In vitro analysis of integrin expression during chondrogenic differentiation of mesenchymal stem cells and chondrocytes upon dedifferentiation in cell culture

Goessler, Ulrich Reinhart; Bugert, Peter; Heller, Tobias Christian; Sadick, Haneen; Hörmann, K.; Riedel, Frank

doi:10.3892/ijmm.17.2.301

Cited by 52 publications

(61 citation statements)

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“…Only 9 genes displayed opposing trends in the 2 settings. Genes up-regulated in both scenarios included the proteases cathepsin S (Ctss), reelin (Reln), and Mmp13, as well as other genes previously implicated in chondrocyte differentiation, including transglutaminase 2 (Tgm2) (48), extracellular matrix protein 1 (Ecm1) (49), Fc receptor, IgG, low affinity III (Fcgr3) (50), latent transforming growth factor ␤ binding protein 2 (Ltbp2) (51), and phosphate regulating endopeptidase homolog, X-linked (Phex) (52).…”

Section: Resultsmentioning

confidence: 99%

Global analyses of gene expression in early experimental osteoarthritis

Appleton¹,

Pitelka²,

Henry³

et al. 2007

Arthritis & Rheumatism

222

224

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Objective. To analyze genome-wide changes in chondrocyte gene expression in a surgically induced model of early osteoarthritis (OA) in rats, to assess the similarity of this model to human OA, and to identify genes and mechanisms leading to OA pathogenesis.Methods. OA was surgically induced in 5 rats by anterior cruciate ligament transection and partial medial meniscectomy. Sham surgery was performed in 5 additional animals, which were used as controls. Both groups underwent 4 weeks of forced mobilization, 3 times per week. RNA was extracted directly from articular chondrocytes in the OA (operated), contralateral, and sham-operated knees. Affymetrix GeneChip expression arrays were used to assess genome-wide changes in gene expression. Expression patterns of selected dysregulated genes, including Col2a1, Mmp13, Adamts5, Ctsc, Ptges, and Cxcr4, were validated by real-time polymerase chain reaction, immunofluorescence, or immunohistochemistry 2, 4, and 8 weeks after surgery.Results. After normalization, comparison of OA and sham-operated samples showed 1,619 differentially expressed probe sets with changes in their levels of expression >1.5-fold, 722 with changes >2-fold, 135 with changes >4-fold, and 20 with changes of 8-fold. Dysregulated genes known to be involved in human OA included Mmp13, Adamts5, and Ptgs2, among others. Several dysregulated genes (e.g., Reln, Phex, and Ltbp2) had been identified in our earlier microarray study of hypertrophic chondrocyte differentiation. Other genes involved in cytokine and chemokine signaling, including Cxcr4 and Ccl2, were identified. Changes in gene expression were also observed in the contralateral knee, validating the sham operation as the appropriate control.Conclusion. Our results demonstrate that the animal model mimics gene expression changes seen in human OA, supporting the relevance of newly identified genes and pathways to early human OA. We propose new avenues for OA pathogenesis research and potential targets for novel OA treatments, including cathepsins and cytokine, chemokine, and growth factor signaling pathways, in addition to factors controlling the progression of chondrocyte differentiation.

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Section: Resultsmentioning

confidence: 99%

Global analyses of gene expression in early experimental osteoarthritis

Appleton¹,

Pitelka²,

Henry³

et al. 2007

Arthritis & Rheumatism

222

224

View full text Add to dashboard Cite

show abstract

“…Similarly, Goessler et al have suggested that TGF-b1/2 might not be involved in the chondrocyte dedifferentiation process. 33 In addition, IGF-1, a member of the IGF family of peptide hormones believed to induce chondrocyte differentiation and proliferation, 34 was found to inhibit the dedifferentiation and apoptosis of articular chondrocytes induced by nitric oxide. 35 Consistent with this finding, our data revealed that IGF-1 was down-regulated in chondrocytes with increasing passage (and dedifferentiation), suggesting that IGF-1 gene expression in passaged monolayer chondrocytes may play a role in protecting the differentiated phenotype human chondrocytes.…”

Section: Gene Expression Profiles Of Chondrocytesmentioning

confidence: 99%

Gene expression profiles of human chondrocytes during passaged monolayer cultivation

Lin

Fitzgerald

et al. 2008

Journal Orthopaedic Research

178

149

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Chondrocyte phenotype has been shown to dedifferentiate during passaged monolayer cultivation. Hence, we have investigated the expression profile of 27 chondrocyte-associated genes from both osteoarthritic cartilage tissue and healthy passaged human articular chondrocytes by quantitative real-time PCR. Our results indicate that the gene expression levels of matrix proteins and proteases in chondrocytes from monolayer culture decrease compared with those from cartilage tissue, while monolayer cultured chondrocytes from normal and osteoarthritic cartilage exhibit similar gene expression patterns. However, chondrocytic gene expression profiles were differentially altered at various stages of passage. The expression of the matrix proteins aggrecan, type II collagen, and fibromodulin inversely correlated with increasing passage number, while fibronectin and link protein exhibited a marked increase with passage. The expression of matrix proteinases MMP-3/9/13 and ADAMTS-4/5 decreased with passage, whereas proteinase inhibitors TIMP-2/3 were elevated. The cytokine IL-1 also showed increased expression with monolayer chondrocyte culture, while IGF-1 expression levels were diminished. No significant changes in TGF-b, or the chondrogenic transcription factors Sox-9, c-fos, or c-jun were observed. Our data indicates that cultured chondrocytes undergo dedifferentiation during monolayer culture, although the gene expression level of transcription factors necessary for chondrogenesis remains unchanged. This data may prove important for the future development of more specific and efficacious cultivation techniques for human articular chondrocyte-based therapies. ß

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“…In contrast, expression of the ubiquitous matrix component fi bronectin and the unspecifi c matrix protein collagen type I, which are both known to be produced in increasing amounts by dedifferentiating chondrocytes (Martin et al 1999;Goessler et al 2005Goessler et al , 2006, decreased in all treatment courses after the second day of Chondrocytes deriving from two different cartilage donors were either adenovirally transduced using an IL-10 overexpression vector (adv/IL-10) or remained non-transduced (co: control) before they were embedded in alginate 24 hrs later. IL-10 secretion in the culture supernatant was measured during two weeks using a human IL-10 ELISA.…”

Section: Discussionmentioning

confidence: 99%

Cartilage-Specific Matrix and Integrin Expression in Three-Dimensional Articular Chondrocyte Cultures Overexpressing Human Interleukin-10

Müller

John

Kohl

et al. 2008

Clinical medicine. Arthritis and musculoskeletal disorders

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Interleukin (IL)-10 overexpression inhibits joint infl ammation, however the effect of high local concentrations of IL-10 on chondrocyte homeostasis remains unclear. The aim of this study was to determine the effects of IL-10 overexpression on cartilage matrix production in three-dimensional (3D) chondrocyte cultures. Human articular chondrocytes were transduced with adenoviral vectors alone (adv/empty) or by vectors either overexpressing enhanced green fl uorescence protein (adv/EGFP) or human IL-10 (adv/hIL-10) before their transfer to a 3D culture system. Non-transduced chondrocytes were used as controls. The expression of IL-10 or EGFP was confi rmed using ELISA or fl ow cytometry. Chondrocytes synthesis of collagen types II and I, aggrecan, fi bronectin and β1-integrin was determined over a period of 14 days post transduction using fl ow cytometry or immunohistochemistry. adv/EGFP or adv/IL-10 transduced chondrocytes expressed EGFP or secreted IL-10 detectable over the 2 weeks culture period. No suppression of collagen type II, aggrecan or β1-integrin synthesis by IL-10 overexpression was found and the deposition of collagen type I and fi bronectin remained unaffected compared to the controls. IL-10 overexpression does not impair key features of chondrocytes differentiated phenotype (e.g. collagen type II and aggrecan expression) suggesting the potential use of IL-10 for gene therapeutic approaches in the joint.

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In vitro analysis of integrin expression during chondrogenic differentiation of mesenchymal stem cells and chondrocytes upon dedifferentiation in cell culture

Cited by 52 publications

References 0 publications

Global analyses of gene expression in early experimental osteoarthritis

Global analyses of gene expression in early experimental osteoarthritis

Gene expression profiles of human chondrocytes during passaged monolayer cultivation

Cartilage-Specific Matrix and Integrin Expression in Three-Dimensional Articular Chondrocyte Cultures Overexpressing Human Interleukin-10

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