Electroporation Mechanisms: The Role of Lipid Orientation in the Kinetics of Pore Formation.

Electroporation is a well-established technique used to stimulate cells, enhancing membrane permeability. Although the biological phenomena occurring after the poration process have been widely studied, the physical mechanisms of pore formation are not clearly understood. In this work we investigated by means of molecular dynamics simulations the kinetics of pore formation, linking the different stages of poration to specific arrangements of lipid membrane domains.Clinical Relevance-The approach followed in this study aims to shed light on the molecular mechanisms at the basis of the electroporation technique, nowadays used to enhance the entrance of poorly permeant anticancer drugs into tumor cells, for gene electrotransfer and all the other applications exploiting the modulation of cell membrane properties.

Geometrical Characterization of an Electropore from Water Positional Fluctuations.

We present here a new method for calculating the radius of a transmembrane pore in a phospholipid bilayer. To compare size-related properties of pores in bilayers of various compositions, generated and maintained under different physical and chemical conditions, reference metrics are needed. Operational metrics can be associated with some observed behavior. For example, pore size can be defined by the largest object that will pass through the length of the pore. The novelty of the present approach resides in the characterization of electropore geometry via a statistical approach, based on essential dynamics rules. We define the pore size geometrically with an algorithm for determining the pore radius. In particular, we extract the radius from the tri-dimensional surface of a defined pore region. The method is applied to a pore formed in a phospholipid bilayer by application of an external electric field. Although the details described here are specific for lipid pores in molecular dynamics simulations, the method can be generalized for any kind of pores for which appropriate structural information is available.

Effects of nanosecond pulsed electric fields on the activity of a Hodgkin and Huxley neuron model.

The cell membrane poration is one of the main assessed biological effects of nanosecond pulsed electric fields (nsPEF). This structural change of the cell membrane appears soon after the pulse delivery and lasts for a time period long enough to modify the electrical activity of excitable membranes in neurons. Inserting such a phenomenon in a Hodgkin and Huxley neuron model by means of an enhanced time varying conductance resulted in the temporary inhibition of the action potential generation. The inhibition time is a function of the level of poration, the pore resealing time and the background stimulation level of the neuron. Such results suggest that the neuronal activity may be efficiently modulated by the delivery of repeated pulses. This opens the way to the use of nsPEFs as a stimulation technique alternative to the conventional direct electric stimulation for medical applications such as chronic pain treatment.

Signal transduction on enzymes: the effect of electromagnetic field stimuli on superoxide dismutase (SOD).

Protein functions and characteristics can highly differ from physiological conditions in presence of chemical, mechanical or electromagnetic stimuli. In this work we provide a rigorous picture of electric field effects on proteins behavior investigating, at atomistic details, the possible ways in which an external signal can be transduced into biochemical effects. Results from molecular dynamics (MD) simulations of a single superoxidismutase (SOD) enzyme in presence of high exogenous alternate electric fields will be discussed.