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Analysis of the ectopic osteogenic potential of adipose-derived mesenchymal stem cells in mice based on the relative gene expression of osteogenic markers

dc.contributor.advisorNajman, Stevo
dc.contributor.otherStamenković-Radak, Marina
dc.contributor.otherNikčević, Gordana
dc.creatorCvetković, Vladimir J.
dc.date.accessioned2017-02-08T10:33:46Z
dc.date.available2017-02-08T10:33:46Z
dc.date.available2020-07-03T08:05:21Z
dc.date.issued2016-09-26
dc.identifier.urihttp://eteze.bg.ac.rs/application/showtheses?thesesId=4416
dc.identifier.urihttp://nardus.mpn.gov.rs/handle/123456789/7482
dc.identifier.urihttps://fedorabg.bg.ac.rs/fedora/get/o:14396/bdef:Content/download
dc.identifier.urihttps://fedorabg.bg.ac.rs/fedora/get/o:14561/bdef:Izvestaj/download
dc.identifier.urihttp://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=1025140914
dc.description.abstractOsnovni cilj disertacije je da se uporedi i analizira osteogeni potencijal sveže izolovanih mezenhimskih matičnih ćelija belog masnog tkiva (ADSC) i in vitro osteoindukovanih ADSC na modelu ektopične implantacije kod Balb/c miševa. ADSC su nakon izolacije okarakterisane imunocitohemijskom analizom. Osteogena diferencijacija ADSC ispitana je analizom relativne ekspresije gena osteriksa (Sp7), osteokalcina (Bglap), alfa 1 lanca kolagena tipa I (Col1a1) i alkalne fosfataze (Alpl), vizuelizacijom fosfata i analizom ekspresije osteopontina. Miševima su subkutano implantirani implantati sastavljeni od sveže izolovanih ADSC, mineralnog koštanog matriksa (BMM) i plazme obogaćene trombocitima (PRP) (SPN grupa), osteoindukovanih ADSC, BMM i PRP (OPN grupa), PRP i BMM (PN kontrola) i samo od BMM (N kontrola). Osteogeneza u implantatima je ispitana analizom relativne ekspresije Sp7, Bglap, Col1a1 i Alpl, histomorfološkom, histomorfometrijskom i imunohistohemijskom analizom. Nađena je pozitivna CD29 i negativna CD14 ekspresija kod ćelija, što je karakteristika ADSC. Nakon petnaestodnevne osteoindukcije ADSC, zabeležena je najjača ekspresija markera osteogene diferencijacije. Najviša ekspresija gena markera osteogeneze, prisustvo ćelija nalik osteoblastima i osteoklastima, resorpcija BMM, visok procenat infiltriranog tkiva i prisustvo tkiva nalik osteoidu, ukazuju na napredovanje osteogeneze u OPN grupi nakon osme nedelje. U SPN grupi, nađeni su izraženi znaci inicijacije osteogeneze posle druge nedelje, a od četvrte ekspresija gena i ostalih markera opada što ukazuje na slabljenje osteogeneze.Rezultati doprinose razumevanju osteogenog potencijala različito pripremljenih ADSC što treba imati u vidu pri izboru načina i uslova za njihovu primenu u tretmanu koštanih defekata.sr
dc.description.abstractThe main subject of this doctoral dissertation was to compare and evaluate the osteogenic potential of freshly isolated, non-treated adipose-derived mesenchymal stem cells (ADSCs) and the potential of in vitro osteoinduced ADSCs in the ectopic boneforming model in Balb/c mice. The ADSCs were characterized after isolation by immunocytochemical analysis. Osteogenic differentiation of ADSCs was evaluated using relative gene expression analysis of osterix (Sp7), osteocalcin (Bglap), collagen type I alpha 1 chain (Col1a1) and alkaline phosphatase (Alpl), along with an assay for detection of phosphate deposits and an immunoexpression analysis of osteopontin. The implants composed of freshly isolated ADSCs, bone mineral matrix (BMM) and platelet-rich plasma (PRP) (SPN group), osteoinduced ADSCs, BMM and PRP (OPN group), BMM and PRP (PN control) and BMM-only (N control), were implanted subcutaneously in mice. The implants were analyzed using relative gene expression analysis of Sp7, Bglap, Col1a1 and Alpl, histochemical, histomorfometrical and immunohistochemical analyses. Positive immunoexpression of CD29 and negative of CD14 was found in the cells, which is a characteristic of ADSCs. After fifteen days of osteoinduction of ADSCs, the expression of the markers of osteogenic differentiation was the highest. The highest level of the bone-related gene expression, the presence of osteoblast-like and osteoclast-like cells, the resorption of BMM, a high percentage of infiltrated tissue and the presence of osteoid-like tissue, all suggest the progress of the osteogenesis in OPN implants after eight weeks. Strongly expressed signs of the initiation of osteogenesis were found in SPN implants after two weeks, but from the fourth week the expression of bone-related genes, as well as other analyzed markers decreased. The obtained results represent a contribution to the understanding of differences in the osteogenic potential of differently prepared ADSCs which is very important because of their utilization for the treatment of bone defects.en
dc.formatapplication/pdf
dc.languagesr
dc.publisherУниверзитет у Београду, Биолошки факултетsr
dc.relationinfo:eu-repo/grantAgreement/MESTD/Integrated and Interdisciplinary Research (IIR or III)/41017/RS//
dc.rightsopenAccessen
dc.sourceУниверзитет у Београдуsr
dc.subjectosteogenezasr
dc.subjectosteogenesisen
dc.subjectadipose-derived mesenchymal stem cellsen
dc.subjectplatelet-rich plasmaen
dc.subjectsubcutaneous implantationen
dc.subjectmiceen
dc.subjectgene expressionen
dc.subjectosteogenic markersen
dc.subjecthistochemistryen
dc.subjecthistomorphometryen
dc.subjectimmunohistochemistryen
dc.subjectmezenhimske matične ćelije masnog tkivasr
dc.subjectplazma obogaćena trombocitimasr
dc.subjectsubkutana implantacijasr
dc.subjectmišsr
dc.subjectekspresija genasr
dc.subjectmarkeri osteogenezesr
dc.subjecthistomorfologijasr
dc.subjecthistomorfometrijasr
dc.subjectimunohistohemijasr
dc.titleAnaliza ektopičnog osteogenog potencijala mezenhimskih matičnih ćelija belog masnog tkiva kod miša na osnovu relativne ekspresije gena markera osteogenezesr
dc.title.alternativeAnalysis of the ectopic osteogenic potential of adipose-derived mesenchymal stem cells in mice based on the relative gene expression of osteogenic markersen
dc.typedoctoralThesis
dc.rights.licenseBY-NC-ND
dcterms.abstractНајман, Стево; Никчевић, Гордана; Стаменковић-Радак, Марина; Цветковић, Владимир Ј.; Aнализа ектопичног остеогеног потенцијала мезенхимских матичних ћелија белог масног ткива код миша на основу релативне експресије гена маркера остеогенезе; Aнализа ектопичног остеогеног потенцијала мезенхимских матичних ћелија белог масног ткива код миша на основу релативне експресије гена маркера остеогенезе;
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/1035/Disertacija.pdf
dc.identifier.fulltexthttp://nardus.mpn.gov.rs/bitstream/id/1036/IzvestajKomisije.pdf


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