Search Results
Conductive Gas Plasma Treatment Augments Tumor Toxicity of Ringer’s Lactate Solutions in a Model of Peritoneal Carcinomatosis
2022, Miebach, Lea, Freund, Eric, Cecchini, Alessandra Lourenço, Bekeschus, Sander
Reactive species generated by medical gas plasma technology can be enriched in liquids for use in oncology targeting disseminated malignancies, such as metastatic colorectal cancer. Notwithstanding, reactive species quantities depend on the treatment mode, and we recently showed gas plasma exposure in conductive modes to be superior for cancer tissue treatment. However, evidence is lacking that such a conductive mode also equips gas plasma-treated liquids to confer augmented intraperitoneal anticancer activity. To this end, employing atmospheric pressure argon plasma jet kINPen-treated Ringer’s lactate (oxRilac) in a CT26-model of colorectal peritoneal carcinomatosis, we tested repeated intraabdominal injection of such remotely or conductively oxidized liquid for antitumor control and immunomodulation. Enhanced reactive species formation in conductive mode correlated with reduced tumor burden in vivo, emphasizing the advantage of conduction over the free mode for plasma-conditioned liquids. Interestingly, the infiltration of lymphocytes into the tumors was equally enhanced by both treatments. However, significantly lower levels of interleukin (IL)4 and IL13 and increased levels of IL2 argue for a shift in intratumoral T-helper cell subpopulations correlating with disease control. In conclusion, our data argue for using conductively over remotely prepared plasma-treated liquids for anticancer treatment.
Influence of redox stress on crosstalk between fibroblasts and keratinocytes
2021, Bhartiya, Pradeep, Masur, Kai, Shome, Debarati, Kaushik, Neha, Nguyen, Linh N., Kaushik, Nagendra Kumar, Choi, Eun Ha
Although the skin is constantly subjected to endogenous and exogenous stress, it maintains a homeostatic state through wound repair and regeneration pathways. Treatment for skin diseases and injury requires a significant understanding of the various mechanisms and interactions that occur within skin cells. Keratinocytes and fibroblasts interact with each other and act as key players in the repair process. Although fibroblasts and keratinocytes are widely studied in wound healing and skin remodeling under different conditions, the influence of redox stress on keratinocyte-fibroblast crosstalk has not been thoroughly investigated. In this study, we used cold atmospheric plasma (CAP) to generate and deliver oxidative stress to keratinocytes and fibroblasts and to assess its impact on their interactions. To this end, we used a well-established in vitro 3D co-culture model imitating a realistic scenario. Our study shows that low CAP exposure is biocompatible and does not affect the viability or energetics of fibroblasts and keratinocytes. Exposure to low doses of CAP enhanced the proliferation rate of cells and stimulated the expression of key genes (KGF, MMP2, GMCSF, IL-6, and IL-8) in fibroblasts, indicating the activation and initiation of the skin repair process. Additionally, enhanced migration was observed under co-culture conditions under the given redox stress conditions, and expression of the upstream regulator and the effectors of the Hippo pathway (YAP and CYR61, respectively), which are associated with enhanced migration, were elevated. Overall, this study reinforces the application of CAP and redox stress in skin repair physiology.
Understanding Surface Modifications Induced via Argon Plasma Treatment through Secondary Electron Hyperspectral Imaging
2021, Farr, Nicholas, Thanarak, Jeerawan, Schäfer, Jan, Quade, Antje, Claeyssens, Frederik, Green, Nicola, Rodenburg, Cornelia
Understanding the effects that sterilization methods have on the surface of a biomaterial is a prerequisite for clinical deployment. Sterilization causes alterations in a material's surface chemistry and surface structures that can result in significant changes to its cellular response. Here we compare surfaces resulting from the application of the industry standard autoclave sterilisation to that of surfaces resulting from the use of low-pressure Argon glow discharge within a novel gas permeable packaging method in order to explore a potential new biomaterial sterilisation method. Material surfaces are assessed by applying secondary electron hyperspectral imaging (SEHI). SEHI is a novel low-voltage scanning electron microscopy based characterization technique that, in addition to capturing topographical images, also provides nanoscale resolution chemical maps by utilizing the energy distribution of emitted secondary electrons. Here, SEHI maps are exploited to assess the lateral distributions of diverse functional groups that are effected by the sterilization treatments. This information combined with a range of conventional surface analysis techniques and a cellular metabolic activity assay reveals persuasive reasons as to why low-pressure argon glow discharge should be considered for further optimization as a potential terminal sterilization method for PGS-M, a functionalized form of poly(glycerol sebacate) (PGS).
Contact-dependent signaling triggers tumor-like proliferation of CCM3 knockout endothelial cells in co-culture with wild-type cells
2022, Rath, Matthias, Schwefel, Konrad, Malinverno, Matteo, Skowronek, Dariush, Leopoldi, Alexandra, Pilz, Robin A., Biedenweg, Doreen, Bekeschus, Sander, Penninger, Josef M., Dejana, Elisabetta, Felbor, Ute
Cerebral cavernous malformations (CCM) are low-flow vascular lesions prone to cause severe hemorrhage-associated neurological complications. Pathogenic germline variants in CCM1, CCM2, or CCM3 can be identified in nearly 100% of CCM patients with a positive family history. In line with the concept that tumor-like mechanisms are involved in CCM formation and growth, we here demonstrate an abnormally increased proliferation rate of CCM3-deficient endothelial cells in co-culture with wild-type cells and in mosaic human iPSC-derived vascular organoids. The observation that NSC59984, an anticancer drug, blocked the abnormal proliferation of mutant endothelial cells further supports this intriguing concept. Fluorescence-activated cell sorting and RNA sequencing revealed that co-culture induces upregulation of proangiogenic chemokine genes in wild-type endothelial cells. Furthermore, genes known to be significantly downregulated in CCM3−/− endothelial cell mono-cultures were upregulated back to normal levels in co-culture with wild-type cells. These results support the hypothesis that wild-type ECs facilitate the formation of a niche that promotes abnormal proliferation of mutant ECs. Thus, targeting the cancer-like features of CCMs is a promising new direction for drug development.
Gas Plasma Technology Augments Ovalbumin Immunogenicity and OT-II T Cell Activation Conferring Tumor Protection in Mice
2021, Clemen, Ramona, Freund, Eric, Mrochen, Daniel, Miebach, Lea, Schmidt, Anke, Rauch, Bernhard H., Lackmann, Jan‐Wilm, Martens, Ulrike, Wende, Kristian, Lalk, Michael, Delcea, Mihaela, Bröker, Barbara M., Bekeschus, Sander
Reactive oxygen species (ROS/RNS) are produced during inflammation and elicit protein modifications, but the immunological consequences are largely unknown. Gas plasma technology capable of generating an unmatched variety of ROS/RNS is deployed to mimic inflammation and study the significance of ROS/RNS modifications using the model protein chicken ovalbumin (Ova vs oxOva). Dynamic light scattering and circular dichroism spectroscopy reveal structural modifications in oxOva compared to Ova. T cells from Ova-specific OT-II but not from C57BL/6 or SKH-1 wild type mice presents enhanced activation after Ova addition. OxOva exacerbates this activation when administered ex vivo or in vivo, along with an increased interferon-gamma production, a known anti-melanoma agent. OxOva vaccination of wild type mice followed by inoculation of syngeneic B16F10 Ova-expressing melanoma cells shows enhanced T cell number and activation, decreased tumor burden, and elevated numbers of antigen-presenting cells when compared to their Ova-vaccinated counterparts. Analysis of oxOva using mass spectrometry identifies three hot spots regions rich in oxidative modifications that are associated with the increased T cell activation. Using Ova as a model protein, the findings suggest an immunomodulating role of multi-ROS/RNS modifications that may spur novel research lines in inflammation research and for vaccination strategies in oncology.
Comprehensive characterization of osseous tissues from impedance measurements by effective medium approximation
2021, Wei, Wenzuo, Shi, Fukun, Zhuang, Jie, Kolb, Juergen F.
A unified mixing (UM) model was developed to derive microstructural information of trabecular bone, i.e., bone volume fraction (BV/TV), from electrical impedance spectroscopy. A distinct advantage of the UM-model over traditional methods, such as equivalent circuit models and multivariate analysis, is that the influence of both the environment (hydroxyapatite) and different inclusions (water, fat, and air) can be taken into account simultaneously. In addition, interactions between the different components such as interfacial polarization can be addressed by a dedicated fitting parameter v. Accordingly, values of BV/TV for different bone samples, e.g., including or not including water, were determined in the higher frequency range of 1-5 MHz. Results showed good agreement with experimental data obtained by micro-computer tomography. In particular, predictions for dielectric parameters that were derived for 3 and 4 MHz were found most promising for the assessment and distinction of osteopathic conditions and differences. This was shown by a clear differentiation of osseous tissues, e.g., the greater trochanter, femoral head, and femoral neck.
Efficiency of cold atmospheric plasma, cleaning powders and their combination for biofilm removal on two different titanium implant surfaces
2022, Kamionka, Julia, Matthes, Rutger, Holtfreter, Birte, Pink, Christiane, Schlüter, Rabea, von Woedtke, Thomas, Kocher, Thomas, Jablonowski, Lukasz
Objectives: Biofilm removal is the decisive factor for the control of peri-implantitis. Cold atmospheric pressure plasma (CAP) can become an effective aid due to its ability to destroy and to inactivate bacterial biofilm residues. This study evaluated the cleaning efficiency of CAP, and air-polishing with glycine (APG) or erythritol (APE) containing powders alone or in combination with CAP (APG + CAP, APE + CAP) on sandblasted/acid etched, and anodised titanium implant surface. Materials and methods: On respective titanium discs, a 7-day ex vivo human biofilm was grown. Afterwards, the samples were treated with CAP, APG, APE, APG + CAP, and APE + CAP. Sterile and untreated biofilm discs were used for verification. Directly after treatment and after 5 days of incubation in medium at 37 °C, samples were prepared for examination by fluorescence microscopy. The relative biofilm fluorescence was measured for quantitative analyses. Results: Air-polishing with or without CAP removed biofilms effectively. The combination of air-polishing with CAP showed the best cleaning results compared to single treatments, even on day 5. Immediately after treatment, APE + CAP showed insignificant higher cleansing efficiency than APG + CAP. Conclusions: CAP supports mechanical cleansing and disinfection to remove and inactivate microbial biofilm on implant surfaces significantly. Here, the type of the powder was not important. The highest cleansing results were obtained on sandblasted/etched surfaces. Clinical relevance. Microbial residuals impede wound healing and re-osseointegration after peri-implantitis treatment. Air-polishing treatment removes biofilms very effectively, but not completely. In combination with CAP, microbial free surfaces can be achieved. The tested treatment regime offers an advantage during treatment of peri-implantitis.
Combined In Vitro Toxicity and Immunogenicity of Cold Plasma and Pulsed Electric Fields
2022, Wolff, Christina M., Kolb, Juergen F., Bekeschus, Sander
In modern oncology, therapies are based on combining monotherapies to overcome treatment resistance and increase therapy precision. The application of microsecond-pulsed electric fields (PEF) is approved to enhance local chemotherapeutic drug uptake within combination electrochemotherapy regimens. Reactive oxygen species (ROS) have been implicated in anticancer effects, and cold physical plasma produces vast amounts of ROS, which have recently been shown to benefit head and neck cancer patients. PEF and cold plasma technology have been linked to immunogenic cell death (ICD) induction, a regulated cell death accompanied by sterile inflammation that promotes antitumor immunity. To this end, we investigated the combined effect of both treatments regarding their intracellular ROS accumulation, toxicity, ICD-related marker expression, and optimal exposure sequence in a leukemia model cell line. The combination treatment substantially increased ROS and intracellular glutathione levels, leading to additive cytotoxic effects accompanied by a significantly increased expression of ICD markers, such as the eat-me signal calreticulin (CRT). Preconditioned treatment with cold plasma followed by PEF exposure was the most potent treatment sequence. The results indicate additive effects of cold plasma and PEF, motivating further studies in skin and breast tumor models for the future improvement of ECT in such patients.
Application of scanning electrochemical microscopy for topography imaging of supported lipid bilayers
2022, Nasri, Zahra, Memari, Seyedali, Striesow, Johanna, Weltmann, Klaus-Dieter, von Woedtke, Thomas, Wende, Kristian
Oxidative stress in cellular environments may cause lipid oxidation and membrane degradation. Therefore, studying the degree of lipid membrane morphological changes by reactive oxygen and nitrogen species will be informative in oxidative stress-based therapies. This study introduces the possibility of using scanning electrochemical microscopy as a powerful imaging technique to follow the topographical changes of a solid-supported lipid bilayer model induced by reactive species produced from gas plasma. The introduced strategy is not limited to investigating the effect of reactive species on the lipid bilayer but could be extended to understand the morphological changes of the lipid bilayer due to the action of membrane proteins or antimicrobial peptides.
Functional thin films synthesized from liquid precursors by combining mist chambers and atmospheric-pressure plasma polymerization
2021, Barillas, Laura, Makhneva, Ekaterina, An, Sehoon, Fricke, Katja
For the creation of thin films, the use of precursors in liquid phase offers a viable alternative when these chemicals are sensitive to high temperatures and phase changes. However, it requires appropriate liquid handling and deposition technologies capable of dispensing the fluid homogeneously to produce a uniform thin film. We report different tailor-made mist chamber designs integrated in an atmospheric-pressure plasma polymerization process for the synthesis of functional thin polymer films from liquid precursors. A systematic investigation, evaluated by performance indicators, is presented on the characteristics and suitability of metallic 3D-printed mist chambers depending on inner volume, geometry and surface post-treatment, for the deposition of a thin liquid monomer film. To assess the quality of the subsequently obtained plasma-polymerized (pp) films, their properties were characterized in terms of thickness, chemical composition, surface morphology and stability in aqueous environment. It was found that the specification of the mist chambers along with the plasma process parameters influences the pp film’s thickness, surface morphology and degree of monomer conversion. This study is one of the first demonstrations of a controllable process able to tune the cross-linked polymeric chains of plasma-polymers at atmospheric pressure, highlighting the opportunities of using mist chambers and plasma technology to discover tailored organic thin films to materials sciences and life sciences.