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- ItemNaturally drug-loaded chitin: Isolation and applications(Basel : MDPI, 2019) Kovalchuk, Valentine; Voronkina, Alona; Binnewerg, Björn; Schubert, Mario; Muzychka, Liubov; Wysokowski, Marcin; Tsurkan, Mikhail V.; Bechmann, Nicole; Petrenko, Iaroslav; Fursov, Andriy; Martinovic, Rajko; Ivanenko, Viatcheslav N.; Fromont, Jane; Smolii, Oleg B.; Joseph, Yvonne; Giovine, Marco; Erpenbeck, Dirk; Gelinsky, Michael; Springer, Armin; Guan, Kaomei; Bornstein, Stefan R.; Ehrlich, HermannNaturally occurring three-dimensional (3D) biopolymer-based matrices that can be used in different biomedical applications are sustainable alternatives to various artificial 3D materials. For this purpose, chitin-based structures from marine sponges are very promising substitutes. Marine sponges from the order Verongiida (class Demospongiae) are typical examples of demosponges with well-developed chitinous skeletons. In particular, species belonging to the family Ianthellidae possess chitinous, flat, fan-like fibrous skeletons with a unique, microporous 3D architecture that makes them particularly interesting for applications. In this work, we focus our attention on the demosponge Ianthella flabelliformis (Linnaeus, 1759) for simultaneous extraction of both naturally occurring (“ready-to-use”) chitin scaffolds, and biologically active bromotyrosines which are recognized as potential antibiotic, antitumor, and marine antifouling substances. We show that selected bromotyrosines are located within pigmental cells which, however, are localized within chitinous skeletal fibers of I. flabelliformis. A two-step reaction provides two products: treatment with methanol extracts the bromotyrosine compounds bastadin 25 and araplysillin-I N20 sulfamate, and a subsequent treatment with acetic acid and sodium hydroxide exposes the 3D chitinous scaffold. This scaffold is a mesh-like structure, which retains its capillary network, and its use as a potential drug delivery biomaterial was examined for the first time. The results demonstrate that sponge-derived chitin scaffolds, impregnated with decamethoxine, effectively inhibit growth of the human pathogen Staphylococcus aureus in an agar diffusion assay
- ItemElectrochemical approach for isolation of chitin from the skeleton of the black coral cirrhipathes sp. (Antipatharia)(Basel : MDPI, 2020) Nowacki, Krzysztof; Stępniak, Izabela; Langer, Enrico; Tsurkan, Mikhail; Wysokowski, Marcin; Petrenko, Iaroslav; Khrunyk, Yuliya; Fursov, Andriy; Bo, Marzia; Bavestrello, Giorgio; Joseph, Yvonne; Ehrlich, HermannThe development of novel and effective methods for the isolation of chitin, which remains one of the fundamental aminopolysaccharides within skeletal structures of diverse marine invertebrates, is still relevant. In contrast to numerous studies on chitin extraction from crustaceans, mollusks and sponges, there are only a few reports concerning its isolation from corals, and especially black corals (Antipatharia). In this work, we report the stepwise isolation and identification of chitin from Cirrhipathes sp. (Antipatharia, Antipathidae) for the first time. The proposed method, aiming at the extraction of the chitinous scaffold from the skeleton of black coral species, combined a well-known chemical treatment with in situ electrolysis, using a concentrated Na2SO4 aqueous solution as the electrolyte. This novel method allows the isolation of a-chitin in the form of a microporous membrane-like material. Moreover, the extracted chitinous scaffold, with a well-preserved, unique pore distribution, has been extracted in an astoundingly short time (12 h) compared to the earlier reported attempts at chitin isolation from Antipatharia corals. © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
- ItemSpider chitin: An Ultrafast Microwave-Assisted Method for Chitin Isolation from Caribena versicolor Spider Molt Cuticle(Basel : MDPI, 2019) Machałowski, Tomasz; Wysokowski, Marcin; Tsurkan, Mikhail V.; Galli, Roberta; Schimpf, Christian; Rafaja, David; Brendler, Erica; Viehweger, Christine; Zółtowska-Aksamitowska, Sonia; Petrenko, Iaroslav; Czaczyk, Katarzyna; Kraft, Michael; Bertau, Martin; Bechmann, Nicole; Guan, Kaomei; Bornstein, Stefan R.; Voronkina, Alona; Fursov, Andriy; Bejger, Magdalena; Biniek-Antosiak, Katarzyna; Rypniewski, Wojciech; Figlerowicz, Marek; Pokrovsky, Oleg; Jesionowski, Teofil; Ehrlich, HermannChitin, as a fundamental polysaccharide in invertebrate skeletons, continues to be actively investigated, especially with respect to new sources and the development of effective methods for its extraction. Recent attention has been focused on marine crustaceans and sponges; however, the potential of spiders (order Araneae) as an alternative source of tubular chitin has been overlooked. In this work, we focused our attention on chitin from up to 12 cm-large Theraphosidae spiders, popularly known as tarantulas or bird-eating spiders. These organisms “lose” large quantities of cuticles during their molting cycle. Here, we present for the first time a highly effective method for the isolation of chitin from Caribena versicolor spider molt cuticle, as well as its identification and characterization using modern analytical methods. We suggest that the tube-like molt cuticle of this spider can serve as a naturally prefabricated and renewable source of tubular chitin with high potential for application in technology and biomedicine. © 2019 by the authors.
- ItemSurface-Dependent Osteoblasts Response to TiO2 Nanotubes of Dfferent Crystallinity(Basel : MDPI, 2020) Khrunyk, Yuliya Y.; Belikov, Sergey V.; Tsurkan, Mikhail V.; Vyalykh, Ivan V.; Markaryan, Alexandr Y.; Karabanalov, Maxim S.; Popov, Artemii A.; Wysokowski, MarcinOne of the major challenges of implantology is to design nanoscale modifications of titanium implant surfaces inducing osseointegration. The aim of this study was to investigate the behavior of rat osteoblasts cultured on anodized TiO2 nanotubes of different crystallinity (amorphous and anatase phase) up to 24 days. TiO2 nanotubes were fabricated on VT1–0 titanium foil via a two-step anodization at 20 V using NH4F as an electrolyte. Anatase-phase samples were prepared by heat treatment at 500 °C for 1 h. VT1–0 samples with flat surfaces were used as controls. Primary rat osteoblasts were seeded over experimental surfaces for several incubation times. Scanning electron microscopy (SEM) was used to analyze tested surfaces and cell morphology. Cell adhesion and proliferation were investigated by cell counting. Osteogenic differentiation of cells was evaluated by qPCR of runt-related transcription factor 2 (RUNX2), osteopontin (OPN), integrin binding sialoprotein (IBSP), alkaline phosphatase (ALP) and osteocalcin (OCN). Cell adhesion and proliferation, cell morphology and the expression of osteogenic markers were affected by TiO2 nanotube layered substrates of amorphous and anatase crystallinity. In comparison with flat titanium, along with increased cell adhesion and cell growth a large portion of osteoblasts grown on the both nanostructured surfaces exhibited an osteocyte-like morphology as early as 48 h of culture. Moreover, the expression of all tested osteogenic markers in cells cultured on amorphous and anatase TiO2 nanotubes was upregulated at least at one of the analyzed time points. To summarize, we demonstrated that amorphous and anodized TiO2 layered substrates are highly biocompatible with rat osteoblasts and that the surface modification with about 1500 nm length nanotubes of 35 ± 4 (amorphous phase) and 41 ± 8 nm (anatase phase) in diameter is sufficient to induce their osteogenic differentiation. Such results are significant to the engineering of coating strategies for orthopedic implants aimed to establish a more efficient bone to implant contact and enhance bone repair. © 2020 by the author. Licensee MDPI, Basel, Switzerland.
- ItemNew Source of 3D Chitin Scaffolds: The Red Sea Demosponge Pseudoceratina arabica (Pseudoceratinidae, Verongiida)(Basel : MDPI, 2019) Shaala, Lamiaa A.; Asfour, Hani Z.; Youssef, Diaa T.A.; Żółtowska-Aksamitowska, Sonia; Wysokowski, Marcin; Tsurkan, Mikhail; Galli, Roberta; Meissner, Heike; Petrenko, Iaroslav; Tabachnick, Konstantin; Ivanenko, Viatcheslav N.; Bechmann, Nicole; Muzychka, Lyubov V.; Smolii, Oleg B.; Martinović, Rajko; Joseph, Yvonne; Jesionowski, Teofil; Ehrlich, HermannThe bioactive bromotyrosine-derived alkaloids and unique morphologically-defined fibrous skeleton of chitin origin have been found recently in marine demosponges of the order Verongiida. The sophisticated three-dimensional (3D) structure of skeletal chitinous scaffolds supported their use in biomedicine, tissue engineering as well as in diverse modern technologies. The goal of this study was the screening of new species of the order Verongiida to find another renewable source of naturally prefabricated 3D chitinous scaffolds. Special attention was paid to demosponge species, which could be farmed on large scale using marine aquaculture methods. In this study, the demosponge Pseudoceratina arabica collected in the coastal waters of the Egyptian Red Sea was examined as a potential source of chitin for the first time. Various bioanalytical tools including scanning electron microscopy (SEM), fluorescence microscopy, FTIR analysis, Calcofluor white staining, electrospray ionization mass spectrometry (ESI-MS), as well as a chitinase digestion assay were successfully used to confirm the discovery of a-chitin within the skeleton of P. arabica. The current finding should make an important contribution to the field of application of this verongiid sponge as a novel renewable source of biologically-active metabolites and chitin, which are important for development of the blue biotechnology especially in marine oriented biomedicine. © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
- ItemExpress method for isolation of ready-to-use 3D chitin scaffolds from aplysina archeri (aplysineidae: verongiida) demosponge(Basel : MDPI, 2019) Klinger, Christine; Zółtowska-Aksamitowska, Sonia; Wysokowski, Marcin; Tsurkan, Mikhail V.; Galli, Roberta; Petrenko, Iaroslav; Machałowski, Tomasz; Ereskovsky, Alexander; Martinović, Rajko; Muzychka, Lyubov; Smolii, Oleg B.; Bechmann, Nicole; Ivanenko, Viatcheslav; Schupp, Peter J.; Jesionowski, Teofil; Giovine, Marco; Bornstein, Stefan R.; Voronkina, Alona; Ehrlich, HermannSponges are a valuable source of natural compounds and biomaterials for many biotechnological applications. Marine sponges belonging to the order Verongiida are known to contain both chitin and biologically active bromotyrosines. Aplysina archeri (Aplysineidae: Verongiida) is well known to contain bromotyrosines with relevant bioactivity against human and animal diseases. The aim of this study was to develop an express method for the production of naturally prefabricated 3D chitin and bromotyrosine-containing extracts simultaneously. This new method is based on microwave irradiation (MWI) together with stepwise treatment using 1% sodium hydroxide, 20% acetic acid, and 30% hydrogen peroxide. This approach, which takes up to 1 h, made it possible to isolate chitin from the tube-like skeleton of A. archeri and to demonstrate the presence of this biopolymer in this sponge for the first time. Additionally, this procedure does not deacetylate chitin to chitosan and enables the recovery of ready-to-use 3D chitin scaffolds without destruction of the unique tube-like fibrous interconnected structure of the isolated biomaterial. Furthermore, these mechanically stressed fibers still have the capacity for saturation with water, methylene blue dye, crude oil, and blood, which is necessary for the application of such renewable 3D chitinous centimeter-sized scaffolds in diverse technological and biomedical fields. © 2019 by the authors.