Characterization of plasma treated surfaces for food safety by terahertz spectroscopy

A physico-chemical approach to modify surfaces not only for use in medicine, but also for preservation of food is nowadays widely studied to lower the risks of increased number of bacterial pathogens that are in a direct contact with people. Food safety is very important part of preserving sustainability during crises, especially after the enterohaemorrhagic Escherichia coli outbreak in Europe in 2011. One of the possibility how we can protect food against various pathogens is the modification of packing materials that are directly in contact with preserved food. This contribution deals with the characterization of modified surfaces with antibacterial properties via Terahertz spectroscopy. For the purpose of this paper, three monomers were used for grafting onto air radiofrequency plasma activated low density polyethylene surface, which created a brush-like structure. Next, the antibacterial agents, Irgasan and Chlorhexidine, were anchored to these surfaces. These antibacterial agents were selected for supposed effect on two most frequently occurring bacterial strains -Escherichia coli and Staphylococcus aureus. Materials were further tested for the presence of antibacterial agent molecules, in our case by means of terahertz spectroscopy. Each material was tested on two spectroscopes - the SPECTRA and the OSCAT terahertz instruments

Antibacterial activity and cytotoxicity of immobilized glucosamine/chondroitin sulfate on polylactic acid films

Polylactic acid (PLA) is one of the most produced polymeric materials, due to its exceptional chemical and mechanical properties. Some of them, such as biodegradability and biocompatibility, make them attractive for biomedical applications. Conversely, the major drawback of PLA in the biomedical field is their vulnerability to bacterial contamination. This study focuses on the immobilization of saccharides onto the PLA surface by a multistep approach, with the aim of providing antibacterial features and evaluting the synergistic effect of these saccharides. In this approach, after poly (acrylic acid) (PAA) brushes attached non-covalently to the PLA surface via plasma post-irradiation grafting technique, immobilization of glucosamine (GlcN) and chondroitin sulfate (ChS) to the PAA brushes was carried out. To understand the changes in surface properties, such as chemical composition, surface topography and hydrophilicity, the untreated and treated PLA films were analyzed using various characterization techniques (contact angle, scanning electron microscopy, X-ray photoelectron spectroscopy). In vitro cytotoxicity assays were investigated by the methyl tetrazolium test. The antibacterial activity of the PLA samples was tested against Escherichia coli and Staphylococcus aureus bacteria strains. Plasma-treated films immobilized with ChS and GlcN, separately and in combination, demonstrated bactericidal effect against the both bacteria strains and also the results revealed that the combination has no synergistic effect on antibacterial action. © 2019 by the authors.Internal Grant Agency of Tomas Bata University in Zlin [IGA/CPS/2019/004]; Czech Science Foundation [17-05095S]; Slovenian Research Agency [L2-8179

The effect of plasma pretreatment and cross-linking degree on the physical and antimicrobial properties of nisin-coated PVA films

Stable antimicrobial nisin layers were prepared on the carrying medium-polyvinyl alcohol (PVA) films, crosslinked by glutaric acid. Surface plasma dielectric coplanar surface barrier discharge (DCSBD) modification of polyvinyl alcohol was used to improve the hydrophilic properties and to provide better adhesion of biologically active peptide-nisin to the polymer. The surface modification of films was studied in correlation to their cross-linking degree. Nisin was attached directly from the salt solution of the commercial product. In order to achieve a stable layer, the initial nisin concentration and the following release were investigated using chromatographic methods. The uniformity and stability of the layers was evaluated by means of zeta potential measurements, and for the surface changes of hydrophilic character, the water contact angle measurements were provided. The nisin long-term stability on the PVA films was confirmed by tricine polyacrylamide gel electrophoresis (SDS-PAGE) and by antimicrobial assay. It was found that PVA can serve as a suitable carrying medium for nisin with tunable properties by plasma treatment and crosslinking degree. © 2018 by the authors.Czech Science Foundation (Grant Agency of the Czech Republic) [17-10813S

Synthesis and characterization of a bioartificial polymeric system with potential antibacterial activity: Chitosan-polyvinyl alcohol-ampicillin

Bio-artificial polymeric systems are a new class of polymeric constituents based on blends of synthetic and natural polymers, designed with the purpose of producing new materials that exhibit enhanced properties with respect to the individual components. In this frame, a combination of polyvinyl alcohol (PVA) and chitosan, blended with a widely used antibiotic, sodium ampicillin, has been developed showing a moderate behavior in terms of antibacterial properties. Thus, aqueous solutions of PVA at 1 wt.% were mixed with acid solutions of chitosan at 1 wt.%, followed by adding ampicillin ranging from 0.3 to 1.0 wt.% related to the total amount of the polymers. The prepared bio-artificial polymeric system was characterized by FTIR, SEM, DSC, contact angle measurements, antibacterial activity against Staphylococcus aureus and Escherichia coli and antibiotic release studies. The statistical significance of the antibacterial activity was determined using a multifactorial analysis of variance with ρ < 0.05 (ANOVA). The characterization techniques did not show alterations in the ampicillin structure and the interactions with polymers were limited to intermolecular forces. Therefore, the antibiotic was efficiently released from the matrix and its antibacterial activity was preserved. The system disclosed moderate antibacterial activity against bacterial strains without adding a high antibiotic concentration. The findings of this study suggest that the system may be effective against healthcare-associated infections, a promising view in the design of novel antimicrobial biomaterials potentially suitable for tissue engineering applications. © 2018 by the authors.Czech Science Foundation [17-10813S

Exploring the critical factors limiting polyaniline biocompatibility

Today, the application of polyaniline in biomedicine is widely discussed. However, information about impurities released from polyaniline and about the cytotoxicity of its precursors aniline, aniline hydrochloride, and ammonium persulfate are scarce. Therefore, cytotoxicity thresholds for the individual precursors and their combinations were determined (MTT assay) and the type of cell death caused by exposition to the precursors was identified using flow-cytometry. Tests on fibroblasts revealed higher cytotoxicity of ammonium persulfate than aniline hydrochloride. Thanks to the synergic effect, both monomers in combination enhanced their cytotoxicities compared with individual substances. Thereafter, cytotoxicity of polyaniline doped with different acids (sulfuric, nitric, phosphoric, hydrochloric, and methanesulfonic) was determined and correlated with impurities present in respective sample (HPLC). The lowest cytotoxicity showed polyaniline doped with phosphoric acid (followed by sulfuric, methanesulfonic, and nitric acid). Cytotoxicity of polyaniline was mainly attributed to the presence of residual ammonium persulfate and low-molecular-weight polar substances. This is crucial information with respect to the purification of polyaniline and production of its cytocompatible form.Ministry of Education, Youth and Sports of the Czech Republic [LO1504]; Czech Science Foundation [17-05095S]; Tomas Bata University in Zlin [IGA/CPS/2019/004

Anticoagulant polyethylene terephthalate surface by plasma-mediated fucoidan immobilization

Biomaterial-based blood clot formation is one of the biggest drawbacks of blood-contacting devices. To avoid blood clot formation, their surface must be tailored to increase hemocompatibility. Most synthetic polymeric biomaterials are inert and lack bonding sites for chemical agents to bond or tailor to the surface. In this study, polyethylene terephthalate was subjected to direct current air plasma treatment to enhance its surface energy and to bring oxidative functional binding sites. Marine-sourced anticoagulant sulphated polysaccharide fucoidan from Fucus vesiculosus was then immobilized onto the treated polyethylene terephthalate (PET) surface at different pH values to optimize chemical bonding behavior and therefore anticoagulant performance. Surface properties of samples were monitored using the water contact angle; chemical analyses were performed by FTIR and X-ray photoelectron spectroscopy (XPS) and their anticoagulant activity was tested by means of prothrombin time, activated partial thromboplastin time and thrombin time. On each of the fucoidan-immobilized surfaces, anticoagulation activity was performed by extending the thrombin time threshold and their pH 5 counterpart performed the best result compared to others. © 2019 by the authors.Czech Science Foundation [17-10813S]; Ministry of Education, Youth and Sports of the Czech Republic, Program NPU I [LO1504]; internal grant agency of Tomas Bata University in Zlin [IGA/CPS/2019/004

Physical and morphological changes of poly(tetrafluoroethylene) after using non-thermal plasma-treatments

A commercial formulation of poly(tetrafluoroethylene) (PTFE) sheets were surfacemodified by using non-thermal air at 40 kHz frequency (DC) and 13.56 MHz radiofrequency (RF) at different durations and powers. In order to assess possible changes of PTFE surface properties, zeta potential (ζ), isoelectric points (IEPs) determinations, contact angle measurements as well as Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM) imaging were carried out throughout the experimentation. The overall outcome indicated that ζ-potential and surface energy progressively changed after each treatment, the IEP shifting to lower pH values and the implicit differences, which are produced after each distinct treatment, giving new surface topographies and chemistry. The present approach might serve as a feasible and promising method to alter the surface properties of poly(tetrafluoroethylene). © 2018 by the authors.Czech Science Foundation [17-10813S

A new route of fucoidan immobilization on low density polyethylene and its blood compatibility and anticoagulation activity

Beside biomaterials’ bulk properties, their surface properties are equally important to control interfacial biocompatibility. However, due to the inadequate interaction with tissue, they may cause foreign body reaction. Moreover, surface induced thrombosis can occur when biomaterials are used for blood containing applications. Surface modification of the biomaterials can bring enhanced surface properties in biomedical applications. Sulfated polysaccharide coatings can be used to avoid surface induced thrombosis which may cause vascular occlusion (blocking the blood flow by blood clot), which results in serious health problems. Naturally occurring heparin is one of the sulfated polysaccharides most commonly used as an anticoagulant, but its long term usage causes hemorrhage. Marine sourced sulfated polysaccharide fucoidan is an alternative anticoagulant without the hemorrhage drawback. Heparin and fucoidan immobilization onto a low density polyethylene surface after functionalization by plasma has been studied. Surface energy was demonstrated by water contact angle test and chemical characterizations were carried out by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Surface morphology was monitored by scanning electron microscope and atomic force microscope. Finally, their anticoagulation activity was examined for prothrombin time (PT), activated partial thromboplastin time (aPTT), and thrombin time (TT). © 2016 by the authors; licensee MDPI, Basel, Switzerland.Operational Program Research and Development for Innovations; European Regional Development Fund (ERDF); national budget of Czech Republic, within the framework of project Centre of Polymer Systems [CZ.1.05/2.1.00/03.0111]; Ministry of Education, Youth and Sports of the Czech Republic [LO1504

Developing a biomaterial interface based on poly(lactic acid) via plasma-assisted covalent anchorage of D-glucosamine and its potential for tissue regeneration

The aim of this study was to develop the potential tissue engineering applications of D-glucosamine (GlcN) immobilized onto the surface of a biodegradable matrix in order to induce a desired biological effect at biointerfaces. Thus, for sample preparation we used a novel multistep physicochemical approach. In the first step the poly(lactic acid) (PLA) films were exposed to a low pressure plasma in air atmosphere, followed by radical graft copolymerization with acrylic acid to yield a carboxyl-functionalized spacer layer on the PLA surface. The carboxyl groups were then coupled to GlcN molecules via the carbodiimide chemistry. The developed surfaces were characterized by X-ray Photoelectron Spectroscopy (XPS), Contact angle measurements and Atomic Force Microscopy (AFM). A preliminary study on the proliferation of fibroblasts on the developed surfaces was performed using the NIH/3T3 cell line. © 2016 Elsevier B.V.ARRS, Slovenian Research AgencyMinistry of Education, Youth and Sports of the Czech Republic [LO1504]; Ministry of Education, Science, Research and Sport of the Slovak Republic; Slovak Academy of Sciences [2/0199/14]; Slovenian Research Agency [P2-0082

Yarrowia lipolytica adhesion and immobilization onto residual plastics

Research in cell adhesion has important implications in various areas, such as food processing, medicine, environmental engineering, biotechnological processes. Cell surface characterization and immobilization of microorganisms on solid surfaces can be performed by promoting cell adhesion, in a relatively simple, inexpensive, and quick manner. The adhesion of Yarrowia lipolytica IMUFRJ 50682 to different surfaces, especially potential residual plastics (polystyrene, poly(ethylene terephthalate), and poly(tetrafluoroethylene)), and its use as an immobilized biocatalyst were tested. Y. lipolytica IMUFRJ 50682 presented high adhesion to different surfaces such as poly(tetrafluoroethylene) (Teflon), polystyrene, and glass, independent of pH, and low adhesion to poly(ethylene terephthalate) (PET). The adhesion of the cells to polystyrene was probably due to hydrophobic interactions involving proteins or protein complexes. The adhesion of the cells to Teflon might be the result not only of hydrophobic interactions but also of acid-basic forces. Additionally, the present work shows that Y. lipolytica cell extracts previously treated by ultrasound waves (cell debris) maintained their enzymatic activity (lipase) and could be attached to polystyrene and PET and used successfully as immobilized biocatalysts in hydrolysis reactions.CAPES (Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior)/GRICES (Gabinete de Relacoes Internacionais da Ciencia e do Ensino Superior)CAPES [102/03]; CNPqNational Council for Scientific and Technological Development (CNPq); national funds through the FCT/MCTES [UID/CTM/50011/2019]; Tomas Bata University in Zlin Internal Grant Agency [IGA/CPS/2020/001