Intracellular Delivery of Bleomycin by Combined Application of Electroporation and Sonoporation in Vitro
- 244 Downloads
In this study, we aimed to determine whether the combination of electroporation (EP) and ultrasound (US) waves (sonoporation) can result in an increased intracellular delivery of anticancer drug bleomycin. CHO cells were treated with electric pulses (1 or 8 high voltage pulses of 800 or 1200 V/cm, 100 μs or 1 low voltage pulse of 100 or 250 V/cm, 100 ms) and with 880 kHz US of 320 or 500 kPa peak negative pressure, 100 % duty cycle, applied for 2 s in the presence or absence of exogenously added contrast agent microbubbles. Various sequential or simultaneous combinations of EP and sonoporation were used. The results of the study showed that i) sequential treatment of cells by EP and sonoporation enhanced bleomycin electrosonotransfer at the reduced energy of electric field and US; ii) sequential combination of EP and sonoporation induced a summation effect which at some conditions was more prominent when the cells were treated first by EP and then by sonoporation; iii) the most efficient intracellular delivery of bleomycin was achieved by the simultaneous application of cell EP and sonoporation resulting in percentage of reversibly porated cells above the summation level; and iv) compared with sequential application of EP and sonoporation, simultaneous use of electric pulses and US increased cell viability in the absence of bleomycin.
KeywordsElectroporation Sonoporation Drug delivery Electrosonoporation Ultrasound Microbubbles
This work was funded by Research Council of Lithuania project TAP-03/2012 (scientific cooperation between Lithuania, Latvia and Taiwan), the project from National Science Council, Taiwan, NSC 100-2923-B-002-004-MY3. In addition, this work was partially supported by Culture Department of French Embassy in Lithuania, CNRS, Univ. Paris-Sud, Gustave Roussy, Fondation EDF and Agence Nationale de la Recherche through the Project Soundelivery ANR-14-CE17-0008. This work was also partly performed in the scope of the LEA EBAM, the European Associated laboratory Electric pulses in Biology and Medicine.
- Čorović S, Al Sakere B, Haddad V, Miklavčič D, Mir LM (2008) Importance of contact surface between electrodes and treated tissue in electrochemotherapy. TCRT 7:393–399Google Scholar
- Lee EW, Loh CT, Kee ST (2007) Imaging guided percutaneous irreversible electroporation: ultrasound and immunohistological correlation. TCRT 6:287–294Google Scholar
- Tamošiūnas M, Jurkonis R, Mir LM, Lukoševičius A, Venslauskas MS, Šatkauskas S (2012b) Adjustment of ultrasound exposure duration to microbubble sonodestruction kinetics for optimal cell sonoporation in vitro. TCRT 11:375–387Google Scholar
- Yamashita YI, Shimada M, Minagawa R, Tsujita E, Harimoto N, Tanaka S, Shirabe K, Miyazaki JI, Maehara Y (2004) Muscle-targeted interleukin-12 gene therapy of orthotopic hepatocellular carcinoma in mice using in vivo electrosonoporation. Mol Cancer Ther 3:177–182Google Scholar