Growth Stimulation of Durum Wheat and Common Buckwheat by Non-Thermal Atmospheric Pressure Plasma.

The grains of durum wheat (Triticum durum Desf.) and achenes of common buckwheat (Fagopyrum esculentum Moench) were tested after treatment with two sources of non-thermal atmospheric pressure plasma (DCSBD, MSDBD) with different treatment times (0, 3, 5, 10, 20, 30, and 40 s). The effect of these treatments was monitored with regard to the seed surface diagnostics (water contact angle-WCA, chemical changes by Fourier transform infrared spectroscopy-FTIR); twenty parameters associated with germination and initial seed growth were monitored. A study of the wettability confirmed a decrease in WCA values indicating an increase in surface energy and hydrophilicity depending on the type of seed, plasma source, and treatment time. Surface analysis by attenuated total reflectance FTIR (ATR-FTIR) showed no obvious changes in the chemical bonds on the surface of the plasma-treated seeds, which confirms the non-destructive effect of the plasma on the chemical composition of the seed shell. A multivariate analysis of the data showed many positive trends (not statistically significant) in germination and initial growth parameters. The repeated results for germination rate and root/shoot dry matter ratio indicate the tendency of plants to invest in underground organs. Durum wheat required longer treatment times with non-thermal plasma (10 s, 20 s) for germination and early growth, whereas buckwheat required shorter times (5 s, 10 s). The responses of durum wheat grains to the two non-thermal plasma sources used were equal. In contrast, the responses of buckwheat achenes were more favorable to MSDBD treatment than to DCSBD.

Modelling of inactivation kinetics of Escherichia coli, Salmonella Enteritidis and Bacillus subtilis treated with a multi-hollow surface dielectric barrier discharge plasma.

The efficacy of multi-hollow surface dielectric barrier discharge treatment against Escherichia coli, Salmonella Enteritidis and Bacillus subtilis was studied. Ambient air, O2, and N2 were used as working gas with a flow rate of 6 l/m. Power delivered into plasma was 30 W over an area of 2 × 2 cm2. The active species in plasma generated in different gases participating in the inactivation of microorganisms were evaluated by optical emission spectroscopy and Fourier transform infrared spectroscopy. Inactivation curves were fitted to the Bigelow log-linear, the biphasic, and Geeraerd models. According to the results, all plasma treatments inactivated tested microorganisms, depending on a working gas. The most sensitivity of bacteria was observed to the ambient air plasma. Inactivation up to 5 log for E. coli and S. Enteritidis could be achieved within 15 s of plasma treatment. Air plasma exposure of 25 s also led to log10 CFU/ml of B. subtilis from 7.98 to 4.39. S. Enteritidis was slight resistance to plasma treatment with N2. Within 180 s nitrogen plasma treatment, a 2.04 log10 CFU/ml reduction was recorded.

Synthesis of coumarin-containing poly(2-oxazoline)s and light-induced crosslinking for hydrogel formation.

Herein, we present a new route to synthesize poly(2-oxazoline)s (POx) with coumarin moieties as pendant groups that can be crosslinked by irradiation to form hydrogels. The synthesis of a novel coumarin-containing 2-oxazoline monomer, 2-[(4-methyl-7-coumarinyloxy)methyl]oxazoline is described in four steps and further its subsequent homo- and copolymerization with 2-ethyl-2-oxazoline (EtOx) via cationic ring-opening polymerization (CROP). The received polymers with different coumarin content (8, 4 and 2 mol%) were compared with a known copolymer synthesized via postpolymerization modification of partially hydrolyzed poly(2-ethyl-2-oxazoline) (PEtOx) with a coumarin derivative. The thermoresponsive behavior of the POx in aqueous solutions was investigated, showing a clear difference in the cloud point temperature (T cp ) between the polymers synthesized via different strategies. The hydrogel formation was achieved by irradiation with UV light by the following two methods: on the one hand with the pure coumarin-containing POx and on the other hand mixed with poly(ethylene glycol) diacrylate (PEGDA). The obtained hydrogels were compared regarding their swelling degree and their gel content, whereas the swelling degree in water of prepared hydrogels can be tuned by using POx by varying the coumarin content. Supplementary Information: The online version contains supplementary material available at 10.1007/s00706-022-03013-8.

Thiol-Ene Cross-linking of Poly(ethylene glycol) within High Internal Phase Emulsions: Degradable Hydrophilic PolyHIPEs for Controlled Drug Release.

Macroporous polymer monoliths prepared from high internal phase emulsions (HIPEs) can be found in various biomedical applications. While typically water-in-oil HIPEs are applied for polyHIPE preparation, they are not suitable for hydrophilic polyHIPE preparation. Herein, direct oil-in-water emulsions based on water-soluble poly(ethylene glycol)diacrylate or poly(ethylene glycol)dimethacrylate were developed. Furthermore, the incorporation of a hydrophilic water-miscible thiol, ethoxylated trimethylolpropane tris(3-mercaptopropionate) (ETTMP) was reported for the first time within thiol-ene polyHIPEs. Due to the transparency of the emulsions, rapid curing via photopolymerization was feasible. The average pore diameters of the resulting polyHIPEs ranged between 1.2 and 3.6 µm, and porosity of up to 90% was achieved. The water uptake of the materials reached up to 1000% by weight. Drug loading and release were demonstrated, employing salicylic acid as a model drug. Porous profile and biodegradability add to the usefulness of the material for biomedical applications.

Cold Atmospheric Pressure Plasma Treatment of Maize Grains-Induction of Growth, Enzyme Activities and Heat Shock Proteins.

Zea mays L. is one of the most produced crops, and there are still parts of the world where maize is the basic staple food. To improve agriculture, mankind always looks for new, better methods of growing crops, especially in the current changing climatic conditions. Cold atmospheric pressure plasma (CAPP) has already showed its potential to enhance the culturing of crops, but it still needs more research for safe implementation into agriculture. In this work, it was shown that short CAPP treatment of maize grains had a positive effect on the vitality of grains and young seedlings, which may be connected to stimulation of antioxidant and lytic enzyme activities by short CAPP treatment. However, the prolonged treatment had a negative impact on the germination, growth, and production indexes. CAPP treatment caused the increased expression of genes for heat shock proteins HSP101 and HSP70 in the first two days after sowing. Using comet assay it was observed that shorter treatment times (30-120 s) did not cause DNA damage. Surface diagnostics of plasma-treated grains showed that plasma increases the hydrophilicity of the surface but does not damage the chemical bonds on the surface.

Effect of Dexamethasone on Thermoresponsive Behavior of Poly(2-Oxazoline) Diblock Copolymers.

Thermoresponsive polymers play an important role in designing drug delivery systems for biomedical applications. In this contribution, the effect of encapsulated hydrophobic drug dexamethasone on thermoresponsive behavior of diblock copolymers was studied. A small series of diblock copoly(2-oxazoline)s was prepared by combining thermoresponsive 2-n-propyl-2-oxazoline (nPrOx) and hydrophilic 2-methyl-2-oxazoline (MeOx) in two ratios and two polymer chain lengths. The addition of dexamethasone affected the thermoresponsive behavior of one of the copolymers, nPrOx20-MeOx180, in the aqueous medium by shifting the cloud point temperature to lower values. In addition, the formation of microparticles containing dexamethasone was observed during the heating of the samples. The morphology and number of microparticles were affected by the structure and concentration of copolymer, the drug concentration, and the temperature. The crystalline nature of formed microparticles was confirmed by polarized light microscopy, confocal Raman microscopy, and wide-angle X-ray scattering. The results demonstrate the importance of studying drug/polymer interactions for the future development of thermoresponsive drug carriers.

Poly(2-oxazoline)- and Poly(2-oxazine)-Based Self-Assemblies, Polyplexes, and Drug Nanoformulations-An Update.

For many decades, poly(2-oxazoline)s and poly(2-oxazine)s, two closely related families of polymers, have led the life of a rather obscure research topic with only a few research groups world-wide working with them. This has changed in the last five to ten years, presumably triggered significantly by very promising clinical trials of the first poly(2-oxazoline)-based drug conjugate. The huge chemical and structural toolbox poly(2-oxazoline)s and poly(2-oxazine)s has been extended very significantly in the last few years, but their potential still remains largely untapped. Here, specifically, the developments in macromolecular self-assemblies and non-covalent drug delivery systems such as polyplexes and drug nanoformulations based on poly(2-oxazoline)s and poly(2-oxazine)s are reviewed. This highly dynamic field benefits particularly from the extensive synthetic toolbox poly(2-oxazoline)s and poly(2-oxazine)s offer and also may have the largest potential for a further development. It is expected that the research dynamics will remain high in the next few years, particularly as more about the safety and therapeutic potential of poly(2-oxazoline)s and poly(2-oxazine)s is learned.

Physico-Chemical Characterization and Antimicrobial Properties of Hybrid Film Based on Saponite and Phloxine B.

This research was aimed at the preparation of a hybrid film based on a layered silicate saponite (Sap) with the immobilized photosensitizer phloxine B (PhB). Sap was selected because of its high cation exchange capacity, ability to exfoliate into nanolayers, and to modify different surfaces. The X-ray diffraction of the films confirmed the intercalation of both the surfactant and PhB molecules in the Sap film. The photosensitizer retained its photoactivity in the hybrid films, as shown by fluorescence spectra measurements. The water contact angles and the measurement of surface free energy demonstrated the hydrophilic nature of the hybrid films. Antimicrobial effectiveness, assessed by the photodynamic inactivation on hybrid films, was tested against a standard strain and against methicillin-resistant bacteria of Staphylococcus aureus (MRSA). One group of samples was irradiated (green LED light; 2.5 h) and compared to nonirradiated ones. S. aureus strains manifested a reduction in growth from 1-log10 to over 3-log10 compared to the control samples with Sap only, and defects in S. aureus cells were proven by scanning electron microscopy. The results proved the optimal photo-physical properties and anti-MRSA potential of this newly designed hybrid system that reflects recent progress in the modification of surfaces for various medical applications.

The effect of cold atmospheric pressure plasma on Aspergillus ochraceus and ochratoxin A production.

Aspergillus ochraceus is a soil fungus known to produce ochratoxin A, a harmful secondary metabolite. Prevention and control of fungal pathogens mostly rely on chemical fungicides, which is one of the contributing factors in the emergence of the fungal resistance, hence novel methods for fungal eradication have been extensively researched. The cold atmospheric pressure (CAP) plasma generated in ambient air has been recently applied in microbial decontamination. Here we used the diffuse coplanar surface barrier discharge in inactivation of a toxigenic strain A. ochraceus. The plasma-treated conidia and mycelium exhibited morphological changes such as ruptures and desiccation. Mycelium dehydration and changes in the chemical composition of hyphal surface accompanied plasma treatment. The growth of 26 h old mycelia were significantly restricted after 30 s of plasma treatment. The conidial vitality declined 4 logs after 180 s of plasma exposure leading to almost complete decontamination. After shorter plasma treatment of conidia, the ochratoxin A (OTA) production increased at the early stage of cultivation, but the overall level was significantly reduced compared to untreated samples after longer cultivation. Our results indicated that the fungal growth and the OTA production were significantly changed by plasma treatment and underscored CAP plasma as a promising method in the decontamination of A. ochraceus without a risk to generate strains with increased OTA production.

Cold atmospheric pressure plasma: simple and efficient strategy for preparation of poly(2-oxazoline)-based coatings designed for biomedical applications.

Poly(2-oxazolines) (POx) are an attractive material of choice for biocompatible and bioactive coatings in medical applications. To prepare POx coatings, the plasma polymerization represents a fast and facile approach that is surface-independent. However, unfavorable factors of this method such as using the low-pressure regimes and noble gases, or poor control over the resulting surface chemistry limit its utilization. Here, we propose to overcome these drawbacks by using well-defined POx-based copolymers prepared by living cationic polymerization as a starting material. Chemically inert polytetrafluoroethylene (PTFE) is selected as a substrate due to its beneficial features for medical applications. The deposited POx layer is additionally post-treated by non-equilibrium plasma generated at atmospheric pressure. For this purpose, diffuse coplanar surface barrier discharge (DCSBD) is used as a source of "cold" homogeneous plasma, as it is operating at atmospheric pressure even in ambient air. Prepared POx coatings possess hydrophilic nature with an achieved water contact angle of 60°, which is noticeably lower in comparison to the initial value of 106° for raw PTFE. Moreover, the increased fibroblasts adhesion in comparison to raw PTFE is achieved, and the physical and biological properties of the POx-modified surfaces remain stable for 30 days.

Pulsatile Drug Delivery System Triggered by Acoustic Radiation Force.

Since biological systems exhibit a circadian rhythm (24-hour cycle), they are susceptible to the timing of drug administration. Indeed, several disorders require a therapy that synchronizes with the onset of symptoms. A targeted therapy with spatially and temporally precise controlled drug release can guarantee a considerable gain in terms of efficacy and safety of the treatment compared to traditional pharmacological methods, especially for chronotherapeutic disorders. This paper presents a proof of concept of an innovative pulsatile drug delivery system remotely triggered by the acoustic radiation force of ultrasound. The device consists of a case, in which a drug-loaded gel can be embedded, and a sliding top that can be moved on demand by the application of an acoustic stimulus, thus enabling drug release. Results demonstrate for the first time that ultrasound acoustic radiation force (up to 0.1 N) can be used for an efficient pulsatile drug delivery (up to 20 µg of drug released for each shot).

Poly(2-oxazoline)-based magnetic hydrogels: Synthesis, performance and cytotoxicity.

Research on the subject of smart biomaterials has become a cornerstone of tissue engineering and regenerative medicine. Herein, the authors report on developing magnetic hydrogels that combine high biocompatibility and remarkable activity in magnetic fields. We fabricated magnetic hydrogels based on poly(2-ethyl-2-oxazoline) (POx) via living ring-opening cationic polymerization with in-situ embedding of the carbonyl iron (CI) particles. Investigation was made as to the effect exerted by the concentration of CI on magnetic, viscoelastic/magnetorheological properties, the degree of equilibrium swelling, and cytotoxicity. The hydrogels exhibited an open pore structure, as evidenced by computed tomography (CT) imaging. Susceptibility measurements revealed the concentration-dependent field-induced particle restructuration indicating elongation/contraction of the material, thereby determining the potential for magneto-mechanical stimulation of the cells. The POx-based magnetic hydrogels were amphiphilic in character, showing decrease in their capability to hold liquid alongside increase in CI concentration. Viscoelastic measurements suggested that interaction occurred between the particles and matrix based on inconsistency between the experimental storage modulus and the Krieger-Dougherty model. The synthesized materials exhibited excellent biocompatibility toward the 3T3 fibroblast cell line in tests of extract toxicity and direct contact cytotoxicity (ISO standards). The unique combination of properties exhibited by the material - magneto-mechanical activity and biocompatibility - could prove favorable in fields such as biomedicine and biomechanics.

Soft Hydrogel Zwitterionic Coatings Minimize Fibroblast and Macrophage Adhesion on Polyimide Substrates.

Minimizing the foreign body reaction to polyimide-based implanted devices plays a pivotal role in several biomedical applications. In this work, we propose materials exhibiting nonbiofouling properties and a Young's modulus reflecting that of soft human tissues. We describe the synthesis, characterization, and in vitro validation of poly(carboxybetaine) hydrogel coatings covalently attached to polyimide substrates via a photolabile 4-azidophenyl group, incorporated in poly(carboxybetaine) chains at two concentrations of 1.6 and 3.1 mol %. The presence of coatings was confirmed by attenuated total reflectance Fourier transform infrared spectroscopy. White light interferometry was used to evaluate the coating continuity and thickness (between 3 and 6 µm under dry conditions). Confocal laser scanning microscopy allowed us to quantify the thickness of the swollen hydrogel coatings that ranged between 13 and 32 µm. The different hydrogel formulations resulted in stiffness values ranging from 2 to 19 kPa and led to different fibroblast and macrophage responses in vitro. Both cell types showed a minimum adhesion on the softest hydrogel type. In addition, both the overall macrophage activation and cytotoxicity were observed to be negligible for all of the tested material formulations. These results are a promising starting point toward future advanced implantable systems. In particular, such technology paves the way for novel neural interfaces able to minimize the fibrotic reaction, once implanted in vivo, and to maximize their long-term stability and functionality.

Investigation of drug release modulation from poly(2-oxazoline) micelles through ultrasound.

Among external stimuli used to trigger release of a drug from a polymeric carrier, ultrasound has gained increasing attention due to its non-invasive nature, safety and low cost. Despite this attention, there is only limited knowledge about how materials available for the preparation of drug carriers respond to ultrasound. This study investigates the effect of ultrasound on the release of a hydrophobic drug, dexamethasone, from poly(2-oxazoline)-based micelles. Spontaneous and ultrasound-mediated release of dexamethasone from five types of micelles made of poly(2-oxazoline) block copolymers, composed of hydrophilic poly(2-methyl-2-oxazoline) and hydrophobic poly(2-n-propyl-2-oxazoline) or poly(2-butyl-2-oxazoline-co-2-(3-butenyl)-2-oxazoline), was studied. The release profiles were fitted by zero-order and Ritger-Peppas models. The ultrasound increased the amount of released dexamethasone by 6% to 105% depending on the type of copolymer, the amount of loaded dexamethasone, and the stimulation time point. This study investigates for the first time the interaction between different poly(2-oxazoline)-based micelle formulations and ultrasound waves, quantifying the efficacy of such stimulation in modulating dexamethasone release from these nanocarriers.

Cold plasma treatment triggers antioxidative defense system and induces changes in hyphal surface and subcellular structures of Aspergillus flavus.

The cold atmospheric-pressure plasma (CAPP) has become one of the recent effective decontamination technologies, but CAPP interactions with biological material remain the subject of many studies. The CAPP generates numerous types of particles and radiations that synergistically affect cells and tissues differently depending on their structure. In this study, we investigated the effect of CAPP generated by diffuse coplanar surface barrier discharge on hyphae of Aspergillus flavus. Hyphae underwent massive structural changes after plasma treatment. Scanning electron microscopy showed drying hyphae that were forming creases on the hyphal surface. ATR-FTIR analysis demonstrated an increase of signal intensity for C=O and C-O stretching vibrations indicating chemical changes in molecular structures located on hyphal surface. The increase in membrane permeability was detected by the fluorescent dye, propidium iodide. Biomass dry weight determination and increase in permeability indicated leakage of cell content and subsequent death. Disintegration of nuclei and DNA degradation confirmed cell death after plasma treatment. Damage of plasma membrane was related to lipoperoxidation that was determined by higher levels of thiobarbituric acid reactive species after plasma treatment. The CAPP treatment led to rise of intracellular ROS levels detected by fluorescent microscopy using 2',7'-dichlorodihydrofluorescein diacetate. At the same time, antioxidant enzyme activities increased, and level of reduced glutathione decreased. The results in this study indicated that the CAPP treatment in A. flavus targeted both cell surface structures, cell wall, and plasma membrane, inflicting injury on hyphal cells which led to subsequent oxidative stress and finally cell death at higher CAPP doses.

Cold atmospheric pressure ambient air plasma inhibition of pathogenic bacteria on the surface of black pepper.

In present study, the inhibition effect of low temperature plasma on Bacillus subtilis, Escherichia coli, Salmonella Enteritidis and B. subtilis endospores inoculated on the surface of black peppercorns was studied. Plasma was generated by Diffuse Coplanar Surface Barrier Discharge (DCSBD) at atmospheric pressure in ambient air. Plasma treatment time of 300 s led to log10 CFU/g reduction of B. subtilis from 7.36 to 2.30 and B. subtilis endospores from 4.42 to 2.39. Plasma treatment reduced the number of E. coli and Salmonella Enteritidis to below detection level (1.0 log10 CFU/g) from initial populations of 7.45 log10 CFU/g and 7.60 log10 CFU/g, respectively. The inactivation kinetics was explained by Weibull model. Decimal reduction times (D-values) for B. subtilis, E. coli, Salmonella Enteritidis, and B. subtilis endospores were determined as 43 s, 47 s, 58 s, and 142 s, respectively. The surface morphology observed by Scanning Electron Microscopy showed no significant changes after the plasma treatment. The influence of plasma on chemical bonds on the surface and inside the peppercorns was studied by Attenuated Total Reflectance - Fourier Transform Infrared Spectroscopy.

Fast low-temperature plasma reduction of monolayer graphene oxide at atmospheric pressure.

We report on an ultrafast plasma-based graphene oxide reduction method superior to conventional vacuum thermal annealing and/or chemical reduction. The method is based on the effect of non-equilibrium atmospheric-pressure plasma generated by the diffuse coplanar surface barrier discharge in proximity of the graphene oxide layer. As the reduction time is in the order of seconds, the presented method is applicable to the large-scale production of reduced graphene oxide layers. The short reduction times are achieved by the high-volume power density of plasma, which is of the order of 100 W cm-3. Monolayers of graphene oxide on silicon substrate were prepared by a modified Langmuir-Schaefer method and the efficient and rapid reduction by methane and/or hydrogen plasma was demonstrated. The best results were obtained for the graphene oxide reduction in hydrogen plasma, as verified by x-ray photoelectron spectroscopy and Raman spectroscopy.

In vitro study of partially hydrolyzed poly(2-ethyl-2-oxazolines) as materials for biomedical applications.

Polymers based on 2-oxazoline, such as poly(2-ethyl-2-oxazolines) (PETOx), are considered to be a type of 'pseudopeptide' with the ability to form novel biomaterials. The hydrolysis of PETOx was carried out to evaluate its use in biomedical applications. In the present work, PETOx samples with a range of molar masses were prepared by living cationic polymerization. Hydrolysis was carried out at time intervals ranging from 15 to 180 min to prepare copolymers with different amounts of ethylene imine units. (1)H NMR spectroscopy was used to identify the structure of the hydrolyzed polymers. The dependence of in vitro cell viability on the degree of hydrolysis was determined using three different model cell lines, namely, mouse embryonic 3T3 fibroblasts, pancreatic ßTC3 cells, and mouse lymphoid macrophages P388.D1. It was demonstrated that increasing the degree of hydrolysis decreased cell viability for all cell types. Fibroblast cells displayed the highest tolerance; additionally, the effect of polymer size showed no observable significance. Macrophage cells, immune system representatives, displayed the highest sensitivity to contact with hydrolyzed PETOx. The effect of polymer hydrolysis, polymer concentration and the incubation time on cell viability was experimentally observed. Confocal laser-scanning microscopy provided evidence of cellular uptake of pyrene-labeled (co)polymers.