Stimulation of adult chondrocyte metabolism by a thyroid‐derived factor

Jones, Deryk G.; Smith, Robert L.

doi:10.1002/jor.1100080211

Cited by 20 publications

(13 citation statements)

References 33 publications

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“…While human articular chondrocytes subjected to T 3 in monolayer and serum containing medium showed enhanced proliferation [55], earlier studies showed no effect of T 3 or T 4 on adult bovine cartilage cell proliferation or GAG synthesis in high-density cultures in serum-free medium [56]. This is consistent with our findings of no difference between T 3 -treated chondrocytes vs. control in means of DNA and GAG content, and in line with other studies performed in serum-free medium [57].…”

Section: Discussionmentioning

confidence: 99%

The Effect of Dexamethasone and Triiodothyronine on Terminal Differentiation of Primary Bovine Chondrocytes and Chondrogenically Differentiated Mesenchymal Stem Cells

et al. 2013

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The newly evolved field of regenerative medicine is offering solutions in the treatment of bone or cartilage loss and deficiency. Mesenchymal stem cells, as well as articular chondrocytes, are potential cells for the generation of bone or cartilage. The natural mechanism of bone formation is that of endochondral ossification, regulated, among other factors, through the hormones dexamethasone and triiodothyronine. We investigated the effects of these hormones on articular chondrocytes and chondrogenically differentiated mesenchymal stem cells, hypothesizing that these hormones would induce terminal differentiation, with chondrocytes and differentiated stem cells being similar in their response. Using a 3D-alginate cell culture model, bovine chondrocytes and chondrogenically differentiated stem cells were cultured in presence of triiodothyronine or dexamethasone, and cell proliferation and extracellular matrix production were investigated. Collagen mRNA expression was measured by real-time PCR. Col X mRNA and alkaline phosphatase were monitored as markers of terminal differentiation, a prerequisite of endochondral ossification. The alginate culture system worked well, both for the culture of chondrocytes and for the chondrogenic differentiation of mesenchymal stem cells. Dexamethasone led to an increase in glycosaminoglycan production. Triiodothyronine increased the total collagen production only in chondrocytes, where it also induced signs of terminal differentiation, increasing both collagen X mRNA and alkaline phosphatase activity. Dexamethasone induced terminal differentiation in the differentiated stem cells. The immature articular chondrocytes used in this study seem to be able to undergo terminal differentiation, pointing to their possible role in the onset of degenerative osteoarthritis, as well as their potential for a cell source in bone tissue engineering. When chondrocyte-like cells, after their differentiation, can indeed be moved on towards terminal differentiation, they can be used to generate a model of endochondral ossification, but this limitation must be kept in mind when using them in cartilage tissue engineering application.

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Section: Discussionmentioning

confidence: 99%

The Effect of Dexamethasone and Triiodothyronine on Terminal Differentiation of Primary Bovine Chondrocytes and Chondrogenically Differentiated Mesenchymal Stem Cells

et al. 2013

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“…The cells were washed two times in Dulbecco's phosphate buffered saline without Mg 2+ and Ca 2+ , washed once with DMEM, and then plated at 2.6 × 10 4 cells/cm 2 in DMEM containing FBS, gentamicin, and ascorbic acid (25 μg/ml) (Sigma Chemical, St. Louis, MO). High density monolayer cultures were tested in serum‐free medium containing 25 μg/ml gentamicin, 25 μg/ml ascorbic acid, 1 mM selenium, and a liposome supplement [Jones and Smith, 1990].…”

Section: Methodsmentioning

confidence: 99%

Regulation of nitric oxide and bcl‐2 expression by shear stress in human osteoarthritic chondrocytes in vitro

Lee

Trindade

Ikenoue

et al. 2003

J of Cellular Biochemistry

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Onset and progression of cartilage degeneration is associated with shear stress occurring in diarthrodial joints subjected to inappropriate loading. This study tested the hypothesis that shear stress induced nitric oxide is associated with altered expression of regulatory onco-proteins, bcl-2, and Fas (APO-1/CD95) and apoptosis in primary human osteoarthritic chondrocyte cultures. Shear stress induced membrane phosphatidylserine and nucleosomal degradation were taken as evidence of chondrocyte apoptosis. Application of shear stress upregulated nitric oxide in a dose-dependent manner and was associated with increases in membrane phosphatidylserine and nucleosomal degradation. Increasing levels of shear stress decreased expression of the anti-apoptotic factor, bcl-2, from 44 to 10 U/ml. Addition of the nitric oxide antagonists, L-N(5)-(1-iminoethyl) ornithine and Nomega-nitro-L-arginine methyl ester (L-NAME), reduced shear stress induced nucleosomal degradation by 62% and 74%, respectively. Inhibition of shear stress induced nitric oxide release by L-NAME coincided with a 2.7-fold increase of bcl-2, when compared to chondrocytes exposed to shear stress in the absence of L-NAME. These data suggest that shear stress induced nitric oxide is associated with changes in apoptotic regulatory factors that alter chondrocyte metabolism and may contribute to joint degeneration.

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“…Serum-containing medium was removed 24 h prior to loading by washing in PBS ( 3 x ) and serum-free medium (DMEM/F12 supplemented with 30 mM HEPES. 0.01 M selenium, 25 pglml ascorbate, a lipid supplement and gentamicin) was added for an overnight incubation prior to loading [20].…”

Section: Cl7onfirocj~te Isolfltionmentioning

confidence: 99%

Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro

Ikenoue

Trindade

Lee

et al. 2003

Journal Orthopaedic Research

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194

125

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This study addressed the hypothesis that duration and magnitude of applied intermittent hydrostatic pressure (IHP) are critical parameters in regulation of normal human articular chondrocyte aggrecan and type I1 collagen expression. Articular chondrocytes were isolated from knee cartilage and maintained as primary, high-density monolayer cultures. IHP was applied at magnitudes of 1, 5 and 10 MPa at I Hz for durations of either 4 h per day for one day (4 x 1) or 4 h per day for four days (4 x 4). Total cellular RNA was isolated and analyzed for aggrecan and type I1 collagen mRNA signal levels using specific primers and reverse transcription polymerase chain reaction (RT-PCR) nested with beta-actin primers as internal controls. With a 4 x 1 loading regimen, aggrecan mRNA signal levels increased 1.3-and 1.5-fold at 5 and 10 MPa, respectively, relative to beta-actin mRNA when compared to unloaded cultures. Changing the duration of loading to a 4 x 4 regimen increased aggrecan mRNA signal levels by 1.4-, 1.8-and 1.9-fold at loads of 1, 5 and 10 MPa, respectively. In contrast to the effects of IHP on aggrecan, type I1 collagen mRNA signal levels were only upregulated at loads of 5 and 10 MPa with the 4 x 4 loading regimen. Analysis of cell-associated protein by western blotting confirmed that IHP increased aggrecan and type I1 collagen in chondrocyte extracts. These data demonstrate that duration and magnitude of applied IHP differentially alter chondrocyte matrix protein expression. The results show that IHP provides an important stimulus for increasing cartilage matrix anabolism and may contribute to repair and regeneration of damaged or diseased cartilage.

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Stimulation of adult chondrocyte metabolism by a thyroid‐derived factor

Cited by 20 publications

References 33 publications

The Effect of Dexamethasone and Triiodothyronine on Terminal Differentiation of Primary Bovine Chondrocytes and Chondrogenically Differentiated Mesenchymal Stem Cells

The Effect of Dexamethasone and Triiodothyronine on Terminal Differentiation of Primary Bovine Chondrocytes and Chondrogenically Differentiated Mesenchymal Stem Cells

Regulation of nitric oxide and bcl‐2 expression by shear stress in human osteoarthritic chondrocytes in vitro

Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro

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