The Accumulation and Effects of Liposomal Doxorubicin in Tissues Treated by Radiofrequency Ablation and Irreversible Electroporation in Liver: In Vivo Experimental Study on Porcine Models

  • Tomáš Andrašina
  • Josef Jaroš
  • Tomáš JůzaEmail author
  • Tomáš Rohan
  • Dalibor Červinka
  • Michal Crha
  • Vlastimil Válek
  • Nahum S. Goldberg
Laboratory Investigation



To compare the accumulation and effect of liposomal doxorubicin in liver tissue treated by radiofrequency ablation (RFA) and irreversible electroporation (IRE) in in vivo porcine models.

Materials and Methods

Sixteen RFA and 16 IRE procedures were performed in healthy liver of two groups of three pigs. Multi-tined RFA parameters included: 100 W, target temperature 105°C for 7 min. 100 IRE pulses were delivered using two monopolar electrodes at 2250 V, 1 Hz, for 100 µsec. For each group, two pigs received 50 mg liposomal doxorubicin (0.5 mg/kg) as a drip infusion during ablation procedure, with one pig serving as control. Samples were harvested from the central and peripheral zones of the ablation at 24 and 72 h. Immunohistochemical analysis to evaluate the degree of cellular stress, DNA damage, and degree of apoptosis was performed. These and the ablation sizes were compared. Doxorubicin concentrations were also analyzed using fluorescence photometry of homogenized tissue.


RFA treatment zones created with concomitant administration of doxorubicin at 24 h were significantly larger than controls (2.5 ± 0.3 cm vs. 2.2 ± 0.2 cm; p = 0.04). By contrast, IRE treatment zones were negatively influenced by chemotherapy (2.2 ± 0.4 cm vs. 2.6 ± 0.4 cm; p = 0.05). At 24 h, doxorubicin concentrations in peripheral and central zones of RFA were significantly increased in comparison with untreated parenchyma (0.431 ± 0.078 µg/g and 0.314 ± 0.055 µg/g vs. 0.18 ± 0.012 µg/g; p < 0.05). Doxorubicin concentrations in IRE zones were not significantly different from untreated liver (0.191 ± 0.049 µg/g and 0.210 ± 0.049 µg/g vs. 0.18 ± 0.012 µg/g).


Whereas there is an increased accumulation of periprocedural doxorubicin and an associated increase in ablation zone following RFA, a contrary effect is noted with IRE. These discrepant findings suggest that different mechanisms and synergies will need to be considered in order to select optimal adjuvants for different classes of ablation devices.


Liposomal doxorubicin Radiofrequency ablation Irreversible electroporation 



This study was supported by Ministry of Health of the Czech Republic, Grant No. 15-32484a, and supported by funds from the Faculty of Medicine MU to junior researcher Tomáš Andrašina.

Compliance with Ethical Standards

Conflict of interest

Author Nahum S. Goldberg is a consultant to Angiodynamics, Inc. and Cosman Medical Inc.

Ethical Approval

All applicable international, national, and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Informed Consent

For this type of study, informed consent is not required.

Consent for Publication

For this type of study, consent for publication is not required.


  1. 1.
    Yu H, Burke CT. Comparison of percutaneous ablation technologies in the treatment of malignant liver tumors. Semin Interv Radiol. 2014;31:129–37. Scholar
  2. 2.
    Ahmed M, Brace CL, Lee FT, Goldberg SN. Principles of and advances in percutaneous ablation. Radiology. 2011;258:351–69. Scholar
  3. 3.
    van Amerongen MJ, Jenniskens SFM, van den Boezem PB, Fütterer JJ, de Wilt JHW. Radiofrequency ablation compared to surgical resection for curative treatment of patients with colorectal liver metastases—a meta-analysis. HPB. 2017;19:749–56. Scholar
  4. 4.
    Lei JY, Wang WT, Yan LN, Wen TF, Li B. Radiofrequency ablation versus surgical resection for small unifocal hepatocellular carcinomas. Medicine (Baltimore). 2014. Scholar
  5. 5.
    Wang X, Sofocleous CT, Erinjeri JP, Petre EN, Gonen M, Do KG, Brown KT, Covey AM, Brody LA, Alago W, Thornton RH, Kemeny NE, Solomon SB. Margin size is an independent predictor of local tumor progression after ablation of colon cancer liver metastases. Cardiovasc Intervent Radiol. 2013;36:166–75. Scholar
  6. 6.
    Goldberg SN, Kamel IR, Kruskal JB, Reynolds K, Monsky WL, Stuart KE, Ahmed M, Raptopoulos V. Radiofrequency ablation of hepatic tumors: increased tumor destruction with adjuvant liposomal doxorubicin therapy. Am J Roentgenol. 2002;179:93–101. Scholar
  7. 7.
    Tang C, Shen J, Feng W, Bao Y, Dong X, Dai Y, Zheng Y, Zhang J. Combination therapy of radiofrequency ablation and transarterial chemoembolization for unresectable hepatocellular carcinoma: a retrospective study. Medicine (Baltimore). 2016;95:e3754. Scholar
  8. 8.
    Tanaka T, Isfort P, Braunschweig T, Westphal S, Woitok A, Penzkofer T, Bruners P, Kichikawa K, Schmitz-Rode T, Mahnken AH. Superselective particle embolization enhances efficacy of radiofrequency ablation: effects of particle size and sequence of action. Cardiovasc Intervent Radiol. 2013;36:773–82. Scholar
  9. 9.
    Yamakado K, Inaba Y, Sato Y, Yasumoto T, Hayashi S, Yamanaka T, Nobata K, Takaki H, Nakatsuka A. Radiofrequency ablation combined with hepatic arterial chemoembolization using degradable starch microsphere mixed with mitomycin c for the treatment of liver metastasis from colorectal cancer: a prospective multicenter study. Cardiovasc Intervent Radiol. 2017;40:560–7. Scholar
  10. 10.
    Mross K, Niemann B, Massing U, Drevs J, Unger C, Bhamra R, Swenson CE. Pharmacokinetics of liposomal doxorubicin (TLC-D99; Myocet) in patients with solid tumors: an open-label, single-dose study. Cancer Chemother Pharmacol. 2004;54:514–24. Scholar
  11. 11.
    Andriyanov AV, Koren E, Barenholz Y, Goldberg SN. Therapeutic efficacy of combining pegylated liposomal doxorubicin and radiofrequency (rf) ablation: comparison between slow-drug-releasing, non-thermosensitive and fast-drug-releasing thermosensitive nano-liposomes. PLoS ONE. 2014. Scholar
  12. 12.
    Goldberg SN, Girnan GD, Lukyanov AN, Ahmed M, Monsky WL, Gazelle GS, Huertas JC, Stuart KE, Jacobs T, Torchillin VP, Halpern EF, Kruskal JB. Percutaneous tumor ablation: increased necrosis with combined radio-frequency ablation and intravenous liposomal doxorubicin in a rat breast tumor model. Radiology. 2002;222:797–804. Scholar
  13. 13.
    Ahmed M, Moussa M, Goldberg SN. Synergy in cancer treatment between liposomal chemotherapeutics and thermal ablation. Chem Phys Lipids. 2012;165:424–37. Scholar
  14. 14.
    Monsky WL, Kruskal JB, Lukyanov AN, Girnun GD, Ahmed M, Gazelle GS, Huertas JC, Stuart KE, Torchilin VP, Goldberg SN. Radio-frequency ablation increases intratumoral liposomal doxorubicin accumulation in a rat breast tumor model. Radiology. 2002;224:823–9. Scholar
  15. 15.
    Lyu T, Wang X, Su Z, Shangguan J, Sun C, Figini M, Wang J, Yaghmai V, Larson AC, Zhang Z. Irreversible electroporation in primary and metastatic hepatic malignancies. Medicine (Baltimore). 2017. Scholar
  16. 16.
    Deipolyi AR, Golberg A, Yarmush ML, Arellano RS, Oklu R. Irreversible electroporation: evolution of a laboratory technique in interventional oncology. Diagn Interv Radiol. 2014;20:147–54. Scholar
  17. 17.
    Miklavčič D, Serša G, Brecelj E, Gehl J, Soden D, Bianchi G, Ruggieri P, Rossi CR, Campana LG, Jarm T. Electrochemotherapy: technological advancements for efficient electroporation-based treatment of internal tumors. Med Biol Eng Comput. 2012;50:1213–25. Scholar
  18. 18.
    Gehl J, Skovsgaard T, Mir LM. Enhancement of cytotoxicity by electropermeabilization: an improved method for screening drugs. Anticancer Drugs. 1998;9:319–25.CrossRefGoogle Scholar
  19. 19.
    Ben-David E, Appelbaum L, Sosna J, Nissenbaum I, Goldberg SN. Characterization of irreversible electroporation ablation in in vivo porcine liver. Am J Roentgenol. 2012;198:W62–8. Scholar
  20. 20.
    Appelbaum L, Ben-David E, Faroja M, Nissenbaum Y, Sosna J, Goldberg SN. Irreversible electroporation ablation: creation of large-volume ablation zones in in vivo porcine liver with four-electrode arrays. Radiology. 2013;270:416–24. Scholar
  21. 21.
    Rubinsky B, Onik G, Mikus P. Irreversible electroporation: a new ablation modality–clinical implications. Technol Cancer Res Treat. 2007;6:37–48. Scholar
  22. 22.
    Ahmed M, Liu Z, Lukyanov AN, Signoretti S, Horkan C, Monsky WL, Torchilin VP, Goldberg SN. Combination radiofrequency ablation with intratumoral liposomal doxorubicin: effect on drug accumulation and coagulation in multiple tissues and tumor types in animals. Radiology. 2005;235:469–77. Scholar
  23. 23.
    Kruskal JB, Oliver B, Huertas JC, Goldberg SN. Dynamic intrahepatic flow and cellular alterations during radiofrequency ablation of liver tissue in mice. J Vasc Interv Radiol JVIR. 2001;12:1193–201.CrossRefGoogle Scholar
  24. 24.
    Moussa M, Goldberg SN, Tasawwar B, Sawant RR, Levchenko T, Kumar G, Torchilin VP, Ahmed M. Adjuvant liposomal doxorubicin markedly affects radiofrequency (RF) ablation-induced effects on periablational microvasculature. J Vasc Interv Radiol JVIR. 2013;24:1021–33. Scholar
  25. 25.
    Bulvik BE, Rozenblum N, Gourevich S, Ahmed M, Andriyanov AV, Galun E, Goldberg SN. Irreversible electroporation versus radiofrequency ablation: a comparison of local and systemic effects in a small-animal model. Radiology. 2016;280:413–24. Scholar
  26. 26.
    Neal RE, Rossmeisl JH, D’Alfonso V, Robertson JL, Garcia PA, Elankumaran S, Davalos RV. In Vitro and numerical support for combinatorial irreversible electroporation and electrochemotherapy glioma treatment. Ann Biomed Eng. 2014;42:475–87. Scholar
  27. 27.
    Mir LM, Orlowski S. Mechanisms of electrochemotherapy. Adv Drug Deliv Rev. 1999;35:107–18. Scholar
  28. 28.
    Speelmans G, Staffhorst RWHM, Steenbergen HG, de Kruijff B. Transport of the anti-cancer drug doxorubicin across cytoplasmic membranes and membranes composed of phospholipids derived from Escherichia coli occurs via a similar mechanism. Biochim Biophys Acta BBA Biomembr. 1996;1284:240–6. Scholar
  29. 29.
    O’Shaughnessy JA. Pegylated liposomal doxorubicin in the treatment of breast cancer. Clin Breast Cancer. 2003;4:318–28.CrossRefGoogle Scholar
  30. 30.
    Rafiyath SM, Rasul M, Lee B, Wei G, Lamba G, Liu D. Comparison of safety and toxicity of liposomal doxorubicin vs. conventional anthracyclines: a meta-analysis. Exp Hematol Oncol. 2012;1:10. Scholar
  31. 31.
    Faroja M, Ahmed M, Appelbaum L, Ben-David E, Moussa M, Sosna J, Nissenbaum I, Goldberg SN. Irreversible electroporation ablation: Is all the damage nonthermal? Radiology. 2013;266:462–70. Scholar
  32. 32.
    Guo Y, Zhang Y, Klein R, Nijm GM, Sahakian AV, Omary RA, Yang G-Y, Larson AC. Irreversible electroporation therapy in the liver: longitudinal efficacy studies in a rat model of hepatocellular carcinoma. Cancer Res. 2010;70:1555–63. Scholar
  33. 33.
    Solazzo SA, Ahmed M, Schor-Bardach R, Yang W, Girnun GD, Rahmanuddin S, Levchenko T, Signoretti S, Spitz DR, Torchilin V, Goldberg SN. Liposomal doxorubicin increases radiofrequency ablation–induced tumor destruction by increasing cellular oxidative and nitrative stress and accelerating apoptotic pathways1. Radiology. 2010;255:62–74. Scholar
  34. 34.
    Ruarus AH, Vroomen LGPH, Puijk RS, Scheffer HJ, Faes TJC, Meijerink MR. Conductivity rise during irreversible electroporation: True permeabilization or heat? Cardiovasc Intervent Radiol. 2018;41:1257–66. Scholar
  35. 35.
    Elliott MR, Ravichandran KS. The dynamics of apoptotic cell clearance. Dev Cell. 2016;38:147–60. Scholar
  36. 36.
    Al-Sakere B, André F, Bernat C, Connault E, Opolon P, Davalos RV, Rubinsky B, Mir LM. Tumor ablation with irreversible electroporation. PLoS ONE. 2007. Scholar
  37. 37.
    Ahmed M, Monsky WE, Girnun G, Lukyanov A, D’Ippolito G, Kruskal JB, Stuart KE, Torchilin VP, Goldberg SN. Radiofrequency thermal ablation sharply increases intratumoral liposomal doxorubicin accumulation and tumor coagulation. Cancer Res. 2003;63:6327–33.Google Scholar
  38. 38.
    Moussa M, Goldberg SN, Kumar G, Sawant RR, Levchenko T, Torchilin VP, Ahmed M. Nanodrug-enhanced radiofrequency tumor ablation: effect of micellar or liposomal carrier on drug delivery and treatment efficacy. PLoS ONE. 2014;9:e102727. Scholar
  39. 39.
    Cox J, Weinman S. Mechanisms of doxorubicin resistance in hepatocellular carcinoma. Hepatic Oncol. 2016;3:57–9. Scholar
  40. 40.
    Thorn CF, Oshiro C, Marsh S, Hernandez-Boussard T, McLeod H, Klein TE, Altman RB. Doxorubicin pathways: pharmacodynamics and adverse effects. Pharmacogenet Genomics. 2011;21:440–6. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature and the Cardiovascular and Interventional Radiological Society of Europe (CIRSE) 2019

Authors and Affiliations

  • Tomáš Andrašina
    • 1
  • Josef Jaroš
    • 2
  • Tomáš Jůza
    • 1
    Email author
  • Tomáš Rohan
    • 1
  • Dalibor Červinka
    • 3
  • Michal Crha
    • 4
  • Vlastimil Válek
    • 1
  • Nahum S. Goldberg
    • 5
  1. 1.Faculty of Medicine, Department of Radiology and Nuclear MedicineUniversity Hospital Brno and Masaryk UniversityBrnoCzech Republic
  2. 2.Faculty of Medicine, Department of Histology and EmbryologyMasaryk UniversityBrnoCzech Republic
  3. 3.Faculty of Electrical Engineering and Communication, Department of Power Electrical and Electronic EngineeringBrno University of TechnologyBrnoCzech Republic
  4. 4.Faculty of Veterinary Medicine, Department of Surgery and Orthopedics, Small Animal ClinicUniversity of Veterinary and Pharmaceutical Sciences BrnoBrnoCzech Republic
  5. 5.Hadassah Hebrew University Medical CenterJerusalemIsrael

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