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PULSED FIELD ABLATION INDUCES HEMOLYSIS IN VITRO DEPENDING ON THE STRENGTH OF THE ELECTRIC FIELD
Topic: Heart rhythm disorders
Type: Presentation - doctors , Number in the programme: 10
Enlisted in: Young investigator awards
Fišerová I. 1, Fišer O. 2, Novák M. 2, Bačová B. 3, Kvapil D. 4, Kapáková A. 5, Trnka J. 4, Hozman M. 6, Heřman D. 6, Gibalová A. 5, Vrba J. 2, Vrba D. 2, Osmančík P. 7

1 Ústav biochemie, buněčné a molekulární biologie, 3. lékařská fakulta, UK, Praha, 2 Katedra biomedicínské techniky, Fakulta biomedicínského inženýrství ČVUT, Kladno, 3 Department of Immunology and Clinical Biochemistry, 3rd Faculty of Medicine, Charles University, Prague 10, 4 Ústav biochemie, buněčné a molekulární biologie, 3. lékařská fakulta UK, Prague, 5 Department of Biochemistry, Cell and Molecular Biology, 3rd Faculty of Medicine, Charles University, Prague 10, 6 Department of Cardiology, 3rd Faculty of Medicine, Charles University, Prague 10, 7 III. interní klinika, Fakultní nemocnice Královské Vinohrady, Prague


Aim:
Several cases of renal failure due to intravascular hemolysis after pulsed field ablation (PFA) has been observed. Red blood cell (RBC) disintegration and increased haptoglobin concentration have also been observed in human patients undergoing PFA. The aim of this study was to investigate the hemolytic effect on plasma samples from healthy volunteers using short high voltage electric pulses with different electric field strength.
Methods:
Anticoagulated blood samples from healthy volunteers were obtained according to the European Union regulation. Blood samples were exposed to different electric field strengths generated by an electric pulse generator (TONAGENA, CZ). The electroporation set-up used a burst consisting of 216 bipolar pulses lasting 2 μs. Each burst was repeated 20 times with a frequency 1 Hz. Applied electric fields ranged from 250 V/cm to 1500 V/cm. Hemolysis was quantified by cell-free hemoglobin concentration and by the number of red blood cell microparticles (RBCμ) from separated plasma.
Results:
All electric fields strengths induce increase of RBCμ. The increase in the number of RBCμ increased exponentially (r2 = 0.96) with linearly increasing electric field. Significant increase in cell-free hemoglobin (0.32 6 0.16 g/L, 2.2 6 0.96 g/L, and 5.7 6 0.39 g/L vs control group – 0.08 g/l; p < 0.01) was observed from electric field strength 1000 V/cm.
Conclusion:
High-voltage electrical impulses cause damage to RBC, leading to haemolysis. A significant increase in cell-free hemoglobin was observed from an electrical filed strength of 1000 V/cm. Higher levels of RBCμ were observed at all electric field strengths (250 - 1500 V/cm). This result may suggest that RBC damage can occur at very low electric field strengths and detection of RBCμ may provide a more sensitive marker of RBC damage than detection of cell-free hemoglobin.