FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W115299416> ?p ?o ?g. }

• W115299416 abstract "Introduction: Human mesenchymal stem cells (hMSC) are easily obtainable from bone mar-row and possess the ability to differentiate into osteoblasts. Therefore, they have been sug-gested as a suitable source for bone regeneration. HMSC are equipped with a variety of in-tegrins that mediate essential cell-matrix interactions. Collagen I represent approximately 90% of the bone protein content. Cell attachment to collagen I is mediated by three members of the integrin receptor family named a1b1, a2b1 and a11b1 integrins. The main aim of this doctoral thesis was to investigate the basal expression of those integrins in hMSC and to func-tionally analyze the knockdown effect of a single collagen I-binding integrin on hMSC behav-ior in vitro.Materials and methods: HMSC were cultured on collagen I-coated surface. A lentiviral trans-fer of a1-, a2- and a11-specific shRNA was applied for downregulation of the corresponding integrin mRNA. Quantitative PCR and western blot analysis were used to assess the basal ex-pression, knockdown efficiency and integrin compensation. Colorimetric adhesion assay was used for estimation of the extent of cells attachment. HMSC spreading and migration was ob-served by time lapse experiments. JC-1 staining was used for investigation of the initiation of apoptosis. Results: Quantitative PCR were used to assess the basal expression of collagen I-binding integrins in three hMSC donors. We found that these integrins are differently expressed as integrin a11 had the highest and integrin a2 the lowest expression. Next, we applied lentiviral delivery of target-specific short hairpin RNA (shRNA) in order to knockdown each of the collagen I-binding integrins and compared them to the hMSC transduced with a sequence against a non-human gene abbreviated as shRNA control. We achieved significant downregulation (> 80%) of the collagen I-binding integrin mRNA and protein. Subsequently to the transduction, we did not noticed pronounce morphological cell changes, however, a clear decrease of a2- and a11-knockdown hMSC numbers was observed during cultivation. Using a quantitative adhesion assay, we estimated that 120 min after plating only 30% of integrin a11-deficent cells were able to attach to collagen I. In contrast, at the same time point, 70% of integrin a2-knockdown hMSC were attached while integrin a1- and shRNA control hMSC have already reached 100% cell adhesion. Furthermore, a time lapse-based investigation showed that integrin a1- and shRNA control hMSC need approximately 35 min to fully spread on collagen I. In contrast, integrin a2- and a11-knockdown hMSC took approximately double more time for spreading in comparison to shRNA control hMSC. Additionally, we analyzed the migration capability of the four different hMSC lines. The average path which integrin a1- and shRNA control hMSC passed was approximately 170 µm with mean speed of 11.5 µm/h. In parallel integrin a2 and a11-deficient hMSC migrated to a distance of approximately 70 µm with a velocity of 5 µm/h. Since it was observed a lost of a2- and a11-deficient hMSC, next we performed JC-1 staining that visualizes mitochondrial leakage, a hallmark of apoptosis. The majority of integrin a2- and a11-knockdown hMSC exhibited mitochondrial leakage whereas integrin a1- and shRNA control hMSC showed intact mitochondria. Finally, we used quantitative PCR to investigate whether there were compensatory effects between the three integrin receptors. We detect that knockdown of integrin a1 led to upregulation of a2 and a11. Similarly, when integrin a2 was downregulated, integrin a1 and a11 expression increased. Interestingly, knockdown of integrin a11 caused only a slight increase in integrin a1 but not in a2 expression. We also observed that upon osteogenic stimulation, integrin a2 and a11-deficient hMSC further reduced in number and did not mineralize the matrix even on a single cell level. Moreover, our preliminary investigation in hMSC-derived from osteoporosis suffering patients showed a tremendous downregulation of integrin a2.Conclusions: Our results strongly suggested that integrins a2b1 and a11b1 mediate an indis-pensible signaling for hMSC. Once these receptors were ablated from cell surface, hMSC re-duced their spreading, adhesion, migration and survival rates. Our integrin knockdown mod-els can be used for further investigations and understanding of the integrins a2b1 and a11b1 importance and signaling in hMSC and hOB since we observed a strong downregulation of integrin a2 expression in osteoporosis." @default.
• W115299416 created "2016-06-24" @default.
• W115299416 creator A5090966575 @default.
• W115299416 date "2011-02-22" @default.
• W115299416 modified "2023-09-27" @default.
• W115299416 title "Characterization of human mesenchymal stem cells by the appearance of integrins and functional analysis of collagen I-binding integrins" @default.
• W115299416 hasPublicationYear "2011" @default.
• W115299416 type Work @default.
• W115299416 sameAs 115299416 @default.
• W115299416 citedByCount "0" @default.
• W115299416 crossrefType "dissertation" @default.
• W115299416 hasAuthorship W115299416A5090966575 @default.
• W115299416 hasConcept C119524353 @default.
• W115299416 hasConcept C1491633281 @default.
• W115299416 hasConcept C153911025 @default.
• W115299416 hasConcept C173396325 @default.
• W115299416 hasConcept C185592680 @default.
• W115299416 hasConcept C190232843 @default.
• W115299416 hasConcept C195687474 @default.
• W115299416 hasConcept C198826908 @default.
• W115299416 hasConcept C28328180 @default.
• W115299416 hasConcept C54355233 @default.
• W115299416 hasConcept C55493867 @default.
• W115299416 hasConcept C81885089 @default.
• W115299416 hasConcept C85789140 @default.
• W115299416 hasConcept C86803240 @default.
• W115299416 hasConcept C95444343 @default.
• W115299416 hasConceptScore W115299416C119524353 @default.
• W115299416 hasConceptScore W115299416C1491633281 @default.
• W115299416 hasConceptScore W115299416C153911025 @default.
• W115299416 hasConceptScore W115299416C173396325 @default.
• W115299416 hasConceptScore W115299416C185592680 @default.
• W115299416 hasConceptScore W115299416C190232843 @default.
• W115299416 hasConceptScore W115299416C195687474 @default.
• W115299416 hasConceptScore W115299416C198826908 @default.
• W115299416 hasConceptScore W115299416C28328180 @default.
• W115299416 hasConceptScore W115299416C54355233 @default.
• W115299416 hasConceptScore W115299416C55493867 @default.
• W115299416 hasConceptScore W115299416C81885089 @default.
• W115299416 hasConceptScore W115299416C85789140 @default.
• W115299416 hasConceptScore W115299416C86803240 @default.
• W115299416 hasConceptScore W115299416C95444343 @default.
• W115299416 hasLocation W1152994161 @default.
• W115299416 hasOpenAccess W115299416 @default.
• W115299416 hasPrimaryLocation W1152994161 @default.
• W115299416 hasRelatedWork W1497145092 @default.
• W115299416 hasRelatedWork W1955701221 @default.
• W115299416 hasRelatedWork W1970738755 @default.
• W115299416 hasRelatedWork W1990370414 @default.
• W115299416 hasRelatedWork W1995857138 @default.
• W115299416 hasRelatedWork W2001640807 @default.
• W115299416 hasRelatedWork W2012122215 @default.
• W115299416 hasRelatedWork W2037166461 @default.
• W115299416 hasRelatedWork W2054408547 @default.
• W115299416 hasRelatedWork W2061732759 @default.
• W115299416 hasRelatedWork W2136293689 @default.
• W115299416 hasRelatedWork W2168856134 @default.
• W115299416 hasRelatedWork W2356502461 @default.
• W115299416 hasRelatedWork W2607759647 @default.
• W115299416 hasRelatedWork W2790826120 @default.
• W115299416 hasRelatedWork W2804606320 @default.
• W115299416 hasRelatedWork W2807683511 @default.
• W115299416 hasRelatedWork W2810327404 @default.
• W115299416 hasRelatedWork W3176746442 @default.
• W115299416 hasRelatedWork W66362127 @default.
• W115299416 isParatext "false" @default.
• W115299416 isRetracted "false" @default.
• W115299416 magId "115299416" @default.
• W115299416 workType "dissertation" @default.