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- ItemPlasma-derived reactive species shape a differentiation profile in human monocytes(Basel : MDPI, 2019) Freund, Eric; Moritz, Juliane; Stope, Matthias; Seebauer, Christian; Schmidt, Anke; Bekeschus, SanderBackground: Monocyte-derived macrophages are key regulators and producers of reactive oxygen and nitrogen species (ROS/RNS). Pre-clinical and clinical studies suggest that cold physical plasma may be beneficial in the treatment of inflammatory conditions via the release of ROS/RNS. However, it is unknown how plasma treatment affects monocytes and their differentiation profile. Methods: Naïve or phorbol-12-myristate-13-acetate (PMA)-pulsed THP-1 monocytes were exposed to cold physical plasma. The cells were analyzed regarding their metabolic activity as well as flow cytometry (analysis of viability, oxidation, surface marker expression and cytokine secretion) and high content imaging (quantitative analysis of morphology. Results: The plasma treatment affected THP-1 metabolisms, viability, and morphology. Furthermore, a significant modulation CD55, CD69, CD271 surface-expression and increase of inflammatory IL1β, IL6, IL8, and MCP1 secretion was observed upon plasma treatment. Distinct phenotypical changes in THP-1 cells arguing for a differentiation profile were validated in primary monocytes from donor blood. As a functional outcome, plasma-treated monocytes decreased the viability of co-cultured melanoma cells to a greater extent than their non-treated counterparts. Conclusions: Our results suggest plasma-derived ROS/RNS shaped a differentiation profile in human monocytes as evidenced by their increased inflammatory profile (surface marker and cytokines) as well as functional outcome (tumor toxicity). © 2019 by the authors.
- ItemTumor cytotoxicity and immunogenicity of a novel V-jet neon plasma source compared to the kINPen(London : Nature Publishing Group, 2021) Miebach, Lea; Freund, Eric; Horn, Stefan; Niessner, Felix; Sagwal, Sanjeev Kumar; von Woedtke, Thomas; Emmert, Steffen; Weltmann, Klaus-Dieter; Clemen, Ramona; Schmidt, Anke; Gerling, Torsten; Bekeschus, SanderRecent research indicated the potential of cold physical plasma in cancer therapy. The plethora of plasma-derived reactive oxygen and nitrogen species (ROS/RNS) mediate diverse antitumor effects after eliciting oxidative stress in cancer cells. We aimed at exploiting this principle using a newly designed dual-jet neon plasma source (Vjet) to treat colorectal cancer cells. A treatment time-dependent ROS/RNS generation induced oxidation, growth retardation, and cell death within 3D tumor spheroids were found. In TUM-CAM, a semi in vivo model, the Vjet markedly reduced vascularized tumors' growth, but an increase of tumor cell immunogenicity or uptake by dendritic cells was not observed. By comparison, the argon-driven single jet kINPen, known to mediate anticancer effects in vitro, in vivo, and in patients, generated less ROS/RNS and terminal cell death in spheroids. In the TUM-CAM model, however, the kINPen was equivalently effective and induced a stronger expression of immunogenic cancer cell death (ICD) markers, leading to increased phagocytosis of kINPen but not Vjet plasma-treated tumor cells by dendritic cells. Moreover, the Vjet was characterized according to the requirements of the DIN-SPEC 91315. Our results highlight the plasma device-specific action on cancer cells for evaluating optimal discharges for plasma cancer treatment.
- ItemOne-Step Liquid Phase Polymerization of HEMA by Atmospheric-Pressure Plasma Discharges for Ti Dental Implants(Basel : MDPI, 2021) Buxadera-Palomero, Judit; Fricke, Katja; Reuter, Stephan; Gil, Francisco Javier; Rodriguez, Daniel; Canal, CristinaDental implants can fail due to various factors, in which bad tissue integration is believed to have a significant role. Specific properties of the implant surface, such as its chemistry and roughness, are of paramount importance to address specific cell responses, such as the adsorption of proteins, as well as the adhesion and differentiation of cells, which are suitable for biomaterial and tissue engineering. In this study, an acrylate-containing coating was produced on titanium surfaces through the atmospheric pressure plasma treatment of a liquid precursor, 2-hydroxyethyl methacrylate. A hydrophilic coating was obtained, showing retention of the monomer chemistry as assessed by FTIR analysis and XPS. Enhanced fibroblast adhesion and decreased Staphylococcus aureus and Escherichia coli adhesion were recorded, showing that this is a suitable method to produce biocompatible coatings with a reduced bacterial adhesion.
- ItemRadiobiological effects of wound fluid on breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic culture([London] : Macmillan Publishers Limited, part of Springer Nature, 2022) Jeibouei, Shabnam; Hojat, Ali; Mostafavi, Ebrahim; Aref, Amir Reza; Kalbasi, Alireza; Niazi, Vahid; Ajoudanian, Mohammad; Mohammadi, Farzaneh; Saadati, Fariba; Javadi, Seyed Mohammadreza; Shams, Forough; Moghaddam, Maryam; Karami, Farshid; Sharifi, Kazem; Moradian, Farid; Akbari, Mohammad Esmaeil; Zali, HakimehIntraoperative radiotherapy (IORT) could abrogate cancer recurrences, but the underlying mechanisms are unclear. To clarify the effects of IORT-induced wound fluid on tumor progression, we treated breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic cell culture systems, respectively. The viability, migration, and invasion of the cells under treatment of IORT-induced wound fluid (WF-RT) and the cells under surgery-induced wound fluid (WF) were compared. Our findings showed that cell viability was increased in spheroids under both WF treatments, whereas viability of the cell lines depended on the type of cells and incubation times. Both WFs significantly increased sub-G1 and arrested the cells in G0/G1 phases associated with increased P16 and P21 expression levels. The expression level of Caspase 3 in both cell culture systems and for both WF-treated groups was significantly increased. Furthermore, our results revealed that although the migration was increased in both systems of WF-treated cells compared to cell culture media-treated cells, E-cadherin expression was significantly increased only in the WF-RT group. In conclusion, WF-RT could not effectively inhibit tumor progression in an ex vivo tumor-on-chip model. Moreover, our data suggest that a microfluidic system could be a suitable 3D system to mimic in vivo tumor conditions than 2D cell culture.
- ItemAuthor Correction: A combination of electrochemistry and mass spectrometry to monitor the interaction of reactive species with supported lipid bilayers (Scientific Reports, (2020), 10, 1, (18683), 10.1038/s41598-020-75514-7)(London : Nature Publishing Group, 2021) Ravandeh, M.; Kahlert, H.; Jablonowski, H.; Lackmann, J.-W.; Striesow, J.; Agmo Hernández, V.; Wende, K.Correction to: Scientific Reports https://doi.org/10.1038/s41598-020-75514-7, published online 29 October 2020
- ItemPlasma-activation of larger liquid volumes by an inductively-limited discharge for antimicrobial purposes(Basel : MDPI, 2019) Schmidt, Michael; Hahn, Veronika; Altrock, Beke; Gerling, Torsten; Gerber, Ioana Cristina; Weltmann, Klaus-Dieter; von Woedtke, ThomasA new configuration of a discharge chamber and power source for the treatment of up to 1 L of liquid is presented. A leakage transformer, energizing two metal electrodes positioned above the liquid, limits the discharge current inductively by utilizing the weak magnetic coupling between the primary and secondary coils. No additional means to avoid arcing (electric short-circuiting), e.g., dielectric barriers or resistors, are needed. By using this technique, exceeding the breakdown voltage leads to the formation of transient spark discharges, producing non-thermal plasma (NTP). These discharges effected significant changes in the properties of the treated liquids (distilled water, physiological saline solution, and tap water). Considerable concentrations of nitrite and nitrate were detected after the plasma treatment. Furthermore, all tested liquids gained strong antibacterial efficacy which was shown by inactivating suspended Escherichia coli and Staphylococcus aureus. Plasma-treated tap water had the strongest effect, which is shown for the first time. Additionally, the pH-value of tap water did not decrease during the plasma treatment, and its conductivity increased less than for the other tested liquids. © 2019 by the authors.
- ItemPhysical plasma-triggered ROS induces tumor cell death upon cleavage of HSP90 chaperone([London] : Macmillan Publishers Limited, part of Springer Nature, 2019) Bekeschus, Sander; Lippert, Maxi; Diepold, Kristina; Chiosis, Gabriela; Seufferlein, Thomas; Azoitei, NinelHSP90 is a ubiquitously expressed molecular chaperone implicated in the correct folding and maturation of a plethora of proteins including protein kinases and transcription factors. While disruption of chaperone activity was associated with augmented cancer cell death and decreased tumor growth both in vitro and in vivo, the regulation of HSP90 is not clearly understood. Here we report that treatment of cancer cells with cold physical plasma, an emerging and less aggressive tumor therapy, resulted in ROS generation which subsequently triggered the cleavage of HSP90. Notably, cleavage of HSP90 was followed by the degradation of PKD2, a crucial regulator of tumor growth and angiogenesis. Pre-sensitization of cancer cells with subliminal doses of PU-H71, an HSP90 inhibitor currently under clinical evaluation, followed by treatment with cold-plasma, synergistically and negatively impacted on the viability of cancer cells. Taken together, cold-plasma can be used in conjunction with pharmacologic treatment in order to target the expression and activity of HSP90 and the downstream client proteins implicated in various cancer cell capabilities.
- ItemCold Atmospheric Pressure Plasma Jet Operated in Ar and He: From Basic Plasma Properties to Vacuum Ultraviolet, Electric Field and Safety Thresholds Measurements in Plasma Medicine(Basel : MDPI, 2022) Nastuta, Andrei Vasile; Gerling, TorstenApplication desired functionality as well as operation expenses of cold atmospheric pressure plasma (CAP) devices scale with properties like gas selection. The present contribution provides a comparative investigation for a CAP system operated in argon or helium at different operation voltages and distance to the surface. Comparison of power dissipation, electrical field strength and optical emission spectroscopy from vacuum ultraviolet over visible up to near infrared ((V)UV-VIS-NIR) spectral range is carried out. This study is extended to safety relevant investigation of patient leakage current, induced surface temperature and species density for ozone (O3) and nitrogen oxides (NOx). It is found that in identical operation conditions (applied voltage, distance to surface and gas flow rate) the dissipated plasma power is about equal (up to 10 W), but the electrical field strength differs, having peak values of 320 kV/m for Ar and up to 300 kV/m for He. However, only for Ar CAP could we measure O3 up to 2 ppm and NOx up to 7 ppm. The surface temperature and leakage values of both systems showed different slopes, with the biggest surprise being a constant leakage current over distance for argon. These findings may open a new direction in the plasma source development for Plasma Medicine.
- ItemOxidatively Modified Proteins: Cause and Control of Diseases(Basel : MDPI, 2020) Clemen, Ramona; Bekeschus, SanderProteins succumb to numerous post-translational modifications (PTMs). These relate to enzymatic or non-enzymatic reactions taking place in either the intracellular or extracellular compartment. While intracellular oxidative changes are mainly due to redox stress, extracellular PTMs may be induced in an inflammatory micro milieu that is rich in reactive species. The increasing recognition of oxidative modifications as a causing agent or side-effect of pathophysiological states and diseases puts oxidative PTMS (oxPTMs) into the spotlight of inflammation research. Pathological hyper-modification of proteins can lead to accumulation, aggregation, cell stress, altered antigenic peptides, and damage-associated molecular pattern (DAMP)-like recognition by host immunity. Such processes are linked to cardiovascular disease and autoinflammation. At the same time, a detailed understanding of the mechanisms governing inflammatory responses to oxPTMs may capitalize on new therapeutic routes for enhancing adaptive immune responses as needed, for instance, in oncology. We here summarize some of the latest developments of oxPTMs in disease diagnosis and therapy. Potential target proteins and upcoming technologies, such as gas plasmas, are outlined for future research that may aid in identifying the molecular basis of immunogenic vs. tolerogenic oxPTMs.
- ItemQuantification of the ozone and singlet delta oxygen produced in gas and liquid phases by a non-thermal atmospheric plasma with relevance for medical treatment([London] : Macmillan Publishers Limited, part of Springer Nature, 2018-8-15) Jablonowski, Helena; Santos Sousa, Joao; Weltmann, Klaus-Dieter; Wende, Kristian; Reuter, StephanIn the field of plasma medicine, the identification of relevant reactive species in the liquid phase is highly important. To design the plasma generated species composition for a targeted therapeutic application, the point of origin of those species needs to be known. The dominant reactive oxygen species generated by the plasma used in this study are atomic oxygen, ozone, and singlet delta oxygen. The species density changes with the distance to the active plasma zone, and, hence, the oxidizing potential of this species cocktail can be tuned by altering the treatment distance. In both phases (gas and liquid), independent techniques have been used to determine the species concentration as a function of the distance. The surrounding gas composition and ambient conditions were controlled between pure nitrogen and air-like by using a curtain gas device. In the gas phase, in contrast to the ozone density, the singlet delta oxygen density showed to be more sensitive to the distance. Additionally, by changing the surrounding gas, admixing or not molecular oxygen, the dynamics of ozone and singlet delta oxygen behave differently. Through an analysis of the reactive species development for the varied experimental parameters, the importance of several reaction pathways for the proceeding reactions was evaluated and some were eventually excluded.