{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,1]],"date-time":"2026-02-01T11:36:30Z","timestamp":1769945790553,"version":"3.49.0"},"reference-count":42,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2018,8,3]],"date-time":"2018-08-03T00:00:00Z","timestamp":1533254400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>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\u2019 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.<\/jats:p>","DOI":"10.3390\/md16080269","type":"journal-article","created":{"date-parts":[[2018,8,3]],"date-time":"2018-08-03T11:03:26Z","timestamp":1533294206000},"page":"269","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":67,"title":["Marine Collagen\/Apatite Composite Scaffolds Envisaging Hard Tissue Applications"],"prefix":"10.3390","volume":"16","author":[{"given":"Gabriela","family":"Diogo","sequence":"first","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Estefania","family":"L\u00f3pez-Senra","sequence":"additional","affiliation":[{"name":"New Materials Group, Department of Applied Physics, Instituto de Investigaci\u00f3n Sanitaria Galicia Sur IISGS, University of Vigo, Campus Lagoas-Marcosende, 36310 Vigo, Spain"}]},{"given":"Rog\u00e9rio","family":"Pirraco","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Raphael","family":"Canadas","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4296-2529","authenticated-orcid":false,"given":"Emanuel","family":"Fernandes","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6187-4025","authenticated-orcid":false,"given":"Julia","family":"Serra","sequence":"additional","affiliation":[{"name":"New Materials Group, Department of Applied Physics, Instituto de Investigaci\u00f3n Sanitaria Galicia Sur IISGS, University of Vigo, Campus Lagoas-Marcosende, 36310 Vigo, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4918-9399","authenticated-orcid":false,"given":"Ricardo","family":"P\u00e9rez-Mart\u00edn","sequence":"additional","affiliation":[{"name":"Instituto de Investigaciones Marinas (CSIC), Eduardo Cabello 6, 36208 Vigo, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0726-4370","authenticated-orcid":false,"given":"Carmen","family":"Sotelo","sequence":"additional","affiliation":[{"name":"Instituto de Investigaciones Marinas (CSIC), Eduardo Cabello 6, 36208 Vigo, Spain"}]},{"given":"Alexandra","family":"Marques","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Pio","family":"Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"New Materials Group, Department of Applied Physics, Instituto de Investigaci\u00f3n Sanitaria Galicia Sur IISGS, University of Vigo, Campus Lagoas-Marcosende, 36310 Vigo, Spain"}]},{"given":"Joana","family":"Moreira-Silva","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8520-603X","authenticated-orcid":false,"given":"Tiago","family":"Silva","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"}]},{"given":"Rui","family":"Reis","sequence":"additional","affiliation":[{"name":"3B\u2019s Research Group\u2014Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, AvePark, Parque de Ci\u00eancia e Tecnologia, 4805-017 Barco, Guimar\u00e3es, Portugal"},{"name":"ICVS\/3B\u2019s\u2014PT Government Associate Laboratory, Braga\/Guimar\u00e3es, Portugal"},{"name":"The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Avepark, 4805-017 Barco, Guimar\u00e3es, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2018,8,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S56","DOI":"10.1016\/j.injury.2011.06.011","article-title":"The use of bone-graft substitutes in large bone defects: Any specific needs?","volume":"42","author":"Calori","year":"2011","journal-title":"Injury"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2385","DOI":"10.1016\/j.compscitech.2005.07.022","article-title":"Development of nanocomposites for bone grafting","volume":"65","author":"Murugan","year":"2005","journal-title":"Compos. Sci. Technol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1016\/j.injury.2011.03.033","article-title":"Scaffolds for bone healing: Concepts, materials and evidence","volume":"42","author":"Lichte","year":"2011","journal-title":"Injury"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1002\/jbm.a.33292","article-title":"Biophysicochemical evaluation of chitosan-hydroxyapatite-marine sponge collagen composite for bone tissue engineering","volume":"100","author":"Pallela","year":"2012","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1179\/1743280412Y.0000000002","article-title":"Materials of marine origin: A review on polymers and ceramics of biomedical interest","volume":"57","author":"Silva","year":"2012","journal-title":"Int. Mater. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/j.foodres.2005.10.010","article-title":"Bioactive compounds from marine processing byproducts\u2014Review","volume":"39","author":"Kim","year":"2006","journal-title":"Food Res. Int."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1016\/j.msec.2011.11.021","article-title":"Biological hydroxyapatite obtained from fish bones","volume":"32","author":"Boutinguiza","year":"2012","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1021\/bm201776r","article-title":"Biomimetically mineralized salmon collagen scaffolds for application in bone tissue engineering","volume":"13","author":"Hoyer","year":"2012","journal-title":"Biomacromolecules"},{"key":"ref_9","unstructured":"Pallela, R., Venkatesan, J., Bhatnagar, I., Shim, Y., and Kim, S. (2013). Applications of Marine Collagen-Based Scaffolds in Bone Tissue Engineering, CRC Press."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5881","DOI":"10.3390\/md12125881","article-title":"Marine origin collagens and its potential applications","volume":"12","author":"Silva","year":"2014","journal-title":"Mar. Drugs"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.1016\/j.foodhyd.2011.02.007","article-title":"Functional and bioactive properties of collagen and gelatin from alternative sources: A review","volume":"25","author":"Montero","year":"2011","journal-title":"Food Hydrocolloid"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.1002\/mabi.201300228","article-title":"Porous hydrogels from shark skin collagen crosslinked under dense carbon dioxide atmosphere","volume":"13","author":"Silva","year":"2013","journal-title":"Macromol. Biosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1016\/j.jsb.2012.03.012","article-title":"Structure, composition, and mechanical properties of shark teeth","volume":"178","author":"Enax","year":"2012","journal-title":"J. Struct. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"035011","DOI":"10.1088\/1748-6041\/11\/3\/035011","article-title":"The improved biological response of shark tooth bioapatites in a comparative in vitro study with synthetic and bovine bone grafts","volume":"11","author":"Serra","year":"2016","journal-title":"Biomed. Mater."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1505","DOI":"10.1007\/s10856-009-3713-5","article-title":"In vivo study of porous strontium-doped calcium polyphosphate scaffolds for bone substitute applications","volume":"20","author":"Tian","year":"2009","journal-title":"J. Mater. Sci. Mater. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1093\/ndt\/gft400","article-title":"Magnesium modulates parathyroid hormone secretion and upregulates parathyroid receptor expression at moderately low calcium concentration","volume":"29","author":"Canalejo","year":"2014","journal-title":"Nephrol. Dial. Transpl."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"E1","DOI":"10.1152\/ajpendo.1997.272.1.E1","article-title":"Effect of magnesium on parathyroid cells: Evidence for two sensing receptors or two intracellular pathways?","volume":"272","author":"Miki","year":"1997","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1728","DOI":"10.1016\/j.biomaterials.2005.10.003","article-title":"Magnesium and its alloys as orthopedic biomaterials: A review","volume":"27","author":"Staiger","year":"2006","journal-title":"Biomaterials"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1002\/jbm.10220","article-title":"Synthesis of functionally graded MgCO3 apatite accelerating osteoblast adhesion","volume":"62","author":"Yamasaki","year":"2002","journal-title":"J. Biomed. Mater. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.dental.2006.01.022","article-title":"Review on fluoride-releasing restorative materials\u2014Fluoride release and uptake characteristics, antibacterial activity and influence on caries formation","volume":"23","author":"Wiegand","year":"2007","journal-title":"Dent. Mater."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1177\/00220345900690S113","article-title":"Chemical and crystallographic events in the caries process","volume":"69","author":"LeGeros","year":"1990","journal-title":"J. Dent. Res."},{"key":"ref_22","first-page":"e91","article-title":"In vivo evaluation of shark teeth-derived bioapatites","volume":"28","author":"Vigo","year":"2017","journal-title":"Clin. Oral Implants Res."},{"key":"ref_23","first-page":"1165","article-title":"Rare-earth-element ordering and structural variations in natural rare-earth\u2019bearing apatites","volume":"76","author":"Hucrrns","year":"1991","journal-title":"Am. Mineral."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.2138\/am.2008.2683","article-title":"The crystal chemistry of whitlockite and merrillite and the dehydrogenation of whitlockite to merrillite","volume":"93","author":"Hughes","year":"2008","journal-title":"Am. Mineral."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1002\/(SICI)1097-4636(19980605)40:3<358::AID-JBM3>3.0.CO;2-E","article-title":"A Study on the Grafting Reaction of Isocyanates with Hydoxyapatite Particles","volume":"40","author":"Liu","year":"1998","journal-title":"J. Biomed. Mater. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.msec.2012.08.014","article-title":"Extraction and characterisation of apatite-and tricalcium phosphate-based materials from cod fish bones","volume":"33","author":"Piccirillo","year":"2013","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.actbio.2010.07.012","article-title":"A review of the mechanical behavior of CaP and CaP\/polymer composites for applications in bone replacement and repair","volume":"7","author":"Johnson","year":"2011","journal-title":"Acta Biomater."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2831","DOI":"10.1016\/j.biomaterials.2003.09.066","article-title":"Influence of different collagen species on physico-chemical properties of crosslinked collagen matrices","volume":"25","author":"Angele","year":"2004","journal-title":"Biomaterials"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.actbio.2016.11.059","article-title":"Fundamental insight into the effect of carbodiimide crosslinking on cellular recognition of collagen-based scaffolds","volume":"49","author":"Bax","year":"2017","journal-title":"Acta Biomater."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2161","DOI":"10.1016\/S0142-9612(03)00044-9","article-title":"Novel bioactive materials with different mechanical properties","volume":"24","author":"Kokubo","year":"2003","journal-title":"Biomaterials"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1440","DOI":"10.1016\/j.biomaterials.2008.10.056","article-title":"The correlation of pore morphology, interconnectivity and physical properties of 3D ceramic scaffolds with bone ingrowth","volume":"30","author":"Jones","year":"2009","journal-title":"Biomaterials"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1002\/jbm.a.31397","article-title":"Preparation of bone-like apatite\u2013collagen nanocomposites by a biomimetic process with phosphorylated collagen","volume":"85","author":"Li","year":"2008","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1016\/j.biotechadv.2012.10.003","article-title":"Review: Development of clinically relevant scaffolds for vascularised bone tissue engineering","volume":"31","author":"Liu","year":"2013","journal-title":"Biotechnol. Adv."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3876","DOI":"10.1016\/j.actbio.2012.07.002","article-title":"Current views on calcium phosphate osteogenicity and the translation into effective bone regeneration strategies","volume":"8","author":"Chai","year":"2012","journal-title":"Acta Biomater."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.actbio.2015.01.031","article-title":"Hydroxyapatite reinforced collagen scaffolds with improved architecture and mechanical properties","volume":"17","author":"Kane","year":"2015","journal-title":"Acta Biomater."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Nam, S., Won, J.-E., Kim, C.-H., and Kim, H.-W. (2011). Odontogenic differentiation of human dental pulp stem cells stimulated by the calcium phosphate porous granules. J. Tissue Eng., 812547.","DOI":"10.4061\/2011\/812547"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"5771","DOI":"10.1016\/j.actbio.2012.10.043","article-title":"Surface properties and ion release from fluoride-containing bioactive glasses promote osteoblast differentiation and mineralization in vitro","volume":"9","author":"Gentleman","year":"2013","journal-title":"Acta Biomater."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1039\/c3tb21371d","article-title":"Development of a bioactive composite of nano fluorapatite and poly (butylene succinate) for bone tissue regeneration","volume":"2","author":"Niu","year":"2014","journal-title":"J. Mater. Chem. B"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1080\/10498850.2013.865283","article-title":"Characterization of collagen from different discarded fish species of the West coast of the Iberian Peninsula","volume":"25","author":"Sotelo","year":"2015","journal-title":"J. Aquat. Food Prod. Technol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1586\/17434440.2014.900436","article-title":"Influence of sterilisation methods on collagen-based devices stability and properties","volume":"11","author":"Delgado","year":"2014","journal-title":"Expert Rev. Med. Devic."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2907","DOI":"10.1016\/j.biomaterials.2006.01.017","article-title":"How useful is SBF in predicting in vivo bone bioactivity?","volume":"27","author":"Kokubo","year":"2006","journal-title":"Biomaterials"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"025001","DOI":"10.1088\/1758-5082\/6\/2\/025001","article-title":"Manufacture of \u03b2-TCP\/alginate scaffolds through a Fab@ home model for application in bone tissue engineering","volume":"6","author":"Diogo","year":"2014","journal-title":"Biofabrication"}],"container-title":["Marine Drugs"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-3397\/16\/8\/269\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:16:31Z","timestamp":1760195791000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-3397\/16\/8\/269"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,8,3]]},"references-count":42,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2018,8]]}},"alternative-id":["md16080269"],"URL":"https:\/\/doi.org\/10.3390\/md16080269","relation":{},"ISSN":["1660-3397"],"issn-type":[{"value":"1660-3397","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,8,3]]}}}