Expression of Prostaglandin-Endoperoxide Synthase 1 and Prostaglandin-Endoperoxide Synthase 2 in Human Osteoblasts

Debrumfernandes, A.J.; Laporte, Stéphane A.; Heroux, Maxime S.; Lora, Maximilien; Patry, Christian; Ménard, Henri A.; Dumais, R.; Leduc, Richard

doi:10.1006/bbrc.1994.1136

Cited by 43 publications

(23 citation statements)

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“…We also provide additional evidence that mESCs do not have the same responses to proinflammatory cytokines as primary osteoblasts, indicating a difference in biochemical response. Although both cell types form bone nodules by day 21, the response of the osteo-mESCs to cytokine stimulation was very different to that of primary osteoblasts, which showed reduced cell viability and marked production of nitrite and [37,38,[50][51][52][53][54][55]. In contrast, the osteo-mESCs showed no reductions in cell viability in response to extended cytokine stimulation nor any significant increases in the production of nitrite or PGE 2 , until the latter stages of the culture period at day 21; the time point at which traditional differentiation protocols indicate as fully differentiated.…”

Section: Discussionmentioning

confidence: 97%

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Sidney

Kirkham

Buttery

2014

Stem Cells and Development

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There are well-established approaches for osteogenic differentiation of embryonic stem cells (ESCs), but few show direct comparison with primary osteoblasts or demonstrate differences in response to external factors. Here, we show comparative analysis of in vitro osteogenic differentiation of mouse ESC (osteo-mESC) and mouse primary osteoblasts. Both cell types formed mineralized bone nodules and produced osteogenic extracellular matrix, based on immunostaining for osteopontin and osteocalcin. However, there were marked differences in the morphology of osteo-mESCs and levels of mRNA expression for osteogenic genes. In response to the addition of proinflammatory cytokines interleukin-1b, tumor necrosis factor-a, and interferon-g to the culture medium, primary osteoblasts showed increased production of nitric oxide (NO) and prostaglandin E 2 (PGE 2 ) at early time points and decreases in cell viability. In contrast, osteo-mESCs maintained viability and did not produce NO and PGE 2 until day 21. The formation of bone nodules by primary osteoblasts was reduced markedly after cytokine stimulation but was unaffected in osteo-mESCs. Cell sorting of osteo-mESCs by cadherin-11 (cad-11) showed clear osteogenesis of cad-11+ cells compared to unsorted osteo-mESCs and cad-11 -cells. Moreover, the cad-11 + cells showed a significant response to cytokines, similar to primary osteoblasts. Overall, these results show that while osteo-mESC cultures, without specific cell sorting, show characteristics of osteoblasts, there are also marked differences, notably in their responses to cytokine stimuli. These findings are relevant to understanding the differentiation of stem cells and especially developing in vitro models of disease, testing new drugs, and developing cell therapies.

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Section: Discussionmentioning

confidence: 97%

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Sidney

Kirkham

Buttery

2014

Stem Cells and Development

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“…HOBs were cultured from these tissues following a previously reported method with minor modifications. (20) Briefly, thin slices of trabecular bone were treated with 2.5 mg/ml of trypsin in phosphate-buffered saline (PBS) for 1 h at 37°C, washed extensively in PBS, and cut into fragments of approximately 2 mm 2 . Fragments were seeded in 100-mm culture dishes containing 5 ml MEM supplemented with 10% FBS, 50 U/ml of penicillin, 150 g/ml of streptomycin, 2.5 g/ml of amphotericin, 2.2 mg/ml of sodium bicarbonate, and 50 g/ml of L-ascorbate.…”

Section: Cell Culturementioning

confidence: 99%

“…These cells have been shown previously to exhibit phenotypic characteristics of osteoblasts such as response to parathyroid hormone (PTH) and production of alkaline phosphatase and osteocalcin. (20) …”

Section: Cell Culturementioning

confidence: 99%

Evidence That Peroxynitrite Affects Human Osteoblast Proliferation and Differentiation

Rocha

Brum‐Fernandes

2002

Journal of Bone and Mineral Research

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“…(17). In all cell types within joint tissues that have been examined, COX-2 expression is dramatically increased by IL-1 (16,17,(37)(38)(39). Regulation of COX-2 expression is likely a critical step in determining levels of PG produced in the joint during inflammation (40).…”

Section: Findings Of Western Blot Analysis Of Nf-kbmentioning

confidence: 99%

Involvement of nuclear factor kB in the regulation of cyclooxygenase‐2 expression by interleukin‐1 in rheumatoid synoviocytes

Crofford

Teng

McCarthy

et al. 1997

Arthritis & Rheumatism

242

165

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Objective. To evaluate involvement of the transcription factor nuclear factor KB (NF-KB) in the increased expression of cyclooxygenase-2 (COX-2) stimulated by interleukin-lp (IL-1p) in primary rheumatoid synoviocytes.Methods. We treated early-passage rheumatoid synoviocytes with IL-lp and examined the time course of NF-KB translocation to the nucleus by Western blot analysis, as well as NF-KB binding to the COX-2 promoter/enhancer by electrophoretic mobility shift assay. We correlated the time course of NF-KB binding with expression of COX-2 messenger RNA (mRNA) and protein. Synoviocytes were then treated with either sense or antisense phosphorothioate-modified oligonucleotides derived from the transcription start site of the human NF-KB p65 RNA. We analyzed NF-KB binding to the COX-2 promoter and COX-2 protein levels after these treatments.Results. IL-1p rapidly stimulated the translocation of the p65, p50, and c-re1 NF-KB subunits from the cytoplasm to the nucleus. Electrophoretic mobility shift assay demonstrated binding to 2 NF-KB sites within the COX-2 promoter/enhancer, with a time course identical to that of nuclear localization of NF-KB. Supershift analysis revealed that binding activity was due primarily to the p65-p50 heterodimer and the p50 homodimer. Submitted for publication June 7, 1996; accepted in revised form August 21, 1996. mRNA and protein were increased. Pretreatment of RA synoviocytes with NF-KB p65 antisense oligonucleotides resulted in decreased binding to the COX-2 promoter and decreased COX-2 protein expression.Conclusion. These data demonstrate that signaling via the NF-KB pathway is involved in regulation of COX-2 expression induced by IL-1p in RA synoviocytes.

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Expression of Prostaglandin-Endoperoxide Synthase 1 and Prostaglandin-Endoperoxide Synthase 2 in Human Osteoblasts

Cited by 43 publications

References 0 publications

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Evidence That Peroxynitrite Affects Human Osteoblast Proliferation and Differentiation

Involvement of nuclear factor kB in the regulation of cyclooxygenase‐2 expression by interleukin‐1 in rheumatoid synoviocytes

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