Jerome Canady

The Effect of Tuning Cold Plasma Composition on Glioblastoma Cell Viability

Previous research in cold atmospheric plasma (CAP) and cancer cell interaction has repeatedly proven that the cold plasma induced cell death. It is postulated that the reactive oxygen species (ROS) and reactive nitrogen species (RNS) play a major role in the CAP cancer therapy. In this paper, we seek to determine a mechanism of CAP therapy on glioblastoma cells (U87) through an understanding of the composition of the plasma, including treatment time, voltage, flow-rate and plasma-gas composition. In order to determine the threshold of plasma treatment on U87, normal human astrocytes (E6/E7) were used as the comparison cell line. Our data showed that the 30 sec plasma treatment caused 3-fold cell death in the U87 cells compared to the E6/E7 cells. All the other compositions of cold plasma were performed based on this result: plasma treatment time was maintained at 30 s per well while other plasma characteristics such as voltage, flow rate of source gas, and composition of source gas were changed one at a time to vary the intensity of the reactive species composition in the plasma jet, which may finally have various effect on cells reflected by cell viability. We defined a term “plasma dosage” to summarize the relationship of all the characteristics and cell viability.

Principles of using Cold Atmospheric Plasma Stimulated Media for Cancer Treatment.

To date, the significant anti-cancer capacity of cold atmospheric plasma (CAP) on dozens of cancer cell lines has been demonstrated in vitro and in mice models. Conventionally, CAP was directly applied to irradiate cancer cells or tumor tissue. Over past three years, the CAP irradiated media was also found to kill cancer cells as effectively as the direct CAP treatment. As a novel strategy, using the CAP stimulated (CAPs) media has become a promising anti-cancer tool. In this study, we demonstrated several principles to optimize the anti-cancer capacity of the CAPs media on glioblastoma cells and breast cancer cells. Specifically, using larger wells on a multi-well plate, smaller gaps between the plasma source and the media, and smaller media volume enabled us to obtain a stronger anti-cancer CAPs media composition without increasing the treatment time. Furthermore, cysteine was the main target of effective reactive species in the CAPs media. Glioblastoma cells were more resistant to the CAPs media than breast cancer cells. Glioblastoma cells consumed the effective reactive species faster than breast cancer cells did. In contrast to nitric oxide, hydrogen peroxide was more likely to be the effective reactive species.

Controlling plasma stimulated media in cancer treatment application

Cold atmospheric plasma (CAP) constitutes a “cocktail” of various reactive species. Accumulating evidence shows the effectiveness of CAP in killing cancer cells and decreasing the tumor size, which provides a solid basis for its potential use in cancer treatment. Currently, CAP is mainly used to directly treat cancer cells and trigger the death of cancer cells via apoptosis or necrosis. By altering the concentration of fetal bovine serum in Dulbeccos modified Eagles medium and the temperature to store CAP stimulated media, we demonstrated controllable strategies to harness the stimulated media to kill glioblastoma cells in vitro. This study demonstrated the significant role of media in killing cancer cells via the CAP treatment.

Synergistic effect of gold nanoparticles and cold plasma on glioblastoma cancer therapy

Gold nanoparticles (AuNPs) have been investigated as a promising reagent for cancer therapy in various fields. In the meantime, cold atmospheric plasma has shown exquisite selectivity towards cancer cells. In this paper, we demonstrate that there is a synergy between gold nanoparticles and cold atmospheric plasma in cancer therapy. Specifically, the concentration of AuNPs plays an important role on plasma therapy. At an optimal concentration, gold nanoparticles can significantly induce glioblastoma (U87) cell death up to a 30% overall increase compared to the control group with the same plasma dosage but no AuNPs applied. The reactive oxygen species (ROS) intensity of the corresponding conditions has a reversed trend compared to cell viability. This matches with the theory that intracellular ROS accumulation results in oxidative stress, which further changes the intracellular pathways, causing damage to the proteins, lipids and DNA. Our results show that this synergy has great potential in improving the efficiency of cancer therapy and reducing harm to normal cells.

Use of cold atmospheric plasma in the treatment of cancer

Cold atmospheric plasma (CAP) is an emerging modality for the treatment of solid tumors. In-vitro experiments have demonstrated that with increasing doses of plasma, tumor cells assays display decreased cell viability. CAP is theorized to induce tumor cells into apoptosis via multiple pathways including reactive oxygen and nitrogen species as well as cell cycle disruption. Studies have shown CAP treatment can decrease mouse model glioblastoma multiforme tumor volume by 56%, increase life span by 60%, and maintain up to 85% viability of normal cells. Emerging evidence suggests that CAP is a viable in-vivo treatment for a number of tumors, including glioblastoma, as it appears to selectively induce tumor cell death while noncancerous cells remain viable.

Electric discharge during electrosurgery

Electric discharge utilized for electrosurgery is studied by means of a recently developed method for the diagnostics of small-size atmospheric plasma objects based on Rayleigh scattering of microwaves on the plasma volume. Evolution of the plasma parameters in the near-electrode sheaths and in the positive column is measured and analyzed. It is found that the electrosurgical system produces a glow discharge of alternating current with strongly contracted positive column with current densities reaching 103 A/cm2. The plasma electron density and electrical conductivities in the channel were found be 1016 cm−3 and (1-2) Ohm−1cm−1, respectively. The discharge interrupts every instance when the discharge-driving AC voltage crosses zero and re-ignites again every next half-wave at the moment when the instant voltage exceeds the breakdown threshold.

A Novel Micro Cold Atmospheric Plasma Device for Glioblastoma Both In Vitro and In Vivo

Cold atmospheric plasma (CAP) treatment is a rapidly expanding and emerging technology for cancer treatment. Direct CAP jet irradiation is limited to the skin and it can also be invoked as a supplement therapy during surgery as it only causes cell death in the upper three to five cell layers. However, the current cannulas from which the plasma emanates are too large for intracranial applications. To enhance efficiency and expand the applicability of the CAP method for brain tumors and reduce the gas flow rate and size of the plasma jet, a novel micro-sized CAP device (µCAP) was developed and employed to target glioblastoma tumors in the murine brain. Various plasma diagnostic techniques were applied to evaluate the physics of helium µCAP such as electron density, discharge voltage, and optical emission spectroscopy (OES). The direct and indirect effects of µCAP on glioblastoma (U87MG-RedFluc) cancer cells were investigated in vitro. The results indicate that µCAP generates short- and long-lived species and radicals (i.e., hydroxyl radical (•OH), hydrogen peroxide (H2O2), and nitrite (NO2−), etc.) with increasing tumor cell death in a dose-dependent manner. Translation of these findings to an in vivo setting demonstrates that intracranial µCAP is effective at preventing glioblastoma tumor growth in the mouse brain. The µCAP device can be safely used in mice, resulting in suppression of tumor growth. These initial observations establish the µCAP device as a potentially useful ablative therapy tool in the treatment of glioblastoma.

Adaptation of Operational Parameters of Cold Atmospheric Plasma for in Vitro Treatment of Cancer Cells.

Cold atmospheric plasma (CAP), an ionized gas operated at near-ambient temperatures, has been introduced as a new therapeutic opportunity for treating cancers. The effectiveness of the therapy has been linked to CAP-generated reactive oxygen and nitrogen species such as hydrogen peroxide and nitrite. In this study, we monitor in real-time cancer cell response to CAP over the course of 48 h. The results demonstrate a correlation between cell viability, exposure time (30, 60, 90, and 180 s), and discharge voltage (3.16 and 3.71 kV), while stressing the likely therapeutic role of plasma-generated reactive species. A 30-60 s increase in CAP exposure time and/or a discharge voltage adjustment from 3.16 to 3.71 kV is consistently accompanied by a significant reduction in cell viability. Comparably, levels of hydrogen peroxide and nitrite vary as a function of voltage with elevated levels detected at the highest tested voltage condition of 3.71 kV. CAP ultimately initiates a reduction in cell viability and triggers apoptosis via damage to the mitochondrial membrane, while also deregulating protein synthesis. The findings presented in this study are discussed in the context of facilitating the development of an adaptive CAP platform which could improve treatment outcomes.

The cutting mechanism of the electrosurgical scalpel

Electrosurgical cutting is a well-known technique for creating incisions often used for the removal of benign and malignant tumors. The proposed mathematical model suggests that incisions are created due to the localized heating of the tissue. The model estimates a volume of tissue heating in the order of 2 10−4 mm3. This relatively small predicted volume explains why the heat generated from the very tip of the scalpel is unable to cause extensive damage to the tissue adjacent to the incision site. The scalpel exposes the target region to an RF field in 60 ms pulses until a temperature of around 100 °C is reached. This process leads to desiccation where the tissue is characterized by a significantly low electrical conductivity, which prevents further heating and charring. Subsequently, the incision is created from the mechanical scraping process that follows.

Temporal Evolution of the Discharge in U.S. Medical Innovations Electrosurgical System SS-200E/Argon-2

A series of instant photographs of a discharge produced by the electrosurgical system SS-200E/Argon-2 by U.S. Medical Innovations is presented and analyzed.