Numerical simulation of molecular uptake via electroporation.

A numerical study of electroporation-mediated molecular delivery is presented. The model consists of the Nernst-Planck equations for species transport, coupled with an asymptotic Smoluchowski equation for membrane permeabilization. The transfer of calcium ions into a Chinese Hamster Ovary cell is simulated. The results reveal important physical insights. First, for this particular case, ion electrophoresis plays an important role, and is an order of magnitude faster than free diffusion on a comparable time scale. Second, the maximum achievable concentration within the cell is reciprocally correlated with the extracellular electrical conductivity. This behavior is mediated by an electrokinetic mechanism known as field-amplified sample stacking. Through this mechanism, the intracellular ion concentration can reach a level higher than the extracellular one provided that the intra-to-extracellular conductivity ratio is greater than unity. The results corroborate well with data in the literature, and offer a mechanistic interpretation to previous experimental observations. This work is a step toward the quantification of molecular delivery via electroporation.

[1]  H. Itoh,et al.  Electroporation of cell membrane visualized under a pulsed-laser fluorescence microscope. , 1988, Biophysical journal.

[2]  J Teissié,et al.  Time courses of mammalian cell electropermeabilization observed by millisecond imaging of membrane property changes during the pulse. , 1999, Biophysical journal.

[3]  K. Schoenbach,et al.  High electrical field effects on cell membranes. , 2007, Bioelectrochemistry.

[4]  Ravindra P. Joshi,et al.  Dynamical modeling of cellular response to short-duration, high-intensity electric fields , 2003 .

[5]  J. Weaver,et al.  Transport lattice approach to describing cell electroporation: use of a local asymptotic model , 2004, IEEE Transactions on Plasma Science.

[6]  M. Bureau,et al.  Importance of association between permeabilization and electrophoretic forces for intramuscular DNA electrotransfer. , 2000, Biochimica et biophysica acta.

[7]  S W Hui,et al.  Local and transient structural changes in stratum corneum at high electric fields: contribution of Joule heating. , 2005, Bioelectrochemistry.

[8]  Muriel Golzio,et al.  Time dependence of electric field effects on cell membranes. A review for a critical selection of pulse duration for therapeutical applications , 2008 .

[9]  E Neumann,et al.  Control by pulse parameters of electric field-mediated gene transfer in mammalian cells. , 1994, Biophysical journal.

[10]  Hao Lin,et al.  The current-voltage relation for electropores with conductivity gradients. , 2010, Biomicrofluidics.

[11]  Damijan Miklavcic,et al.  Kinetics of transmembrane transport of small molecules into electropermeabilized cells. , 2008, Biophysical journal.

[12]  K. H. Schoenbach,et al.  Effects of submicrosecond, high intensity pulsed electric fields on living cells - intracellular electromanipulation , 2003 .

[13]  J. Weaver,et al.  Theory of electroporation: A review , 1996 .

[14]  E. Neumann,et al.  Electroporation of subcutaneous mouse tumors by rectangular and trapezium high voltage pulses. , 2004, Bioelectrochemistry.

[15]  Ring-Ling Chien,et al.  Field amplified sample injection in high-performance capillary electrophoresis , 1991 .

[16]  Véronique Préat,et al.  Transdermal Delivery of Fentanyl by Electroporation I. Influence of Electrical Factors , 1996, Pharmaceutical Research.

[17]  Justin Teissié,et al.  Fluorescence imaging in the millisecond time range of membrane electropermeabilisation of single cells using a rapid ultra-low-light intensifying detection system , 1998, European Biophysics Journal.

[18]  Bipartite expressions for diffusional mass transport in biomembranes. , 2006, Biophysical journal.

[19]  J. Weaver,et al.  The Spatially Distributed Dynamic Transmembrane Voltage of Cells and Organelles due to 10 ns Pulses: Meshed Transport Networks , 2006, IEEE Transactions on Plasma Science.

[20]  Yu Zhang,et al.  Zeta potential: a surface electrical characteristic to probe the interaction of nanoparticles with normal and cancer human breast epithelial cells , 2008, Biomedical microdevices.

[21]  U. Pliquett,et al.  Mechanism for the conductivity changes caused by membrane electroporation of CHO cell-pellets , 2004 .

[22]  K. Schoenbach,et al.  Self-consistent simulations of electroporation dynamics in biological cells subjected to ultrashort electrical pulses. , 2001, Physical review. E, Statistical, nonlinear, and soft matter physics.

[23]  Efficiency of the delivery of small charged molecules into cells in vitro. , 2010, Bioelectrochemistry.

[24]  W Krassowska,et al.  Modeling electroporation in a single cell. II. Effects Of ionic concentrations. , 1999, Biophysical journal.

[25]  E. Neumann,et al.  Electroporation and Electrofusion in Cell Biology , 1989, Springer US.

[26]  J. Teissié,et al.  Osmotically induced membrane tension facilitates the triggering of living cell electropermeabilization. , 2004, Bioelectrochemistry.

[27]  W Krassowska,et al.  Theoretical modeling of the effects of shock duration, frequency, and strength on the degree of electroporation. , 2000, Bioelectrochemistry.

[28]  R. Bharadwaj,et al.  Dynamics of field-amplified sample stacking , 2005, Journal of Fluid Mechanics.

[29]  Damijan Miklavcic,et al.  Quantitative model of small molecules uptake after in vitro cell electropermeabilization. , 2003, Bioelectrochemistry.

[30]  James C Weaver,et al.  Active mechanisms are needed to describe cell responses to submicrosecond, megavolt-per-meter pulses: cell models for ultrashort pulses. , 2008, Biophysical journal.

[31]  F. Yuan,et al.  Mechanistic Analysis of Electroporation-Induced Cellular Uptake of Macromolecules , 2008, Experimental biology and medicine.

[32]  L. Chernomordik,et al.  Electroporation and electrophoretic DNA transfer into cells. The effect of DNA interaction with electropores. , 1992, Biophysical journal.

[33]  L. Chernomordik,et al.  Electrically induced DNA uptake by cells is a fast process involving DNA electrophoresis. , 1991, Biophysical journal.

[34]  J Teissié,et al.  Electropermeabilization of mammalian cells to macromolecules: control by pulse duration. , 1998, Biophysical journal.

[35]  W. Krassowska,et al.  Modeling electroporation in a single cell. , 2007, Biophysical journal.

[36]  D. Miklavčič,et al.  Determination of the lipid bilayer breakdown voltage by means of linear rising signal. , 2007, Bioelectrochemistry.

[37]  W. Krassowska,et al.  Modeling electroporation in a single cell. I. Effects Of field strength and rest potential. , 1999, Biophysical journal.

[38]  M. Rols,et al.  Direct visualization at the single-cell level of electrically mediated gene delivery , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[39]  U. Zimmermann,et al.  Reversible Electropermeabilization of Mammalian Cells by High-Intensity, Ultra-Short Pulses of Submicrosecond Duration , 2001, The Journal of Membrane Biology.

[40]  Véronique Préat,et al.  Skin electroporation for transdermal and topical delivery. , 2004, Advanced drug delivery reviews.

[41]  U. Zimmermann,et al.  Effect of medium conductivity and composition on the uptake of propidium iodide into electropermeabilized myeloma cells. , 1996, Biochimica et biophysica acta.

[42]  Mounir Tarek,et al.  Membrane electroporation: a molecular dynamics simulation. , 2005, Biophysical journal.

[43]  Wanda Krassowska,et al.  Asymptotic model of electroporation , 1999 .

[44]  K. Schoenbach,et al.  Simulations of electroporation dynamics and shape deformations in biological cells subjected to high voltage pulses , 2002 .

[45]  P. Thomas Vernier,et al.  Life Cycle of an Electropore: Field-Dependent and Field-Independent Steps in Pore Creation and Annihilation , 2010, The Journal of Membrane Biology.

[46]  K. Schoenbach,et al.  Electroporation dynamics in biological cells subjected to ultrafast electrical pulses: a numerical simulation study. , 2000, Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics.

[47]  W. Briels,et al.  Free energy of a trans-membrane pore calculated from atomistic molecular dynamics simulations. , 2006, The Journal of chemical physics.

[48]  I. Vattulainen,et al.  Pore formation coupled to ion transport through lipid membranes as induced by transmembrane ionic charge imbalance: atomistic molecular dynamics study. , 2005, Journal of the American Chemical Society.

[49]  Boris Rubinsky,et al.  Mass Transfer Model for Drug Delivery in Tissue Cells with Reversible Electroporation. , 2008, International journal of heat and mass transfer.

[50]  Edward L Cussler,et al.  Diffusion: Mass Transfer in Fluid Systems , 1984 .

[51]  L. Mir,et al.  Efficient DNA electrotransfer into tumors. , 2000, Bioelectrochemistry.

[52]  A. T. Esser,et al.  Towards Solid Tumor Treatment by Irreversible Electroporation: Intrinsic Redistribution of Fields and Currents in Tissue , 2007, Technology in cancer research & treatment.

[53]  I. Lamprecht,et al.  Dielectric properties of yeast cells as determined by electrorotation. , 1992, Biochimica et biophysica acta.

[54]  B. Lindholm-Sethson Supported Lipid Membranes for Reconstitution of Membrane Proteins , 2001 .

[55]  L. Mir,et al.  Therapeutic perspectives of in vivo cell electropermeabilization. , 2001, Bioelectrochemistry.

[56]  U. Zimmermann,et al.  Measurement of the permeability and resealing time constant of the electroporated mammalian cell membranes , 2004 .

[57]  U. Zimmermann,et al.  Electrotransfection of anchorage-dependent mammalian cells. , 2003, Experimental cell research.

[58]  P. Århem,et al.  Role of individual surface charges of voltage-gated K channels. , 1999, Biophysical journal.

[59]  Karl H. Schoenbach,et al.  Stimulation of Capacitative Calcium Entry in HL-60 Cells by Nanosecond Pulsed Electric Fields* , 2004, Journal of Biological Chemistry.

[60]  G. T. Martin,et al.  Theoretical analysis of localized heating in human skin subjected to high voltage pulses. , 2002, Bioelectrochemistry.

[61]  E. Neumann,et al.  Gene transfer into mouse lyoma cells by electroporation in high electric fields. , 1982, The EMBO journal.

[62]  M. Rols,et al.  Mechanisms of cell membrane electropermeabilization: a minireview of our present (lack of ?) knowledge. , 2005, Biochimica et biophysica acta.

[63]  W. Krassowska,et al.  Modeling postshock evolution of large electropores. , 2003, Physical review. E, Statistical, nonlinear, and soft matter physics.

[64]  James C Weaver,et al.  Electrical behavior and pore accumulation in a multicellular model for conventional and supra-electroporation. , 2006, Biochemical and biophysical research communications.

[65]  R. O. Price,et al.  Plasma membrane voltage changes during nanosecond pulsed electric field exposure. , 2006, Biophysical journal.

[66]  J. Gehl,et al.  Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research. , 2003, Acta physiologica Scandinavica.

[67]  J. Keizer,et al.  A simple numerical model of calcium spark formation and detection in cardiac myocytes. , 1998, Biophysical journal.

[68]  Damijan Miklavcic,et al.  Numerical study of the electroporation pulse shape effect on molecular uptake of biological cells , 2010, Radiology and oncology.

[69]  Juergen F Kolb,et al.  Selective field effects on intracellular vacuoles and vesicle membranes with nanosecond electric pulses. , 2005, Biophysical journal.

[70]  A. T. Esser,et al.  Microdosimetry for conventional and supra-electroporation in cells with organelles. , 2006, Biochemical and biophysical research communications.

[71]  M. Golzio,et al.  Control by membrane order of voltage-induced permeabilization, loading and gene transfer in mammalian cells. , 2001, Bioelectrochemistry.

[72]  H. Itoh,et al.  Membrane conductance of an electroporated cell analyzed by submicrosecond imaging of transmembrane potential. , 1991, Biophysical journal.

[73]  V. Préat,et al.  Mechanisms of a Phosphorothioate Oligonucleotide Delivery by Skin Electroporation , 1998 .

[74]  Marie-Pierre Rols,et al.  What is (Still not) Known of the Mechanism by Which Electroporation Mediates Gene Transfer and Expression in Cells and Tissues , 2009, Molecular biotechnology.

[75]  Wanda Krassowska,et al.  Model of creation and evolution of stable electropores for DNA delivery. , 2004, Biophysical journal.

[76]  Sadhana Talele,et al.  Non-linear time domain model of electropermeabilization: Response of a single cell to an arbitrary applied electric field , 2007 .

[77]  D Peter Tieleman,et al.  BMC Biochemistry BioMed Central Research article The molecular basis of electroporation , 2004 .

[78]  Siewert J Marrink,et al.  Molecular dynamics simulations of hydrophilic pores in lipid bilayers. , 2004, Biophysical journal.

[79]  T. Reese,et al.  Changes in membrane structure induced by electroporation as revealed by rapid-freezing electron microscopy. , 1990, Biophysical journal.

[80]  James C Weaver,et al.  Model of a confined spherical cell in uniform and heterogeneous applied electric fields. , 2006, Bioelectrochemistry.

[81]  V. F. Pastushenko,et al.  247 - Electric breakdown of bilayer lipid membranes II. Calculation of the membrane lifetime in the steady-state diffusion approximation , 1979 .

[82]  V. Préat,et al.  Transdermal Delivery of Metoprolol by Electroporation , 1994, Pharmaceutical Research.

[83]  James C. Weaver,et al.  Electroporation: a unified, quantitative theory of reversible electrical breakdown and mechanical rupture in artificial planar bilayer membranes☆ , 1991 .

[84]  A. T. Esser,et al.  Membrane electroporation: The absolute rate equation and nanosecond time scale pore creation. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[85]  W. Webb,et al.  Optical imaging of cell membrane potential changes induced by applied electric fields. , 1986, Biophysical journal.

[86]  J. Weaver,et al.  Electroporation of mammalian skin: a mechanism to enhance transdermal drug delivery. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[87]  J. Weaver,et al.  Theory of electroporation of planar bilayer membranes: predictions of the aqueous area, change in capacitance, and pore-pore separation. , 1994, Biophysical journal.