高压纳秒脉冲电场对同源细胞的杀伤差异性研究

doi:10.19595/j.cnki.1000-6753.tces.231473

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Abstract In terms of biological effects caused by the pulsed electric fields, the duration of the nanosecond pulse is less than the charging time constant of the cell membrane (~1 μs), which has more profound effects on the intracellular organelles, known as the intracellular electron-manipulation. Based on this, the nanosecond pulsed electric fields (nsPEFs) have unique bioelectromagnetic properties in the field of tumor ablation. Therefore, the killing effects of homologous cells under the action of nsPEFs are studied to investigate the difference between homologous cells and high-voltage nsPEFs.
The homologous cells, human immortal keratinocytes (HaCaT), human melanoma cells (A375), and human melanoma chemoresistant cells (A375/R) are chosen as research objects. The homemade pulse generator can output high-voltage nanosecond pulses with flexible and adjustable parameters. The selected pulse parameters are 5 kV/cm to 15 kV/cm field intensity, 300 ns pulse width, 1 Hz repetition frequency, and 80 pulse number. Cell viability assay based on CCK-8 is used to study the survival rate of homologous cells after pulse treatment. The ablation effect of nanosecond pulses is further studied based on hydrogel-simulated tissue. The lethal threshold for a specific ablation area is calculated. Furthermore, the electroporation response of the single cell to the nanosecond pulse is simulated by the finite element simulation model.
The survival rate of the three cells decreases with the increase of the electric field intensity of nsPEFs. The cell viability assay shows that with the same nsPEFs parameters, the survival rate of normal cells is the highest, followed by tumor cells, and tumor chemoresistant cells have the lowest survival rate. The results of the hydrogel simulated tissue ablation experiment show that with the same pulse parameters, the ablation area of tumor chemoresistant cells is the largest, reaching (20.28±0.85) mm², and the ablation area of normal cells is the smallest, only (10.26±0.49) mm². Meanwhile, the lethal threshold of normal and chemoresistant cells is (7.15±0.65) kV/cm and (4.75±0.15) kV/cm, respectively, with a difference of approximately 2.4 kV/cm. Significant differences exist in the sensitivity of normal, tumor, and tumor chemoresistant cells to the high- voltage nanosecond pulsed electric fields. The nsPEFs have the least effect on normal cells and the strongest effect on tumor chemoresistant cells.
To analyze the potential reasons for the killing difference of cells by nsPEFs, the cell size and nucleus size of HaCaT, A375, and A375/R cells are measured and recorded by fluorescence microscopy. The differences between HaCaT, A375, and A375/R are significant in the nucleus size. Based on the real cell geometry, the electroporation area and the pore number in the nuclear membrane are simulated. The results suggest that the difference in cell response to nsPEFs may be related to the nucleus size. Hence, tumor chemoresistant cells with larger nuclei are more susceptible to nsPEFs. High-voltage nanosecond pulsed electric fields have potential clinical applications in tumor ablation without affecting surrounding normal tissues.

Key words： Nanosecond pulsed electric fields homologous cells cell viability hydrogel scaffolds killing difference

Received: 06 September 2023

PACS:	TM836
	Q64

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	Peng Wencheng
	Tang Xiao
	Liu Hongmei
	Dong Shoulong
	Yao Chenguo

Cite this article:

Peng Wencheng,Tang Xiao,Liu Hongmei等. Study on the Killing Difference of Homologous Cells by High-Voltage Nanosecond Pulsed Electric Fields[J]. Transactions of China Electrotechnical Society, 2024, 39(20): 6544-6552.

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https://dgjsxb.ces-transaction.com/EN/10.19595/j.cnki.1000-6753.tces.231473 OR https://dgjsxb.ces-transaction.com/EN/Y2024/V39/I20/6544

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