Calcium electroporation is a novel anti-cancer treatment investigated in clinical trials. We explored cell sensitivity to calcium electroporation and electroporation with bleomycin, using viability assays at different time and temperature points, as well as heat calorimetry, lipidomics, and flow cytometry. Three cell lines: HT29 (colon cancer), MDA-MB231 (breast cancer), and HDF-n (normal fibroblasts) were investigated for; (a) cell survival dependent on time of addition of drug relative to electroporation (1.2 kV/cm, 8 pulses, 99 µs, 1 Hz), at different temperatures (37 °C, 27 °C, 17 °C); (b) heat capacity profiles obtained by differential scanning calorimetry without added calcium; (c) lipid composition by mass spectrometry; (d) phosphatidylserine in the plasma membrane outer leaflet using flow cytometry. Temperature as well as time of drug administration affected treatment efficacy in HT29 and HDF-n cells, but not MDA-MB231 cells. Interestingly the HT29 cell line displayed a higher phase transition temperature (approximately 20 °C) versus 14 °C (HDF-n) and 15 °C (MDA-MB231). Furthermore the HT29 cell membranes had a higher ratio of ethers to esters, and a higher expression of phosphatidylserine in the outer leaflet. In conclusion, lipid composition and heat capacity of the membrane might influence permeabilisation of cells and thereby the effect of calcium electroporation and electrochemotherapy.

We investigated melting transitions in biological membranes in their native state that include their membrane proteins. These membranes originated from E. coli, B. subtilis, lung surfactant and nerve tissue from the spinal cord of several mammals. For some preparations, we studied the pressure, pH and ionic strength dependence of the transition. For porcine spine, we compared the transition of the native membrane to that of the extracted lipids. All preparations displayed melting transitions of 10-20 degrees below physiological or growth temperature, independent of the organism of origin and the respective cell type. The position of transitions in E. coli membranes depends on the growth temperature. We discuss these findings in the context of the thermodynamic theory of membrane fluctuations that leads to largely altered elastic constants, an increase in fluctuation lifetime and in membrane permeability associated with the transitions. We also discuss how to distinguish lipid transitions from protein unfolding transitions. Since the feature of a transition slightly below physiological temperature is conserved even when growth conditions change, we conclude that the transitions are likely to be of major biological importance for the survival and the function of the cell.

In the electromechanical theory of nerve stimulation, the nerve impulse consists of a traveling region of solid membrane in a liquid environment. Therefore, the free energy necessary to stimulate a pulse is directly related to the free energy difference necessary to induce a phase transition in the nerve membrane. It is a function of temperature and pressure, and it is sensitively dependent on the presence of anesthetics which lower melting transitions. We investigate the free energy difference of solid and liquid membrane phases under the influence of anesthetics. We calculate stimulus-response curves of electromechanical pulses and compare them to measured stimulus-response profiles in lobster and earthworm axons. We also compare them to stimulus-response experiments on human median nerve and frog sciatic nerve published in the literature.