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- ItemGas-discharge plasma-assisted functionalization of titanium implant surfaces(Baech : Trans Tech Publications Ltd., 2010) Schröder, Karsten; Finke, Birgit; Polak, Martin; Lüthen, Frank; Nebe, Barbara; Rychly, Joachim; Bader, Rainer; Lukowski, Gerold; Walschus, Uwe; Schlosser, Michael; Ohl, Andreas; Weltmann, Klaus DieterA crucial factor for in-growth of metallic implants in the bone stock is the rapid cellular acceptance whilst prevention of bacterial adhesion on the surface. Such contradictorily adhesion events could be triggered by surface properties. There already exists fundamental knowledge about the influence of physicochemical surface properties like roughness, titanium dioxide modifications, cleanness, and (mainly ceramic) coatings on cell and microbial behavior in vitro and in vivo. The titanium surface can be equipped with antimicrobial properties by plasma-based copper implantation, which allows the release and generation of small concentrations of copper ions during contact with water-based biological liquids. Additionally, the titanium surface was equipped with amino groups by the deposition of an ultrathin plasma polymer. This coating on the one hand does not significantly reduce the generation of copper ions, and on the other hand improves the adhesion and spreading of osteoblast cells. The process development was accompanied by physicochemical surface analyses like XPS, FTIR, contact angle, SEM, and AFM. Very thin modified layers were created, which are resistant to hydrolysis and delamination. These titanium surface functionalizations were found to have either an antimicrobial activity or cell-adhesive properties. Intramuscular implantation of titanium samples coated with the cell-adhesive plasma polymer in rats revealed a reduced inflammation reaction compared to uncoated titanium. © (2010) Trans Tech Publications.
- ItemA size dependent evaluation of the cytotoxicity and uptake of nanographene oxide(London [u.a.] : RSC, 2015) Mendes, Rafael Gregorio; Koch, Britta; Bachmatiuk, Alicja; Ma, Xing; Sanchez, Samuel; Damm, Christine; Schmidt, Oliver G.; Gemming, Thomas; Eckert, Jürgen; Rümmeli, Mark H.Graphene oxide (GO) has attracted great interest due to its extraordinary potential for biomedical application. Although it is clear that the naturally occurring morphology of biological structures is crucial to their precise interactions and correct functioning, the geometrical aspects of nanoparticles are often ignored in the design of nanoparticles for biological applications. A few in vitro and in vivo studies have evaluated the cytotoxicity and biodistribution of GO, however very little is known about the influence of flake size and cytotoxicity. Herein, we aim at presenting an initial cytotoxicity evaluation of different nano-sized GO flakes for two different cell lines (HeLa (Kyoto) and macrophage (J7742)) when they are exposed to samples containing different sized nanographene oxide (NGO) flakes (mean diameter of 89 and 277 nm). The obtained data suggests that the larger NGO flakes reduce cell viability as compared to smaller flakes. In addition, the viability reduction correlates with the time and the concentration of the NGO nanoparticles to which the cells are exposed. Uptake studies were also conducted and the data suggests that both cell lines internalize the GO nanoparticles during the incubation periods studied.
- ItemBlood platelet enrichment in mass-producible surface acoustic wave (SAW) driven microfluidic chips(Cambridge : RSC, 2019) Richard, Cynthia; Fakhfouri, Armaghan; Colditz, Melanie; Striggow, Friedrich; Kronstein-Wiedemann, Romy; Tonn, Torsten; Medina-Sánchez, Mariana; Schmidt, Oliver G.; Gemming, Thomas; Winkler, AndreasThe ability to separate specific biological components from cell suspensions is indispensable for liquid biopsies, and for personalized diagnostics and therapy. This paper describes an advanced surface acoustic wave (SAW) based device designed for the enrichment of platelets (PLTs) from a dispersion of PLTs and red blood cells (RBCs) at whole blood concentrations, opening new possibilities for diverse applications involving cell manipulation with high throughput. The device is made of patterned SU-8 photoresist that is lithographically defined on the wafer scale with a new proposed methodology. The blood cells are initially focused and subsequently separated by an acoustic radiation force (ARF) applied through standing SAWs (SSAWs). By means of flow cytometric analysis, the PLT concentration factor was found to be 7.7, and it was proven that the PLTs maintain their initial state. A substantially higher cell throughput and considerably lower applied powers than comparable devices from literature were achieved. In addition, fully coupled 3D numerical simulations based on SAW wave field measurements were carried out to anticipate the coupling of the wave field into the fluid, and to obtain the resulting pressure field. A comparison to the acoustically simpler case of PDMS channel walls is given. The simulated results show an ideal match to the experimental observations and offer the first insights into the acoustic behavior of SU-8 as channel wall material. The proposed device is compatible with current (Lab-on-a-Chip) microfabrication techniques allowing for mass-scale, reproducible chip manufacturing which is crucial to push the technology from lab-based to real-world applications. © The Royal Society of Chemistry.
- ItemImpact of plasma jet vacuum ultraviolet radiation on reactive oxygen species generation in bio-relevant liquids([S.l.] : American Institute of Physics, 2015) Jablonowski, H.; Bussiahn, R.; Hammer, M.U.; Weltmann, K.-D.; von Woedtke, T.; Reuter, S.Plasma medicine utilizes the combined interaction of plasma produced reactive components. These are reactive atoms, molecules, ions, metastable species, and radiation. Here, ultraviolet (UV, 100–400 nm) and, in particular, vacuum ultraviolet (VUV, 10–200 nm) radiation generated by an atmospheric pressure argon plasma jet were investigated regarding plasma emission, absorption in a humidified atmosphere and in solutions relevant for plasma medicine. The energy absorption was obtained for simple solutions like distilled water (dH2O) or ultrapure water and sodium chloride (NaCl) solution as well as for more complex ones, for example, Rosewell Park Memorial Institute (RPMI 1640) cell culture media. As moderate stable reactive oxygen species, hydrogen peroxide (H2O2) was studied. Highly reactive oxygen radicals, namely, superoxide anion (O2•−) and hydroxyl radicals (•OH), were investigated by the use of electron paramagnetic resonance spectroscopy. All species amounts were detected for three different treatment cases: Plasma jet generated VUV and UV radiation, plasma jet generated UV radiation without VUV part, and complete plasma jet including all reactive components additionally to VUV and UV radiation. It was found that a considerable amount of radicals are generated by the plasma generated photoemission. From the experiments, estimation on the low hazard potential of plasma generated VUV radiation is discussed.
- ItemRedox Stimulation of Human THP-1 Monocytes in Response to Cold Physical Plasma(Austin, Tex. : Landes Bioscience, 2015) Bekeschus, Sander; Schmidt, Anke; Bethge, Lydia; Masur, Kai; von Woedtke, Thomas; Hasse, Sybille; Wende, KristianIn plasma medicine, cold physical plasma delivers a delicate mixture of reactive components to cells and tissues. Recent studies suggested a beneficial role of cold plasma in wound healing. Yet, the biological processes related to the redox modulation via plasma are not fully understood. We here used the monocytic cell line THP-1 as a model to test their response to cold plasma in vitro. Intriguingly, short term plasma treatment stimulated cell growth. Longer exposure only modestly compromised cell viability but apparently supported the growth of cells that were enlarged in size and that showed enhanced metabolic activity. A significantly increased mitochondrial content in plasma treated cells supported this notion. On THP-1 cell proteome level, we identified an increase of protein translation with key regulatory proteins being involved in redox regulation (hypoxia inducible factor 2α), differentiation (retinoic acid signaling and interferon inducible factors), and cell growth (Yin Yang 1). Regulation of inflammation is a key element in many chronic diseases, and we found a significantly increased expression of the anti-inflammatory heme oxygenase 1 (HMOX1) and of the neutrophil attractant chemokine interleukin-8 (IL-8). Together, these results foster the view that cold physical plasma modulates the redox balance and inflammatory processes in wound related cells.
- ItemThe LEGATO cross-disciplinary integrated ecosystem service research framework: an example of integrating research results from the analysis of global change impacts and the social, cultural and economic system dynamics of irrigated rice production(Heidelberg : Springer Verlag, 2017) Spangenberg, J.H.; Beaurepaire, A.L.; Bergmeier, E.; Burkhard, B.; van Chien, H.; Cuong, L.Q.; Görg, C.; Grescho, V.; Hai, L.H.; Heong, K.L.; Horgan, F.G.; Hotes, S.; Klotzbücher, A.; Klotzbücher, T.; Kühn, I.; Langerwisch, F.; Marion, G.; Moritz, R.F.A.; Nguyen, Q.A.; Ott, J.; Sann, C.; Sattler, C.; Schädler, M.; Schmidt, A.; Tekken, V.; Thanh, T.D.; Thonicke, K.; Türke, M.; VáclavÃk, T.; Vetterlein, D.; Westphal, C.; Wiemers, M.; Settele, J.In a cross-disciplinary project (LEGATO) combining inter- and transdisciplinary methods, we quantify the dependency of rice-dominated socio-ecological systems on ecosystem functions (ESF) and the ecosystem services (ESS) the integrated system provides. In the collaboration of a large team including geo- and bioscientists, economists, political and cultural scientists, the mutual influences of the biological, climate and soil conditions of the agricultural area and its surrounding natural landscape have been analysed. One focus was on sociocultural and economic backgrounds, another on local as well as regional land use intensity and biodiversity, and the potential impacts of future climate and land use change. LEGATO analysed characteristic elements of three service strands defined by the Millennium Ecosystem Assessment (MA): (a) provisioning services: nutrient cycling and crop production; (b) regulating services: biocontrol and pollination; and (c) cultural services: cultural identity and aesthetics. However, in line with much of the current ESS literature, what the MA called supporting services is treated as ESF within LEGATO. As a core output, LEGATO developed generally applicable principles of ecological engineering (EE), suitable for application in the context of future climate and land use change. EE is an emerging discipline, concerned with the design, monitoring and construction of ecosystems and aims at developing strategies to optimise ecosystem services through exploiting natural regulation mechanisms instead of suppressing them. Along these lines LEGATO also aims to create the knowledge base for decision-making for sustainable land management and livelihoods, including the provision of the corresponding governance and management strategies, technologies and system solutions.
- ItemPeriodic Exposure of Keratinocytes to Cold Physical Plasma: An In Vitro Model for Redox-Related Diseases of the Skin(London: Hindawi, 2016) Schmidt, Anke; von Woedtke, Thomas; Bekeschus, SanderOxidative stress illustrates an imbalance between radical formation and removal. Frequent redox stress is critically involved in many human pathologies including cancer, psoriasis, and chronic wounds. However, reactive species pursue a dual role being involved in signaling on the one hand and oxidative damage on the other. Using a HaCaT keratinocyte cell culture model, we investigated redox regulation and inflammation to periodic, low-dose oxidative stress after two, six, eight, ten, and twelve weeks. Chronic redox stress was generated by recurrent incubation with cold physical plasma-treated cell culture medium. Using transcriptome microarray technology, we identified both acute ROS-stress responses as well as numerous adaptions after several weeks of redox challenge. We determined a differential expression (2-fold, FDR < 0.01, p < 0.05) of 260 genes that function in inflammation and redox homeostasis, such as cytokines (e.g., IL-6, IL-8, and IL-10), growth factors (e.g., CSF2, FGF, and IGF-2), and antioxidant enzymes (e.g., HMOX, NQO1, GPX, and PRDX). Apoptotic signaling was affected rather modestly, especially in p53 downstream targets (e.g., BCL2, BBC3, and GADD45). Strikingly, the cell-protective heat shock protein HSP27 was strongly upregulated (p < 0.001). These results suggested cellular adaptions to frequent redox stress and may help to better understand the inflammatory responses in redox-related diseases.
- ItemRaman-spectroscopy based cell identification on a microhole array chip(Basel : MDPI AG, 2014) Neugebauer, U.; Kurz, C.; Bocklitz, T.; Berger, T.; Velten, T.; Clement, J.H.; Krafft, C.; Popp, J.Circulating tumor cells (CTCs) from blood of cancer patients are valuable prognostic markers and enable monitoring responses to therapy. The extremely low number of CTCs makes their isolation and characterization a major technological challenge. For label-free cell identification a novel combination of Raman spectroscopy with a microhole array platform is described that is expected to support high-throughput and multiplex analyses. Raman spectra were registered from regularly arranged cells on the chip with low background noise from the silicon nitride chip membrane. A classification model was trained to distinguish leukocytes from myeloblasts (OCI-AML3) and breast cancer cells (MCF-7 and BT-20). The model was validated by Raman spectra of a mixed cell population. The high spectral quality, low destructivity and high classification accuracy suggests that this approach is promising for Raman activated cell sorting.
- ItemRolled-up functionalized nanomembranes as three-dimensional cavities for single cell studies(Washington, DC : American Chemical Society, 2014) Xi, W.; Schmidt, C.K.; Sanchez, S.; Gracias, D.H.; Carazo-Salas, R.E.; Jackson, S.P.; Schmidt, O.G.We use micropatterning and strain engineering to encapsulate single living mammalian cells into transparent tubular architectures consisting of three-dimensional (3D) rolled-up nanomembranes. By using optical microscopy, we demonstrate that these structures are suitable for the scrutiny of cellular dynamics within confined 3D-microenvironments. We show that spatial confinement of mitotic mammalian cells inside tubular architectures can perturb metaphase plate formation, delay mitotic progression, and cause chromosomal instability in both a transformed and nontransformed human cell line. These findings could provide important clues into how spatial constraints dictate cellular behavior and function.
- ItemContractile cell forces deform macroscopic cantilevers and quantify biomaterial performance(London : Royal Soc. of Chemistry, 2015) Allenstein, U.; Mayr, S.G.; Zink, M.Cells require adhesion to survive, proliferate and migrate, as well as for wound healing and many other functions. The strength of contractile cell forces on an underlying surface is a highly relevant quantity to measure the affinity of cells to a rigid surface with and without coating. Here we show with experimental and theoretical studies that these forces create surface stresses that are sufficient to induce measurable bending of macroscopic cantilevers. Since contractile forces are linked to the formation of focal contacts, results give information on adhesion promoting qualities and allow a comparison of very diverse materials. In exemplary studies, in vitro fibroblast adhesion on the magnetic shape memory alloy Fe–Pd and on the L-lysine derived plasma-functionalized polymer PPLL was determined. We show that cells on Fe–Pd are able to induce surface stresses three times as high as on pure titanium cantilevers. A further increase was observed for PPLL, where the contractile forces are four times higher than on the titanium reference. In addition, we performed finite element simulations on the beam bending to back up the calculation of contractile forces from cantilever bending under non-homogenous surface stress. Our findings consolidate the role of contractile forces as a meaningful measure of biomaterial performance.