Bulletin of the American Physical Society
67th Annual Gaseous Electronics Conference
Volume 59, Number 16
Sunday–Friday, November 2–7, 2014; Raleigh, North Carolina
Session DT3: Effects of Plasmas on Biological Cells |
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Chair: Mounir Laroussi, Old Dominion University Room: State D |
Tuesday, November 4, 2014 10:00AM - 10:30AM |
DT3.00001: Application of atmospheric plasma sources in growth and differentiation of plant and mammalian stem cells Invited Speaker: Nevena Puac The expansion of the plasma medicine and its demand for in-vivo treatments resulted in fast development of various plasma devices that operate at atmospheric pressure. These sources have to fulfill all demands for application on biological samples. One of the sources that meet all the requirements needed for treatment of biological material is plasma needle. Previously, we have used this device for sterilization of planctonic samples of bacteria, MRSA biofilm, for improved differentiation of human periodontal stem cells into osteogenic line and for treatment of plant meristematic cells. It is well known that plasma generates reactive oxygen species (ROS) and reactive nitrogen species (RNS) that strongly affect metabolism of living cells. One of the open issues is to correlate external plasma products (electrons, ions, RNS, ROS, photons, strong fields etc.) with the immediate internal response which triggers or induces effects in the living cell. For that purpose we have studied the kinetics of enzymes which are typical indicators of the identity of reactive species from the plasma created environment that can trigger signal transduction in the cell and ensue cell activity.\\[4pt] In collaboration with Suzana Zivkovicm, Institute for Biological Research ``Sinisa Stankovic,'' University of Belgrade; Nenad Selakovic, Institute of Physics, University of Belgrade; Milica Milutinovic, Jelena Boljevic, Institute for Biological Research ``Sinisa Stankovic,'' University of Belgrade; and Gordana Malovic, Zoran Lj. Petrovic, Institute of Physics, University of Belgrade. [Preview Abstract] |
Tuesday, November 4, 2014 10:30AM - 11:00AM |
DT3.00002: Minimally-Invasive Gene Transfection by Chemical and Physical Interaction of Atmospheric Pressure Plasma Flow Invited Speaker: Toshiro Kaneko Non-equilibrium atmospheric pressure plasma irradiated to the living-cell is investigated for medical applications such as gene transfection, which is expected to play an important role in molecular biology, gene therapy, and creation of induced pluripotent stem (iPS) cells. However, the conventional gene transfection using the plasma has some problems that the cell viability is low and the genes cannot be transferred into some specific lipid cells, which is attributed to the unknown mechanism of the gene transfection using the plasma. Therefore, the time-controlled atmospheric pressure plasma flow is generated and irradiated to the living-cell suspended solution for clarifying the transfection mechanism toward developing highly-efficient and minimally- invasive gene transfection system. In this experiment, fluorescent dye YOYO-1 is used as the simulated gene and LIVE/DEAD Stain is simultaneously used for cell viability assay. By the fluorescence image, the transfection efficiency is calculated as the ratio of the number of transferred and surviving cells to total cell count. It is clarified that the transfection efficiency is significantly increased by the short-time (\textless 4 sec) and short-distance (\textless 40 mm) plasma irradiation, and the high transfection efficiency of 53{\%} is realized together with the high cell viability (\textgreater 90{\%}). This result indicates that the physical effects such as the electric field caused by the charged particles arriving at the surface of the cell membrane, and chemical effects associated with plasma-activated products in solution act synergistically to enhance the cell-membrane transport with low-damage. [Preview Abstract] |
Tuesday, November 4, 2014 11:00AM - 11:15AM |
DT3.00003: Impact of plasma induced liquid chemistry and charge on bacteria loaded aerosol droplets David Rutherford, David McDowell, Davide Mariotti, Charles Mahony, Declan Diver, Hugh Potts, Euan Bennet, Paul Maguire The introduction of living organisms, such as bacteria, into atmospheric pressure microplasmas offers a unique opportunity to study the local chemical and electrical effects on cell structure and viability. Individual bacteria, each encapsulated in an aerosol droplet, were successfully transmitted through a non-thermal equilibrium RF coaxial plasma, using a custom-design concentric double gas shroud interface and via adjustment of transit times and plasma parameters, we can control cell viability. Plasma electrical characteristics (n$_{\mathrm{e}}$ $\sim$ 10$^{13}$ cm$^{-3}$), droplet velocity profiles and aspects of plasma-induced droplet chemistry were determined in order to establish the nature of the bacteria in droplet environment. Plasma-exposed viable E coli cells were subsequently cultured and the growth rate curves (lag and exponential phase gradient) used to explore the effect of radical chemistry and electron bombardment on cell stress. The extent and nature of membrane disruption in viable and non-viable cells were investigated through genomic and protein/membrane lipid content estimation. We will also compare our results with simulations [1] of the effect of bacterial presence on plasma induced droplet charging and evaporation.\\[4pt] [1] E Bennet et al., New J. Physics (submitted). [Preview Abstract] |
Tuesday, November 4, 2014 11:15AM - 11:30AM |
DT3.00004: Quantitative inactivation-mechanisms of P. digitatum and A. niger spores based on atomic oxygen dose Masafumi Ito, Hiroshi Hashizume, Takayuki Ohta, Masaru Hori We have investigated inactivation mechanisms of Penicillium digitatum and Asperguills niger spores using atmospheric-pressure radical source quantitatively. The radical source was specially developed for supplying only neutral radicals without charged species and UV-light emissions. Reactive oxygen radical densities such as grand-state oxygen atoms, excited-state oxygen molecules and ozone were measured using VUV and UV absorption spectroscopies. The measurements and the treatments of spores were carried out in an Ar-purged chamber for eliminating the influences of OH, NOx and so on. The results revealed that the inactivation of spores can be explained by atomic-oxygen dose under the conditions employing neutral ROS irradiations. On the basis of the dose, we have observed the changes of intracellular organelles and membrane functions using TEM, SEM and confocal- laser fluorescent microscopy. From these results, we discuss the detail inactivation-mechanisms quantitatively based on atomic-oxygen dose. [Preview Abstract] |
Tuesday, November 4, 2014 11:30AM - 11:45AM |
DT3.00005: Treatment of prostate cancer cell lines and primary cells using low temperature plasma Deborah O'Connell, Adam Hirst, Fiona F. Frame, Norman J. Maitland The mechanisms of cell death after plasma treatment of both benign and cancerous prostate epithelial cells are investigated. Prostate cancer tissue was obtained with patient consent from targeted needle core biopsies following radical prostatectomy. Primary cells were cultured from cancer tissue and plated onto a chamber slide at a density of 10,000 cells per well in 200 microliter of stem cell media (SCM). The treated sample was previously identified as Gleason grade 7 cancer through tissue histo-pathology. A dielectric barrier discharge (DBD) jet configuration, with helium as a carrier gas, and 0.3\% O2 admixture was used for treating the cells. Reactive oxygen and nitrogen species (RONS) produced by the plasma are believed to be the main mediators of the plasma-cell interaction and response. We found the concentration of reactive oxygen species (ROS) induced inside the cells increased with plasma exposure. Exposure to the plasma for $>$3 minutes showed high levels of DNA damage compared to untreated and hydrogen peroxide controls. Cell viability and cellular recovery are also investigated and will be presented. All findings were common to both cell lines, suggesting the potential of LTP therapy for both benign and malignant disease. [Preview Abstract] |
Tuesday, November 4, 2014 11:45AM - 12:00PM |
DT3.00006: Power source effects of soft plasma jet and the differential response of skin cancer and normal cells Nathaniel Taylor, Danil Dobrynin, Alexander Fridman, Eun Ha Choi The effects of pulsed power direct current energy sources were compared using an indirect discharge plasma jet applied to treat cancerous and normal skin cells. Two power supplies with different voltage and current profiles were compared and optimized through the measurement of physical parameters and evaluated through the treatment of skin cells using an atmospheric pressure nitrogen gas plasma jet. Plasma density and temperature, power output, gas output temperature, and reactive species production were measured. Cell morphology, viability, and ROS generation were investigated using staining. A differential response has been shown between the normal and cancerous cell lines. The cancer cells viability reduced while normal cells did not over the same treatment time. [Preview Abstract] |
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