{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T16:15:17Z","timestamp":1777565717450,"version":"3.51.4"},"reference-count":46,"publisher":"IOP Publishing","issue":"5","license":[{"start":{"date-parts":[[2023,8,11]],"date-time":"2023-08-11T00:00:00Z","timestamp":1691712000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2023,8,11]],"date-time":"2023-08-11T00:00:00Z","timestamp":1691712000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/iopscience.iop.org\/info\/page\/text-and-data-mining"}],"funder":[{"name":"PESCANOVA S.A, Spain"},{"name":"FCT","award":["PD\/BD\/143044\/2018"],"award-info":[{"award-number":["PD\/BD\/143044\/2018"]}]},{"name":"The German Research Foundation"},{"name":"Fundaci\u00f3n CETMAR"}],"content-domain":{"domain":["iopscience.iop.org"],"crossmark-restriction":false},"short-container-title":["Biomed. Mater."],"published-print":{"date-parts":[[2023,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>In the past decade, there has been significant progress in 3D printing research for tissue engineering (TE) using biomaterial inks made from natural and synthetic compounds. These constructs can aid in the regeneration process after tissue loss or injury, but achieving high shape fidelity is a challenge as it affects the construct\u2019s physical and biological performance with cells. In parallel with the growth of 3D bioprinting approaches, some marine-origin polymers have been studied due to their biocompatibility, biodegradability, low immunogenicity, and similarities to human extracellular matrix components, making them an excellent alternative to land mammal-origin polymers with reduced disease transmission risk and ethical concerns. In this research, collagen from shark skin, chitosan from squid pens, and fucoidan from brown algae were effectively blended for the manufacturing of an adequate biomaterial ink to achieve a printable, reproducible material with a high shape fidelity and reticulated using four different approaches (phosphate-buffered saline, cell culture medium, 6% CaCl<jats:sub>2<\/jats:sub>, and 5 mM Genipin). Materials characterization was composed by filament collapse, fusion behavior, swelling behavior, and rheological and compressive tests, which demonstrated favorable shape fidelity resulting in a stable structure without deformations, and interesting shear recovery properties around the 80% mark. Additionally, live\/dead assays were conducted in order to assess the cell viability of an immortalized human mesenchymal stem cell line, seeded directly on the 3D printed constructs, which showed over 90% viable cells. Overall, the Roswell Park Memorial Institute cell culture medium promoted the adequate crosslinking of this biopolymer blend to serve the TE approach, taking advantage of its capacity to hamper pH decrease coming from the acidic biomaterial ink. While the crosslinking occurs, the pH can be easily monitored by the presence of the indicator phenol red in the cell culture medium, which reduces costs and time.<\/jats:p>","DOI":"10.1088\/1748-605x\/acecec","type":"journal-article","created":{"date-parts":[[2023,8,2]],"date-time":"2023-08-02T22:48:15Z","timestamp":1691016495000},"page":"055017","update-policy":"https:\/\/doi.org\/10.1088\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Assessing non-synthetic crosslinkers in biomaterial inks based on polymers of marine origin to increase the shape fidelity in 3D extrusion printing"],"prefix":"10.1088","volume":"18","author":[{"given":"Duarte Nuno","family":"Carvalho","sequence":"first","affiliation":[]},{"given":"Sophie","family":"Dani","sequence":"additional","affiliation":[]},{"given":"Carmen G","family":"Sotelo","sequence":"additional","affiliation":[]},{"given":"Ricardo I","family":"P\u00e9rez-Mart\u00edn","sequence":"additional","affiliation":[]},{"given":"Rui L","family":"Reis","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8520-603X","authenticated-orcid":true,"given":"Tiago H","family":"Silva","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9075-5121","authenticated-orcid":true,"given":"Michael","family":"Gelinsky","sequence":"additional","affiliation":[]}],"member":"266","published-online":{"date-parts":[[2023,8,11]]},"reference":[{"key":"bmmacececbib1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1088\/1758-5090\/aa90e2","article-title":"Assessing bioink shape fidelity to aid material development in 3D bioprinting","volume":"10","author":"Ribeiro","year":"2017","journal-title":"Biofabrication"},{"key":"bmmacececbib2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1088\/1758-5090\/8\/1\/013001","article-title":"Biofabrication: reappraising the definition of an evolving field","volume":"8","author":"Groll","year":"2016","journal-title":"Biofabrication"},{"key":"bmmacececbib3","doi-asserted-by":"publisher","first-page":"1742","DOI":"10.1002\/adhm.201500168","article-title":"3D printing of scaffolds for tissue regeneration applications","volume":"4","author":"Do","year":"2015","journal-title":"Adv. Healthcare Mater."},{"key":"bmmacececbib4","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1177\/1947603516665445","article-title":"Three-dimensional bioprinting and its potential in the field of articular cartilage regeneration","volume":"8","author":"Mouser","year":"2017","journal-title":"Cartilage"},{"key":"bmmacececbib5","doi-asserted-by":"publisher","first-page":"636","DOI":"10.1016\/j.bioactmat.2020.04.017","article-title":"3D printing of multilayered scaffolds for rotator cuff tendon regeneration","volume":"5","author":"Jiang","year":"2020","journal-title":"Bioact. Mater."},{"key":"bmmacececbib6","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/polym12051002","article-title":"Marine-derived polymeric materials and biomimetics: an overview","volume":"12","author":"Claverie","year":"2020","journal-title":"Polymers"},{"key":"bmmacececbib7","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/polym14102026","article-title":"A design of experiments (DoE) approach to optimize cryogel manufacturing for tissue engineering applications","volume":"14","author":"Carvalho","year":"2022","journal-title":"Polymers"},{"key":"bmmacececbib8","doi-asserted-by":"publisher","first-page":"2059","DOI":"10.1007\/s12541-012-0271-4","article-title":"Comparative analysis of collagens extracted from different animal sources for application of cartilage tissue engineering","volume":"13","author":"Park","year":"2012","journal-title":"Int. J. Precis. Eng. Manuf."},{"key":"bmmacececbib9","doi-asserted-by":"publisher","first-page":"4031","DOI":"10.3390\/molecules24224031","article-title":"Hydrolyzed collagen-sources and applications","volume":"24","author":"Leon-Lopez","year":"2019","journal-title":"Molecules"},{"key":"bmmacececbib10","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/md17080467","article-title":"Marine collagen as a promising biomaterial for biomedical applications","volume":"17","author":"Lim","year":"2019","journal-title":"Mar. Drugs"},{"key":"bmmacececbib11","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/md20060366","article-title":"Suitability of marine- and porcine-derived collagen type I hydrogels for bioprinting and tissue engineering scaffolds","volume":"20","author":"Maher","year":"2022","journal-title":"Mar. Drugs"},{"key":"bmmacececbib12","doi-asserted-by":"publisher","first-page":"6718","DOI":"10.1039\/D1BM00809A","article-title":"Marine origin materials on biomaterials and advanced therapies to cartilage tissue engineering and regenerative medicine","volume":"9","author":"Carvalho","year":"2021","journal-title":"Biomater. Sci."},{"key":"bmmacececbib13","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.msec.2020.110963","article-title":"Marine collagen and its derivatives: versatile and sustainable bio-resources for healthcare","volume":"113","author":"Salvatore","year":"2020","journal-title":"Mater. Sci. Eng. C"},{"key":"bmmacececbib14","first-page":"543","article-title":"Seaweed polysaccharides as sustainable building blocks for biomaterials in tissue engineering","author":"Carvalho","year":"2020"},{"key":"bmmacececbib15","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/polym10060664","article-title":"Preparation and properties of 3D printed alginate-chitosan polyion complex hydrogels for tissue engineering","volume":"10","author":"Liu","year":"2018","journal-title":"Polymers"},{"key":"bmmacececbib16","doi-asserted-by":"publisher","first-page":"1496","DOI":"10.1021\/acs.chemrev.5b00303","article-title":"Strategies and molecular design criteria for 3D printable hydrogels","volume":"116","author":"Jungst","year":"2016","journal-title":"Chem. Rev."},{"key":"bmmacececbib17","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1016\/j.actbio.2015.07.030","article-title":"A hydrogel bioink toolkit for mimicking native tissue biochemical and mechanical properties in bioprinted tissue constructs","volume":"25","author":"Skardal","year":"2015","journal-title":"Acta Biomater."},{"key":"bmmacececbib18","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/jfb9040063","article-title":"Development and application of an additively manufactured calcium chloride nebulizer for alginate 3D-bioprinting purposes","volume":"9","author":"Raddatz","year":"2018","journal-title":"J. Funct. Biomater."},{"key":"bmmacececbib19","doi-asserted-by":"publisher","first-page":"973","DOI":"10.1016\/j.ijbiomac.2019.02.010","article-title":"Development of genipin-crosslinked and fucoidan-adsorbed nano-hydroxyapatite\/hydroxypropyl chitosan composite scaffolds for bone tissue engineering","volume":"128","author":"Lu","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"bmmacececbib20","doi-asserted-by":"publisher","first-page":"589","DOI":"10.1002\/bab.1123","article-title":"Effect of cross-linkers in fabrication of carrageenan-alginate matrices for tissue engineering application","volume":"60","author":"Ki","year":"2013","journal-title":"Biotechnol. Appl. Biochem."},{"key":"bmmacececbib21","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/md16080269","article-title":"Marine collagen\/apatite composite scaffolds envisaging hard tissue applications","volume":"16","author":"Diogo","year":"2018","journal-title":"Mar. Drugs"},{"key":"bmmacececbib22","doi-asserted-by":"publisher","first-page":"1410","DOI":"10.1016\/j.ijbiomac.2016.05.024","article-title":"Extracellular matrix-based cryogels for cartilage tissue engineering","volume":"93","author":"Han","year":"2016","journal-title":"Int. J. Biol. Macromol."},{"key":"bmmacececbib23","doi-asserted-by":"publisher","first-page":"366","DOI":"10.1016\/j.ijbiomac.2017.05.067","article-title":"Chitosan-co-hyaluronic acid porous cryogels and their application in tissue engineering","volume":"103","author":"Kutlusoy","year":"2017","journal-title":"Int. J. Biol. Macromol."},{"key":"bmmacececbib24","article-title":"High molecular weight chitosan, process for obtaining and uses thereof\u2014association for the advancement of tissues engineering and cell based technologies & therapies (A4TEC)","author":"L\u00f3pez-Cebral","year":"2018"},{"key":"bmmacececbib25","doi-asserted-by":"publisher","first-page":"74","DOI":"10.3390\/md18020074","article-title":"The incorporation of marine coral microparticles into collagen\u2010based scaffolds promotes osteogenesis of human mesenchymal stromal cells via calcium ion signalling","volume":"18","author":"Sheehy","year":"2020","journal-title":"Mar. Drugs"},{"key":"bmmacececbib26","doi-asserted-by":"publisher","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: an open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"bmmacececbib27","article-title":"Cell culture environment\u2014pH levels","author":""},{"key":"bmmacececbib28","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.polymer.2018.01.070","article-title":"Chemical modification and printability of shear-thinning hydrogel inks for direct-write 3D printing","volume":"152","author":"Smith","year":"2018","journal-title":"Polymer"},{"key":"bmmacececbib29","doi-asserted-by":"publisher","first-page":"1743","DOI":"10.1021\/acsbiomaterials.6b00158","article-title":"3D printing of shear-thinning hyaluronic acid hydrogels with secondary cross-linking","volume":"2","author":"Ouyang","year":"2016","journal-title":"ACS Biomater. Sci. Eng."},{"key":"bmmacececbib30","doi-asserted-by":"publisher","first-page":"6922","DOI":"10.1021\/acs.iecr.6b00523","article-title":"Extraction of collagen\/gelatin from the marine demosponge chondrosia reniformis (Nardo, 1847) using water acidified with carbon dioxide\u2014process optimization","volume":"55","author":"Silva","year":"2016","journal-title":"Ind. Eng. Chem. Res."},{"key":"bmmacececbib31","doi-asserted-by":"publisher","first-page":"1105","DOI":"10.1122\/1.1392299","article-title":"A slotted plate device for measuring static yield stress","volume":"45","author":"Zhu","year":"2001","journal-title":"J. Rheol."},{"key":"bmmacececbib32","doi-asserted-by":"publisher","first-page":"1521","DOI":"10.1038\/nprot.2017.053","article-title":"Shear-thinning and self-healing hydrogels as injectable therapeutics and for 3D-printing","volume":"12","author":"Loebel","year":"2017","journal-title":"Nat. Protocols"},{"key":"bmmacececbib33","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1515\/arh-2007-0001","article-title":"Rheological behavior of fugitive organic inks for direct-write assembly","volume":"17","author":"Therriault","year":"2007","journal-title":"Appl. Rheol."},{"key":"bmmacececbib34","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1016\/j.tifs.2017.06.018","article-title":"Genipin: a natural blue pigment for food and health purposes","volume":"67","author":"Neri-Numa","year":"2017","journal-title":"Trends Food Sci. Technol."},{"key":"bmmacececbib35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.bioadv.2022.212843","article-title":"Marine origin biomaterials using a compressive and absorption methodology as cell-laden hydrogel envisaging cartilage tissue engineering","volume":"137","author":"Carvalho","year":"2022","journal-title":"Biomater. Adv."},{"key":"bmmacececbib36","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.actbio.2017.07.028","article-title":"Manufacturing of hydrogel biomaterials with controlled mechanical properties for tissue engineering applications","volume":"62","author":"Vedadghavami","year":"2017","journal-title":"Acta Biomater."},{"key":"bmmacececbib37","doi-asserted-by":"publisher","first-page":"1974","DOI":"10.1002\/term.2628","article-title":"A review on gradient hydrogel\/fiber scaffolds for osteochondral regeneration","volume":"12","author":"Khorshidi","year":"2018","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"bmmacececbib38","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/polym10030285","article-title":"3D printing and electrospinning of composite hydrogels for cartilage and bone tissue engineering","volume":"10","author":"De Mori","year":"2018","journal-title":"Polymers"},{"key":"bmmacececbib39","doi-asserted-by":"publisher","first-page":"1090","DOI":"10.1007\/s11661-018-4468-8","article-title":"Theoretical conversions of different hardness and tensile strength for ductile materials based on stress\u2013strain curves","volume":"49","author":"Chen","year":"2018","journal-title":"Metall. Mater. Trans. A"},{"key":"bmmacececbib40","doi-asserted-by":"publisher","first-page":"758","DOI":"10.1007\/s42765-022-00135-w","article-title":"3D printed gelatin scaffold with improved shape fidelity and cytocompatibility by using Antheraea pernyi silk fibroin nanofibers","volume":"4","author":"Zou","year":"2022","journal-title":"Adv. Fiber Mater."},{"key":"bmmacececbib41","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1002\/jbm.a.32869","article-title":"Genipin-cross-linked collagen\/chitosan biomimetic scaffolds for articular cartilage tissue engineering applications","volume":"95","author":"Yan","year":"2010","journal-title":"J. Biomed. Mater. Res. A"},{"key":"bmmacececbib42","doi-asserted-by":"publisher","first-page":"901","DOI":"10.1016\/j.carbpol.2013.10.060","article-title":"Preparation and characterization of genipin cross-linked porous chitosan-collagen-gelatin scaffolds using chitosan-CO2 solution","volume":"102","author":"Gorczyca","year":"2014","journal-title":"Carbohydr. Polym."},{"key":"bmmacececbib43","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/j.supflu.2013.01.004","article-title":"Unleashing the potential of supercritical fluids for polymer processing in tissue engineering and regenerative medicine","volume":"79","author":"Duarte","year":"2013","journal-title":"J. Supercrit. Fluids"},{"key":"bmmacececbib44","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1088\/2057-1976\/ab54a7","article-title":"3D hybrid printing platform for auricular cartilage reconstruction","volume":"6","author":"Chung","year":"2020","journal-title":"Biomed. Phys. Eng. Express"},{"key":"bmmacececbib45","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1088\/1758-5090\/aad36d","article-title":"Bioprinting of mineralized constructs utilizing multichannel plotting of a self-setting calcium phosphate cement and a cell-laden bioink","volume":"10","author":"Ahlfeld","year":"2018","journal-title":"Biofabrication"},{"key":"bmmacececbib46","doi-asserted-by":"publisher","first-page":"1347","DOI":"10.1111\/j.1582-4934.2008.00299.x","article-title":"Introducing a single-cell-derived human mesenchymal stem cell line expressing hTERT after lentiviral gene transfer","volume":"12","author":"Bocker","year":"2008","journal-title":"J. Cell. Mol. Med."}],"container-title":["Biomedical Materials"],"original-title":[],"link":[{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec","content-type":"text\/html","content-version":"am","intended-application":"text-mining"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec\/pdf","content-type":"application\/pdf","content-version":"am","intended-application":"text-mining"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec\/pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec\/pdf","content-type":"application\/pdf","content-version":"am","intended-application":"similarity-checking"},{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,11]],"date-time":"2023-08-11T07:53:12Z","timestamp":1691740392000},"score":1,"resource":{"primary":{"URL":"https:\/\/iopscience.iop.org\/article\/10.1088\/1748-605X\/acecec"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8,11]]},"references-count":46,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,8,11]]},"published-print":{"date-parts":[[2023,9,1]]}},"URL":"https:\/\/doi.org\/10.1088\/1748-605x\/acecec","relation":{},"ISSN":["1748-6041","1748-605X"],"issn-type":[{"value":"1748-6041","type":"print"},{"value":"1748-605X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,8,11]]},"assertion":[{"value":"Assessing non-synthetic crosslinkers in biomaterial inks based on polymers of marine origin to increase the shape fidelity in 3D extrusion printing","name":"article_title","label":"Article Title"},{"value":"Biomedical Materials","name":"journal_title","label":"Journal Title"},{"value":"paper","name":"article_type","label":"Article Type"},{"value":"\u00a9 2023 The Author(s). Published by IOP Publishing Ltd","name":"copyright_information","label":"Copyright Information"},{"value":"2023-04-26","name":"date_received","label":"Date Received","group":{"name":"publication_dates","label":"Publication dates"}},{"value":"2023-08-02","name":"date_accepted","label":"Date Accepted","group":{"name":"publication_dates","label":"Publication dates"}},{"value":"2023-08-11","name":"date_epub","label":"Online publication date","group":{"name":"publication_dates","label":"Publication dates"}}]}}