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Impact of plasma jet vacuum ultraviolet radiation on reactive oxygen species generation in bio-relevant liquids

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.

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Contractile cell forces deform macroscopic cantilevers and quantify biomaterial performance

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.

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Colloidal crystals of compliant microgel beads to study cell migration and mechanosensitivity in 3D

2019, Wagner, Katrin, Girardo, Salvatore, Goswami, Ruchi, Rosso, Gonzalo, Ulbricht, Elke, Müller, Paul, Soteriou, Despina, Träber, Nicole, Guck, Jochen

Tissues are defined not only by their biochemical composition, but also by their distinct mechanical properties. It is now widely accepted that cells sense their mechanical environment and respond to it. However, studying the effects of mechanics in in vitro 3D environments is challenging since current 3D hydrogel assays convolve mechanics with gel porosity and adhesion. Here, we present novel colloidal crystals as modular 3D scaffolds where these parameters are principally decoupled by using monodisperse, protein-coated PAAm microgel beads as building blocks, so that variable stiffness regions can be achieved within one 3D colloidal crystal. Characterization of the colloidal crystal and oxygen diffusion simulations suggested the suitability of the scaffold to support cell survival and growth. This was confirmed by live-cell imaging and fibroblast culture over a period of four days. Moreover, we demonstrate unambiguous durotactic fibroblast migration and mechanosensitive neurite outgrowth of dorsal root ganglion neurons in 3D. This modular approach of assembling 3D scaffolds from mechanically and biochemically well-defined building blocks allows the spatial patterning of stiffness decoupled from porosity and adhesion sites in principle and provides a platform to investigate mechanosensitivity in 3D environments approximating tissues in vitro.