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1.
2.
Effect of peptides' binding on the antimicrobial activity and biocompatibility of ‎protein-based substrates ‎
Maja Kaisersberger Vincek, 2017, doctoral dissertation

Abstract: This work reveals the effect of coupling approach (chemical by using carbodiimide chemistry ‎and grafting-to vs. grafting-from synthesis routes, and enzymatic by using transglutaminase) ‎of a hydrophilic ε-poly-L-lysine (εPL) and an amphiphilic oligo-acyl-lysyl (OAK) derivative (K-7α12-OH) to wool fibers and gelatine (GEL) macromolecules, respectively, and substrates ‎antibacterial activity against Gram-negative E. coli and Gram-positive S. aureus bacteria ‎after 1–24 h of exposure, as well as their cytotoxicity. Different spectroscopic (ultraviolet-‎visible, infrared, fluorescence and electron paramagnetic resonance) and separation ‎techniques (size-exclusion chromatography and capillary zone electrophoresis) as well as ‎zeta potential and potentiometric titration analysis, were performed to confirm the covalent ‎coupling of εPL/OAK, and to determine the amount and orientation of its immobilisation.‎ The highest and kinetically the fastest level of bacterial reduction was achieved with ‎wool/GEL functionalised with εPL/OAK by chemical grafting-to approach. This effect ‎correlated with both the highest grafting yield and conformationally the highly-flexible ‎(brush-like) orientation linkage of εPL/OAK, implicating on the highest amount of accessible ‎amino groups interacting with bacterial membrane. However, OAK`s amphipathic structure, ‎the cationic charge and the hydrophobic moieties, resulted to relatively high reduction of S. ‎aureus for grafting-from and the enzymatic coupling approaches using OAK-functionalised ‎GEL. ‎ The εPL/OAK-functionalised GEL did not induce toxicity in human osteoblast cells, even at ‎‎~25-fold higher concentration than bacterial minimum inhibitory (MIC) concentration of ‎εPL/OAK, supporting their potential usage in biomedical applications.‎ It was also shown that non-ionic surfactant adsorbs strongly onto the wool surface during ‎the process of washing, thereby blocking the functional sites of immobilized εPL and ‎decreases its antibacterial efficiency. ‎ ‎
Keywords: wool, gelatine, antimicrobial peptides, ε-poly-L-lysine, oligo-acyl-lysyl, grafting chemistry, ‎grafting approach, peptide orientation, antibacterial activity, cytotoxicity‎
Published: 17.08.2017; Views: 967; Downloads: 94
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3.
Influence of the temperature on the efficiency of cellulose treatment using copolymer chitosan-eugenol
Olivera Šauperl, Jasna Tompa, Julija Volmajer Valh, 2014, original scientific article

Abstract: In order to achieve effective antimicrobial protection of textile materials against microorganisms, a natural compound called chitosan has become very interesting. In regard to the antimicrobial protection of textile materials, functionalization with chitosan does not affect some other properties, such as anti-oxidative or any other action. For this reason, it seems appropriate for chitosan to be combined with any natural antimicrobial active compound, such as eugenol, an extract of clove oil. During this research viscose as a representative of cellulose fibers was used, because it can be functionalized relatively easily. In terms of functionalization, the drying temperature of viscose, after antimicrobial compound application onto substrate, as well as the successful synthesis of copolymer chitosan/eugenol is also important. FTIR spectroscopy was used to evaluate the efficiency of synthesizing a chitosan/eugenol graft copolymer. The spectrophotometric method Acid Orange 7 was chosen as a means for determining the proportion of available antimicrobial active amino groups. In addition, microbiological testing of selected pathogenic micro-organisms was also performed. The results were compared with the results for viscose functionalized by a 1% solution of chitosan.
Keywords: chitosan, eugenol, functionalization, viscose, FTIR spectroscopy, Acid orange 7, antimicrobial
Published: 02.08.2017; Views: 547; Downloads: 328
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4.
Chemical binding of chitosan and chitosan nanoparticles onto oxidized cellulose
Olivera Šauperl, Mirjana Kostić, Jovana Milanovic, Lidija Fras Zemljič, 2015, original scientific article

Abstract: The aim of this study was to analyze binding of chitosan and chitosan nanoparticles onto cellulose via oxidized cellulose. The ability of chitosan and chitosan nanoparticles to be adsorbed onto surfaces was determined by the use of the XPS spectroscopy which provided information about chemical composition of the fiber surface. On the other hand, the gravimetric method was also used by which the amount of chitosan and chitosan nanoparticles bounded onto surface was calculated based on the difference in masses before and after functionalization. The most important was to study the influence of aldehyde groups on the stability of chitosan binding onto cellulose. Thus, desorption of chitosan/chitosan nanoparticles from the fiber surfaces was evaluated by the presence of total nitrogen (TN) in desorption bath as well as by polyelectrolyte titrations. Together with these two methods, desorption was evaluated also by gravimetric method, where the extent of desorption was evaluated on the basis of the differences in the masses of fibers before and after desorption. It is concluded that the chitosan and chitosan nanoparticles are more efficiently bounded onto oxidized cellulose in comparison with the non-oxidized (reference) ones. Despite the binding of the positively-charged amino groups with the negative groups of cellulose and consequently smaller amount of available/residual protonated amino groups that are responsible for bioactivity, such functionalized fibers are still specifically antimicrobial.
Keywords: cellulose, oxidized cellulose, oxidation, chitosan, chitosan nanoparticles, FTIR, XPS, antimicrobial functionalization
Published: 02.08.2017; Views: 684; Downloads: 378
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5.
Surface modification of silicone with polysaccharides for the development of antimicrobial urethral catheters
Matej Bračič, 2016, doctoral dissertation

Abstract: In this work, alternative polysaccharide-based coatings were used to improve the antimicrobial and antifouling properties of silicone surfaces used for urethral catheters. The introduction of a catheter in the urethra is commonly connected with a high risk of microbial infections which often result in long-term health damage. Polysaccharide-based coatings like chitosan, carboxymethyl chitosan, and a synergistic formulation between hyaluronic acid and a natural lysine-based surfactant, were used to treat silicone surfaces to overcome the infection problems as an alternative to conventional approaches, which include the administration of antibiotics or coatings with metal ions. The polysaccharide-based macromolecular solutions and dispersions were firstly characterised by means of pH-titrations, dynamic light scattering and scanning electron microscopy in order to determine the size of particles in dispersions and their pH dependant charging behaviour. The knowledge gained from this was used to thoroughly study the interactions of the polysaccharide-based solutions and dispersions with the model silicone surfaces. The model surfaces were ultra-thin films prepared by dissolution of silicone in toluene and subsequent spin-coating on quartz crystals. The influence of pH, salt concentration, and various surface activation processes on the adsorption behaviour was evaluated by means of a very precise quartz crystal microbalance with dissipation. This knowledge was transferred to the application of the coatings on real systems i.e. casted silicone sheets and silicone tubes. The surface morphology, surface chemistry, as well as the mechanical and chemical stability of the coatings were further characterised on both model and real systems. For this purpose different microscopy and spectroscopy methods, pH-potentiometric titrations and methods for evaluation of mechanical properties were used. Finally the antimicrobial and antifouling properties were evaluated. The antimicrobial properties were tested against gram-positive and gram-negative bacteria as well as fungi, which are commonly found in infected urine, while the antifouling properties were tested by measuring the adhesion of bovine serum albumin, fibrinogen, and lysozyme biomolecules onto functionalised model films using the quartz crystal microbalance. The results showed that homogeneous and stable coatings can be achieved by adsorption from dispersions of the polysaccharide-based nanoparticles of 200-300 nm in size, which are formed by precipitation; i.e. careful pH adjustments of chitosan to pH = 6.5, carboxymethyl chitosan to pH = 7, and by mixing the hyaluronic acid and natural lysine-based surfactant solutions at concentrations of 2.5 x 10-4 mol/L and 1.25 x 10- 3 mol/L for the hyaluronic acid and 5.0 x 10-4 mol/L and 1.2 x 10-3 mol/L for the surfactant, respectively . The mass of the coatings on model silicone films can be increased by a 3-step adsorption, which directly influences the antimicrobial properties of the coatings that are improved with increasing coating mass, reaching values of up to 90 % in reduction of microorganism growth. It was also shown that the hyaluronic acid-natural surfactant formulation is superior to the chitosan coatings. The same conclusions were drawn from the antifouling evaluation where the zwitterionic nature of the formulation between natural based lysine surfactant and HA successfully suppressed the adhesion of biomolecules on silicone surfaces, while the chitosan coatings only moderately prevented the adhesion of proteins. One can conclude that the polysaccharide-based coatings can be successfully introduced to silicone surfaces from dispersion and as such successfully prevent biomolecule adhesion and reduce the growth of pathogen microorganisms which can be found in the urine during urethral infections.
Keywords: Urethral catheters, Antimicrobial coatings, Silicone, Polysaccharides, Natural surfactants
Published: 24.10.2016; Views: 1276; Downloads: 196
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6.
Hemicelluloses application for synthetic polymer surfaces functionalisation
Nena Dimitrušev, 2016, doctoral dissertation

Abstract: The main aim of this thesis was development of thin functional layers from hemicelluloses xylans on the polyethylene terephthalate (PET) surfaces. Hemicelluloses, xylans, as renewable polymers, were chemically modified in order to introduce anionic and cationic functional groups. Two types of chemical modifications were performed: carboxymethylation in order to increase anionic nature of xylans and improve their hydrophilic character and cationization for introducing of amino groups and antimicrobial characteristics. Both types of modifications were successful, which was proved by ATR FTIR and raman techniques, elemental analysis, total bound nitrogen determination, size exclusion chromatography and polyelectrolyte titrations. Polyelectrolyte titration results showed increased amounts of deprotonated carboxyl groups in carboxymethylated xylans as well as increased amounts of protonated groups in cationized xylans. Antimicrobial activity of xylans was investigated by the determination of minimal inhibitory concentration (MIC) against S. aureus, E. coli, and C. albicans and it was found out that the samples with higher amounts of active amino groups showed lower MIC. Cationised glucuronoxylan showed significantly higher antimicrobial activities against S. aureus in comparison to cationised arabinoxylan and nonmodified xylan samples. However, none of xylan samples was active against fungi. In order to analyze surface properties of solid surfaces, films from xylan (nonmodified and modified) water solution was formed by casting method. The surface chemical composition of films were investigated by x-ray photoelectron spectroscopy (XPS), and the results showed that films made from carboxymethylated xylans had significantly higher amounts of carbon fraction involved in O=C-O bonds, compared to nonmodified xylans. Such surface chemical structure caused higher surface free energy with higher electron-donor contribution and thus high hydrophilicity of these films. Films made by cationized xylans had higher amount of carbon involved in C-C and C-H bonds compared to nonmodified and lower surface free energy with increase of dispersive Lifshitz Van der Waals contribution. In order to thoroughly investigate the adsorption of xylans onto synthetic surfaces Quartz crystal microbalance with dissipation unit (QCM-D) was used. For these measurements model films were prepared from PET by spin coating technique. Adsorption studies were performed at different conditions, such as pH, concentration and ionic strength of xylan solutions. For all the chemically modified xylans the adsorption was improved at pH 5 and with increased ionic strength with divalent ions. The adsorption increased as well with increasing of xylan solution concentration. In order to improve binding of adsorbed xylans so-called anchoring polymers were applied. When anchoring polymers were applied, better adsorption and fixation of adsorbed layer was confirmed, thus the adsorbed masses of xylans after rinsing with water were significantly higher in comparison to the adsorption without immediate anchoring layer. On the basis of these results, real PET fabric surfaces were treated using chemically modified xylans. The xylan solutions were applied onto PET fabric samples using spray coating technique, which is the best approximate to the large-scale procedures. In the first step, PET fabric was activated by alkaline hydrolysis and after that, anchoring agents and carboxymethylated and/or cationized xylans were adsorbed. The success of these treatments was evaluated by the determination of negative and positive charge of the treated PET fabric samples by titration techniques, methylene blue and acid orange 7 adsorption methods, water contact angles and wettability determination. From the potentiometric titrations results it was clearly seen that each new adsorbed layer onto PET fabric totally screened the charge of the former one. FESEM images showed rather thick layers covering the
Keywords: hemicellulose, polyethylene terephthalate, glucuronoxylan, arabinoxylan, carboxymethylation, cationization, PET model films, quartz crystal microbalance, PET fabric, surface free energy, wettability, antimicrobial properties
Published: 11.03.2016; Views: 1238; Downloads: 83
.pdf Full text (4,53 MB)

7.
Viscose functionalisation with a combination of chitosan/BTCA using microwaves
Olivera Šauperl, Julija Volmajer Valh, 2013, original scientific article

Abstract: Improved hygiene and health care standards have a great impact on the development of hygiene and health care products. For this purpose, viscose is a very popular substrate. One of the most promising anti-microbial compounds of modern times is chitosan. The anti-microbial action of this polysaccharide depends on the amino group amount, which is crucial for ensuring the effectiveness of anti-microbial treated material. In textile finishing, 1,2,3,4-buthanetetracarboxylic acid (BTCA) is usually used as a non-formaldehyde crease-resistant reagent. But, on the other hand, the negatively charged carboxyl groups of BTCA can be explored as additional binding sites for positively-charged protonated amino groups of chitosan. When using microwaves, polar materials (e.g. chitosan) orient and reorient themselves according to the direction of the electro-magnetic field, which means that chitosan chain-bending may have taken place during the drying with microwaves. This could result in a higher specific surface of the chitosan and, consequently, in a higher proportion of available amino groups. It is concluded that the combination chitosan/BTCA supported by microwaves drying represents an ideal combination to increase the proportion of available amino groups.
Keywords: viskoza, protimikrobna zaščita, viscose, chitosan/BTCA, microwawes, Acid Orange VII, methylene blue, antimicrobial activity
Published: 10.07.2015; Views: 944; Downloads: 30
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Antioxidant and antimicrobial activity of PGSStm micronized curcuma powder
Željko Knez, Tina Perko, Mojca Škerget, 2012, published scientific conference contribution

Keywords: antioxidant, antimicrobial activity, curcumin extract
Published: 10.07.2015; Views: 701; Downloads: 20
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