{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,16]],"date-time":"2026-03-16T04:45:48Z","timestamp":1773636348952,"version":"3.50.1"},"reference-count":45,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2021,11,15]],"date-time":"2021-11-15T00:00:00Z","timestamp":1636934400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["FIS PI18\/0332"],"award-info":[{"award-number":["FIS PI18\/0332"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["FIS PI18\/0331"],"award-info":[{"award-number":["FIS PI18\/0331"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["FIS PI21\/0980"],"award-info":[{"award-number":["FIS PI21\/0980"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["ICI19\/00024"],"award-info":[{"award-number":["ICI19\/00024"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Consejer\u00eda de Salud y Familias","award":["PI-0442-2019"],"award-info":[{"award-number":["PI-0442-2019"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Polymers"],"abstract":"<jats:p>Critical defects of the mandibular bone are very difficult to manage with currently available materials and technology. In the present work, we generated acellular and cellular substitutes for human bone by tissue engineering using nanostructured fibrin-agarose biomaterials, with and without adipose-tissue-derived mesenchymal stem cells differentiated to the osteogenic lineage using inductive media. Then, these substitutes were evaluated in an immunodeficient animal model of severely critical mandibular bone damage in order to assess the potential of the bioartificial tissues to enable bone regeneration. The results showed that the use of a cellular bone substitute was associated with a morpho-functional improvement of maxillofacial structures as compared to negative controls. Analysis of the defect site showed that none of the study groups fully succeeded in generating dense bone tissue at the regeneration area. However, the use of a cellular substitute was able to improve the density of the regenerated tissue (as determined via CT radiodensity) and form isolated islands of bone and cartilage. Histologically, the regenerated bone islands were comparable to control bone for alizarin red and versican staining, and superior to control bone for toluidine blue and osteocalcin in animals grafted with the cellular substitute. Although these results are preliminary, cellular fibrin-agarose bone substitutes show preliminary signs of usefulness in this animal model of severely critical mandibular bone defect.<\/jats:p>","DOI":"10.3390\/polym13223939","type":"journal-article","created":{"date-parts":[[2021,11,15]],"date-time":"2021-11-15T08:19:20Z","timestamp":1636964360000},"page":"3939","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Usefulness of a Nanostructured Fibrin-Agarose Bone Substitute in a Model of Severely Critical Mandible Bone Defect"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4639-3175","authenticated-orcid":false,"given":"Miguel-Angel","family":"Martin-Piedra","sequence":"first","affiliation":[{"name":"Tissue Engineering Group, Department of Histology, Faculty of Medicine, University of Granada, E18016 Granada, Spain"},{"name":"Instituto de Investigaci\u00f3n Biosanitaria ibs.GRANADA, E18012 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bel\u00e9n","family":"Giron\u00e9s-Camarasa","sequence":"additional","affiliation":[{"name":"Division of Pediatric Surgery, University Hospital Virgen de las Nieves, E18014 Granada, Spain"},{"name":"Doctoral Program in Biomedicine, University of Granada, E18071 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio","family":"Espa\u00f1a-L\u00f3pez","sequence":"additional","affiliation":[{"name":"Craniofacial Malformations and Cleft Lip and Palate Management Unit, University Hospital Virgen de las Nieves, E18014 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ricardo","family":"Fern\u00e1ndez-Valad\u00e9s G\u00e1mez","sequence":"additional","affiliation":[{"name":"Division of Oral and Maxillofacial Surgery, San Pedro Hospital, La Rioja, E26006 Logro\u00f1o, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0992-0113","authenticated-orcid":false,"given":"Cristina","family":"Blanco-Elices","sequence":"additional","affiliation":[{"name":"Tissue Engineering Group, Department of Histology, Faculty of Medicine, University of Granada, E18016 Granada, Spain"},{"name":"Instituto de Investigaci\u00f3n Biosanitaria ibs.GRANADA, E18012 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ingrid","family":"Garz\u00f3n","sequence":"additional","affiliation":[{"name":"Tissue Engineering Group, Department of Histology, Faculty of Medicine, University of Granada, E18016 Granada, Spain"},{"name":"Instituto de Investigaci\u00f3n Biosanitaria ibs.GRANADA, E18012 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4876-2672","authenticated-orcid":false,"given":"Miguel","family":"Alaminos","sequence":"additional","affiliation":[{"name":"Tissue Engineering Group, Department of Histology, Faculty of Medicine, University of Granada, E18016 Granada, Spain"},{"name":"Instituto de Investigaci\u00f3n Biosanitaria ibs.GRANADA, E18012 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ricardo","family":"Fern\u00e1ndez-Valad\u00e9s","sequence":"additional","affiliation":[{"name":"Instituto de Investigaci\u00f3n Biosanitaria ibs.GRANADA, E18012 Granada, Spain"},{"name":"Division of Pediatric Surgery, University Hospital Virgen de las Nieves, E18014 Granada, Spain"},{"name":"Craniofacial Malformations and Cleft Lip and Palate Management Unit, University Hospital Virgen de las Nieves, E18014 Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1097\/ID.0000000000000910","article-title":"Reconstruction and Implant-Supported Rehabilitation of an Iatrogenically Caused Maxillary Alveolar Defect","volume":"28","author":"Meral","year":"2019","journal-title":"Implant. Dent."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1177\/0022034510385236","article-title":"Inflammation and Uncoupling as Mechanisms of Periodontal Bone Loss","volume":"90","author":"Graves","year":"2011","journal-title":"J. Dent. Res."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Bernardi, S., Macchiarelli, G., and Bianchi, S. (2020). Autologous Materials in Regenerative Dentistry: Harvested Bone, Platelet Concentrates and Dentin Derivates. Molecules, 25.","DOI":"10.3390\/molecules25225330"},{"key":"ref_4","unstructured":"Taqi, M., and Raju, S. (2021). Fibula Free Flaps. StatPearls, StatPearls Publishing."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e31","DOI":"10.1016\/j.oooo.2016.02.018","article-title":"Complications with Allogeneic, Cancellous Bone Blocks in Vertical Alveolar Ridge Augmentation: Prospective Clinical Case Study and Review of the Literature","volume":"122","author":"Draenert","year":"2016","journal-title":"Oral. Surg Oral. Med. Oral. Pathol. Oral. Radiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"109858","DOI":"10.1016\/j.msec.2019.109858","article-title":"Advanced Biomaterials for Repairing and Reconstruction of Mandibular Defects","volume":"103","author":"Zhang","year":"2019","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Zimina, A., Senatov, F., Choudhary, R., Kolesnikov, E., Anisimova, N., Kiselevskiy, M., Orlova, P., Strukova, N., Generalova, M., and Manskikh, V. (2020). Biocompatibility and Physico-Chemical Properties of Highly Porous PLA\/HA Scaffolds for Bone Reconstruction. Polymers, 12.","DOI":"10.3390\/polym12122938"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"8037","DOI":"10.1016\/j.actbio.2013.06.014","article-title":"Calcium Phosphate Ceramics in Bone Tissue Engineering: A Review of Properties and Their Influence on Cell Behavior","volume":"9","author":"Samavedi","year":"2013","journal-title":"Acta Biomater."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2015","DOI":"10.1039\/C7RA11278E","article-title":"Biological Properties of Calcium Phosphate Biomaterials for Bone Repair: A Review","volume":"8","author":"Lu","year":"2018","journal-title":"Rsc Adv."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1016\/j.bjoms.2020.03.016","article-title":"Systematic Scoping Review of Mandibular Bone Tissue Engineering","volume":"58","author":"Basyuni","year":"2020","journal-title":"Br. J. Oral Maxillofac. Surg."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e360605","DOI":"10.1590\/acb360605","article-title":"Comparison of Two Different Biomaterials in the Bone Regeneration (15, 30 and 60 Days) of Critical Defects in Rats","volume":"36","author":"Brassolatti","year":"2021","journal-title":"Acta Cir. Bras."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"055015","DOI":"10.1088\/2057-1976\/ac1878","article-title":"Bone Regeneration Using Wollastonite\/\u03b2-TCP Scaffolds Implants in Critical Bone Defect in Rat Calvaria","volume":"7","year":"2021","journal-title":"Biomed. Phys. Eng. Express"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1055\/s-0034-1393724","article-title":"Current Concepts of Bone Tissue Engineering for Craniofacial Bone Defect Repair","volume":"8","author":"Fishero","year":"2015","journal-title":"Craniomaxillofac. Trauma Reconstr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1097\/00001665-200205000-00014","article-title":"Human Segmental Mandibular Defects Treated with Naturally Derived Bone Morphogenetic Proteins","volume":"13","author":"Ferretti","year":"2002","journal-title":"J. Craniofac. Surg."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"S20","DOI":"10.1016\/j.injury.2005.07.029","article-title":"Bone Substitutes: An Update","volume":"36","author":"Giannoudis","year":"2005","journal-title":"Injury"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3311","DOI":"10.1167\/iovs.05-1647","article-title":"Construction of a Complete Rabbit Cornea Substitute Using a Fibrin-Agarose Scaffold","volume":"47","author":"Alaminos","year":"2006","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2142","DOI":"10.1002\/term.2951","article-title":"Successful Development and Clinical Translation of a Novel Anterior Lamellar Artificial Cornea","volume":"13","author":"Punzano","year":"2019","journal-title":"J. Tissue Eng. Regen Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1111\/jre.12247","article-title":"Sequential Keratinocytic Differentiation and Maturation in a Three-Dimensional Model of Human Artificial Oral Mucosa","volume":"50","author":"Oliveira","year":"2015","journal-title":"J. Periodont. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"501","DOI":"10.3389\/fncel.2018.00501","article-title":"In Vivo Evaluation of Nanostructured Fibrin-Agarose Hydrogels With Mesenchymal Stem Cells for Peripheral Nerve Repair","volume":"12","author":"Campos","year":"2018","journal-title":"Front. Cell Neurosci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1159\/000330682","article-title":"Epithelial and Stromal Developmental Patterns in a Novel Substitute of the Human Skin Generated with Fibrin-Agarose Biomaterials","volume":"196","author":"Carriel","year":"2012","journal-title":"Cells Tissues Organs"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1007\/978-981-13-0445-3_16","article-title":"Fibrin-Based Biomaterial Applications in Tissue Engineering and Regenerative Medicine","volume":"1064","author":"Park","year":"2018","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Oliver, J.D., Madhoun, W., Graham, E.M., Hendrycks, R., Renouard, M., and Hu, M.S. (2020). Stem Cells Regenerating the Craniofacial Skeleton: Current State-Of-The-Art and Future Directions. J. Clin. Med., 9.","DOI":"10.3390\/jcm9103307"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1097\/01.prs.0000204566.13979.c1","article-title":"Repair of Palatal Bone Defects Using Osteogenically Differentiated Fat-Derived Stem Cells","volume":"117","author":"Conejero","year":"2006","journal-title":"Plast Reconstr. Surg."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2573","DOI":"10.1002\/jbm.a.34929","article-title":"Effect of the Hydration on the Biomechanical Properties in a Fibrin-Agarose Tissue-like Model","volume":"102","author":"Scionti","year":"2014","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1177\/0885328209360425","article-title":"Pluripotential Differentiation Capability of Human Adipose-Derived Stem Cells in a Novel Fibrin-Agarose Scaffold","volume":"25","author":"Serrato","year":"2011","journal-title":"J. Biomater Appl."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1907","DOI":"10.1002\/term.2088","article-title":"Generation of a Bioengineered Autologous Bone Substitute for Palate Repair: An in Vivo Study in Laboratory Animals","volume":"11","author":"Oliveira","year":"2017","journal-title":"J. Tissue Eng. Regen Med."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Alfonso-Rodr\u00edguez, C.-A., Garz\u00f3n, I., Garrido-G\u00f3mez, J., Oliveira, A.-C.-X., Mart\u00edn-Piedra, M.-\u00c1., Scionti, G., Carriel, V., Hern\u00e1ndez-Cort\u00e9s, P., Campos, A., and Alaminos, M. (2014). Identification of Histological Patterns in Clinically Affected and Unaffected Palm Regions in Dupuytren\u2019s Disease. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0112457"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1002\/term.1949","article-title":"Differential Expression of GAP-43 and Neurofilament during Peripheral Nerve Regeneration through Bio-Artificial Conduits","volume":"11","author":"Carriel","year":"2017","journal-title":"J. Tissue Eng. Regen Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1007\/s10237-018-1067-y","article-title":"Effects of Hydration and Mineralization on the Deformation Mechanisms of Collagen Fibrils in Bone at the Nanoscale","volume":"18","author":"Fielder","year":"2019","journal-title":"Biomech. Model. Mechanobiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9933331","DOI":"10.1155\/2021\/9933331","article-title":"Improving Fibrin Hydrogels\u2019 Mechanical Properties, through Addition of Silica or Chitosan-Silica Materials, for Potential Application as Wound Dressings","volume":"2021","author":"Becerra","year":"2021","journal-title":"Int. J. Biomater."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1412","DOI":"10.1002\/term.2039","article-title":"In Vitro Characterization of a Nanostructured Fibrin Agarose Bio-Artificial Nerve Substitute","volume":"11","author":"Carriel","year":"2017","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Veremeev, A., Bolgarin, R., Nesterenko, V., Andreev-Andrievskiy, A., and Kutikhin, A. (2020). Native Bovine Hydroxyapatite Powder, Demineralised Bone Matrix Powder, and Purified Bone Collagen Membranes Are Efficient in Repair of Critical-Sized Rat Calvarial Defects. Materials, 13.","DOI":"10.3390\/ma13153393"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"55","DOI":"10.4252\/wjsc.v12.i1.55","article-title":"Early Therapeutic Effect of Platelet-Rich Fibrin Combined with Allogeneic Bone Marrow-Derived Stem Cells on Rats\u2019 Critical-Sized Mandibular Defects","volume":"12","author":"Awadeen","year":"2020","journal-title":"World J. Stem. Cells"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"534","DOI":"10.2334\/josnusd.18-0458","article-title":"Transplantation of Dedifferentiated Fat Cells Combined with a Biodegradable Type I Collagen-Recombinant Peptide Scaffold for Critical-Size Bone Defects in Rats","volume":"61","author":"Tateno","year":"2019","journal-title":"J. Oral. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.archoralbio.2019.01.010","article-title":"Evaluation of Bone Regeneration in a Critical Size Cortical Bone Defect in Rat Mandible Using MicroCT and Histological Analysis","volume":"101","author":"Medina","year":"2019","journal-title":"Arch. Oral. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Dong, Q.N., Kanno, T., Bai, Y., Sha, J., and Hideshima, K. (2019). Bone Regeneration Potential of Uncalcined and Unsintered Hydroxyapatite\/Poly l-Lactide Bioactive\/Osteoconductive Sheet Used for Maxillofacial Reconstructive Surgery: An In Vivo Study. Materials, 12.","DOI":"10.3390\/ma12182931"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Gandhimathi, C., Quek, Y.J., Ezhilarasu, H., Ramakrishna, S., Bay, B.-H., and Srinivasan, D.K. (2019). Osteogenic Differentiation of Mesenchymal Stem Cells with Silica-Coated Gold Nanoparticles for Bone Tissue Engineering. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20205135"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1007\/s00418-016-1489-5","article-title":"Development of a Multilayered Palate Substitute in Rabbits: A Histochemical Ex Vivo and in Vivo Analysis","volume":"147","author":"Alaminos","year":"2017","journal-title":"Histochem. Cell Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"512","DOI":"10.3389\/fimmu.2020.00512","article-title":"Versican-A Critical Extracellular Matrix Regulator of Immunity and Inflammation","volume":"11","author":"Wight","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3068","DOI":"10.2741\/2296","article-title":"Signaling and Transcriptional Regulation in Osteoblast Commitment and Differentiation","volume":"12","author":"Huang","year":"2007","journal-title":"Front. Biosci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1177\/1947603518764262","article-title":"A Standardized Method of Applying Toluidine Blue Metachromatic Staining for Assessment of Chondrogenesis","volume":"10","author":"Bergholt","year":"2019","journal-title":"Cartilage"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"025021","DOI":"10.1088\/1748-605X\/aa9ad2","article-title":"Generation of Genipin Cross-Linked Fibrin-Agarose Hydrogel Tissue-like Models for Tissue Engineering Applications","volume":"13","author":"Campos","year":"2018","journal-title":"Biomed. Mater."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Lopez-Lopez, M.T., Scionti, G., Oliveira, A.C., Duran, J.D.G., Campos, A., Alaminos, M., and Rodriguez, I.A. (2015). Generation and Characterization of Novel Magnetic Field-Responsive Biomaterials. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0133878"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1186\/s12951-020-00732-0","article-title":"Generation of a Novel Human Dermal Substitute Functionalized with Antibiotic-Loaded Nanostructured Lipid Carriers (NLCs) with Antimicrobial Properties for Tissue Engineering","volume":"18","author":"Vairo","year":"2020","journal-title":"J. Nanobiotechnol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3047","DOI":"10.1016\/j.transproceed.2019.08.014","article-title":"Transplant of Tissue-Engineered Artificial Autologous Human Skin in Andalusia: An Example of Coordination and Institutional Collaboration","volume":"51","author":"Carmona","year":"2019","journal-title":"Transpl. Proc."}],"container-title":["Polymers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4360\/13\/22\/3939\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:30:21Z","timestamp":1760167821000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4360\/13\/22\/3939"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,15]]},"references-count":45,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["polym13223939"],"URL":"https:\/\/doi.org\/10.3390\/polym13223939","relation":{},"ISSN":["2073-4360"],"issn-type":[{"value":"2073-4360","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,15]]}}}