Oxygen Tension Is a Determinant of the Matrix-Forming Phenotype of Cultured Human Meniscal Fibrochondrocytes 英文参考文献.docVIP

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Oxygen Tension Is a Determinant of the Matrix-Forming Phenotype of Cultured Human Meniscal Fibrochondrocytes 英文参考文献.doc

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Oxygen Tension Is a Determinant of the Matrix-Forming Phenotype of Cultured Human Meniscal Fibrochondrocytes 英文参考文献

OxygenTensionIsaDeterminantoftheMatrix-Forming PhenotypeofCulturedHumanMeniscal Fibrochondrocytes AdetolaB.Adesida*,AilletteMulet-Sierra,LeilaLaouar,NadrM.Jomha DepartmentofSurgery,UniversityofAlberta,Edmonton,Alberta,Canada Abstract Background:Meniscalcartilagedisplaysapoorrepaircapacity,especiallywheninjuryislocatedintheavascularregionof thetissue.Cell-basedtissueengineeringstrategiestogeneratefunctionalmeniscussubstitutesisapromisingapproachto treatmeniscusinjuries.Meniscusfibrochondrocytes(MFC)canbeusedinthisapproach.However,MFCareunabletoretain theirphenotypewhenexpandedinculture.Inthisstudy,weexploredtheeffectofoxygentensiononMFCexpansionand ontheirmatrix-formingphenotype. Methodology/PrincipalFindings:MFCwereisolatedfromhumanmeniscifollowedbybasicfibroblastgrowthfactor(FGF- 2) mediated cell expansion in monolayer culture under normoxia (21%O2) or hypoxia (3%O2). Normoxia and hypoxia expandedMFCwereseededontoacollagenscaffold.TheMFCseededscaffolds(constructs)wereculturedinaserumfree chondrogenic medium for 3 weeks under normoxia and hypoxia. Constructs containing normoxia-expanded MFC were subsequentlyculturedundernormoxiawhilethoseformedfromhypoxia-expandedMFCweresubsequentlyculturedunder hypoxia.After3weeksofinvitroculture,theconstructswereassessedbiochemically,histologicallyandforgeneexpression viareal-timereversetranscription-PCRassays.Theresultsshowedthatconstructsundernormoxiaproducedamatrixwith enhanced mRNA ratio (3.5-fold higher; p,0.001) of collagen type II to I. This was confirmed by enhanced deposition of collagenIIusingimmuno-histochemistry.Furthermore,theconstructsunderhypoxiaproducedamatrixwithhighermRNA ratio of aggrecan to versican (3.5-fold, p,0.05). However, both constructs had the same capacity to produce aglycosaminoglycan(GAG)–specificextracellularmatrix. Conclusions: Our data provide evidence that oxygen tension is a key player in determining the matrix phenotype of cultured MFC. These findings suggest that the use of normal and low