Fiserova Ivana, Fiser Ondrej, Novak Marek, Trnka Jan, Gibalova Antonia, Kvapil David, Bacova Barbora, Hozman Marek, Herman Dalibor, Benesova Klara, Osmancik Pavel
Department of Biochemistry, Cell and Molecular Biology, Charles University, Prague, Czech Republic.
Department of Biomedical Technology, Faculty of Biomedical Engineering, Czech Technical University in Prague, Prague, Czech Republic.
Heart Rhythm. 2025 Feb;22(2):466-474. doi: 10.1016/j.hrthm.2024.08.019. Epub 2024 Aug 13.
Pulsed field ablation (PFA) of atrial fibrillation is a new method in clinical practice. Despite a favorable safety profile of PFA in atrial fibrillation ablation, rare cases of renal failure, probably due to hemolysis, have recently been reported.
The aim of this study was to determine the rate of hemolysis and cardiac cell death during in vitro PFA with different electric field intensities.
Blood samples from healthy volunteers and mouse HL-1 cardiomyocyte cell lines were subjected to in vitro irreversible electroporation using 216 bipolar pulses, each lasting 2 μs with intervals of 5 μs, repeated 20 times at a frequency of 1 Hz. These pulses varied from 500 V to 1500 V. Cell-free hemoglobin levels were assessed spectrophotometrically, and red blood cell microparticles were evaluated by flow cytometry. Cardiomyocyte death was quantified with propidium iodide.
Pulsed field energy (1000 V/cm, 1250 V/cm, and 1500 V/cm) was associated with a significant increase in cell-free hemoglobin (0.32 ± 0.16 g/L, 2.2 ± 0.96 g/L, and 5.7 ± 0.39 g/L; P < .01) and similar increase in the concentration of red blood cell microparticles. Significant rates of cardiomyocyte death were observed at electric field strengths of 750 V/cm, 1000 V/cm, 1250 V/cm, and 1500 V/cm (26.5% ± 5.9%, 44.3% ± 6.2%, 55.5% ± 6.9%, and 74.5% ± 17.8% of cardiomyocytes; P < .01).
The most effective induction of cell death in vitro was observed at 1500 V/cm. This intensity was also associated with a significant degree of hemolysis.
心房颤动的脉冲场消融术（PFA）是临床实践中的一种新方法。尽管PFA在心房颤动消融术中具有良好的安全性，但最近有报道称，罕见的肾衰竭病例可能是由于溶血所致。
本研究旨在确定不同电场强度下体外PFA期间的溶血率和心肌细胞死亡率。
从健康志愿者采集的血样和小鼠HL-1心肌细胞系，使用216个双相脉冲进行体外不可逆电穿孔，每个脉冲持续2微秒，间隔5微秒，以1赫兹的频率重复20次。这些脉冲的电压从500伏到1500伏不等。通过分光光度法评估无细胞血红蛋白水平，通过流式细胞术评估红细胞微粒。用碘化丙啶对心肌细胞死亡进行定量。
脉冲场能量（1000伏/厘米、1250伏/厘米和1500伏/厘米）与无细胞血红蛋白显著增加（分别为0.32±0.16克/升、2.2±0.96克/升和5.7±0.39克/升；P<.01）以及红细胞微粒浓度的类似增加相关。在750伏/厘米、1000伏/厘米、1250伏/厘米和1500伏/厘米的电场强度下观察到显著的心肌细胞死亡率（分别为心肌细胞的26.5%±5.9%、44.3%±6.2%、55.5%±6.9%和74.5%±17.8%；P<.01）。
在体外，1500伏/厘米时观察到最有效的细胞死亡诱导。该强度也与显著程度的溶血相关。
