The influence of electroporation on the Photofrin uptake and distribution was evaluated in the breast adenocarcinoma cells (MCF-7) and normal Chinese hamster ovary cells (CHO) lacking voltage-dependent channels in vitro. Photofrin was used at a concentration of 5 and 25 μM. The uptake of Photofrin was assessed using flow cytometry and fluorescence microscopy methods. Cells viability was evaluated with crystal violet assay. Our results indicated that electropermeabilization of cells, in the presence of Photofrin, increased the uptake of the photosensitizer. Even at the lowest electric field intensity (700 V/cm) Photofrin transport was enhanced. Flow cytometry results for MCF-7 cells revealed ~1.7 times stronger fluorescence emission intensity for cells exposed to Photofrin and electric field of 700 V/cm than cells treated with Photofrin alone. Photofrin was effective only when irradiated with blue light. Our studies on combination of photodynamic reaction with electroporation suggested improved effectiveness of the treatment and showed intracellular distribution of Photofrin. This approach may be attractive for cancer treatment as enhanced cellular uptake of Photofrin in MCF-7 cells can help to reduce effective dose of the photosensitizer and exposure time in this type of cancer, diminishing side effects of the therapy.

No abstract

Single metastable nanopores, appearing before the actual electroporation under constant-current conditions, are used to characterize the onset of electroporation. Unlike the long-lived electropores typical of the current controlled methods, these pores survive for milliseconds and observing them is possible due to slow development of electroporation, provided by the gradual accumulation of charges on a planar membrane. Analysis of the metastable pore appearance frequency and lifetime shows the first introductory stage of electroporation. During this stage two species of metastable pores open, the majority of very low conductance that seem not fully developed as hydrophilic electropores. The experiments reveal that voltage value defines the electroporation onset while the current value affects the rate of electroporation. Membrane capacitance has a great impact on the membrane susceptibility to the pore appearance, related to its thickness and integrity. Pores of nonperfect membranes appear more easily, but they do not live any longer than others.