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1.
Novel magnetic iron oxide-dextran sulphate nanocomposites as potential anticoagulants: Investigating interactions with blood components and assessing cytotoxicity
Olivija Plohl, Lidija Fras Zemljič, Boštjan Vihar, Alenka Vesel, Sašo Gyergyek, Uroš Maver, Irena Ban, Matej Bračič, 2024, izvirni znanstveni članek

Opis: Examining the critical role of anticoagulants in medical practice, particularly their central function in preventing abnormal blood clotting, is of the utmost importance. However, the study of interactions between blood proteins and alternative anticoagulant nano-surfaces is still understood poorly. In this study, novel approach involving direct functionalisation of magnetic iron oxide nanoparticles (MNPs) as carriers with sulphated dextran (s-dext) is presented, with the aim of evaluating the potential of magnetically-responsive MNPs@s-dext as anticoagulants. The physicochemical characterisation of the synthesised MNPs@s-dext includes crystal structure analysis, morphology study, surface and electrokinetic properties, thermogravimetric analysis and magnetic properties` evaluation, which confirms the successful preparation of the nanocomposite with sulfonate groups. The anticoagulant potential of MNPs@s-dext was investigated using a standardised activated partial thromboplastin time (APTT) test and a modified APTT test with a quartz crystal microbalance with dissipation (QCM-D) which confirmed the anticoagulant effect. Time-resolved solid-liquid interactions between the MNPs@s-dext and model blood proteins bovine serum albumin and fibrinogen were also investigated, to gain insight into their hemocompatibility, and revealed protein-repellence of MNPs@s-dext against blood proteins. The study also addressed comprehensive cytotoxicity studies of prepared nanocomposites, and provided valuable insights into potential applicability of MNPs@s-dext as a promising magnetic anticoagulant in biomedical contexts.
Ključne besede: dextran sulphate, magnetic nanoparticles, blood protein interactions, clot formation, anticoagulants, cytotoxicity studies
Objavljeno v DKUM: 25.07.2024; Ogledov: 107; Prenosov: 15
.pdf Celotno besedilo (892,02 KB)
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2.
Synthesis of betaine, choline and carnitine containing polymers for dermal wound healing
Lucija Jurko, 2024, doktorska disertacija

Opis: In this study, we explored the development of cationized 2-hydroxyethyl cellulose (HEC) and succinylated polyallylamine (PAA) in conjunction with polyvinyl alcohol (PVA) for potential applications as antimicrobial wound dressings. Quaternary ammonium compounds are known for their broad-spectrum antimicrobial properties against both Gram-negative and Gram-positive bacteria, but their non-selectivity can lead to high cytotoxicity. To improve the biocompatibility of the cationic materials investigates two distinct approaches. The first approach involves reducing the cationic nature of protonated PAA at a pH below 8.3 through amidation reaction with succinic anhydride. Gradual reduction of the cationic charge is achieved by varying the molar ratios of succinic anhydride during the reaction process. Notably, this reaction can be carried out in an aqueous solution, eliminating potential issues associated with solvent removal. The second part of this thesis focuses on introducing cationic charge by covalently binding naturally occurring quaternary ammonium compounds, such as betaine hydrochloride (BET HCl), choline chloride (ChCl), and carnitine hydrochloride (carnitine). Using 1,1′-carbonyldiimidazole (CDI), we quaternized HEC in anhydrous DMSO, with a detailed investigation of the reaction mechanism by isolating and characterizing intermediate products. This same procedure was applied to form a cationic dimer between BET HCl and ChCl. The chemical structures of the resulting materials were characterized using attenuated total reflectance infrared spectroscopy (ATR-IR) and nuclear magnetic resonance (NMR). The quantification of cationic and total charge was determined through polyelectrolyte and potentiometric titration, respectively. While succinylated PAA exhibited the anticipated biological properties associated with the reduction of cationic characteristics, the same behaviour was not observed for quaternized materials. High cytotoxicity and low antimicrobial properties in the derivatized HEC may be attributed to inadequate purification and the sensitivity of fibroblasts. We produced electrospun non-woven mats incorporating PVA and these derived materials using a single-needle electrospinning machine. The average diameter of the fibres was determined through scanning electron microscopy (SEM) images. Despite the cytotoxicity and the absence of significant antimicrobial properties in these materials, they exhibit potential as effective cationic flocculants for wastewater treatment. Further investigations are pending to address purification issues and make slight adjustments to the procedure to potentially scale up production. These materials still hold promise for applications in both biomedical and environmental chemistry.
Ključne besede: cellulose, quaternary ammonium compounds, antimicrobial, cytotoxicity, electrospinning
Objavljeno v DKUM: 11.07.2024; Ogledov: 127; Prenosov: 32
.pdf Celotno besedilo (16,36 MB)

3.
Efficient and Green Isolation of Keratin from Poultry Feathers by Subcritical Water
Mojca Škerget, Maja Čolnik, Lidija Fras Zemljič, Lidija Gradišnik, Tanja Živković Semren, Blanka Tariba Lovaković, Uroš Maver, 2023, izvirni znanstveni članek

Opis: The isolation of keratin from poultry feathers using subcritical water was studied in a batch reactor at temperatures (120–250 °C) and reaction times (5–75 min). The hydrolyzed product was characterized by FTIR and elemental analysis, while the molecular weight of the isolated product was determined by SDS-PAGE electrophoresis. To determine whether disulfide bond cleavage was followed by depolymerization of protein molecules to amino acids, the concentration of 27 amino acids in the hydrolysate was analyzed by GC/MS. The optimal operating parameters for obtaining a high molecular weight protein hydrolysate from poultry feathers were 180 °C and 60 min. The molecular weight of the protein hydrolysate obtained under optimal conditions ranged from 4.5 to 12 kDa, and the content of amino acids in the dried product was low (2.53% w/w). Elemental and FTIR analyses of unprocessed feathers and dried hydrolysate obtained under optimal conditions showed no significant differences in protein content and structure. Obtained hydrolysate is a colloidal solution with a tendency for particle agglomeration. Finally, a positive influence on skin fibroblast viability was observed for the hydrolysate obtained under optimal processing conditions for concentrations below 6.25 mg/mL, which makes the product interesting for various biomedical applications.
Ključne besede: poultry feathers, subcritical water hydrolysis, keratin, physico-chemical characterization, cytotoxicity
Objavljeno v DKUM: 28.11.2023; Ogledov: 456; Prenosov: 153
.pdf Celotno besedilo (2,78 MB)
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4.
Effect of peptides' binding on the antimicrobial activity and biocompatibility of ‎protein-based substrates ‎
Maja Kaisersberger Vincek, 2017, doktorska disertacija

Opis: 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. ‎ ‎
Ključne besede: wool, gelatine, antimicrobial peptides, ε-poly-L-lysine, oligo-acyl-lysyl, grafting chemistry, ‎grafting approach, peptide orientation, antibacterial activity, cytotoxicity‎
Objavljeno v DKUM: 17.08.2017; Ogledov: 1931; Prenosov: 175
.pdf Celotno besedilo (3,98 MB)

5.
Native cellulose nanofibrills induce immune tolerance in vitro by acting on dendritic cells
Sergej Tomić, Vanja Kokol, Dušan Mihajlović, Aleksandar Mirčić, Miodrag Čolić, 2016, izvirni znanstveni članek

Opis: Cellulose nanofibrills (CNFs) are attractive biocompatible, natural nanomaterials for wide biomedical applications. However, the immunological mechanisms of CNFs have been poorly investigated. Considering that dendritic cells (DCs) are the key immune regulatory cells in response to nanomaterials, our aim was to investigate the immunological mechanisms of CNFs in a model of DC-mediated immune response. We found that non-toxic concentrations of CNFs impaired the differentiation, and subsequent maturation of human monocyte-derived (mo)-DCs. In a co-culture with CD4+T cells, CNF-treated mo-DCs possessed a weaker allostimulatory and T helper (Th)1 and Th17 polarizing capacity, but a stronger capacity to induce Th2 cells and CD4+CD25hiFoxP3hi regulatory T cells. This correlated with an increased immunoglobulin-like transcript-4 and indolamine dioxygenase-1 expression by CNF-treated mo-DCs, following the partial internalization of CNFs and the accumulation of CD209 and actin bundles at the place of contacts with CNFs. Cumulatively, we showed that CNFs are able to induce an active immune tolerance by inducing tolerogenic DCs, which could be beneficial for the application of CNFs in wound healing and chronic inflammation therapies.
Ključne besede: nanocellulose, cytotoxicity, T-cell proliferation, cytokine production, in vitro
Objavljeno v DKUM: 23.06.2017; Ogledov: 1538; Prenosov: 383
.pdf Celotno besedilo (2,87 MB)
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6.
Antimicrobial medical textiles based on chitosan nanoparticles for gynaecological treatment
Tijana Ristić, 2014, doktorska disertacija

Opis: The aim of this dissertation was to develop a novel medical tampon for alternative gynaecological treatment using chitosan nanoparticles as an antimicrobial agent or as a drug delivery system. For this purpose viscose tampon band was used and functionalized with chitosan and trimethyl chitosan nanoparticles. A comprehensive of interactions between chitosan and cellulose as well as characterization of prepared materials were done. At the beginning, chitosan (CS) and trimethyl chitosan (TMC) solutions, as well as nanoparticles synthesised by ionic gelation were studied. Their characterization was focused on determining the charge and antimicrobial properties against common pathogenic microorganism. The influence of cationic charge on the inhibition of microbial growth was confirmed. Since CS and TMC solutions and nanoparticles dispersions exhibited antibacterial activity against Lactobacillus, a detailed investigation in chitosan’s antimicrobial mode of action was performed using a novel diffusion nuclear magnetic resonance (D-NMR). D-NMR allowed the monitoring of intra- and extracellular water exchange from the cells indicating the membrane alteration and leakage of intracellular constituencies. Further, in order to study the adsorption phenomena and molecular interactions between CS/TMC (solution or nanoparticles) and cellulose material, model cellulose surfaces were used, and adsorption was studied by quartz crystal microbalance with dissipation. CS and TMC were favourably deposited onto cellulose model surface at higher ionic strength, higher pH values, i.e. factors causing lower solubility, where the presence of electrostatic interactions was negligible and non-electrostatic interactions were dominant. The knowledge gained from the model surfaces was extremely helpful in characterization of real systems, i.e. functionalized cellulose fibres and for understanding the obtained results. Immobilization of CS and TMC (in the form of solution and/or nanoparticles) onto cellulose viscose fibres was confirmed with several analytical methods. The attachment of chitosan onto fibres was reversible, as endorsed with desorption studies mimicking the conditions of vaginal usage. Evaluation of antimicrobial properties was performed using two different techniques, both revealing a high inhibition of the tested microorganism. In addition, Lactobacillus susceptibility testing has shown that chitosan-coated fibres do not have any negative influence on the resident microbiota. Assessment of in-vitro cytotoxicity demonstrated that samples do not cause a cytotoxic effect in direct contact. Additionally, model drug was incorporated into chitosan nanoparticles and subsequently attached onto fibres in order to create modern, vaginal drug delivery systems. Antimicrobial medical textiles investigated in the scope of this dissertation show the potential for their exploitation in gynaecological field as preventive or curative treatment without triggering any adverse effects for the user.
Ključne besede: chitosan, N, N, N-trimethyl chitosan, nanoparticles, antimicrobial activity, cellulose model films, regenerated cellulose fibres, diffusion nuclear magnetic resonance, quartz crystal microbalance, vaginal infections, drug delivery systems, cytotoxicity
Objavljeno v DKUM: 13.02.2014; Ogledov: 3877; Prenosov: 188
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