Non-invasive technique for Cell Membrane Permeabilization using pulsed electromagnetic fields (2018-2021) |
| Head: | Prof. Damijan Miklavčič, University of Ljubljana, Faculty of Electrical Engineering |
| Partner: | / |
| Funding: | Slovenian Research Agency (ARRS), Slovenia |
| Code: | J2-9225 |
The project is funded by the Slovenian Research Agency (ARRS).
- Member of University of Ljubljana
- Code
- J2-9225
- Project
- Non-invasive technique for Cell Membrane Permeabilization using pulsed electromagnetic fields
- Period
- 1.7.2018 - 30.6.2021
- Amount of financing
- 1.3 FTE
- Head
- Research activity
- Engineering sciences and technologies - Systems and cybernetics
- Research Organisation
- Abstract
An exposure of a cell to an electric field of an adequate strength and duration leads to a transient increase of cell membrane permeability. This phenomenon, termed electroporation, allows various otherwise nonpermeant molecules to cross the membrane and enter the cell. Both in vitro and in vivo, electroporation allows for internalization of a wide range of substances, including chemotherapeutics and DNA. All electroporation applications require direct contact between the electrodes and the treated object, that is either via plate electrodes, which embrace the tissue, or using invasive needle electrodes, which are inserted into the tissue. The use of invasive electrodes, such as needle electrodes, which are most effective in electroporation treatment of a variety of tissues have a number of drawbacks common to all invasive procedures, e.g., assuring sterile incisions and causing trauma to tissues by incision. There are additional side effects due to application of electric pulses.
Recently, a new cell membrane permeabilization method by pulsed electromagnetic fields has been proposed. Pulsed electromagnetic fields induced increase of the cell membrane permeability is similar to electroporation with the important difference of non-invasive establishment of electric field by exposing a treated tissue to a time-varying magnetic field. Currently, however, the electric field that is induced by the time-varying magnetic field is lower (by several orders of magnitude) than the electric field causing membrane permeabilization in conventional electroporation experiments, resulting in inferior efficacy of the pulsed electromagnetic fields treatment to conventional electroporation. Still, the proof of concept has been confirmed both in vitro and in vivo for the pulsed electromagnetic fields mediated transport of small molecules and large functional molecules.
The non-straightforward dependence of the pulsed electromagnetic fields induced permeabilization on the treatment parameters requires further research of different parameters of applied electromagnetic field. The proposed project is the first attempt to systematically explore the phenomenon of cell membrane permeabilization using magnetic pulses through in vitro and in vivo experimentation combined with theoretical and numerical analysis. In the project, we will explore a range of the magnetic pulse parameters, develop new magnetic pulse applicators and determine possibility of different molecules being loaded into cells by means of pulsed electromagnetic fields.
In the project, we will first determine both magnetic and electric field established during application of pulsed electromagnetic fields in cell suspensions and in tissues. Determination will be based on a numerical model which we will build from the geometry of the applicator and treated object, i.e. either cell suspensions or tissues. Then, we will focus on the development of new geometries of magnetic pulse applicators. We will evaluate different geometrical arrangement of coils for the applicators used in in vitro experiments as well for in vivo. New geometries will be developed through numerical modelling and most the appropriate coils will be built and used in experiments in the next stages of the project. We will start by performing experiments in vitro by evaluating the cell membrane permeabilization of cell suspensions treated with pulsed electromagnetic fields delivered by newly developed magnetic pulse applicator. Using magnetic pulse parameters, that will enable highest permeabilization of cells, we will perform the delivery of cytotoxic compound and nucleic acids into cells. In the last part of the project, we will analyze and improve antitumor effectiveness of pulsed electromagnetic fields as a drug delivery system for cytotoxic compounds to murine subcutaneous tumors and explore the feasibility of pulsed electromagnetic fields in gene electrotransfer for small and large molecules in tissues in vivo.- Researchers
- The phases of the project and their realization
The aim of the project is to systematically explore the phenomenon of membrane permeabilization using pulsed electromagnetic fields through in vitro (WP 1) and in vivo (WP 2) experimentation combined with theoretical and numerical analysis (WP 3). Since the phenomenon of non-invasive membrane permeabilization is still not very known, we will take special care to disseminate achieved results of the project to the scientific community through different communication channels (WP 4).
In Work Package 1 (WP 1), we will systematically explore the phenomenon of cell membrane permeabilization using pulsed electromagnetic fields through in vitro experimentation. We will start by performing experiments on cell suspension mixed with fluorescent dyes that are commonly used for detection of cell permeabilization. We will evaluate the cell membrane permeabilization of cell suspensions treated with pulsed electromagnetic fields and determine the magnetic pulse parameters that will enable highest permeabilization of cells and survival. Using determined magnetic pulse parameters, we will perform the delivery of cytotoxic compound and nucleic acids into cells.
In Work Package 2 (WP 2), we will apply new and improved applicator that will be determined in WP 3 for drug and gene delivery in vivo using pulsed electromagnetic fields and magnetic pulses parameters that enabled most effective cell membrane permeabilization (determined in WP 1). First, we will analyze and improve antitumor effectiveness of pulsed electromagnetic fields as a drug delivery system for cytotoxic compounds to murine subcutaneous tumors using new applicator. Mouse bearing the tumor will be treated with intravenously injection of cisplatin, pulsed electromagnetic fields or with the combination of both therapies. We will evaluate antitumor effectiveness of treatments by tumor growth delay assay and by measuring cytotoxic compound uptake in tumors. In the second part of WP 2, we will explore the feasibility of pulsed electromagnetic fields in gene electrotransfer for small and large molecules in tissues in vivo.
In Work Package 3 (WP 3), we will first determine both magnetic and electric field established during application of pulsed electromagnetic fields in cell suspensions from WP 1 and in tissues from WP 2. Determination will be based on a numerical model which we will build from the geometry of the applicator and treated object, i.e. either cell suspensions or tissues. We will also perform validation of the model by comparison of calculated values with measurements. In the second part of WP 3, we will focus on the development of new geometries of magnetic pulse applicators. We will evaluate different geometrical arrangement of coils for the applicators used in in vitro experiments as well for in vivo. New geometries will be developed through numerical modelling and most the appropriate coils will be built and applied in experiments in WP 1 and WP 2.
In Work Package 4 (WP 4), we will protect intellectual property and disseminate achieved results of the project to interested scientific community. Since the phenomenon of non-invasive cell membrane permeabilization is still not very known, we will take special care to use various communication channels, such as scientific journals and magazines, conferences and congresses. In order to exchange knowledge and expertise related to non-invasive technique for cell permeabilization, we will prepare a training course in the scope of the international scientific workshop and postgraduate course Electroporation based Technologies and Treatment.
- Citations for bibliographic records
Novickij V, Kranjc M, Staigvila G, Dermol-Černe J, Meleško J, Novickij J, Miklavčič D. High pulsed electromagnetic field generator for contactless permeabilization of cells in vitro.
IEEE Trans. Magn.: 1-6, 2020. 2020 IEEE DOI 10.1109/TMAG.2020.2979120Miklavčič D, Novickij V, Kranjc M, Polajžer T, Haberl Meglič S, Batista Napotnik T, Romih R, Lisjak D. Contactless electroporation induced by high intensity pulsed electromagnetic fields via distributed nanoelectrodes.
Bioelectrochemistry 132: 1-9, 2019. 2019 The Authors DOI 10.1016/j.bioelechem.2019.107440Kranjc M, Kranjc Brezar S, Serša G, Miklavčič D. Contactless delivery of plasmid encoding EGFP in vivo by high-intensity pulsed electromagnetic field.
Bioelectrochemistry 141: 107847, 2021. 2021 The Authors DOI 10.1016/j.bioelechem.2021.10784Kranjc Brezar S, Kranjc M, Čemažar M, Buček S, Serša G, Miklavčič D. Electrotransfer of siRNA to silence enhanced green fluorescent protein in tumor mediated by a high intensity pulsed electromagnetic field.
Vaccines 2020, 8: 1-15, 2020. 2020 The Authors DOI 10.3390/vaccines8010049Hu Q, Joshi RP, Miklavcic D. Calculations of cell transmembrane voltage induced by time-varying magnetic fields.
IEEE Trans. Plasma Sci.: 1-8, 2020. 2020 IEEE DOI 10.1109/TPS.2020.2975421