Permeability Changes of Cationic Liposomes Loaded with Carbonic Anhydrase Induced by Millimeter Waves Radiation

The interaction of millimeter wave radiation, in the 30-300 GHz range, with biological systems is a topic of great interest as many of the vibrational dynamics that occur in biochemical reactions of large macromolecules in living organisms fall in the 1-100 GHz range. Membranes and cellular organelles may have different ways of interacting with this radiation as well. In this article, we investigate the influence of 53.37 GHz of radiation on lipid membrane permeability by using cationic liposomes that contain dipalmitoylphosphatidylcholine (DPPC), cholesterol and stearylamine. Carbonic anhydrase (CA) is loaded inside the liposome and the substrate pnitrophenyl acetate (p-NPA) is added in the bulk aqueous phase. Upon permeation across the lipid bilayer, the trapped CA catalyzes the conversion of the p-NPA molecules into products. Because the self-diffusion rate of p-NPA across intact liposomes is very low, the CA reaction rate expressed as DA/min is used to track membrane permeability changes. A highly significant (P , 0.0001) enhancement of the CA reaction rate, typically from DA/min ¼ 0.0043 6 0.0017 (n ¼ 26) to DA/min ¼ 0.0100 6 0.0020 (n ¼ 32) resulted at a lowlevel density power of 0.1 mW/cm2. The enhancement of the CA reaction rate was observed at a lesser extent on liposomes with a larger diameter and, in turn with leaflets less bent. The different packing of the phospholipid bilayer--due to the higher curvature--could be a critical factor in eliciting membrane permeability changes indicating a possible role for water molecules bound to functional groups in the glycerol region. Since numerical dosimetry indicates that the temperature rise during the exposure was negligible, the observed effects cannot be attributed to heating of the samples