{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T05:09:19Z","timestamp":1773292159590,"version":"3.50.1"},"reference-count":41,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2019,7,15]],"date-time":"2019-07-15T00:00:00Z","timestamp":1563148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2013\/20554-0 and 2013\/09055-2"],"award-info":[{"award-number":["2013\/20554-0 and 2013\/09055-2"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Materials"],"abstract":"<jats:p>The use of porous scaffolds created by additive manufacturing is considered a viable approach for the regeneration of critical-size bone defects. This paper investigates the xenotransplantation of polycaprolactone (PCL) tissue constructs seeded with differentiated and undifferentiated human adipose-derived mesenchymal stem cells (hADSCs) to treat calvarial critical-sized defect in Wistar rats. PCL scaffolds without cells were also considered. In vitro and in vivo biological evaluations were performed to assess the feasibility of these different approaches. In the case of cell seeded scaffolds, it was possible to observe the presence of hADSCs in the rat tissue contributing directly (osteoblasts) and indirectly (stimulation by paracrine factors) to tissue formation, organization and mineralization. The presence of bone morphogenetic protein-2 (BMP-2) in the rat tissue treated with cell-seeded PCL scaffolds suggests that the paracrine factors of undifferentiated hADSC cells could stimulate BMP-2 production by surrounding cells, leading to osteogenesis. Moreover, BMP-2 acts synergistically with growth factors to induce angiogenesis, leading to higher numbers of blood vessels in the groups containing undifferentiated and differentiated hADSCs.<\/jats:p>","DOI":"10.3390\/ma12142268","type":"journal-article","created":{"date-parts":[[2019,7,15]],"date-time":"2019-07-15T04:55:27Z","timestamp":1563166527000},"page":"2268","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Tissue Constructs with Human Adipose-Derived Mesenchymal Stem Cells to Treat Bone Defects in Rats"],"prefix":"10.3390","volume":"12","author":[{"given":"Guilherme","family":"Caetano","sequence":"first","affiliation":[{"name":"Department of Internal Medicine, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo (USP), Ribeir\u00e3o Preto 14040-900, SP, Brazil"},{"name":"Graduate Program in Biomedical Sciences, University Centre of Herm\u00ednio Ometto Foundation, Araras 13607339, SP, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8959-329X","authenticated-orcid":false,"given":"Weiguang","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Mechanical, Aerospace and Civil Engineering, University of Manchester, Manchester M13 9PL, UK"}]},{"given":"Adriana","family":"Murashima","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Otolaryngology and Head and Neck Surgery, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo (USP), Ribeir\u00e3o Preto 14040-900, SP, Brazil"}]},{"suffix":"Jr.","given":"Jos\u00e9 Roberto","family":"Passarini","sequence":"additional","affiliation":[{"name":"Graduate Program in Biomedical Sciences, University Centre of Herm\u00ednio Ometto Foundation, Araras 13607339, SP, Brazil"}]},{"given":"Leonardo","family":"Bagne","sequence":"additional","affiliation":[{"name":"Graduate Program in Biomedical Sciences, University Centre of Herm\u00ednio Ometto Foundation, Araras 13607339, SP, Brazil"}]},{"given":"Marcel","family":"Leite","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo (USP), Ribeir\u00e3o Preto 14040-900, SP, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9688-782X","authenticated-orcid":false,"given":"Miguel","family":"Hyppolito","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Otolaryngology and Head and Neck Surgery, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo (USP), Ribeir\u00e3o Preto 14040-900, SP, Brazil"}]},{"given":"Salem","family":"Al-Deyab","sequence":"additional","affiliation":[{"name":"Department of Chemistry, College of Science, King Saud University, Riyadh 11451, Saudi Arabia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4265-0701","authenticated-orcid":false,"given":"Mohamed","family":"El-Newehy","sequence":"additional","affiliation":[{"name":"Department of Chemistry, College of Science, King Saud University, Riyadh 11451, Saudi Arabia"},{"name":"Department of Chemistry, Faculty of Science, Tanta University, Tanta 31527, Egypt"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3683-726X","authenticated-orcid":false,"given":"Paulo","family":"B\u00e1rtolo","sequence":"additional","affiliation":[{"name":"School of Mechanical, Aerospace and Civil Engineering, University of Manchester, Manchester M13 9PL, UK"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2700-5971","authenticated-orcid":false,"given":"Marco Andrey Cipriani","family":"Frade","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Ribeir\u00e3o Preto Medical School, University of S\u00e3o Paulo (USP), Ribeir\u00e3o Preto 14040-900, SP, Brazil"}]}],"member":"1968","published-online":{"date-parts":[[2019,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s40898-017-0003-8","article-title":"Advances in bioprinted cell-laden hydrogels for skin tissue engineering","volume":"2","author":"Pereira","year":"2017","journal-title":"Biomanuf. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1079","DOI":"10.1016\/j.progpolymsci.2011.11.007","article-title":"Additive manufacturing of tissues and organs","volume":"37","author":"Melchels","year":"2012","journal-title":"Prog. Polym. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.cirp.2012.05.005","article-title":"Biomedical production of implants by additive electro-chemical and physical processes","volume":"61","author":"Bartolo","year":"2012","journal-title":"CIRP Ann."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1080\/17452750903476288","article-title":"Biomanufacturing for tissue engineering: Present and future trends","volume":"4","author":"Chua","year":"2009","journal-title":"Virtual Phys. Prototyp."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1016\/j.msec.2013.12.007","article-title":"A simultaneous process of 3D magnesium phosphate scaffold fabrication and bioactive substance loading for hard tissue regeneration","volume":"36","author":"Lee","year":"2014","journal-title":"Mater. Sci. Eng. C Mater. Biol. Appl."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1007\/s40610-015-0022-2","article-title":"Bone tissue engineering","volume":"1","author":"Black","year":"2015","journal-title":"Curr. Mol. Biol. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1507","DOI":"10.1142\/S0192415X16500841","article-title":"Phyllanthus urinaria\u2019s inhibition of human osteosarcoma xenografts growth in mice is associated with modulation of mitochondrial fission\/fusion machinery","volume":"44","author":"Huang","year":"2016","journal-title":"Am. J. Chin. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1016\/j.msec.2019.03.047","article-title":"Engineered 3D printed poly (\u025b-caprolactone)\/graphene scaffolds for bone tissue engineering","volume":"100","author":"Wang","year":"2019","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.msec.2018.12.100","article-title":"Fabrication and characterisation of 3D printed MWCNT composite porous scaffolds for bone regeneration","volume":"98","author":"Huang","year":"2019","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"100","DOI":"10.30612\/hre.v5i9.7644","article-title":"A single-component hydrogel bioink for bioprinting of bioengineered 3D constructs for dermal tissue engineering","volume":"5","author":"Pereira","year":"2018","journal-title":"Mater. Horiz."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1016\/j.msec.2018.08.050","article-title":"Biomechanical performance of hybrid electrospun structures for skin regeneration","volume":"93","author":"Dias","year":"2018","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1089\/3dp.2018.0012","article-title":"3D-Printed Poly (\u025b-caprolactone)\/Graphene Scaffolds Activated with P1-Latex Protein for Bone Regeneration","volume":"5","author":"Caetano","year":"2018","journal-title":"3D Print. Addit. Manuf."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Wang, W., Caetano, G., Ambler, W., Blaker, J., Frade, M., Mandal, P., Diver, C., and B\u00e1rtolo, P. (2016). Enhancing the hydrophilicity and cell attachment of 3D printed PCL\/graphene scaffolds for bone tissue engineering. Materials, 9.","DOI":"10.3390\/ma9120992"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"95","DOI":"10.18063\/IJB.2016.02.009","article-title":"Morphological, mechanical and biological assessment of PCL\/pristine graphene scaffolds for bone regeneration","volume":"2","author":"Wang","year":"2016","journal-title":"Int. J. Bioprint."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"B\u00e1rtolo, P.J., Almeida, H.A., Rezende, R.A., Laoui, T., and Bidanda, B. (2008). Advanced processes to fabricate scaffolds for tissue engineering. Virtual Prototyping & Bio Manufacturing in Medical Applications, Springer.","DOI":"10.1007\/978-0-387-68831-2_8"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1246","DOI":"10.1016\/j.msec.2017.05.017","article-title":"Scaffolds for Bone Tissue Engineering: State of the art and new perspectives","volume":"1","author":"Roseti","year":"2017","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_17","first-page":"1","article-title":"Adipose-derived stem cells for tissue engineering and regenerative medicine applications","volume":"2016","author":"Dai","year":"2016","journal-title":"Stem Cells Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1108\/RPJ-04-2012-0037","article-title":"Fabrication and characterisation of PCL and PCL\/PLA scaffolds for tissue engineering","volume":"20","author":"Domingos","year":"2014","journal-title":"Rapid Prototyp. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1016\/j.ijbiomac.2011.07.022","article-title":"Novel hybrid scaffolds for the cultivation of osteoblast cells","volume":"49","author":"Sasmazel","year":"2011","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.proeng.2015.07.010","article-title":"Osteogenic differentiation of adipose-derived mesenchymal stem cells into Polycaprolactone (PCL) scaffold","volume":"1","author":"Caetano","year":"2015","journal-title":"Procedia Eng."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1016\/j.matlet.2016.05.152","article-title":"Cellularized versus decellularized scaffolds for bone regeneration","volume":"1","author":"Caetano","year":"2016","journal-title":"Mater. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.jmbbm.2019.01.020","article-title":"Assessment of PCL\/carbon material scaffolds for bone regeneration","volume":"93","author":"Wang","year":"2019","journal-title":"J. Mech. Behav. Biomed. Mater."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2075","DOI":"10.1007\/s10439-017-1866-9","article-title":"Carbon nanotube reinforced collagen\/hydroxyapatite scaffolds improve bone tissue formation in vitro and in vivo","volume":"45","author":"Jing","year":"2017","journal-title":"Ann. Biomed. Eng."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"025036","DOI":"10.1088\/1758-5090\/aa747f","article-title":"Repairing a bone defect with a three-dimensional cellular construct composed of a multi-layered cell sheet on electrospun mesh","volume":"9","author":"Ren","year":"2017","journal-title":"Biofabrication"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1007\/s10735-015-9623-6","article-title":"Overexpression of PLAP-1 in bone marrow stromal cells inhibits the rat critical-size skull defect repair","volume":"46","author":"Yu","year":"2015","journal-title":"J. Mol. Histol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1007\/s10735-013-9551-2","article-title":"Healing of periodontal defects and calcitonin gene related peptide expression following inferior alveolar nerve transection in rats","volume":"45","author":"Lv","year":"2014","journal-title":"J. Mol. Histol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1002\/jbm.b.34187","article-title":"Electrical stimulation: Complementary therapy to improve the performance of grafts in bone defects?","volume":"107","author":"Fonseca","year":"2019","journal-title":"J. Biomed. Mater. Res. Part B Appl. Biomater."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1080\/14653240600855905","article-title":"Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement","volume":"8","author":"Dominici","year":"2006","journal-title":"Cytotherapy"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1089\/ten.teb.2011.0327","article-title":"Stem cells in dental pulp of deciduous teeth","volume":"18","author":"Kerkis","year":"2012","journal-title":"Tissue Eng. Part B Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4792","DOI":"10.1016\/j.biomaterials.2014.02.048","article-title":"Osteogenic differentiation and angiogenesis with cocultured adipose-derived stromal cells and bone marrow stromal cells","volume":"35","author":"Kim","year":"2014","journal-title":"Biomaterials"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1590\/1678-775720160037","article-title":"Aging impairs osteoblast differentiation of mesenchymal stem cells grown on titanium by favoring adipogenesis","volume":"24","author":"Abuna","year":"2016","journal-title":"J. Appl. Oral Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1038\/nm.2665","article-title":"Directing mesenchymal stem cells to bone to augment bone formation and increase bone mass","volume":"18","author":"Guan","year":"2012","journal-title":"Nat. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1434","DOI":"10.1007\/s00167-013-2426-y","article-title":"Xenotransplantation of human mesenchymal stem cells for repair of osteochondral defects in rabbits using osteochondral biphasic composite constructs","volume":"22","author":"Jang","year":"2014","journal-title":"Knee Surg. Sports Traumatol. Arthrosc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1007\/s11626-014-9814-6","article-title":"Characterization of adipose tissue-derived stromal vascular fraction for clinical application to cartilage regeneration","volume":"51","author":"Jang","year":"2015","journal-title":"Cell. Dev. Biol. Anim."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1089\/ten.tea.2009.0401","article-title":"Xenotransplantation of human mesenchymal stem cells into immunocompetent rats for calvarial bone repair","volume":"16","author":"Chuang","year":"2009","journal-title":"Tissue Eng. Part A"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"109","DOI":"10.22203\/eCM.v020a10","article-title":"Reconstruction of rat calvarial defects with human mesenchymal stem cells and osteoblast-like cells in poly-lactic-co-glycolic acid scaffolds","volume":"1","author":"Zong","year":"2010","journal-title":"Eur. Cells Mater."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/j.bone.2015.11.011","article-title":"Tissue engineering strategies for promoting vascularized bone regeneration","volume":"1","author":"Almubarak","year":"2016","journal-title":"Bone"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1016\/j.bone.2009.09.019","article-title":"COMP-Ang1, a chimeric form of Angiopoietin 1, enhances BMP2-induced osteoblast differentiation and bone formation","volume":"46","author":"Jeong","year":"2010","journal-title":"Bone"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Choi, H., Jeong, B.C., Hur, S.W., Kim, J.W., Lee, K.B., and Koh, J.T. (2015). The Angiopoietin-1 Variant COMP-Ang1 Enhances BMP2-Induced Bone Regeneration with Recruiting Pericytes in Critical Sized Calvarial Defects. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0140502"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1096\/fj.04-2529fje","article-title":"Endothelial cell modulation of bone marrow stromal cell osteogenic potential","volume":"19","author":"Kaigler","year":"2005","journal-title":"FASEB J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.msec.2016.08.027","article-title":"A new bi-layered scaffold for osteochondral tissue regeneration: In vitro and in vivo preclinical investigations","volume":"70","author":"Sartori","year":"2017","journal-title":"Mater. Sci. Eng. C"}],"container-title":["Materials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1996-1944\/12\/14\/2268\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:05:40Z","timestamp":1760187940000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1996-1944\/12\/14\/2268"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,7,15]]},"references-count":41,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2019,7]]}},"alternative-id":["ma12142268"],"URL":"https:\/\/doi.org\/10.3390\/ma12142268","relation":{},"ISSN":["1996-1944"],"issn-type":[{"value":"1996-1944","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,7,15]]}}}