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An efficient approach for tailoring surface properties of polymers is presented, which enables rapid modification leading to superhydrophilic properties. The approach is based on vacuum-ultraviolet radiation (VUV) pretreatment of the surface to create reactive dangling bonds. This step is followed by a second treatment using neutral oxygen atoms that react with the dangling bonds and form functional groups. The beneficial effect of VUV pretreatment for enhanced functionalization was clearly demonstrated by comparing VUV pretreatment in plasmas created in different gases, i.e., hydrogen, nitrogen, and oxygen, which differ in the intensity of VUV/UV radiation. The emission intensity of VUV radiation for all gases was measured by vacuum ultraviolet spectroscopy. It was shown that VUV has a strong influence on the treatment time and final surface wettability. A superhydrophilic surface was obtained only if using VUV pretreatment. Furthermore, the treatment time was significantly reduced to only a second of treatment. These findings show that such an approach may be used to enhance the surface reaction efficiency for further grafting of chemical groups.

An extensive study on using plant waste aqueous extracts as natural chemicals for in-situ synthesis of zinc oxide (ZnO) on cotton is presented. Reducing agents were prepared from green tea leaves (GT), pomegranate peels (PG), and staghorn sumac leaves (SsL) and drupes (SsD), and the alkaline medium from discarded wood ash. Zinc acetate was found to be more appropriate precursor than zinc nitrate. Formation of ZnO on cotton was confirmed by energy dispersive spectroscopy, X-ray photoelectron spectroscopy and X-ray diffraction analysis (XRD). The inductively coupled plasma mass spectrometry and X-ray fluorescence results showed the highest amount of ZnO on cotton was formed using PG and SsL extracts, which was also confirmed with scanning electron microscopy and UV/visible spectroscopy. The ZnO-functionalised samples exhibited excellent UV-blocking ability and different wetting properties (hydrophilic or hydrophobic) depending on the reducing agent used due to their different total phenolic content. This study shows that by choosing the plant waste source as a reducing agent for ZnO formation directly on cotton, the properties of cotton can be designed to be hydrophilic or hydrophobic with excellent UV-blocking properties. The XRD results of ex-situ synthesis prove that the short reaction time enables the formation of ZnO.

Plasma agriculture is a promising niche of interdisciplinary research where the physics, of non-equilibrium gases meets surface chemistry and biological responses. Despite numerous scientific papers, the interaction of gaseous plasma with seeds is not understood enough to make the technique useful in practical agriculture. An obstacle is an improper methodology adopted by different authors. In this paper, we show that the surface wettability does not depend on discharge parameters such as power and pressure, but rather on the fluence of oxygen atoms onto the seed surface. The proper methodology is demonstrated for the case of corn seeds. The surface activation, which enables improved water uptake or good adhesion of a coating, progresses relatively linearly up to the O-atom fluence of 3 × 1024 m−3 and remains constant thereafter. The minimal water contact angle achievable using oxygen plasma treatment is a few degrees.

Efficient and selective internalization of nanoscale diamonds (also termed nanodiamonds, NDs) by living cells is of fundamental importance for their bionanotechnological applications. The biocompatibility of NDs is well established and has been suggested to arise from the limited membrane perturbation during their cellular translocation. However, the latter may be affected when cells are subjected to external stress. This study shows that the oxidative stress generated by atmospheric pressure cold plasmas (APCP) alters cell sensitivity to NDs, and their cytotoxicity profile. Both positively and negatively charged NDs are nontoxic to cells, here Saccharomyces cerevisiae and human cell lines, i.e., near-normal human mammary epithelial cells (MCF-10A) and breast cancer cells (MDA-MB-468 and T47D), unless the APCP stress is introduced. A brief exposure of the cells to APCP leads to a significant increase in their ND affinity (uptake and/or surface attachment) and intracellular ROS accumulation, particularly for positively charged NDs and both yeast and cancer cells. A concomitant decrease in cell viability and yeast cell growth, reflected by longer lag phases and lower cell density after 24 h of incubation, demonstrates a considerably enhanced ND toxicity to these cells. These results suggest that chemo-radiative stress, such as that produced by plasma, may influence the toxicity of nanoparticles to different cells, with specificity achieved through controlling particle charges. Moreover, since oxidative stress is not only associated with the use of APCP but can arise unintentionally within an organism and/or in the environment, these findings may have broader implications for the use of nontoxic nanoparticles in bionanotechnology in general.

The literature on plasma-stimulated modification of seed wettability and germination is reviewed and analyzed. The results reported by different authors are scattered, but there is a positive correlation between the change in the water contact angle and the germination improvement. There is hardly any correlation between the germination and the power density used for sustaining gaseous plasma. The wettability, on the other hand, exhibits optimal values at the power density of roughly 1 W cm−3, but the results differ significantly. In fact, a super hydrophilic surface finish was only reported at such moderate power densities. Both the wettability and germination increase with treatment time, but saturate at a certain level, depending on the discharge power. Unlike for most polymers, the hydrophobic recovery does not appear to be a considerable obstacle, so the plasma treatment may be performed well before sowing, which makes the technique applicable in agricultural practice. The scattering of results indicates that the commonly reported parameters such as the type and mode of discharge, the discharge power or power density, the type of gas and its pressure, the treatment time and the type of the seeds, are not decisive parameters governing wettability and germination. Based on the observations of the reviewing authors, conclusions and scientific challenges in this scientific niche are summarized.

Seeds of common bean (Phaseolus vulgaris L.), of the Etna variety, were treated with low-pressure oxygen plasma sustained by an inductively coupled radiofrequency discharge in the H-mode for a few seconds. The high-intensity treatment improved seed health in regard to fungal contamination. Additionally, it increased the wettability of the bean seeds by altering surface chemistry, as established by X-ray photoelectron spectroscopy, and increasing surface roughness, as seen with a scanning electron microscope. The water contact angle at the seed surface dropped to immeasurably low values after a second of plasma treatment. Hydrophobic recovery within a month returned those values to no more than half of the original water contact angle, even for beans treated for the shortest time (0.5 s). Increased wettability resulted in accelerated water uptake. The treatment increased the bean radicle length, which is useful for seedling establishment in the field. These findings confirm that even a brief plasma treatment is a useful technique for the disinfection and stimulation of radicle growth. The technique is scalable to large systems due to the short treatment times.

Cold atmospheric plasma is an ionized gas that shows promise in regenerative medical treatments, yet the mechanisms underlying its effects are still poorly understood. Plasma treatment promotes cell growth or cell death depending on the cell type and exposure parameters. To date, no early cell response to plasma, such as stress granule (SG) formation has been addressed. Cytoplasmic SGs are formed as an immediate cell response to acute stress stimuli by recruitment of over 140 proteins intertwined with cytoplasmic RNAs that leads to transient suspension of protein translation. Encouraged by the plasma effects in regenerative medicine and oncology, the atmospheric pressure plasma jet with argon gas flow is being utilized to treat SH-SY5Y cells with an inducible expression of the stress granule marker G3BP1, to gain an insight into early cell response to plasma and SG formation dynamics. Plasma effectively induces SG formation in the exposed cells in a flow/time-dependent manner, with the SG assembly clearly prompted by plasma-induced oxidative stress. Plasma causes SG formation via eIF2α-signaling, which is repressed with the SG formation inhibitor ISRIB. This insight into the early cell response to plasma treatment may lead to improved therapies in regenerative medicine and cancer treatment.

Improving the biocompatibility of polyethylene terephthalate (PET) vascular grafts is an important task for avoiding thrombus formation. Therefore, SO2 plasma at various treatment periods were used to modify PET surface properties by forming sulfate functional groups. These groups were shown to act antithrombogenically, ensuring good hemocompatibility of the materials, although the biocompatibility of such materials still remains a mystery. For this reason, the adhesion and viability of HUVEC cells on SO2 plasma-modified PET surfaces were studied, and the possible toxicity of the tested material was determined using two different assays, MTT (metabolic activity assay) and SRB (in-vitro toxicology assay). Changes in chemical composition, morphology and wettability were determined as well. Improved endothelialization was observed for all plasma-treated samples, with the most optimal being the sample treated for 80 s, which can be explained by it having the best combination of surface functionalization, roughness and morphology. Furthermore, toxicity was observed to some extent on the sample treated for 160 s, indicating the lowest cell density among the plasma-treated samples. X-ray photoelectron spectroscopy showed increased oxygen and sulfur content on the surface, which was independent on treatment time. Surface roughness of the plasma-treated samples increased, reaching its maximum after 80 s of treatment, and decreased thereafter.

Seeds of common bean (Phaseolus vulgaris L.), of the Etna variety, were treated with low-pressure oxygen plasma sustained by an inductively coupled radiofrequency discharge in the H-mode for a few seconds. The high-intensity treatment improved seed health in regard to fungal contamination. Additionally, it increased the wettability of the bean seeds by altering surface chemistry, as established by X-ray photoelectron spectroscopy, and increasing surface roughness, as seen with a scanning electron microscope. The water contact angle at the seed surface dropped to immeasurably low values after a second of plasma treatment. Hydrophobic recovery within a month returned those values to no more than half of the original water contact angle, even for beans treated for the shortest time (0.5 s). Increased wettability resulted in accelerated water uptake. The treatment increased the bean radicle length, which is useful for seedling establishment in the field. These findings confirm that even a brief plasma treatment is a useful technique for the disinfection and stimulation of radicle growth. The technique is scalable to large systems due to the short treatment times.

Achieving optimal hydrophobicity of polymer materials especially polymer–matrix composites is important for many material applications. Herein the interplay of factors determining hydrophobic surface is presented during CF4 plasma treatments which lead to functionalization as well as selective polymer–matrix etching. The continuous exposure to plasma reactive species induces functionalization and etching on the surface, which decides the surface morphology and surface chemistry. Consequently, exothermic processes during the plasma–surface interactions are another important factor which influences the surface chemistry and etching rate of the material. The results demonstrate that despite etching and increasing surface roughness, the major contribution to hydrophobic character is dependent on the number of carbon atoms populated with fluorine, whereas the temperature is a deciding factor for type of created bonds. Copyright © 2016 John Wiley & Sons, Ltd.

The gold nanoparticles and cold plasma have been widely investigated separately in cancer therapy. In this paper, the two agents were combined in order to achieve a more profound understanding of their interactions in glioblastoma (U87) and normal human astrocytes (E6/E7). Gold nanoparticles were endocytosed at an accelerated rate in the U87 cell membrane due to the plasma treatment. This caused many divots across the cell membrane which made it more porous. On the contrary, in E6/E7 cells, there was no significant difference for penetration of gold nanoparticles or the cell membrane morphology. As proven in previous experiments the addition of the nanoparticles decrease cell viability, allowing the cold plasma to be more efficient. Present work demonstrated the synergistic effect of the two agents, which can be correlated in order to maximize the efficiency.

Production of ethanol by the yeast Saccharomyces cerevisiae is a process of global importance. In these processes, productivities and yields are pushed to their maximum possible values leading to cellular stress. Transient and lasting enhancements in tolerance and performance have been obtained by genetic engineering, forced evolution, and exposure to moderate levels of chemical and/or physical stimuli, yet the drawbacks of these methods include cost, and multi-step, complex and lengthy treatment protocols. Here, plasma agitation is shown to rapidly induce desirable phenotypic changes in S. cerevisiae after a single treatment, resulting in improved conversion of glucose to ethanol. With a complex environment rich in energetic electrons, highly-reactive chemical species, photons, and gas flow effects, plasma treatment simultaneously mimics exposure to multiple environmental stressors. A single treatment of up to 10 minutes performed using an atmospheric pressure plasma jet was sufficient to induce changes in cell membrane structure, and increased hexokinase 2 activity and secondary metabolite production. These results suggest that plasma treatment is a promising strategy that can contribute to improving metabolic activity in industrial microbial strains, and thus the practicality and economics of industrial fermentations.

The performance of electrical insulating polymers measured by comparative tracking index can be improved by flame retardant fillers in the polymer–matrix, but is not necessary effective due to persisting surface layers. We demonstrated a method for selective etching of polyphenol–matrix composites which leads to improvement of the tracking properties. Selective etching was performed by low pressure inductively coupled oxygen plasma generated at 35 Pa. Species created in discharge were monitored in situ during the treatment by optical emission spectroscopy (OES). Their surface reactions were established indirectly from X-ray photoelectron spectroscopy (XPS) and energy dispersive X-ray spectroscopy (EDXS) after plasma treatment. The effective etching was done by oxygen species which removed off surface polymer layer within 10 to 15 s during plasma pulse operation to prevent surface overheating. The selective polymer removal improved tracking properties of the 70% glass filler reinforced composites up to 56%, whereas the performance level decreased with the decrease in filler content. This technology provides an important enhancement of polymer–matrix composites' tracking properties and resistance of composites to electrical short-circuit fire.

Samples of polymer polyethylene terephthalate were exposed to a weakly ionized gaseous plasma to modify the polymer surface properties for better cell cultivation. The gases used for treatment were sulfur dioxide and oxygen of various partial pressures. Plasma was created by an electrodeless radio frequency discharge at a total pressure of 60 Pa. X-ray photoelectron spectroscopy showed weak functionalization of the samples' surfaces with the sulfur, with a concentration around 2.5 at %, whereas the oxygen concentration remained at the level of untreated samples, except when the gas mixture with oxygen concentration above 90% was used. Atomic force microscopy revealed highly altered morphology of plasma-treated samples; however, at high oxygen partial pressures this morphology vanished. The samples were then incubated with human umbilical vein endothelial cells. Biological tests to determine endothelialization and possible toxicity of the plasma-treated polyethylene terephthalate samples were performed. Cell metabolic activity (MTT) and in vitro toxic effects of unknown compounds (TOX) were assayed to determine the biocompatibility of the treated substrates. The biocompatibility demonstrated a well-pronounced maximum versus gas composition which correlated well with development of the surface morphology.

Plasma treatment is often used to alter cell interaction with polymer surfaces used in biomedical application. The influence of surface hydrophilicity/hydrophobicity on human mammary epithelial cell (HMEC) proliferation and adhesion of protein albumin to plasma treated polystyrene (PS) was studied. The PS surface was made hydrophilic or hydrophobic by treatment either in O2 or CF4 plasma. The rate of protein adhesion was studied by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) after incubation of PS in albumin solution for different periods, while cell viability and morphology was studied by MTT assay and scanning electron microscope (SEM). XPS measurements have shown that the quantity of adsorbed protein was higher for both plasma treated samples than for the untreated one. No significant difference regarding protein adhesion on hydrophilic or hydrophobic plasma treated surface was found by XPS. Contrary, the results for cell proliferation showed much better proliferation on hydrophilic surface.

Polystyrene (PS) foils were exposed to oxygen plasma in order to improve the ability of human osteosarcoma cells (HOS) proliferation. Plasma was created in an electrodeless radio-frequency (RF) discharge in pure oxygen. Plasma treatment time of 30 s allowed for saturation of the surface with oxygen rich functional groups. Incubation of proteins from cell culture media was studied versus incubation time for untreated and plasma treated PS by X-ray photoelectron spectroscopy (XPS). Then HOS cells were cultivated according to a standard procedure, applied on the PS substrates and incubated. The proliferation was studied qualitatively by optical microscopy, scanning electron microscopy (SEM) and quantitatively by MTT assay. All techniques revealed significant differences in HOS cells proliferation on untreated and oxygen plasma treated PS. The differences were most pronounced after 24 h of incubation when the cells have already stared to proliferate on plasma treated PS, while they were still in quiescence on untreated PS. The confluency was achieved till the 6th day on plasma treated PS, while on untreated PS such confluency has never been achieved. The cell viability assay revealed that HOS cells grew better on plasma treated PS for various incubation times. Similar trend was observed regarding the adhesion of proteins. They adhered better on plasma treated PS causing better environment for further cell proliferation. The results clearly show that oxygen plasma treatment allows for much better proliferation of HOS cells on plasma treated PS as compared to the untreated PS substrates.

Films of proteins fibrinogen and albumin are exposed to neutral reactive particles in order to study their etching rates in early and late oxygen afterglows of microwave plasma. The etching rates are determined at different pressures between 50 and 400 Pa. The etching rates for both proteins first increase with the increasing pressure, while later they reach a constant value. Furthermore, the etching rates are higher in the early than in the late afterglow. The concentration of all available species in the afterglow, i.e. neutral oxygen atoms in the ground state, neutral molecules in the first excited state, and ozone is determined either by experimental measurements or theoretically using a well-established model. The etching rates are explained by the synergistic effects of atoms and metastable molecules.

Faculty of Mechanical Engineering, University of Maribor, 2000 Maribor, SloveniaReceived 23 May 2013; revised 21 July 2013; accepted 7 August 2013Published online in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.a.34911Abstract: The formation of endothelial cell monolayer onprosthetic implants has not sufficiently explored. The mainreasons leading to the development of thrombosis and=orintimal hyperplasia is the lack of endothelialization. In thepresent work, we have studied the influence of oxygen andfluorine plasma treatment of polyethylene terephthalate (PET)polymers on human microvascular endothelial cell adhesionand proliferation. We characterized the polymer surface, wett-ability, and oxidation potential upon plasma treatment. More-over, binding of serum and media compounds on PETsurface was monitored by Quartz crystal microbalancemethod, X-ray photoelectron spectroscopy, and atomic forcemicroscopy. Cell adhesion and morphology was assessed bylight and scanning electron microscopy. The influence ofplasma treatment on induction of cellular oxidative stressand cell proliferation was evaluated. The results obtainedshowed that treatment with oxygen plasma decreased theoxidation potential of the PET surface and revealed the high-est affinity for binding of serum components. Accordingly,the cells reflected the best adhesion and morphological prop-erties on oxygen-treated PET polymers. Moreover, treatmentwith oxygen plasma did not induce intracellular reactive oxy-gen species production while it stimulated endothelial cellproliferation by 25% suggesting the possible use of oxygenplasma treatment to enhance endothelialization of syntheticvascular grafts.

Thin films of human serum albumin (HSA) were immobilized on polystyrene (PS) substrates previously functionalized either with polar or nonpolar functional groups. The functionalization was performed by treatment with cold gaseous plasma created in pure oxygen and tetrafluoromethane (CF4) plasmas, respectively. Samples were examined with soft X-rays in the photon energy range of 520 to 710 eV in the ANTARES beam line at SOLEIL Synchrotron. NEXAFS spectra of O K-edge and F K-edge were collected at different spots of the sample, and measurements at each spot were repeated many times. A strong degradation of the HSA protein was observed. The weakly irradiated samples exhibited strong absorption at 531.5 eV associated with the O 1s→π*amide transitions, and a broad non distinctive peak at 540 eV was attributed to the O 1s→σ*C–O transitions. Both peaks decreased with increasing irradiation time until they were completely replaced by a broad non-distinctive peak at around 532 eV, indicating the destruction of the original protein conformation. The shortage of the amide groups indicated breakage of the peptide bonds.

Inducing selective or targeted cell apoptosis without affecting large number of neighbouring cells remains a challenge. A plausible method for treatment of posterior capsular opacification (PCO) due to remaining lens epithelial cells (LECs) by reactive chemistry induced by localized single electrode microplasma discharge at top of a needle-like glass electrode with spot size ~3 μm is hereby presented. The focused and highly-localized atmospheric pressure microplasma jet with electrode discharge could induce a dose-dependent apoptosis in selected and targeted individual LECs, which could be confirmed by real-time monitoring of the morphological and structural changes at cellular level. Direct cell treatment with microplasma inside the medium appeared more effective in inducing apoptosis (caspase 8 positivity and DNA fragmentation) at a highly targeted cell level compared to treatment on top of the medium (indirect treatment). Our results show that single cell specific micropipette plasma can be used to selectively induce demise in LECs which remain in the capsular bag after cataract surgery and thus prevent their migration (CXCR4 positivity) to the posterior lens capsule and PCO formation.

The atomic force microscope (AFM) is broadly used to study the morphology of cells. The morphological characteristics and differences of the cell membrane between normal human astrocytes and glial tumor cells are not well explored. Following treatment with cold atmospheric plasma, evaluation of the selective effect of plasma on cell viability of tumor cells is poorly understood and requires further evaluation. Using AFM we imaged morphology of glial cells before and after cold atmospheric plasma treatment. To look more closely at the effect of plasma on cell membrane, high resolution imaging was used. We report the differences between normal human astrocytes and human glioblastoma cells by considering the membrane surface details. Our data, obtained for the first time on these cells using atomic force microscopy, argue for an architectural feature on the cell membrane, i.e. brush layers, different in normal human astrocytes as compared to glioblastoma cells. The brush layer disappears from the cell membrane surface of normal E6/E7 cells and is maintained in the glioblastoma U87 cells after plasma treatment.

Protein adhesion and cell response to plasma-treated polymer surfaces were studied. The polymer polyethylene terephthalate (PET) was treated in either an oxygen plasma to make the surface hydrophilic, or a tetrafluoromethane CF(4) plasma to make the surface hydrophobic. The plasma source was radiofrequency (RF) discharge. The adsorption of albumin and other proteins from a cell-culture medium onto these surfaces was studied using a quartz crystal microbalance (QCM), X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The cellular response to plasma-treated surfaces was studied as well using an MTT assay and scanning electron microscopy (SEM). The fastest adsorption rate was found on the hydrophilic oxygen plasma-treated sample, and the lowest was found on the pristine untreated sample. Additionally, the amount of adsorbed proteins was higher for the oxygen-plasma-treated surface, and the adsorbed layer was more viscoelastic. In addition, cell adhesion studies support this finding because the best cell adhesion was observed on oxygen-plasma-treated substrates.

In the present study the biological response to various nanotopographic features after gaseous plasma treatment were studied. The usefulness of nanostructured surfaces for implantable materials has already been acknowledged, while less is known on the combined effect of nanostructured plasma modified surfaces. In the present work the influence of oxygen plasma treatment on nanostructured titanium oxide (TiO2) surfaces was studied. Characterization of the TiO2 surface chemical composition and morphological features was analyzed after plasma modification by x-ray photoelectron spectroscopy and by scanning electron microscopy while surface wettability was studied with measuring the water contact angle. Cell adhesion and morphology was assessed from images taken with scanning electron microscopy, whereas cell viability was measured with a calorimetric assay. The obtained results showed that oxygen plasma treatment of TiO2 nanotube surfaces significantly influences the adhesion and morphology of osteoblast-like cells in comparison to untreated nanostructured surfaces. Marked changes in surface composition of plasma treated surfaces were observed, as plasma treatment removed hydrocarbon contamination and removed fluorine impurities, which were present due to the electrochemical anodization process. However no differences in wettability of untreated and plasma treated surfaces were noticed. Treatment with oxygen plasma stimulated osteoblast-like cell adhesion and spreading on the nanostructured surface, suggesting the possible use of oxygen plasma surface treatment to enhance osteoblast-like cell response.

Some applications of extremely non-equilibrium oxygen plasma for tailoring the surface properties of organic as well as inorganic materials are presented. Plasma of low or moderate ionization fraction and very high dissociation fraction is created by high frequency electrodeless discharges created in chambers made from a material of low recombination coefficient. The O atom density often exceeds 1021 m−3 which allows for rapid functionalization of carbon-containing materials. Surface saturation with polar oxygen-rich groups is achieved in a fraction of a second and further exposure leads to etching. The etching is often non-uniform and results in nano-structuring of surface morphology. A combination of rich morphology and saturation with polar functional groups allows for a super-hydrophilic character of originally hydrophobic materials. Polymer composites are etched selectively so the polymer component is removed from the sample surface, leading to modified surface properties. Furthermore, such a treatment allows for distinguishing the distribution and orientation of fillers inside the polymer matrix. The exposure of inorganic materials to non-equilibrium oxygen plasma causes one-dimensional growth of metal oxide nanoparticles, thus representing a unique technique for the rapid catalyser-free growth of nanowires.