization of pulse amplitudes and electrode positions (depth of insertion, inter-electrode distance, orientation) for gene electrotransfer in skeletal muscle. We calculate volumes of reversibly and irreversibly electroporated tissue. Our finite-element model combined with genetic algorithms is generic and can be applied to other tissues. The results can be used as a guideline for researchers in selecting optimal parameters for gene therapy.
COBISS.SI-ID: 8218708
We demonstrated for the first time that electrophoresis is key for the efficient electrogene transfection in cases where plasmid DNA concentrations are relatively low (e.g. in vivo) and that use of low DNA concentrations allows better transfer of conclusions from in vitro to in vivo conditions. We also first used LV+HV and HV+LV pulsing protocols and by this demonstrated that electrophoresis is crucial for insertion into permeabilized cell membrane. Presented results are important for understanding of the mechanisms of electrotransfer and for improvement of the protocols.
COBISS.SI-ID: 6679380
We performed a series of in vitro studies where we showed that there is no direct correlation between increase of Mg2+ ions and transfection efficiency. This is in contrast with some other studies and can be explained by the effect of Mg2+ on stability of DNA in the cytoplasm and its binding to the cell membrane. We also found increased permeabilization of the cells with increased Mg2+ ions, which shows that the mechanisms of transfer for electrotransfection are different from transfer of small molecules.
COBISS.SI-ID: 7666260
We describe ion and molecular diffusion during electroporation and show that the process can be explained with two populations of pores.We show that the existing theoretical models, which describe formation of transient pores in the membrane, cannot describe long-lived pores that are key for molecular transport and successful electrogene transfection. We calculate the fraction of the stable pores with respect to the electric field and number of pulses and show that larger number of pulses is vital for the pore stabilization which is in agreement with experiments of electrotransfer.
COBISS.SI-ID: 6443604
We are first to analyse gene electrotransfer in cells embedded in collagen gels, which represent a complex system similar to realistic environment. 3-D collagen models are very important since they enable us to to anaylse different protocols. We obtained that also in such 3-D model is electropermeabilization crucial for electrotransfer. We obtained that longer several ms pulses are needed for efficient electrotransfer , while in vitro also short pulses are efficient.
COBISS.SI-ID: 7803220