Effectiveness of tumor electrochemotherapy as a function of electric pulse strength and duration

The aim of this study was to evaluate the effectiveness of electrochemotherapy (ECT) as a function of various combinations of pulse strength and duration. C57Bl mice bearing LLC tumors were injected i.p. with bleomycin (BLM) at doses 5 mg/kg in 0.2 ml of physiological saline. Thirty minutes later, tumors were positioned between plate electrodes and were pulsed with eight-square wave electric pulses with an individual pulse strength of 900, 1100, 1300 or 1500 V/cm and duration of 0.1, 0.25, 0.5 or 1 ms. Effectiveness of ECT was estimated by measuring inhibition of tumor growth and by estimating extent of necrosis in histological slices of the treated tumors. At pulse strength of 900 V/cm and duration of 0.1 ms, electrochemotherapy was ineffective. Noticeable inhibition of tumor growth (threshold of ECT) was obtained when pulse duration at this field strength was increased up to 0.25 ms. Further increase of pulse strength and/or duration resulted in progressive enhancement of antitumor effects. Using tumor doubling time (DT) as a criteria, we showed that the same efficacy of ECT could be achieved using various pairs of values for pulse strength and duration. Largest antitumor efficacy of ECT was obtained at pulse strength of 1500 V/cm and duration of 1 ms. These pulse conditions applied alone neither significantly suppressed tumor growth nor induced noticeable side effects of the surrounding tissues. The results of this study thus suggest that the effectiveness of electrochemotherapy can be enhanced (in comparison to widely accepted conditions of electrochemotherapy--8 pulses of 1300 V/cm, 0.1 ms) if 1500-V/cm, 1-ms electric pulses are used. Our study also implicates that other pulse conditions could be found for this enhanced ECT.