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The synthesis, surface analysis, and cellular response of titania and titanium oxynitride nanotube arrays prepared on TiAl6V4 for potential biomedical applications
Katja Andrina Kravanja, Luka Suhadolnik, Marjan Bele, Uroš Maver, Jan Rožanc, Željko Knez, Maša Knez Marevci, Matjaž Finšgar, 2023, original scientific article

Abstract: Titania nanotubes are gaining prominence in the biomedical field as implant materials due to their mechanical durability, nano-rough properties, and positive influence on cellular response. This work aimed to synthesize titania and titanium oxynitride (TieOeN) nano- tubular arrays on TiAl6V4 substrates using an anodic oxidation process followed by annealing in air or by additional nitridation in NH3 atmosphere. Different nanotubular layers of unique morphology and structure were fabricated and investigated using advanced surface analysis and biocompatibility tests. In-depth surface analysis was per- formed by field emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM), 3D profilometry, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and time-of-flight secondary ion mass spectroscopy (ToF-SIMS). Cell testing using adipose- derived mesenchymal stem cells and human fetal osteoblasts demonstrated good cell viability, high proliferative capacity, and a favorable overall effect on cell morphology for the TieOeN nanotubes.
Keywords: surface analysis, XPS, AFM, tandem ToF-SIMS, titania nanotubes, Ti-O-N nanotubes, anodization
Published in DKUM: 07.05.2024; Views: 102; Downloads: 8
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Multilayer methacrylate-based wound dressing as a therapeutic tool for targeted pain relief
Tanja Zidarič, Kristijan Skok, Kristjan Orthaber, Matevž Pristovnik, Lidija Gradišnik, Tina Maver, Uroš Maver, 2023, original scientific article

Abstract: This study presents an innovative wound dressing system that offers a highly effective therapeutic solution for treating painful wounds. By incorporating the widely used non-steroidal anti-inflammatory drug diclofenac, we have created an active wound dressing that can provide targeted pain relief with ease. The drug was embedded within a biocompatible matrix composed of polyhydroxyethyl methacrylate and polyhydroxypropyl methacrylate. The multilayer structure of the dressing, which allows for sustained drug release and an exact application, was achieved through the layer-by-layer coating technique and the inclusion of superparamagnetic iron platinum nanoparticles. The multilayered dressings’ physicochemical, structural, and morphological properties were characterised using various methods. The synergistic effect of the incorporated drug molecules and superparamagnetic nanoparticles on the surface roughness and release kinetics resulted in controlled drug release. In addition, the proposed multilayer wound dressings were found to be biocompatible with human skin fibroblasts. Our findings suggest that the developed wound dressing system can contribute to tailored therapeutic strategies for local pain relief.
Keywords: wound dressings, pain relief, superparamagnetic nanoparticles, methacrylate
Published in DKUM: 19.04.2024; Views: 401; Downloads: 463
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The development of an electropolymerized, molecularly imprinted polymer (MIP) sensor for insulin determination using single-drop analysis
Tanja Zidarič, David Majer, Tina Maver, Matjaž Finšgar, Uroš Maver, 2023, original scientific article

Abstract: An electrochemical sensor for the detection of insulin in a single drop (50 μL) was developed based on the concept of molecularly imprinted polymers (MIP). The synthetic MIP receptors were assembled on a screen-printed carbon electrode (SPCE) by the electropolymerization of pyrrole (Py) in the presence of insulin (the protein template) using cyclic voltammetry. After electropolymerization, insulin was removed from the formed polypyrrole (Ppy) matrix to create imprinting cavities for the subsequent analysis of the insulin analyte in test samples. The surface characterization, before and after each electrosynthesis step of the MIP sensors, was performed using atomic force microscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. The performance of the developed MIP–SPCE sensor was evaluated using a single drop of solution containing K3Fe(CN)6 and the square-wave voltammetry technique. The MIP–SPCE showed a linear concentration range of 20.0–70.0 pM (R2 = 0.9991), a limit of detection of 1.9 pM, and a limit of quantification of 6.2 pM. The rapid response time to the protein target and the portability of the developed sensor, which is considered a disposable MIP-based system, make this MIP–SPCE sensor a promising candidate for point-of-care applications. In addition, the MIP–SPCE sensor was successfully used to detect insulin in a pharmaceutical sample. The sensor was deemed to be accurate (the average recovery was 108.46%) and precise (the relative standard deviation was 7.23%).
Keywords: electrochemical sensors, electropolymerisation, electron microscopy
Published in DKUM: 14.03.2024; Views: 237; Downloads: 22
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Novel in vitro model for toxicological analysis of drugs
Eneko Jose Madorran Esteiro, 2024, doctoral dissertation

Abstract: Despite there being a substantial range of available drugs on the market, many patients still lack effective and safe medications for their specific conditions, showing a constant need to develop new ones. Toxicological models enable us to assess their safety to various degrees, but all such models have certain limitations. Considering the advantages and drawbacks of these models, we have developed a novel viability method, Membrane Potential Cell Viability Assay (MPCVA), and a novel cell-based in vitro liver model to overcome some of the limitations of these models. The MPCVA evaluates the changes in the membrane potential, so we tested the membrane potential and cell viability relation with a cell-by-cell analysis. The MPCVA was further tested in case of cell exposure to various toxic agents that induce different cell death pathways. Finally, we used MPCVA to determine the viability of our novel cell-based in vitro liver model, which we built by combining four different types of liver cells (hepatocytes, hepatic stellate cells, Kupffer cells, and liver sinusoidal endothelial cells). Along with the MPCVA, we used clinical instrumentation to test the toxicity of three hepatotoxic drugs (5-fluorouracil, ibuprofen, and rifampicin). We observed that, under the given conditions, the MPCVA had a similar toxicity assessment as standard viability methods. We also observed that the toxicity assessment of hepatotoxic drugs is similar to the in vivo toxicity evaluation (clinical data). Therefore, the suggested approach can potentially improve existing toxicological models. However, we understand its translation to clinical practice warrants further experimentation in exposures to other drugs/toxins and by also using other cell types.
Keywords: Keywords: Toxicological model, translational analysis, liver in vitro model, cell viability.
Published in DKUM: 23.02.2024; Views: 271; Downloads: 11
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Single-strain probiotic lactobacilli for the treatment of atopic dermatitis in children : a systematic review and meta-analysis
Sabina Fijan, Nina Kolč, Metka Hrašovec, Gro Jamtvedt, Maja Šikić Pogačar, Dušanka Mičetić-Turk, Uroš Maver, 2023, review article

Abstract: Probiotics are known for their positive effects on the gut microbiota. There is growing evidence that the infant gut and skin colonization have a role in the development of the immune system, which may be helpful in the prevention and treatment of atopic dermatitis. This systematic review focused on evaluating the effect of single-strain probiotic lactobacilli consumption on treating children's atopic dermatitis. Seventeen randomized placebo-controlled trials with the primary outcome of the Scoring Atopic Dermatitis (SCORAD) index were included in the systematic review. Clinical trials using single-strain lactobacilli were included. The search was conducted until October 2022 using PubMed, ScienceDirect, Web of Science, Cochrane library and manual searches. The Joanna Briggs Institute appraisal tool was used to assess the quality of the included studies. Meta-analyses and sub meta-analyses were performed using Cochrane Collaboration methodology. Due to different methods of reporting the SCORAD index, only 14 clinical trials with 1124 children were included in the meta-analysis (574 in the single-strain probiotic lactobacilli group and 550 in the placebo group) and showed that single-strain probiotic lactobacilli statistically significantly reduced the SCORAD index compared to the placebo in children with atopic dermatitis (mean difference [MD]: -4.50; 95% confidence interval [CI]: -7.50 to -1.49; Z = 2.93; p = 0.003; heterogeneity I-2 = 90%). The subgroup meta-analysis showed that strains of Limosilactobacillus fermentum were significantly more effective than strains of Lactiplantibacillus plantarum, Lacticaseibacillus paracasei or Lacticaseibacillus rhamnosus. A longer treatment time and younger treatment age statistically significantly reduced symptoms of atopic dermatitis. The result of this systematic review and meta-analysis shows that certain single-strain probiotic lactobacilli are more successful than others in reducing atopic dermatitis severity in children. Therefore, careful consideration to strain selection, treatment time and the age of the treated patients are important factors in enhancing the effectiveness of reducing atopic dermatitis in children when choosing probiotic single-strain lactobacilli.
Keywords: probiotics, lactobacilli, atopic dermatitis, children, meta-analysis, systematic review
Published in DKUM: 16.02.2024; Views: 314; Downloads: 13
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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, original scientific article

Abstract: 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.
Keywords: poultry feathers, subcritical water hydrolysis, keratin, physico-chemical characterization, cytotoxicity
Published in DKUM: 28.11.2023; Views: 336; Downloads: 142
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Amoxicillin doped hyaluronic acid/fucoidan multifunctional coatings for medical grade stainless steel orthopedic implants
Matej Bračič, Sanja Potrč, Matjaž Finšgar, Lidija Gradišnik, Uroš Maver, Hanna Budasheva, Dorota Korte, Mladen Franko, Lidija Fras Zemljič, 2023, original scientific article

Abstract: This work investigated the potential of amoxicillin-doped hyaluronic acid/fucoidan multifunctional coatings on medical grade stainless steel as biocompatible, osteointegration enhancing, antimicrobial, and bacterial biofilm inhibiting coatings for orthopedic implants. The coatings were prepared by layer-by-layer spin coating and confirmed by optical contact angle goniometry and infrared spectroscopy. Time-of-flight secondary ion mass spectrometry showed a homogeneous distribution of the individual layers. In contrast, thermal diffusivity, and thermal conductivity measurements by photothermal beam deflection spectrometry showed diffusion of the amoxicillin into the hyaluronic acid and fucoidan layers. In vitro release of amoxicillin showed complete release within one hour, as reflected by the formation of inhibition zones against Staphylococcus aureus and Escherichia coli. Synergistic effects were observed between the hyaluronic acid and amoxicillin in inhibiting S.Aureus biofilm, and between the fucoidan and amoxicillin in improving the antioxidant properties by an ABTS radical scavenging assay. Biocompatibility was determined with human osteoblasts, confirming the potential of such multifunc- tional coatings to enhance the bioactivity of steel-based orthop edic implants.
Keywords: orthopedic implant, medical grade stainless steel, amoxicillin, hyaluronic acid, fucoidan, multifunctional coating
Published in DKUM: 18.08.2023; Views: 480; Downloads: 86
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