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Antithrombogenic polysaccharide coatings to improve hemocompatibility, protein-repellence, and endothelial cell response
Matej Bračič, Bence M. Nagy, Olivija Plohl, Florian Lackner, Tobias Alexander Steindorfer, Roland Fischer, Thomas Heinze, Andrea Olschewski, Karin Stana-Kleinschek, Chandran Nagaraj, Tamilselvan Mohan, 2024, original scientific article

Abstract: Polyester biomaterials play a crucial in vascular surgery, but suffer from unspecific protein adsorption, thrombogenicity, and inadequate endothelial cell response, which limit their success. To address these issues, we investigated the functionalization of polyester biomaterials with antithrombogenic polysaccharide coatings. A two-step and water-based method was used to coat cationized polycaprolactone with different sulfated polysaccharides (SPS), which resulted in long-term stability, tunable morphology, roughness, film thickness, chemical compositions, zeta potential, and water content. The coatings significantly increased the anticoagulant activity and reduced the thrombogenicity of polycaprolactone, particularly with highly sulfated heparin and cellulose sulfate. Less SPS, such as chondroitin sulfate, fucoidan, and carrageenan, despite showing reduced anticoagulant activity, also exhibited lower fibrinogen adsorption. The adhesion and viability of human primary endothelial cells cultured on modified polycaprolactone correlated with the type and sulfate content of the coatings.
Keywords: polyester biomaterials, vascular surgery, antithrombogenic polysaccharide coatings
Published in DKUM: 10.09.2024; Views: 65; Downloads: 10
.pdf Full text (5,07 MB)
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3.
Covalent modification of chitosan surfaces with a sugar amino acid and lysine analogues
Tobias Dorn, Matjaž Finšgar, Karin Stana-Kleinschek, Tobias Alexander Steindorfer, Martin Simon Thonhofer, Tanja Wrodnigg, Rupert Kargl, 2024, original scientific article

Abstract: This work explores the modifcation and characterization of chitosan thin flms as a model for functionalized polysaccharide interfaces. The solid–liquid interface of oligo- and polysaccharides is crucial for various biological processes such as cell adhesion and recognition. By covalent surface modifcation of the chitosan via amide formation with diferent small molecules containing carboxylic acids, e.g. specially designed glycoside hydrolase inhibitors, interactions with biomolecules and living cells could potentially be controlled in the future. As a frst step towards this aim, three fuorescent compounds were conjugated onto nanometric chitosan thin flms. The layers were analysed by fuorescence spectroscopy, X-ray photoelectron spectroscopy, time-of-fight secondary ion mass spectrometry, and atomic force microscopy, to proof the covalent attachment of the target molecules. By this analysis, a uniform and chemically stable covalent attachment of the target molecules on the chitosan thin flms could be demonstrated under various conditions. This publication serves as a proof-of-concept-study for further biofunctionalization, pattering, and interaction studies involving polysaccharide interfaces, glycosidase inhibitors, proteins, or living cells.
Keywords: carbohydrates, fluorescence spectroscopy, amino acid, chitosan, thin flm modifcation, C-Glycosides
Published in DKUM: 07.08.2024; Views: 127; Downloads: 9
.pdf Full text (2,03 MB)

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Functional 3D printed polysaccharide derivative scaffolds for vascular graft application : doctoral disertation
Fazilet Gürer, 2023, doctoral dissertation

Abstract: Tissue engineering (TE) is an interdisciplinary field that aims towards replacement, healing or reconstruction of damaged tissue and organs. Incurable diseases are currently treated with organ transplantation, that have the disadvantages of insufficient donors, immune response, and organ rejection after transplantation. TE imitate the functions of extracellular matrix (ECM) to develop biocompatible/biodegradable scaffolds with appropriate features which are utilized to provide mechanical support, cellular infiltration, migration, and tissue formation, and to mimic the biochemical and biophysical cues of cells. Several fabrication methods have been introduced to mimic the 3D structure of ECM and 3D printing is one of the additive manufacturing techniques, widely used in TE because of its feasibility to build complex tissue constructs and control over fabrication and cell distribution. The polysaccharide-peptide conjugate has gained enormous interest in recent years owing to its biocompatibility, degradability, flexibility, and structural matching to natural proteoglycans. In this context, we reported here on investigation of biocompatibility with HUVECs, surface modification of 3D printed PCL scaffolds with an amine group and chemically crosslinked oxidized HA-amino acid/peptide conjugates (OHACs) was used to develop a novel biomaterial for use as a tissue engineered vascular graft. Modified polysaccharides were characterized with respect to their chemical structure, charge, UV and fluorescence properties and cytotoxicity. The successful conjugation was demonstrated by XPS, and a decrease in the free amine peaks on the surface was observed after conjugation. In addition, the water contact angle measurements showed improved wetting, an indication that the conjugation to the PCL-A surface was successful. Finally, the biocompatibility of the novel scaffolds was characterized by the MTS and the live- dead assay. In both assays, proliferation of cells was observed after 7 days and cell spreading on the surface was detected by phalloidin staining of actin filaments. In conclusion, it was possible to prepare surface-active scaffolds by combining the advantages of biocompatibility and mechanical strength of polysaccharides and polyesters, respectively.
Keywords: 3D tiskanje, karboksimetilceluloza, hialuronska kislina, polikaprolakton, kemija karbodiimida, kemija Shiffove baze, endotelizacija 3D printing, carboxymethyl cellulose, hyaluronic acid, polycaprolactone, carbodiimide chemistry, shiff-base chemistry, endothelialization
Published in DKUM: 06.10.2023; Views: 522; Downloads: 53
.pdf Full text (7,97 MB)

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Preparation of Three Dimensional Structures of Polysaccharide Derivatives for Application in Regenerative Medicine : doctoral disertation
Andreja Dobaj-Štiglic, 2022, doctoral dissertation

Abstract: Biocompatible polysaccharide scaffolds with controllable pore size, good mechanical properties, and no hazardous chemical crosslinkers are desirable for long-term tissue engineering applications. Despite decades of development of novel scaffolds, there are still many challenges to be solved regarding their production and optimization for specifically engineered tissues. Herein, we have fabricated several three-dimensional (3D) scaffolds using polysaccharide or polysaccharide-protein composite hydrogels or inks for 3D printing, featuring strong shear thinning behavior and adequate printability. The inks, composed of various combinations of chitosan, nanofibrillated cellulose, carboxymethyl cellulose, collagen, and citric acid, were 3D printed, freeze-dried, and dehydrothermally heat-treated to obtain dimensionally and mechanically stable scaffolds. The heat-assisted step induced the formation of covalent amide and ester bonds between the functional groups of chosen polysaccharides and protein collagen. Citric acid was chosen as a non-hazardous and „green” crosslinker to further tailor the mechanical properties and long-term stability of the scaffolds. We have investigated how the complexation conditions, charge ratio, dehydrothermal treatment, and degree of crosslinking influence the scaffolds' chemical, surface, swelling, and degradation properties in the dry and hydrated states. The compressive strength, elastic modulus, dimensional stability and shape recovery of the (crosslinked) scaffolds increased significantly with balanced charge ratio, dehydrothermal treatment, and increased concentrations of citric acid crosslinker and collagen concentrations. The prepared crosslinked scaffolds promoted (clustered) cell adhesion and showed no cytotoxic effects, as determined by cell viability assays and live/dead staining with human bone tissue-derived osteoblasts and human adipose tissue-derived mesenchymal stem cells. The water-based and non-hazardous crosslinking methods presented here can be extended to all polysaccharide- or polysaccharide-protein-based materials to develop cell-friendly scaffolds with tailored properties suitable for various tissue engineering applications.
Keywords: chitosan, carboxymethyl cellulose, nanofibrillated cellulose, citric acid, collagen, freeze drying, 3D printing, dehydrothermal treatment
Published in DKUM: 11.10.2022; Views: 675; Downloads: 155
.pdf Full text (33,63 MB)

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Design, Characterisation and Applications of Cellulose-Based Thin Films, Nanofibers and 3D Printed Structures : A Laboratory Manual
Tanja Pivec, Tamilselvan Mohan, Rupert Kargl, Manja Kurečič, Karin Stana-Kleinschek, 2021, other educational material

Abstract: The introduction of the Laboratory Manual gives the theoretical bases on cellulose and its derivatives, which are used as starting polymers for the preparation of multifunctional polymers with three different advanced techniques - spin coating, electrospinning and 3D printing. In the following, each technique is presented in a separate Lab Exercise. Each exercise covers the theoretical basics on techniques for polymer processing and methods for their characterisation, with an emphasis on the application of prepared materials. The experimental sections contain all the necessary information needed to implement the exercises, while the added results provide students with the help to implement correct and successful exercises and interpret the results.
Keywords: multifunctional polymers, polysaccharides, cellulose, electrospun, spin coating, 3D printing, nanofibers, thin films, multifunctional materials, laboratory manuals
Published in DKUM: 09.03.2021; Views: 965; Downloads: 25
URL Link to file

10.
Development of polymeric materials with rutin and polyrutin for healing of chronic leg ulcers
Tanja Pivec, 2018, doctoral dissertation

Abstract: In this work, the development of cellulose wound dressing materials with rutin (RU) and polyrutin (PR) for the healing of chronic leg ulcers is presented as a new approach of local treatment of this wound type. The flavonoid rutin is a known antioxidant substance of plant origin with wound healing promoting properties. Despite the proven beneficial properties of rutin, its potential application in wound healing is limited due to its low water solubility. This limitation can be overcome by polymerization of rutin into polyrutin. In this work an enzymatic polymerization of rutin in water without addition of organic solvents was performed to obtain a water-soluble polymer polyrutin. The chemical structure of rutin and polyrutin were investigated using UV-Vis spectroscopy, nuclear magnetic resonance spectroscopy, Fourier transform infrared spectroscopy, size-exclusion chromatography and potentiometric titrations. Biological activity related to a desired positive influence on chronic leg ulcers was investigated through the determination of the antioxidant activity, iron-chelation ability, cell viability, determination of cell proliferation and through use of the so called “scratch assay” to measure in vitro wound healing performance. Results indicate that rutin and polyrutin have a positive influence on the healing of chronic wounds. Rutin and polyrutin water dispersions at different pH and ionic strength were further characterised by means of dynamic light scattering in order to determine the size of particles and their pH dependent ζ-potential. The knowledge gained from these measurements aided the systematic interaction studies of rutin and polyrutin dispersions with cellulose-based surfaces via model and real wound healing systems. The model cellulose surfaces were thin films prepared by dissolution of trimethylsilyl cellulose in tetrahydrofuran, spin-coating of this solution on sensors of a quartz crystal microbalance (QCM-D) and subsequent regeneration of trimethylsilyl cellulose to cellulose with acid vapors. The influence of pH, salt concentration, and rutin/polyrutin concentration on the interaction with cellulose thin films was evaluated by means of a quartz crystal microbalance with dissipation. This knowledge was transferred to the application of the coatings on real wound healing systems i.e. cellulose non-wovens. The surface morphology was further characterised on model and real wound healing systems. The antioxidant activity and release kinetics were investigated for a real wound healing system, similar to the clinically used, cellulose based wound dressing materials. The main results showed that a higher solubility of polyrutin at low ionic strength contributes to the formation of continuous layers of polyrutin on cellulose surface, while the low solubility of rutin and reduced solubility of polyrutin at higher ionic strengths contribute to deposition of particles of rutin and polyrutin on the cellulose surface. The presence of particles on the surface of non-woven cellulose fibres led to a faster initial release of rutin and polyrutin. On the contrary, a continuous layer of the well soluble polyrutin contributes to a prolonged release. Namely, adsorption of the water soluble polyrutin at pH 2 without the addition of salt results in higher masses of attached polyrutin that release slower and over longer time periods. Since wound dressing materials for chronic leg ulcers often require a lower frequency of dressing change, the latter could provide an efficient therapeutic approach to their treatment.
Keywords: Chronic wounds, Wound dressings, Cellulose, Polymerization of flavonoids, Rutin, Polyrutin
Published in DKUM: 11.06.2018; Views: 1802; Downloads: 250
.pdf Full text (7,34 MB)

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