Spatio-temporal dynamics of calcium electrotransfer during cell membrane permeabilization

Abstract

Pulsed electric fields (PEFs) are applied as physical stimuli for DNA/drug delivery, cancer therapy, gene transformation, and microorganism eradication. Meanwhile, calcium electrotransfer offers an interesting approach to treat cancer, as it induces cell death easier in malignant cells than in normal cells. Here, we study the spatial and temporal cellular responses to 10 μs duration PEFs; by observing real-time, the uptake of extracellular calcium through the cell membrane. The experimental setup consisted of an inverted fluorescence microscope equipped with a color high-speed framing camera and a specifically designed miniaturized pulsed power system. The setup allowed us to accurately observe the permeabilization of HeLa S3 cells during application of various levels of PEFs ranging from 0.27 to 1.80 kV/cm. The low electric field experiments confirmed the threshold value of transmembrane potential (TMP). The high electric field observations enabled us to retrieve the entire spatial variation of the permeabilization angle (θ). The temporal observations proved that after a minimal permeabilization of the cell membrane, the ionic diffusion was the prevailing mechanism of the delivery to the cell cytoplasm. The observations suggest 0.45 kV/cm and 100 pulses at 1 kHz as an optimal condition to achieve full calcium concentration in the cell cytoplasm. The results offer precise levels of electric fields to control release of the extracellular calcium to the cell cytoplasm for inducing minimally invasive cancer calcium electroporation, an interesting affordable method to treat cancer patients with minimum side effects.

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Abbreviations

PEFs:

Pulsed electric fields

TMP:

Transmembrane potential

MEM:

Minimum essential medium

PBS:

Phosphate buffer saline

EDTA:

Ethylene diamine tetra acetic acid

FBS:

Fetal bovine serum

HBSS:

Hank’s balanced salt solution

MOSFET:

Metal oxide semiconductor field effect transistor

VDCC:

Voltage-dependent calcium channels

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Acknowledgements

The authors would like to thank Ms. M. Ota, Mr. R. Matsushima, and Mr. Mr. N. Ohnishi for their help in conducting the experiments.

Funding

This work was supported in part by Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (17K06163).

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Correspondence to Hamid Hosseini.

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Guionet, A., Moosavi Nejad, S., Teissié, J. et al. Spatio-temporal dynamics of calcium electrotransfer during cell membrane permeabilization. Drug Deliv. and Transl. Res. 8, 1152–1161 (2018). https://doi.org/10.1007/s13346-018-0533-5

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Keywords

  • Calcium electrotransfer
  • Pulsed electric fields
  • Permeabilization angle
  • Transmembrane potential
  • HeLa S3 cells