Lucifer Yellow uptake by CHO cells exposed to magnetic and electric pulses
Background. The cell membrane acts as a barrier that hinders free entrance of most hydrophilic molecules into the cell. Due to numerous applications in medicine, biology and biotechnology, the introduction of impermeant molecules into biological cells has drawn considerable attention in the past years. One of the most famous methods in this field is electroporation, in which electric pulses with high intensity and short duration are applied to the cells. The aim of our study was to investigate the effect of time-varying magnetic field with different parameters on transmembrane molecular transport.
Materials and methods. ‘Moreover, a comparison was made between the uptake results due to magnetic pulse exposure and electroporation mediated uptake.’ at the end of Background part. The Chinese hamster ovary (CHO) cells were exposed to magnetic pulses of 2.2 T peak strength and 250 μs duration delivered by Magstim stimulator and double 70 mm coil. Three different frequencies of 0.25, 1 and 10 Hz pulses with 112, 56 and 28 number of pulses were applied (altogether nine experimental groups) and Lucifer Yellow uptake was measured in each group. Moreover, maximum uptake of Lucifer Yellow obtained by magnetic pulses was compared to the measured uptake due to electroporation with typical parameters of 8 pulses of 100 μs, repetition frequency of 1 Hz and electric field intensities of 200 to 600 V/cm.
Results and conclusions. Our results show that time-varying magnetic field exposure increases transmembrane molecular transport and this uptake is greater for lower frequencies and larger number of pulses. Besides, the comparison shows that electroporation is more effective than pulsed magnetic field, but the observed uptake enhancement due to magnetic exposure is still considerable.
Barnes FS, Greenebaum B. Biological and medical aspects of electromagnetic fields. Handbook of biological effects of electromagnetic fields. Third edition. Boca Raton: Taylor and Francis group, CRC Press; 2006.
Cameron IL, Short NJ, Markov MS. Safe alternative cancer therapy using electromagnetic fields The Environmentalist 2007; 27: 453-6.
Pope J. Medical physics: imaging. Portsmouth: Heinemann; 1999.
Adey WR. International encyclopedia of neuroscience. Third edition. New York: Elsevier; 2003.
Neumann E, Schaefer-Ridder M, Wang Y, Hofschneider PH. Gene transfer into mouse lyoma cells by electroporation in high electric field. EMBO J 1982; 1: 841-5.
Neumann, E, Kakorin S, Tönsing K. Fundamentals of electroporative delivery of drugs and genes. Bioelectrochem Bioenerg 1999; 48: 3-16.
Teissié J, Rols MP. An experimental evaluation of the critical potential difference inducing cell membrane electropermeabilization. Biophys J 1993; 65: 409-13.
Kotnik T, Pucihar G, Rebersek M, Mir LM, Miklavcic D. Role of pulse shape in cell membrane electropermeabilization. Biochim Biophys Acta 2003; 1614: 193-200.
Valic B, Golzio M, Pavlin M, Schatz A, Faurie C, Gabriel B, et al. Effect of electric field induced transmembrane potential on spheroidal cells: theory and experiment. Eur Biophys J 2003; 32: 519-28.
Miklavcic D, Towhidi L. Numerical study of the electroporation pulse shape effect on molecular uptake of biological cells. Radiol Oncol 2010; 44: 34-41.
Faurie C, Golzio M, Phez E, Teissié J, Rols MP. Electric field induced cell membrane permeabilization and gene transfer: theory and experiments. Eng Life Sci 2005; 5: 179-86.
Teissie J, Eynard N, Vernhes MC, Bénichou A, Ganeva V, Galutzov B, et al. Recent biotechnological developments of electropulsation. A prospective review. Bioelectrochem 2002; 55: 107-12.
Sersa G, Miklavcic D, Cemazar M, Rudolf Z, Pucihar G, Snoj M. Electrochemotherapy in treatment of tumours. Eur J Surg Oncol 2008; 34: 232-40.
Mir LM. Bases and rationale of the electrochemotherapy. Eur J Cancer Suppl 2006; 4: 38-44.
Miklavčič D, Snoj M, Županič A, Kos B, Čemažar M, Kropivnik M, et al. Towards treatment planning and treatment of deep-seated solid tumors by electrochemotherapy. Biomed Eng Online 2010; 9: 10.
He J, Wang X, Guan H, Chen W, Wang M, Wu H, et al. Clinical efficacy of local targeted chemotherapy for triple-negative breast cancer. Radiol Oncol 2011; 45: 123-8.
Chiarella P, Fazio VM, Signori E. Application of electroporation in DNA vaccination protocols. Curr Gene Ther 2010; 10: 281-6.
Prud'homme GJ, Glinka Y, Khan AS, Draghia-Akli R. Electroporation-enhanced nonviral gene transfer for the prevention or treatment of immunological, endocrine and neoplastic diseases. Curr Gene Ther 2006; 6: 243-73.
Cemazar M, Golzio M, Sersa G, Rols MP, Teissié J. Electrically-assisted nucleic acids delivery to tissues in vivo: where do we stand? Curr Pharm Design 2006; 12: 3817-25.
Andre FM, Mir LM. Nucleic acids electrotransfer in vivo: mechanisms and practical aspects. Curr Gene Ther 2010; 10: 267-80.
Pavlin D, Cemazar M, Cör A, Sersa G, Pogacnik A, Tozon N. Electrogene therapy with interleukin-12 in canine mast cell tumors. Radiol Oncol 2011; 45: 30-9.
Böckmann RA, Groot BL, Kakorin S, Neumann E, Grubmüller H. Kinetics, statistics, and energetics of lipid membrane electroporation studied by molecular dynamics simulations. Biophys J 2008; 95: 1837-50.
Rosemberg Y, Korenstein R. Incorporation of macromolecules into cells and vesicles by low electric fields: induction of endocytotic-like process. Bioelectrochem Bioenerg 1997; 42: 275-81.
Antov Y, Barbul A, Korenstein R. Electroendocytosis: stimulation of adsorptive and fluid-phase uptake by pulsed low electric fields. Exp Cell Res 2004; 297: 348-62.
Antov Y, Barbul A, Mantsur H, Korenstein R. Electroendocytosis: exposure of cells to pulsed low electric fields enhances adsorption and uptake of macromolecules. Biophys J 2005; 88: 2206-22.
Mahrour N, Pologea-Moraru R, Moisescu MG, Orlowski S, Leveque P, Mir LM. In vitro increase of the fluid-phase endocytosis induced by pulsed radiofrequency electromagnetic fields: importance of the electric field component. Biochim Biophys Acta 2005; 1668: 126-37.
Marszalek P, Tsong TY. Cell fission and formation of mini cell bodies by high frequency alternating electric field. Biophys J 1995; 68: 1218-21.
Hallett M. Transcranial magnetic stimulation and the human brain. Nature 2000; 406: 147-50.
Sinclair C, Faulkner D, Hammond G. Flexible real-time control of MagStim 200(2) units for use in transcranial magnetic stimulation studies. J Neurosci Meth 2006; 158: 133-6.
Rossi S, Hallett M, Rossini P, Pascual-Leone A. Safety, ethical considerations, and application guidelines for the use of transcranial magnetic stimulation in clinical practice and research. Clin Neurophys 2009; 120: 2008-39.
Jalinous R. Technical and practical aspects of magnetic nerve stimulation. J Clin Neurophysiol 1991; 8: 10-25.
Roth y, Zangen A, Hallett M. A coil design for transcranial magnetic stimulation of deep brain regions. J Clin Neurophysiol 2002; 19: 361-70.
Ravazzani P, Ruohonen J, Grandori F, Tognola G. Magnetic stimulation of the nervous system: induced electric field in unbounded, semi-infinite, spherical, and cylindrical media. Ann Biomed Eng 1996; 24: 606-16.
Salinas FS, Lancaster JL, Fox PT. Detailed 3D models of the induced electric field of transcranial magnetic stimulation coils. Phys Med Biol 2007; 52: 2879-92.
Macek-Lebar A, Sersa G, Kranjc S, Groselj A, Miklavcic D. Optimisation of pulse parameters in vitro for in vivo electrochemotherapy. Anticancer Res 2002; 22: 1731-6.
Towhidi L, Kotnik T, Pucihar G, Firoozabadi SMP, Mozdarani H, Miklavcic D. Variability of the minimal transmembrane voltage resulting in detectable membrane electroporation. Electromagn Biol Med 2008; 27: 372-85.
Marjanovič I, Haberl S, Miklavčič D, Kandušer M, Pavlin M. Analysis and comparison of electrical pulse parameters for gene electrotransfer of two different cell lines. J Membrane Bio 2010; 236: 97-105.
Usaj M, Torkar D, Kanduser Mm Miklavcic D. Cell counting tool parameters optimization approach for electroporation efficiency determination of attached cells in phase contrast images. J Microscopy 2011; 241: 303-14.
Stewart WW. Functional connexions between cells as revealed by dye coupling with a highly fluorescent naphthalamide tracer. Cell 1978; 14: 741-59.
Stewart WW. Lucifer dyes highly fluorescent dyes for biological tracing. Nature 1981; 292: 17-21.
Puc M, Kotnik T, Mir LM, Miklavčič D. Quantitative model of small molecules uptake after in vitro cell electropermeabilization. Bioelectrochemistry 2003; 60: 1-10.
Pucihar G, Kotnik T, Kanduser M, Miklavcic D. The influence of medium conductivity on electropermeabilization and survival of cells in vitro. Bioelectrochemistry 2001; 54: 107-15.
Kotnik T, Pucihar G, Miklavčič D. Induced transmembrane voltage and its correlation with electroporation-mediated molecular transport. J Membrane Biol 2010; 236: 3-13.
Pucihar G, Krmelj J, Reberšek M, Batista Napotnik T, Miklavčič D. Equivalent pulse parameters for electroporation. IEEE T Biomed Eng 2011; 58: 3279-88.
Rols MP, Femenia P, Teissié J. Long-lived macropinocytosis takes place in electropermeabilized mammalian cells. Biochem Biophys Res Commun 1995; 208: 26-38.
Zimmermann U, Schnettler R, Klöck G, Watzka H, Donath E, Glaser RW. Mechanisms of electrostimulated uptake of macromolecules into living cells. Naturwissenschaften 1990; 77: 543-5.
Glogauer M, Lee W, McCulloch CA. Induced endocytosis in human fibroblasts by electrical fields. Exp Cell Res 1993; 208: 232-40.
Escoffre JM, Dean DS, Hubert M. Rols MP, Favard C. Membrane perturbation by an external electric field: a mechanism to permit molecular uptake. Eur Biophys J 2007; 36: 973-83.
Timothy EV, Weaver JC. Molecular change due to biomagnetic stimulation and transient magnetic fields: mechanical interference constraints on possible effects by cell membrane pore creation via magnetic particles. Bioelectrochem Bioenerg 1998; 46: 121-8.