Suchergebnisse

Collagens are the most abundant high molecular weight proteins in both invertebrate and vertebrate organisms, including mammals, and possess mainly a structural role, existing different types according with their specific organization in distinct tissues. From this, they have been elected as one of the key biological materials in tissue regeneration approaches. Also, industry is constantly searching for new natural sources of collagen and upgraded methodologies for their production. The most common sources are from bovine and porcine origin, but other ways are making their route, such as recombinant production, but also extraction from marine organisms like fish. Different organisms have been proposed and explored for collagen extraction, allowing the sustainable production of different types of collagens, with properties depending on the kind of organism (and their natural environment) and extraction methodology. Such variety of collagen properties has been further investigated in different ways to render a wide range of applications. The present review aims to shed some light on the contribution of marine collagens for the scientific and technological development of this sector, stressing the opportunities and challenges that they are and most probably will be facing to assume a role as an alternative source for industrial exploitation. ; The authors would like to acknowledge the funding from ERDF through Atlantic Area Transnational Cooperation Programme Project 2011-1/164 MARMED, POCTEP Projects 0330_IBEROMARE_1_P and 0687_NOVOMAR_1_P and from European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreements nr. KBBE-2010-266033-SPECIAL and nr. REGPOT-CT2012-316331-POLARIS. The Portuguese Foundation for Science and Technology is also acknowledged for the Post-Doc fellowship SFRH/BPD/70230/2010 ...

Sponges occur ubiquitously in the marine realm and in some deep-sea areas they dominate the benthic communities forming complex biogenic habitats â sponge grounds, aggregations, gardens and reefs. However, deep-sea sponges and spongegrounds are still poorly investigated with regards to biotechnological potential in support of a Blue growth strategy. Under the scope of this study, five dominant North Atlantic deep-sea sponges, were characterized to elucidate promising applications in human health, namely for bone tissue engineering approaches. Geodia barretti (Gb), Geodia atlantica (Ga), Stelletta normani (Sn), Phakellia ventilabrum (Pv), and Axinella infundibuliformis (Ai), were morphologically characterized to assess macro and microstructural features, as well as chemical composition of the skeletons, using optical and scanning electron microscopy, energy dispersive x-ray spectroscopy and microcomputed tomography analyses. Moreover, compress tests were conducted to determine the mechanical properties of the skeletons. Results showed that all studied sponges have porous skeletons with porosity higher than 68%, pore size superior than 149 mm and higher interconnectivity (>96%), thus providing interesting models for the development of scaffolds for tissue engineering. Besides that, EDS analyses revealed that the chemical composition of sponges, pointed that demosponge skeletons are mainly constituted by carbon, silicon, sulfur, and oxygen combined mutually with organic and inorganic elements embedded its internal architecture that can be important features for promoting bone matrix quality and bone mineralization. Finally, the morphological, mechanical, and chemical characteristics here investigated unraveled the potential of deep-sea sponges as a source of biomaterials and biomimetic models envisaging tissue engineering applications for bone regeneration. ; The authors would like to acknowledge the funding from the European Union Framework Program for Research and Innovation Horizon 2020 through project ...

An Open Access paper submitted under the category "Uncovered". ; We acknowledge the financial support from European Union H2020 programme (SponGES – 679849, CHEM2NATURE – 692333 and FORECAST – 668983) and FP7 programme (ERC Advanced Grant COMPLEXITE, ERC- 2012-ADG-20120216-321266).

In this study, the stability and biocompatibility of methacrylated gellan gum hydrogels, obtained either by ionic- (iGG-MA) or photo-crosslinking (phGGMA), were evaluated in vitro and in vivo. Size exclusion chromatography analysis of the methacrylated gellan gum (GG-MA) powder revealed that molecular weight is lower as compared to the non-modifi ed material, i.e., low acyl gellan gum. The water uptake and swelling of iGG-MA and phGGMA hydrogels were investigated in phosphate-buffered saline solution (pH 7.4). The biocompatibility of the hydrogels was fi rstly evaluated by producing cell-laden hydrogels. The in vitro cells encapsulation study showed that lung fi broblast cells (L929 cells) and human intervertebral disc (hIVD) cells are viable when cultured within both hydrogels, up to 21 days of culturing. The iGG-MA and phGG-MA hydrogels were also subcutaneously implanted in Lewis rats for 10 and 18 days. Tissue response to the hydrogels implantation was determined by histological analysis (haematoxylin-eosin staining). A thin fi brous capsule was observed around the implanted hydrogels. No necrosis, calcifi cation, and acute infl ammatory reaction were observed. The results presented in this study demonstrate that iGG-MA and phGG-MA hydrogels are stable in vitro and in vivo, support L929 and hIVD cells' encapsulation and viability, and were found to be well-tolerated and non-toxic in vivo. ; The authors thank the funds provided by Portuguese Foundation for Science and Technology (FCT) through POCTI and FEDER programmes. This work was also supported by the European Union funded Collaborative Project Disc Regeneration ...

Demosponges are a well-known source of a plethora of bioactive compounds. In particular, they are able to form a skeleton by direct deposition of silica in a process catalysed by silicatein. Herein, we isolated biosilicas from five different Atlantic deep-sea sponges Geodia atlantica (GA), Geodia barretti (GB), Stelletta normani (SN), Axinella infundibuliformis (AI) and Phakellia ventilabrum (PV) to explore the bioactivity and osteogenic capacity of its silica-based materials. We chemically characterized the isolated biosilicas and evaluated them for their bioactivity to deposit Ca and P on their surface (by immersion in simulated body fluid, SBF). GB-, SN-, AI- and PV-based biosilicas did not generate a stable calcium phosphate (CaP) layer over time in the presence of SBF, however, the GA-derived one was able to form a CaP surface layer (at a Ca/P ratio of ~1.7, similar to the one observed for hydroxyapatite), that was stable during the 28 days of testing. In addition, no cytotoxicity towards L929 and SaOs2 cells was observed for the GA-based biosilica up to a concentration of 10 mg/mL. Overall, the GA-based biosilica presents the characteristics to be used in the development of biomaterials for bone tissue engineering (BTE). ; This research has been performed with funding from the European Union's Horizon 2020 research and innovation programme under Grant Agreements No. 679849 (SponGES) and No. 668983 (FORECAST). Funding from the European Regional Development Fund, through Atlantic Area Program, under the scope of BlueHuman project (EAPA_151/2016) is also acknowledged. JRX research was also supported by National Funds through FCT, the Portuguese Foundation for Science and Technology within the scope of UIDB/04423/2020, UIDP/04423/2020, and ...

"Publication Date (Web): December 23, 2014" ; Marine sponges are extremely rich in natural products and are considered a promising biological resource. The major objective of this work is to couple a green extraction process with a natural origin raw material to obtain sponge origin collagen/gelatin for biomedical applications. Marine sponge collagen has unique physicochemical properties, but its application is hindered by the lack of availability due to inefficient extraction methodologies. Traditional extraction methods are time consuming as they involve several operating steps and large amounts of solvents. In this work, we propose a new extraction methodology under mild operating conditions in which water is acidified with carbon dioxide (CO2) to promote the extraction of collagen/gelatin from different marine sponge species. An extraction yield of approximately 50% of collagen/gelatin was achieved. The results of Fourier transformed infrared spectroscopy (FTIR), circular dichroism (CD), and differential scanning calorimetry (DSC) spectra suggest a mixture of collagen/gelatin with high purity, and the analysis of the amino acid composition has shown similarities with collagen from other marine sources. Additionally, in vitro cytotoxicity studies did not demonstrate any toxicity effects for three of the extracts. ; The authors are grateful for financial support of FCT through Grant EXP/QEQ:EPS/0745/2012, SWIMS (Subcritical Water Isolation of compounds from Marine Sponges). The funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement numbers REGPOT-CT2012-316331-POLARIS and KBBE-2010-266033 (project SPECIAL), as well as from ERDF under the project "Novel smart and biomimetic materials for innovative regenerative medicine approaches" RLI-ABMR-NORTE-01-0124-FEDER-000016), cofinanced by North Portugal Regional Operational Programme (ON.2,O Novo Norte), under the National Strategic Reference Framework (NSRF) are also gratefully ackowledged. The authors are also truly ...

Marine biodiversity is expressed through the huge variety of vertebrate and invertebrate species inhabiting intertidal to deep-sea environments. The extraordinary variety of â forms and functionsâ  exhibited by marine animals suggests they are a promising source of bioactive molecules and provides potential inspiration for different biomimetic approaches. This diversity is familiar to biologists and has led to intensive investigation of metabolites, polysaccharides, and other compounds. However, marine collagens are less well-known. This review will provide detailed insight into the diversity of collagens present in marine species in terms of their genetics, structure, properties, and physiology. In the last part of the review the focus will be on the most common marine collagen sources and on the latest advances in the development of innovative materials exploiting, or inspired by, marine collagens. ; The authors are grateful for the financial support from European Union, under the scope of European Regional Development Fund((ERDF) through the POCTEP project 0687_NOVOMAR_1_P and Structured Project NORTE-01- 0145-FEDER-000021 and from the Portuguese Foundation for Science and Technology (FCT), under the scope of the BiogenInk project (M-ERA-NET2/0022/2016) and from the European Cooperation in Science & Technology program (EU COST). Grant title: "Stem cells of marine/aquatic inverte brates: from basic research to innovative applications" (MARISTEM). MSR acknowledges FCT for the Ph.D. scholarship ...

Peripheral nerve injuries have produced major concerns in regenerative medicine for several years, as the recovery of normal nerve function continues to be a significant clinical challenge. Chitosan (CHT), because of its good biocompatibility, biodegradability and physicochemical properties, has been widely used as a biomaterial in tissue engineering scaffolding. In this study, CHT membranes were produced with three different Degrees of Acetylation (DA), envisioning its application in peripheral nerve regeneration. The three CHT membranes (DA I: 1%, DA II: 2%, DA III: 5%) were extensively characterized and were found to have a smooth and flat surface, with DA III membrane having slightly higher roughness and surface energy. All the membranes presented suitable mechanical properties and did not show any signs of calcification after SBF test. Biodegradability was similar for all samples, and adequate to physically support neurite outgrowth. The in vitro cell culture results indicate selective cell adhesion. The CHT membranes favoured Schwann cells invasion and proliferation, with a display of appropriate cytoskeletal morphology. At the same time they presented low fibroblast infiltration. This fact may be greatly beneficial for the prevention of fibrotic tissue formation, a common phenomenon impairing peripheral nerve regeneration. The great deal of results obtained during this work permitted to select the formulation with the greatest potential for further biological tests. ; This work has received funding from the European Community's Seventh Framework Programme (FP7-HEALTH-2011) under grant agreement no 278612 (BIOHYBRID). This study was also funded by European Union's FP7 Programme under grant agreement no REGPOT-CT2012-316331-POLARIS.The authors thank the chitosan raw material provided by Altakitin S.A., (Lisboa, Portugal). We are further thankful to Silke Fischer, Natascha Heidrich, Kerstin Kuhlemann, Jennifer Metzen, Hildegard Streich and Maike Wesemann (all from the Institute of Neuroanatomy, Hannover ...

The possibility to fabricate marine collagen porous structures crosslinked with genipin under high pressure carbon dioxide is investigated. Collagen from shark skin is used to prepare prescaffolds by freeze-drying. The poor stability of the structures and low mechanical properties require crosslinking of the structures. Under dense CO2 atmosphere, crosslinking of collagen pre-scaffolds is allowed for 16 h. Additionally, the hydrogels are foamed and the scaffolds obtained present a highly porous structure. In vitro cell culture tests performed with a chondrocyte-like cell line show good cell adherence and proliferation, which is a strong indication of the potential of these scaffolds to be used in tissue cartilage tissue engineering. ; The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. KBBE-2010-266033 (project SPECIAL) and from FEDER through POCTEP Project 0330_IBEROMARE_1_P. Portuguese Foundation for Science and Technology is also gratefully acknowledged for post-doc grants of J. Moreira-Silva and T.H. ...

Collagen is the most abundant protein found in mammals and it exhibits a low immunogenicity, high biocompatibility and biodegradability when compared with others natural polymers. For this reason, it has been explored for the development of biologically instructive biomaterials with applications for tissue substitution and regeneration. Marine origin collagen has been pursued as an alternative to the more common bovine and porcine origins. This study focused on squid (Teuthoidea: Cephalopoda), particularly the Antarctic squid Kondakovia longimana and the Sub-Antarctic squid Illex argentinus as potential collagen sources. In this study, collagen has been isolated fromthe skins of the squids using acid-based and pepsin-based protocols, with the higher yield being obtained from I. argentinus in the presence of pepsin. The produced collagen has been characterized in terms of physicochemical properties, evidencing an amino acid profile similar to the one of calf collagen, but exhibiting a less preserved structure, with hydrolyzed portions and a lower melting temperature. Pepsin-soluble collagen isolated fromI. argentinus was selected for further evaluation of biomedical potential, exploring its incorporation on poly-ε-caprolactone (PCL) 3D printed scaffolds for the development of hybrid scaffolds for tissue engineering, exhibiting hierarchical features. ; This work was partially funded by ERDF through POCTEP Project 0687_NOVOMAR_1_P and by the European Union Seventh Framework Programme for research, technological development and demonstration under grant agreement on ERC-2012-ADG 20120216-321266 (ComplexiTE). The Portuguese Foundation for Science and Technology (FCT) is also acknowledged for post-doctoral fellowships of JMS (SFRH/BPD/70230/2010) and RPP (SFRH/BPD/101886/2014), financed by POPH/FSE, and FCT Investigator grant of JX (IF/00616/2013). The authors also want to thank Dr. Julio Maroto (Fundación CETMAR, Spain) for the kind offer of the samples of skins of I. argentinus, to Dr. Dario Fassini for the ...

One of the major scientific challenges that tissue engineering and regenerative medicine (TERM) faces to move from benchtop to bedside regards biomaterials development, despite the latest advances in polymer processing technologies. A variety of scaffolds processing techniques have been developed and include solvent casting and particles leaching, compression molding and particle leaching, thermally induced phase separation, rapid prototyping, among others. Supercritical fluids appear as an interesting alternative to the conventional methods for processing biopolymers as they do not require the use of large amounts of organic solvents and the processes can be conducted at mild temperatures. However, this processing technique has only recently started to receive more attention from researchers. Different processing methods based on the use of supercritical carbon dioxide have been proposed for the creation of novel architectures based on natural and synthetic polymers and these will be unleashed in this paper. ; The research leading to these results has recieved funding from the European Union Seventh Framework Programme (FP7) under grant agreement no. KBBE-2010-266033 (project SPECIAL), no. NMP4-SL-2009-229292 (project Find & Bind), from FEDER through POCTEP Project 0330_IBEROMARE_1_P, from the Sixth Framework Programme (FP6) under grant agreement NMP3-CT-2004-500283 (project EXPERTISSUES), Portuguese Foundation fo Science and Technology (FCT) is also acknowledged. for PhD and Post-Doc fellowships of Ana Rits C. Duarte, Vitor E. Santo, Anabela Alves, Simone S. Silva, Joana Moreira-Silva and Tiage H. Silva.Ana Rita C. Duarte would like to acknowledgs Fundacao Luso-Americana para o Desenvolvimento for the travel grant awarded to present this work at the international Symposium of Supercritical Fluids - ISSF ...

The high prevalence of bone defects has become a worldwide problem. Despite the significant amount of research on the subject, the available therapeutic solutions lack efficiency. Autografts, the most commonly used approaches to treat bone defects, have limitations such as donor site morbidity, pain and lack of donor site. Marine resources emerge as an attractive alternative to extract bioactive compounds for further use in bone tissue-engineering approaches. On one hand they can be isolated from by-products, at low cost, creating value from products that are considered waste for the fish transformation industry. One the other hand, religious constraints will be avoided. We isolated two marine origin materials, collagen from shark skin (Prionace glauca) and calcium phosphates from the teeth of two different shark species (Prionace glauca and Isurus oxyrinchus), and further proposed to mix them to produce 3D composite structures for hard tissue applications. Two crosslinking agents, 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride/N-Hydroxysuccinimide (EDC/NHS) and hexamethylene diisocyanate (HMDI), were tested to enhance the scaffolds' properties, with EDC/NHS resulting in better properties. The characterization of the structures showed that the developed composites could support attachment and proliferation of osteoblast-like cells. A promising scaffold for the engineering of bone tissue is thus proposed, based on a strategy of marine by-products valorisation. ; This work was funded by INTERREG under the POCTEP Project 0687_NOVOMAR_1_P and the Atlantic Area Transnational Cooperation Programme Project MARMED (2011-1/164), as well as by European Union FP7 under the project POLARIS (REGPOT-CT2012-316331). The authors gratefully acknowledge the funding support received from European Union through INTERREG—POCTEP Project 0687_NOVOMAR_1_P and Atlantic Area Transnational Cooperation Programme Project MARMED (2011-1/164)—and Research and Development Framework Programme FP7–project POLARIS (REGPOT-CT2012-316331). The authors acknowledge also the Centro Tecnológico del Mar (CETMAR, Vigo, Spain) and COPEMAR SA (fishing company, Spain) for the supply of fish by-products. J.M.S. and R.P. thank FCT respectively for the Post-Doctoral fellowship financed by POPH/FSE (SFRH/BPD/70230/2010) and for the contract IF/00347/2015. G.S.D. thanks Norte2020 by the PhD scholarship (NORTE-08-5369-F SE-000044). ; Peer reviewed