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    First Report on Chitin in a Non-Verongiid Marine Demosponge: The Mycale euplectellioides Case
    (Basel : MDPI, 2018) Żółtowska-Aksamitowska, Sonia; Shaala, Lamiaa A.; Youssef, Diaa T.A.; Elhady, Sameh S.; Tsurkan, Mikhail V.; Petrenko, Iaroslav; Wysokowski, Marcin; Tabachnick, Konstantin; Meissner, Heike; Ivanenko, Viatcheslav N.; Bechmann, Nicole; Joseph, Yvonne; Jesionowski, Teofil; Ehrlich, Hermann
    Sponges (Porifera) are recognized as aquatic multicellular organisms which developed an effective biochemical pathway over millions of years of evolution to produce both biologically active secondary metabolites and biopolymer-based skeletal structures. Among marine demosponges, only representatives of the Verongiida order are known to synthetize biologically active substances as well as skeletons made of structural polysaccharide chitin. The unique three-dimensional (3D) architecture of such chitinous skeletons opens the widow for their recent applications as adsorbents, as well as scaffolds for tissue engineering and biomimetics. This study has the ambitious goal of monitoring other orders beyond Verongiida demosponges and finding alternative sources of naturally prestructured chitinous scaffolds; especially in those demosponge species which can be cultivated at large scales using marine farming conditions. Special attention has been paid to the demosponge Mycale euplectellioides (Heteroscleromorpha: Poecilosclerida: Mycalidae) collected in the Red Sea. For the first time, we present here a detailed study of the isolation of chitin from the skeleton of this sponge, as well as its identification using diverse bioanalytical tools. Calcofluor white staining, Fourier-transform Infrared Spcetcroscopy (FTIR), electrospray ionization mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and fluorescence microscopy, as well as a chitinase digestion assay were applied in order to confirm with strong evidence the finding of a-chitin in the skeleton of M. euplectellioides. We suggest that the discovery of chitin within representatives of the Mycale genus is a promising step in their evaluation of these globally distributed sponges as new renewable sources for both biologically active metabolites and chitin, which are of prospective use for pharmacology and biomaterials oriented biomedicine, respectively.
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    Spider 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, Hermann
    Chitin, 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.
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    Express 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, Hermann
    Sponges 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.
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    Electrochemical 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, Hermann
    The 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/).
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    Naturally prefabricated marine biomaterials: Isolation and applications of flat chitinous 3D scaffolds from Ianthella labyrinthus (demospongiae: Verongiida)
    (Basel : Molecular Diversity Preservation International, 2019) Schubert, Mario; Binnewerg, Björn; Voronkina, Alona; Muzychka, Lyubov; Wysokowski, Marcin; Petrenko, Iaroslav; Kovalchuk, Valentine; Tsurkan, Mikhail; Martinovic, Rajko; Bechmann, Nicole; Ivanenko, Viatcheslav N.; Fursov, Andriy; Smolii, Oleg B.; Fromont, Jane; Joseph, Yvonne; Bornstein, Stefan R.; Giovine, Marco; Erpenbeck, Dirk; Guan, Kaomei; Ehrlich, Hermann
    Marine sponges remain representative of a unique source of renewable biological materials. The demosponges of the family Ianthellidae possess chitin-based skeletons with high biomimetic potential. These three-dimensional (3D) constructs can potentially be used in tissue engineering and regenerative medicine. In this study, we focus our attention, for the first time, on the marine sponge Ianthella labyrinthus Bergquist & Kelly-Borges, 1995 (Demospongiae: Verongida: Ianthellidae) as a novel potential source of naturally prestructured bandage-like 3D scaffolds which can be isolated simultaneously with biologically active bromotyrosines. Specifically, translucent and elastic flat chitinous scaffolds have been obtained after bromotyrosine extraction and chemical treatments of the sponge skeleton with alternate alkaline and acidic solutions. For the first time, cardiomyocytes differentiated from human induced pluripotent stem cells (iPSC-CMs) have been used to test the suitability of I. labyrinthus chitinous skeleton as ready-to-use scaffold for their cell culture. Results reveal a comparable attachment and growth on isolated chitin-skeleton, compared to scaffolds coated with extracellular matrix mimetic Geltrex®. Thus, the natural, unmodified I. labyrinthus cleaned sponge skeleton can be used to culture iPSC-CMs and 3D tissue engineering. In addition, I. labyrinthus chitin-based scaffolds demonstrate strong and efficient capability to absorb blood deep into the microtubes due to their excellent capillary effect. These findings are suggestive of the future development of new sponge chitin-based absorbable hemostats as alternatives to already well recognized cellulose-based fabrics. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
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    New 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, Hermann
    The 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/).