Synergistic Effects between Nano-Pulse and Cold Plasma to Improve Cancer Treatment
College
The Graduate School
Graduate Level
Doctoral
Graduate Program/Concentration
Biomedical Sciences
Presentation Type
Poster Presentation
Abstract
A high-power pulse technology, compressing electrical energy into nanosecond electric pulses (Nano-Pulse) or pulsed electric fields, has been developed at our research center to treat cancer successfully in many animal cancer models and a few clinical trials. However, due to the complexity of animal and human tumors (irregular shapes, heterogeneous physical properties, etc.) and the limitations of required high electric fields, the improvement of this bioelectric technology is necessary to increase the efficacy of tumor ablation and reduce side effects. We propose the combination of Nano-Pulse and cold plasma to address this issue. Our preliminary results support the feasibility of this novel idea. Utilizing Pan02 mouse pancreatic cancer cells, we have shown that a low dose of Nano-Pulse can sensitize cancer cells, making them more susceptible to cold plasma. The synergistic effects on Pan02 pancreatic cancer cells in vitro were assessed by the combination of cold plasma (pulse parameters: 9 kV, 200 ns pulses, 2 kHz and treatment times varied) and Nano-Pulse (pulse parameters:60 ns 50 kV/cm, 1 Hz and pulse numbers varied). Cell cytotoxicity and membrane permeabilization were examined by WST-1 assay and transcellular electrical resistance (ECIS). Results indicated that exposure to cold plasma for 3 minutes led to 42.45% of Pan02 cell death. Applying Nano-Pulse alone, specifically 20 pulses, resulted in 15.32% of Pano2 cell death. Although the cytotoxicity from individual treatments was moderate, the combined application of both treatments demonstrated an 80% of Pan02 cell death—a significant 2.7-to-5.3-fold increase compared to individual treatments. Additionally, ECIS assays showed a significantly more increase in cell permeabilization from the combination treatment than that from single treatments. The combination treatment using the same parameters revealed a 13% increase in overall cytoplasmic reactive oxygen species (ROS) generation measured by H2DCFDA, and a 7% increase in superoxide species measured by MitoSox (mitochondrial superoxide indicator). In contrast, individual treatments with Nano pulse and Cold plasma showed more modest effects, with Nano pulse leading to a 1.66% increase in superoxide species and 5.81% in ROS species, and Cold plasma resulting in a 2.66% increase in superoxide species and 6.71% in ROS species. The examination of mitochondrial membrane potential revealed a 65% greater reduction during combination treatments than single treatments. The notable reduction in membrane potential and elevated ROS levels observed in combination treatment implies the potential initiation of diverse cell death pathways.
Ongoing research is dedicated to elucidating the mechanisms underlying the synergistic effects between Nano-Pulse and cold plasma. We will investigate how these two treatments induce intracellular changes, especially cell death pathways, the upregulation or downregulation of multiple cellular pathway proteins, etc. Unraveling these mechanisms will enhance our comprehension of this innovative combination therapy, offering potential avenues for refining and optimizing its effectiveness in cancer treatment..
Synergistic Effects between Nano-Pulse and Cold Plasma to Improve Cancer Treatment
A high-power pulse technology, compressing electrical energy into nanosecond electric pulses (Nano-Pulse) or pulsed electric fields, has been developed at our research center to treat cancer successfully in many animal cancer models and a few clinical trials. However, due to the complexity of animal and human tumors (irregular shapes, heterogeneous physical properties, etc.) and the limitations of required high electric fields, the improvement of this bioelectric technology is necessary to increase the efficacy of tumor ablation and reduce side effects. We propose the combination of Nano-Pulse and cold plasma to address this issue. Our preliminary results support the feasibility of this novel idea. Utilizing Pan02 mouse pancreatic cancer cells, we have shown that a low dose of Nano-Pulse can sensitize cancer cells, making them more susceptible to cold plasma. The synergistic effects on Pan02 pancreatic cancer cells in vitro were assessed by the combination of cold plasma (pulse parameters: 9 kV, 200 ns pulses, 2 kHz and treatment times varied) and Nano-Pulse (pulse parameters:60 ns 50 kV/cm, 1 Hz and pulse numbers varied). Cell cytotoxicity and membrane permeabilization were examined by WST-1 assay and transcellular electrical resistance (ECIS). Results indicated that exposure to cold plasma for 3 minutes led to 42.45% of Pan02 cell death. Applying Nano-Pulse alone, specifically 20 pulses, resulted in 15.32% of Pano2 cell death. Although the cytotoxicity from individual treatments was moderate, the combined application of both treatments demonstrated an 80% of Pan02 cell death—a significant 2.7-to-5.3-fold increase compared to individual treatments. Additionally, ECIS assays showed a significantly more increase in cell permeabilization from the combination treatment than that from single treatments. The combination treatment using the same parameters revealed a 13% increase in overall cytoplasmic reactive oxygen species (ROS) generation measured by H2DCFDA, and a 7% increase in superoxide species measured by MitoSox (mitochondrial superoxide indicator). In contrast, individual treatments with Nano pulse and Cold plasma showed more modest effects, with Nano pulse leading to a 1.66% increase in superoxide species and 5.81% in ROS species, and Cold plasma resulting in a 2.66% increase in superoxide species and 6.71% in ROS species. The examination of mitochondrial membrane potential revealed a 65% greater reduction during combination treatments than single treatments. The notable reduction in membrane potential and elevated ROS levels observed in combination treatment implies the potential initiation of diverse cell death pathways.
Ongoing research is dedicated to elucidating the mechanisms underlying the synergistic effects between Nano-Pulse and cold plasma. We will investigate how these two treatments induce intracellular changes, especially cell death pathways, the upregulation or downregulation of multiple cellular pathway proteins, etc. Unraveling these mechanisms will enhance our comprehension of this innovative combination therapy, offering potential avenues for refining and optimizing its effectiveness in cancer treatment..