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dc.contributor.authorMc Garrigle, M.J.
dc.contributor.authorMullen, Conleth A.
dc.contributor.authorHaugh, Matthew G.
dc.contributor.authorVoisin, Muriel C.
dc.contributor.authorMcNamara, Laoise M.
dc.date.accessioned2016-12-12T12:43:26Z
dc.date.available2016-12-12T12:43:26Z
dc.date.issued2016-05-26
dc.identifier.citationMc Garrigle, MJ,Mullen, CA,Haugh, MG,Voisin, MC,McNamara, LM (2016) 'OSTEOCYTE DIFFERENTIATION AND THE FORMATION OF AN INTERCONNECTED CELLULAR NETWORK IN VITRO'. European Cells & Materials, 31 :323-340. doi:10.22203/eCM.v031a21en_IE
dc.identifier.issn1473-2262
dc.identifier.urihttp://hdl.handle.net/10379/6220
dc.description.abstractExtracellular matrix (ECM) stiffness and cell density can regulate osteoblast differentiation in two dimensional environments. However, it is not yet known how osteoblast-osteocyte differentiation is regulated within a 3D ECM environment, akin to that existing in vivo. In this study we test the hypothesis that osteocyte differentiation is regulated by a 3D cell environment, ECM stiffness and cell density. We encapsulated MC3T3-E1 pre-osteoblastic cells at varied cell densities (0.25, 1 and 2 x 10(6) cells/mL) within microbial transglutaminase (mtgase) gelatin hydrogels of low (0.58 kPa) and high (1.47 kPa) matrix stiffnesses. Cellular morphology was characterised from phalloidin-FITC and 4',6-diamidino-2-phenylindole (DAPI) dilactate staining. In particular, the expression of cell dendrites, which are phenotypic of osteocyte differentiation, were identified. Immunofluorescent staining for the osteocytes specific protein DMP-1 was conducted. Biochemical analyses were performed to determine cell number, alkaline phosphatase activity and mineralisation at 2.5 hours, 3, 21 and 56 days. We found that osteocyte differentiation and the formation of an interconnected network between dendritic cells was significantly increased within low stiffness 3D matrices, compared to cells within high stiffness matrices, at high cell densities. Moreover we saw that this network was interconnected, expressed DMP-1 and also connected with osteoblast-like cells at the matrix surface. This study shows for the first time the role of the 3D physical nature of the ECM and cell density for regulating osteocyte differentiation and the formation of the osteocyte network in vitro. Future studies could apply this method to develop 3D tissue engineered constructs with an osteocyte network in place.en_IE
dc.description.sponsorshipThis project was supported by the European Research Council Grant 258992 (BONEMECHBIO). The authors acknowledge the facilities and scientific and technical assistance of the Centre for Microscopy & Imaging at the National University of Ireland Galway (www.imaging. nuigalway.ie), a facility that is funded by NUIG and the Irish Government’s Programme for Research in Third Level Institutions, Cycles 4 and 5, National Development Plan 2007-2013.en_IE
dc.formatapplication/pdfen_IE
dc.language.isoenen_IE
dc.publisherAO Research Institute Davosen_IE
dc.relation.ispartofEuropean Cells & Materialsen
dc.subjectOsteoblasten_IE
dc.subjectMC3T3-E1 CELLSen_IE
dc.subjectBiomedical engineeringen_IE
dc.subjectOsteocyteen_IE
dc.subjectInterconnected networken_IE
dc.subjectThree dimensionalen_IE
dc.subjectCell densityen_IE
dc.subjectMatrix stiffnessen_IE
dc.subjectIn vitroen_IE
dc.subjectDentin matrix protein-1en_IE
dc.subjectMesenchymal stem cellsen_IE
dc.subjectCollagen glycosaminoglycan scaffoldsen_IE
dc.subjectBone tissue regenerationen_IE
dc.subjectStrain amplificationen_IE
dc.subjectOsteoblastic differentiationen_IE
dc.subjectOsteogenic differentiationen_IE
dc.subjectSubstrate stiffnessen_IE
dc.subjectEndothelial cellsen_IE
dc.titleOsteocyte differentiation and the formation of an interconnected cellular network in vitroen_IE
dc.typeArticleen_IE
dc.date.updated2016-12-07T14:49:37Z
dc.identifier.doi10.22203/eCM.v031a21
dc.local.publishedsourcehttp:/dx.doi.org/10.22203/eCM.v031a21en_IE
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funder|~|
dc.internal.rssid11623198
dc.local.contactLaoise Mcnamara, Biomedical Engineering, Eng-3038, New Engineering Building, Nui Galway. 2251 Email: laoise.mcnamara@nuigalway.ie
dc.local.copyrightcheckedNo
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