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1.
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, izvirni znanstveni članek

Opis: 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.
Ključne besede: wound dressings, pain relief, superparamagnetic nanoparticles, methacrylate
Objavljeno v DKUM: 19.04.2024; Ogledov: 57; Prenosov: 0
.pdf Celotno besedilo (1,61 MB)
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2.
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: 245; Prenosov: 18
.pdf Celotno besedilo (2,78 MB)
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3.
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, izvirni znanstveni članek

Opis: 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.
Ključne besede: orthopedic implant, medical grade stainless steel, amoxicillin, hyaluronic acid, fucoidan, multifunctional coating
Objavljeno v DKUM: 18.08.2023; Ogledov: 400; Prenosov: 80
.pdf Celotno besedilo (10,18 MB)
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4.
Biomaterials and host versus graft response : a short review
Tomaž Velnar, Gorazd Bunc, Robert Klobucar, Lidija Gradišnik, 2016, pregledni znanstveni članek

Opis: Biomaterials and biotechnology are increasing becoming an important area in modern medicine. The main aim in this area is the development of materials, which are biocompatible to normal tissue. Tissue-implant interactions with molecular, biological and cellular characteristics at the implant-tissue interface are important for the use and development of implants. Implantation may cause an inflammatory and immune response in tissue, foreign body reaction, systemic toxicity and imminent infection. Tissue-implant interactions determine the implant life-period. The aims of the study are to consider the biological response to implants. Biomaterials and host reactions to implants and their mechanisms are also briefly discussed.
Ključne besede: host versus graft disease, GVHD, biomaterial, wound healing, transplant, tissue, prosthetic, implants, biological response, complications
Objavljeno v DKUM: 03.08.2017; Ogledov: 1333; Prenosov: 187
.pdf Celotno besedilo (815,30 KB)
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5.
Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)
Jakob Naranđa, Lidija Gradišnik, Mario Gorenjak, Matjaž Vogrin, Uroš Maver, 2017, izvirni znanstveni članek

Opis: BACKGROUND: Cartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue. METHODS: Isolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production. RESULTS: Cartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well. DISCUSSION: In this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very frequently performed orthopaedic surgery during which both femoral condyles are removed. The latter present the ideal source for a simple and relatively cheap isolation of chondrocytes as was confirmed in our study.
Ključne besede: aggrecan, collagen 2, gene expression, human articular chondrocytes, isolation protocol, phenotype preservation, TKA, total knee arthroplasty
Objavljeno v DKUM: 02.08.2017; Ogledov: 1584; Prenosov: 639
.pdf Celotno besedilo (42,50 MB)
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6.
Single-cell analysis reveals IGF-1 potentiation of inhibition of the TGF-ß/Smad pathway of fibrosis in human keratocytes in vitro
Tomislav Šarenac, Martin Trapečar, Lidija Gradišnik, Marjan Rupnik, Dušica Pahor, 2016, izvirni znanstveni članek

Opis: Corneal wound healing is often affected by TGF-β–mediated fibrosis and scar formation. Guided fibrosis with IGF-1 and antifibrotic substances might maintain corneal transparency. Primary human corneal keratocytes under serum-free conditions were used as a model of corneal stromal wounding, with markers of corneal fibrosis and opacity studied under TGF-β2 stimulation. Single-cell imaging flow cytometry was used to determine nuclearization of Smad3, and intracellular fluorescence intensity of Smad7 and the corneal crystallin aldehyde dehydrogenase 3A1. Extracellular matrix proteoglycans keratocan and biglycan were quantified using ELISAs. On the TGF-β2 background, the keratocytes were treated with IGF-1, and suberoylanilidehydroxamic acid (SAHA) or halofuginone ± IGF-1. IGF-1 alone decreased Smad3 nuclearization and increased aldehyde dehydrogenase 3A1 expression, with favorable extracellular matrix proteoglycan composition. SAHA induced higher Smad7 levels and inhibited translocation of Smad3 to the nucleus, also when combined with IGF-1. Immunofluorescence showed that myofibroblast transdifferentiation is attenuated and appearance of fibroblasts is favored by IGF-1 alone and in combination with the antifibrotic substances. The TGF-β/Smad pathway of fibrosis and opacity was inhibited by IGF-1, and further with SAHA in particular, and with halofuginone. IGF-1 is thus a valid aid to antifibrotic treatment, with potential for effective and transparent corneal wound healing.
Ključne besede: cornea, wounds, treatment, antifibrotic treatment, keratocytes
Objavljeno v DKUM: 23.06.2017; Ogledov: 1317; Prenosov: 172
.pdf Celotno besedilo (1,89 MB)
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7.
Polyester type polyHIPE scaffolds with an interconnected porous structure for cartilage regeneration
Jakob Naranđa, Maja Sušec, Uroš Maver, Lidija Gradišnik, Mario Gorenjak, Andreja Vukasović, Alan Ivković, Marjan Rupnik, Matjaž Vogrin, Peter Krajnc, 2016, izvirni znanstveni članek

Opis: Development of artificial materials for the facilitation of cartilage regeneration remains an important challenge in orthopedic practice. Our study investigates the potential for neocartilage formation within a synthetic polyester scaffold based on the polymerization of high internal phase emulsions. The fabrication of polyHIPE polymer (PHP) was specifically tailored to produce a highly porous (85%) structure with the primary pore size in the range of 50–170 μm for cartilage tissue engineering. The resulting PHP scaffold was proven biocompatible with human articular chondrocytes and viable cells were observed within the materials as evaluated using the Live/Dead assay and histological analysis. Chondrocytes with round nuclei were organized into multicellular layers on the PHP surface and were observed to grow approximately 300 μm into the scaffold interior. The accumulation of collagen type 2 was detected using immunohistochemistry and chondrogenic specific genes were expressed with favorable collagen type 2 to 1 ratio. In addition, PHP samples are biodegradable and their baseline mechanical properties are similar to those of native cartilage, which enhance chondrocyte cell growth and proliferation.
Ključne besede: polyester, polymerization, polyHIPE
Objavljeno v DKUM: 23.06.2017; Ogledov: 1804; Prenosov: 398
.pdf Celotno besedilo (1,24 MB)
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8.
Novel chitosan/diclofenac coatings on medical grade stainless steel for hip replacement applications
Matjaž Finšgar, Amra Perva-Uzunalić, Janja Stergar, Lidija Gradišnik, Uroš Maver, 2016, izvirni znanstveni članek

Opis: Corrosion resistance, biocompatibility, improved osteointegration, as well the prevention of inflammation and pain are the most desired characteristics of hip replacement implants. In this study we introduce a novel multi-layered coating on AISI 316LVM stainless steel that shows promise with regard to all mentioned characteristics. The coating is prepared from alternating layers of the biocompatible polysaccharide chitosan and the non-steroid anti-inflammatory drug (NSAID), diclofenac. Electrochemical methods were employed to characterize the corrosion behavior of coated and uncoated samples in physiological solution. It is shown that these coatings improve corrosion resistance. It was also found that these coatings release the incorporated drug in controlled, multi-mechanism manner. Adding additional layers on top of the as-prepared samples, has potential for further tailoring of the release profile and increasing the drug dose. Biocompatibility was proven on human-derived osteoblasts in several experiments. Only viable cells were found on the sample surface after incubation of the samples with the same cell line. This novel coating could prove important for prolongation of the application potential of steel-based hip replacements, which are these days often replaced by more expensive ceramic or other metal alloys.
Ključne besede: corrosion, corrosion resistance, chitosan, biocompatibility, biomaterials, biomedical materials, coatings, stainless steel
Objavljeno v DKUM: 23.06.2017; Ogledov: 1885; Prenosov: 370
.pdf Celotno besedilo (2,73 MB)
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9.
Platelet-rich plasma, especially when combined with a TGF-ß inhibitor promotes proliferation, viability and myogenic differentiation of myoblasts in vitro
Robi Kelc, Martin Trapečar, Lidija Gradišnik, Marjan Rupnik, Matjaž Vogrin, 2015, izvirni znanstveni članek

Opis: Regeneration of skeletal muscle after injury is limited by scar formation, slow healing time and a high recurrence rate. A therapy based on platelet-rich plasma (PRP) has become a promising lead for tendon and ligament injuries in recent years, however concerns have been raised that PRP-derived TGF-β could contribute to fibrotic remodelling in skeletal muscle after injury. Due to the lack of scientific grounds for a PRP -based muscle regeneration therapy, we have designed a study using human myogenic progenitors and evaluated the potential of PRP alone and in combination with decorin (a TGF-β inhibitor), to alter myoblast proliferation, metabolic activity, cytokine profile and expression of myogenic regulatory factors (MRFs). Advanced imaging multicolor single-cell analysis enabled us to create a valuable picture on the ratio of quiescent, activated and terminally committed myoblasts in treated versus control cell populations. Finally high-resolution confocal microscopy validated the potential of PRP and decorin to stimulate the formation of polynucleated myotubules. PRP was shown to down-regulate fibrotic cytokines, increase cell viability and proliferation, enhance the expression of MRFs, and contribute to a significant myogenic shift during differentiation. When combined with decorin further synergistc effects were identified. These results suggest that PRP could not only prevent fibrosis but could also stimulate muscle commitment, especially when combined with a TGF-β inhibitor.
Ključne besede: muscles, skeletal, injuries, TGF-beta, plasma, thrombocytes, myoblasts, fibrosis, prevention, regeneration
Objavljeno v DKUM: 19.06.2017; Ogledov: 1272; Prenosov: 158
.pdf Celotno besedilo (2,07 MB)
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10.
A co-culture model of the developing small intestine offers new insight in the early immunomodulation of enterocytes and macrophages by Lactobacillus spp. through STAT1 and NF-kB p65 translocation
Martin Trapečar, Aleš Goropevšek, Mario Gorenjak, Lidija Gradišnik, Marjan Rupnik, 2014, izvirni znanstveni članek

Opis: The early establishment of a complete microbiome has been shown to play an integral part in the development and maintenance of an intact intestine and its immune system, although much remains unknown about the specific mechanisms of immune modulation in newborns. In our study we show in a co-culture model of the undeveloped small intestine that members of Lactobacillus spp. influence STAT1 and NF-kB p65 nuclear translocation in both intestinal epithelial cells as well as underlying macrophages. Moreover, by using imaging flow cytometry we were able to monitor each individual cell and create a framework of the percentage of cells in which translocation occurred in challenged versus control cell populations. We also observed a significant difference in baseline translocation in intestinal cells when cultured alone versus those in a co-culture model, underpinning the importance of 3D models over monolayer set-ups in epithelial in vitro research. In conclusion, our work offers new insights into the potential routes by which the commensal microbiome primes the early immune system to fight pathogens, and shows how strain-specific these mechanisms really are.
Ključne besede: microbiome, Lactobacillus, immune system, pathogens
Objavljeno v DKUM: 19.06.2017; Ogledov: 1362; Prenosov: 359
.pdf Celotno besedilo (2,86 MB)
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