Investigation of a direct effect of nanosecond pulse electric fields on mitochondria

Larry E. Estlack; Caleb C. Roth; Cesario Z. Cerna; Gerald J. Wilmink; Bennett L. Ibey

doi:10.1117/12.2041981

13 March 2014 Investigation of a direct effect of nanosecond pulse electric fields on mitochondria

Larry E. Estlack, Caleb C. Roth, Cesario Z. Cerna, Gerald J. Wilmink, Bennett L. Ibey

Proceedings Volume 8941, Optical Interactions with Tissue and Cells XXV; and Terahertz for Biomedical Applications; 89411S (2014) https://doi.org/10.1117/12.2041981
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

The unique cellular response to nanosecond pulsed electric field (nsPEF) exposure, as compared to longer pulse exposure, has been theorized to be due to permeabilization of intracellular organelles including the mitochondria. In this investigation, we utilized a high-throughput oxygen and pH sensing system (Seahorse® XF24 extracellular flux analyzer) to assess the mitochondrial activity of Jurkat and U937 cells after nsPEF. The XF Analyzer uses a transient micro-chamber of only a few μL in specialized cell culture micro-plates to enable oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) to be monitored in real-time. We found that for nsPEF exposures of 10 pulses at 10-ns pulse width and at 50 kV/cm e-field, we were able to cause an increase in OCR in both U937 and Jurkat cells. We also found that high pulse numbers (>100) caused a significant decrease in OCR. Higher amplitude 150 kV/cm exposures had no effect on U937 cells and yet they had a deleterious effect on Jurkat cells, matching previously published 24 hour survival data. These results suggest that the exposures were modulating metabolic activity in cells possibly due to direct effects on the mitochondria themselves. To validate this hypothesis, we isolated mitochondria from U937 cells and exposed them similarly and found no significant change in metabolic activity for any pulse number. In a final experiment, we removed calcium from the buffer solution that the cells were exposed in and found that no significant enhancement in metabolic activity was observed. These results suggest that direct permeabilization of the mitochondria is unlikely a primary effect of nsPEF exposure and calcium-mediated intracellular pathway activation is likely responsible for observed pulse-induced mitochondrial effects.

Citation Download Citation

Larry E. Estlack, Caleb C. Roth, Cesario Z. Cerna, Gerald J. Wilmink, and Bennett L. Ibey "Investigation of a direct effect of nanosecond pulse electric fields on mitochondria", Proc. SPIE 8941, Optical Interactions with Tissue and Cells XXV; and Terahertz for Biomedical Applications, 89411S (13 March 2014); https://doi.org/10.1117/12.2041981

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