Real-time quantitative PCR analysis of Vegf knockdown on expression of extracellular matrix and adhension molecules in male and female primary murine osteoblasts
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PROVIDER: PRJNA691882 | ENA |
REPOSITORIES: ENA
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Project description:Primary murine osteoblasts, isolated from the long bones of 4-day old male and female Vegffl/fl mice, were cultured prior to transduction with adenovirus-Cre-GFP at an multiplicity of infection of 100, to knockdown Vegf expression (OBVEGFKO) in vitro for six days. Control cells were transduced with an adenovirus expressing GFP (WT). Following this, cells were stepped down in low serum media containing 1% FBS for 24 hours, after which the conditioned media was collected for knockdown validation using an VEGF ELISA. We used the Qiagen Extracellular Matrix and Adhension Molecule RT2 Profiler PCR Array to assay the sexually dimorphic effects of Vegf on the expression of genes associated with bone matrix formation and mineralisation, remodelling and angiogenesis.
2021-02-28 | GSE164735 | GEO
Project description:To identify molecules regulated by glucosylceramide synthase in osteoblasts, we conducted microarray analysis using samles treated with the inhibitors of glucosylceramide synthase.
2021-08-30 | GSE174323 | GEO
Project description:Identify molecules regulated by glucosylceramide synthase in mouse osteoblasts.
| PRJNA729440 | ENA
Project description:Osteoblasts are adherent cells. Under physiological conditions they attach to mineralized and non-mineralized osseous surfaces. However, how exactly osteoblasts respond to these different osseous surfaces is largely unknown. Our hypothesis was that the state of matrix mineralization provides a functional signal to osteoblasts. To assess the osteoblast response to mineralized compared to demineralized osseous surfaces, we assessed the transcriptom.
2018-09-30 | GSE114237 | GEO
Project description:Osteoarthritis (OA) is the most common joint disease and this is a major cause of joint pain and disability in the aging population. Its etiology is multifactorial (i.e., age, obesity, joint injury, genetic predisposition), and the pathophysiologic process affects the entirety of the joint (Martel-Pelletier J et al. Osteoarthritis. Nature reviews Disease primers. 2016;2:16072). Although it is not yet clear if it precedes or occurs subsequently to cartilage damage, subchondral bone sclerosis is an important feature in OA pathophysiology (Goldring SR et al. Changes in the osteochondral unit during osteoarthritis: structure, function and cartilage-bone crosstalk. Nat Rev Rheumatol. 2016;12:632-44). It is characterized by local bone resorption and the accumulation of weakly mineralized osteoid substance (Bailey AJ et al. Phenotypic expression of osteoblast collagen in osteoarthritic bone: production of type I homotrimer. Int J Biochem Cell Biol. 2002;34:176-82). Subchondral bone sclerosis is suspected to be linked to cartilage degradation, not only by modifying the mechanical stresses transmitted to the cartilage, but also by releasing biochemical factors with an activity on cartilage metabolism (Sanchez C et al. Osteoblasts from the sclerotic subchondral bone downregulate aggrecan but upregulate metalloproteinases expression by chondrocytes. This effect is mimicked by interleukin-6, -1beta and oncostatin M pre-treated non-sclerotic osteoblasts. Osteoarthritis Cartilage. 2005;13:979-87; Sanchez C et al. Subchondral bone osteoblasts induce phenotypic changes in human osteoarthritic chondrocytes. Osteoarthritis Cartilage. 2005;13:988-97; Westacott CI et al J. Alteration of cartilage metabolism by cells from osteoarthritic bone. Arthritis Rheum. 1997;40:1282-91. We have previously demonstrated that osteoblasts isolated from subchondral OA bone exhibited an altered phenotype. More precisely, we showed that osteoblasts coming from the thickening (called sclerotic, SC) of subchondral bone located just below a cartilage lesion produced higher levels of alkaline phosphatase, interleukin (IL)-6, IL-8, prostaglandinE2, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-9 and transforming growth factor(TGF)-β1 and type I collagen than osteoblasts coming from the non-thickening neighboring area (called non-sclerotic area, NSC) (Sanchez C et al. Phenotypic characterization of osteoblasts from the sclerotic zones of osteoarthritic subchondral bone. Arthritis Rheum. 2008;58:442-55; Sanchez C et al. Regulation of subchondral bone osteoblast metabolism by cyclic compression. Arthritis Rheum. 2012;64:1193-203.) To compare secretome of cells living in different in vivo conditions is useful, not only to better understand the pathological mechanisms underlying changes in OA subchondral bone, but also to identify soluble biomarkers potentially reflecting these changes. Using our well-characterised human subchondral osteoblast culture model, we compared the secretome of osteoblasts coming from sclerotic and non sclerotic OA subchondral bone. This approach allowed to identify changes in secretome that contribute to explain some subchondral bone abnormalities in OA and to propose osteomodulin and fibulin-3 as potential biomarkers of OA subchondral bone remodelling.
2018-03-20 | PXD008494 | Pride
Project description:Matrix mineralization controls gene expression in osteoblasts
| PRJNA470642 | ENA
Project description:Osteoblasts and osteoclasts are two critical cell populations in bone remodelling. FasL is known from osteoblast-induced osteoclast apoptosis. Notably, FasL deficient (gld) mice display increased bone mass pointing to non-apoptotic functions of FasL. Some key extracellular matrix associated molecules, such as osteoprotegerin, osteocalcin, osteopontin and alkaline phosphatase were reported to be affected by FasL deficiency in osteoblasts but a complex insight has been missing. To fill this gap and provide a robust dataset, gld calva-derived cells were differentiated along with controls and subjected to RNASequencing. The analysis revealed statistically significant changes in expression of 1,710 genes. 15 % (263) out of them were associated with the extracellular matrix. This applied for the collagen components, but also non-collagenous proteoglycans including versican and aggrecan. This report provides enrichment analysis of statistically significant expression changes in calva-derived osteoblastic cells with focus on extracellular matrix components. The dataset further provides a solid background for further functional analyses revealing the exact mechanisms behind and consequences of such dysregulations in pathophysiology.
2025-10-16 | GSE250608 | GEO
Project description:The aim of this project was to investigate how genome-wide gene expression patterns change when the expression of VEGF-A is modulated using different lentivirally delivered shRNA molecules that are complementary to VEGF-A promoter region. To study these changes, 12 gene expression array experiments were conducted with mouse endothelial cell line (C166). The C166 cells were transduced (20% confluency, MOI 10) with lentiviral vectors that all express GFP marker gene and two of them express additional shRNA molecule that is complementary to different regions in the VEGF-A promoter and either up-regulate (VEGF-up) or downregulate (VEGF-down) VEGF-A expression. Fresh medium was changed 72h after transduction. Cells were collected at day 7.
2015-08-06 | E-GEOD-47444 | biostudies-arrayexpress
Project description:RNA sequencing of HUVECs stimulated with VEGF, OSM+VEGF, or CNTF+CNTFR+VEGF
| PRJNA807293 | ENA
Project description:The aim of this project was to investigate how genome-wide gene expression patterns change when the expression of VEGF-A is modulated using different lentivirally delivered shRNA molecules that are complementary to VEGF-A promoter region. To study these changes, 12 gene expression array experiments were conducted with mouse endothelial cell line (C166).
2015-08-06 | GSE47444 | GEO