Bystander effect of metal byproducts released from electroporated cells after electroporation in vitro

Electrodes dissolution during electroporation releases metal ions into the medium, altering the microenvironment of electroporated cells and allowing metal ions to penetrate cell membrane. During cell membrane repair, homeostasis restoration or activation of cell death pathways, cells eliminate excess metals from the cytoplasm and membrane. This study assessed the effects of post-electroporation metal byproducts on untreated (non-electroporated) cells in vitro. CHO and HCT116 cells were electroporated with three pulse protocols (unipolar: 100 μs, 5 ms; bipolar: 2 μs) using either aluminum or stainless-steel electrodes. After electroporation, cells were transferred to fresh growth medium and incubated for 2 or 4 h. Incubation period allowed either cell recovery or the activation of cell death pathways, leading to the accumulation of metal byproducts in the incubation medium. Stainless-steel electrodes with the 5 ms pulse protocol caused a considerable increase in iron, chromium and nickel ions in incubation medium compared to aluminum electrodes or other protocols. Metal ions in incubation medium caused toxicity in non-electroporated cells, disrupting cell cycle function or inducing cell death. The observed toxicity results from combined effects of metal ions on cellular functions and the mechanisms the cells use to protect themselves from metal overload.