RESUMO
Cold atmospheric plasma (CAP), an ambient temperature ionized gas, is gaining extensive interest as a promising addition to anti-tumor therapy primarily due to the ability to generate and control delivery of electrons, ions, excited molecules, UV photons, and reactive species such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) to a specific site. The heterogeneous composition of CAP offers the opportunity to mediate several signaling pathways that regulate tumor cells. Consequently, the array of CAP generated products has limited the identification of the mechanisms of action on tumor cells. The aim of this work is to assess the cell death response of human myeloid leukemia cells by remote exposure to CAP generated RNS by utilizing a novel resistive barrier discharge system that primarily produces RNS. The effect of variable treatments of CAP generated RNS was tested in THP-1 cell (human monocytic leukemia cell line), a model for hematological malignancy. The number of viable cells was evaluated with erythrosine-B staining, while apoptosis and necrosis was assessed by endonuclease cleavage observed by agarose gel electrophoresis and detection of cells with the exclusionary dye propidium iodide and fluorescently labeled annexin-V by flow cytometry and fluorescent microscopy. Our observations indicate that treatment dosage levels of 45 s of exposure to CAP emitted RNS-induced apoptotic cell death and for higher dosage conditions of ≥50 s of exposure to CAP induced necrosis. Overall the results suggest that CAP emitted RNS play a significant role in the anti-tumor potential of CAP.
Assuntos
Antineoplásicos/metabolismo , Apoptose , Monócitos/efeitos dos fármacos , Monócitos/fisiologia , Gases em Plasma , Espécies Reativas de Nitrogênio/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citometria de Fluxo , Humanos , Microscopia de Fluorescência , Coloração e RotulagemRESUMO
This research investigated the modulation of cell death through exposure of non-thermal resistive barrier based indirect air plasma on monocytic leukemia cancer cells (THP-1). Specifically, we explored cell death through apoptosis and necrosis, since generally apoptotic cell death has a limited inflammatory response as compared to necrosis. We have demonstrated a preference for apoptosis in plasma treated THP-1 cells, under specific plasma characteristics and dosage levels, using fluorescent dyes conjugated with annexin V followed by identification of the cells through fluorescent microscopy and flowcytometry diagnostics. At much higher plasma dosages, the necrotic morphologies in the THP-1 cells were observed. The presented outcomes in the death morphologies of plasma treated THP-1 cells signify the need for further investigation on the cellular mechanisms induced by the indirect plasma exposure. The results obtained from this research indicate the significant potential for the use of our portable non-thermal resistive barrier based indirect plasma treatment method as an inexpensive and less invasive method for treating leukemia and other cancerous lesions.
Assuntos
Leucemia Monocítica Aguda/patologia , Leucemia Monocítica Aguda/fisiopatologia , Gases em Plasma/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Leucemia Monocítica Aguda/terapia , Resultado do TratamentoRESUMO
In this work an atmospheric pressure non-thermal resistive barrier (RB) plasma jet was constructed, characterized and was applied for biomedical applications. The RB plasma source can operate in both DC (battery) as well as in standard 60/50 Hz low frequency AC excitation, and it functions effectively in both direct and indirect plasma exposure configurations. The characteristics of the RB plasma jet such as electrical properties, plasma gas temperature and nitric oxides concentration were determined using voltage-current characterization, optical emission spectroscopy and gas analyzer diagnostic techniques. Plasma discharge power of 26.33 W was calculated from voltage-current characterization. An optical emission spectroscopy was applied and the gas temperature which is equivalent to the nitrogen rotational (T
Assuntos
Fenômenos Fisiológicos Bacterianos/efeitos dos fármacos , Desinfecção/métodos , Leucemia Monocítica Aguda/tratamento farmacológico , Leucemia Monocítica Aguda/fisiopatologia , Gases em Plasma/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Resultado do TratamentoRESUMO
This research study examined the effect of non-thermal portable atmospheric air plasma system on leukemia cancer cells. Acute monocytic leukemia cells (THP-1) were exposed to atmospheric pressure non-thermal plasma. To assess death caused by plasma exposure, cells were subjected to trypan blue exclusion assays and a kill-curve and assessment of death overtime were compiled using data from the assays. In addition to this, DNA was harvested from treated and untreated samples to determine if apoptotic ladders were present. Results have indicated that non-thermal plasma can cause cell death in THP-1 cells overtime, and the death that occurs corresponds directly to the amount of time that the cells were exposed to ionized plasma. Preliminary fluorescent imaging of the treated cells revealed that higher treatment doses are not only more likely to induce cellular death but are likely to induce necrotic death, while lower treatment doses that are capable of inducing death may induce apoptotic or programmed cellular death. Ideally the results obtained from these experiments will allow for further investigation of the effects of ionized non-thermal plasma on melanoma cell lines and will lead to an inexpensive method for treating early stage skin cancer and cancerous lesions.