Quantitation of chondroitin 4—sulfate and chondroitin 6—sulfate in pathologic joint fluid

Shinmei, Masayuki; Miyauchi, Satoshi; Machida, Akemi; Miyazaki, Kyosuke

doi:10.1002/art.1780351110

Cited by 149 publications

(114 citation statements)

References 12 publications

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“…Variations in the quantities of each CS type have been implicated in disease states, such as osteoarthritis. 33 Intact glycosaminoglycans are too large and heterogeneous to analyze, therefore they are enzymatically depolymerized into smaller chain lengths for analysis. Chondroitin lyase enzymes cleave CS chain N-acetylgalactosamine bonds using an eliminative mechanism to produce a 4,5-unsaturated (Δ-unsaturated) uronic acid on the non-reducing terminus of the products.…”

Section: Resultsmentioning

confidence: 99%

Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

2007

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Although stable isotopic labeling has found widespread use in the proteomics field, its application to carbohydrate quantification has been limited. Herein we report the design, synthesis, and application of a novel series of compounds that allow for the incorporation of isotopic variation within glycan structures. The novel feature of the compounds is the ability to incorporate the isotopes in a controlled manner, allowing for the generation of four tags that vary only in their isotopic content. This allows for the direct comparisons of three samples or triplicate measurements with an internal standard within one mass spectral analysis. Quantitation of partially depolymerized glycosaminoglycan mixtures, as well as N-linked glycans released from fetuin, is used to demonstrate the utility of the tetraplex tagging strategy.

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Section: Resultsmentioning

confidence: 99%

Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

2007

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“…High-performance liquid chromatography (HPLC) of the unsaturated tetrasaccharide of HA (⌬tetra-HA) and the unsaturated hexasaccharide of HA (⌬hexa-HA) was performed according to the method described by Takazono and Tanaka (25) and Shinmei et al (26). The HPLC system used in this study was constructed from 2 pumps (model 880-PU; Japan Spectroscopic, Tokyo, Japan), an autosampling injector (model 851-AS; Japan Spectroscopic), a stainless steel column packed with polyamine-bound silica (YMC gel PA-120; YMC, Kyoto, Japan), a dry reaction bath (DB-3; Shimamura Instrument Company, Tokyo, Japan), a fluoro-monitor (model FP-920; Japan Spectroscopic), and an integrator (model 807-IT; Japan Spectroscopic).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of interleukin‐1β‐stimulated production of matrix metalloproteinases by hyaluronan via CD44 in human articular cartilage

Julovi

Yasuda

Shimizu

et al. 2004

Arthritis & Rheumatism

156

122

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Objective. To investigate the mechanism of the inhibitory action of hyaluronan (HA) on interleukin-1␤ (IL-1␤)-stimulated production of matrix metalloproteinases (MMPs) in human articular cartilage.Methods. IL-1␤ was added to normal and osteoarthritic (OA) human articular cartilage in explant culture to stimulate MMP production. Articular cartilage was incubated or preincubated with a clinically used form of 800-kd HA to assess its effect on IL-1␤-induced MMPs. Levels of secreted MMPs 1, 3, and 13 in conditioned media were detected by immunoblotting; intracellular MMP synthesis in chondrocytes was evaluated by immunofluorescence microscopy. Penetration of HA into cartilage tissue and its binding to CD44 were analyzed by fluorescence microscopy using fluoresceinated HA. Blocking experiments with anti-CD44 antibody were performed to investigate the mechanism of action of HA.Results. Treatment and pretreatment with 800-kd HA at 1 mg/ml resulted in significant suppression of IL-1␤-stimulated production of MMPs 1, 3, and 13 in normal and OA cartilage explant culture. Fluorescence histocytochemistry revealed that HA penetrated cartilage tissue and localized in the pericellular matrix around chondrocytes. HA-binding blocking experiments using anti-CD44 antibody demonstrated that the association of HA with chondrocytes was mediated by CD44. Preincubation with anti-CD44 antibody, which suppressed IL-1␤-stimulated MMPs, reversed the inhibitory effect of HA on MMP production that was induced by IL-1␤ in normal and OA cartilage.Conclusion. This study demonstrates that HA effectively inhibits IL-1␤-stimulated production of MMP-1, MMP-3, and MMP-13, which supports the clinical use of HA in the treatment of OA. The action of HA on IL-1␤ may involve direct interaction between HA and CD44 on chondrocytes.Osteoarthritis (OA) is the most prevalent disease of articular joints and is the major cause of disability in the elderly. Pathophysiologic changes occur in OA cartilage due to the excessive expression of cartilagedegrading proteinases, the resultant progressive breakdown of collagen fibers, and the degradation of proteoglycan, mainly aggrecan (1).Matrix metalloproteinases (MMPs) are zinccontaining, calcium-dependent proteinases, which collectively degrade all components of the extracellular matrix. MMPs are considered to be important in the chondrolytic processes that contribute to the degenerative changes in OA cartilage (2-4). Recent studies have identified the messenger RNA (mRNA) for some MMPs, such as MMP-1, in human OA cartilage (4,5), and other investigators have reported specific MMP proteins and collagenasemediated type II collagen degradation products (6,7). There is a consensus that these enzymes play a critical role in intrinsic chondrocyte-mediated degenerative changes of the cartilage matrix in OA. Proinflammatory cytokines such as interleukin-1 (IL-1) strongly stimulate the expression of MMPs by chondrocytes in arthritis (8).Hyaluronan (HA) is a major component of synovial fluid and cartilage matrix, and it play...

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“…The GAG content was quantified according to a previously described method (15). Cartilage pellets were digested with 3 mg/ml collagenase in 0.25 ml of DMEM for 3 hours at 37°C, and a fraction of the cells was removed.…”

Section: Reverse Transcription-polymerase Chain Reaction (Rt-pcr)mentioning

confidence: 99%

Higher chondrogenic potential of fibrous synovium– and adipose synovium–derived cells compared with subcutaneous fat–derived cells: Distinguishing properties of mesenchymal stem cells in humans

Mochizuki

Muneta

Sakaguchi

et al. 2006

Arthritis & Rheumatism

278

259

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Objective. Mesenchymal stem cells from synovium have a greater proliferation and chondrogenic potential than do those from bone marrow, periosteum, fat, and muscle. This study was undertaken to compare fibrous synovium and adipose synovium (components of the synovium with subsynovium) to determine which is a more suitable source for mesenchymal stem cells, especially for cartilage regeneration, and to examine the features of adipose synovium-derived cells, fibrous synovium-derived cells, and subcutaneous fat-derived cells to determine their similarities.Methods. Human fibrous synovium, adipose synovium, and subcutaneous fat were harvested from 4 young donors and 4 elderly donors. After digestion, the nucleated cells were plated at a density considered proper to expand at a maximum rate without colony-to-colony contact. The surface epitopes, proliferative capacity, cloning efficiency, and chondrogenic, osteogenic, and adipogenic differentiation potentials of the cells were compared.Results. Fibrous synovium-and adipose synovium-derived cells were higher in STRO-1 and CD106 and lower in CD10 compared with subcutaneous fat-derived cells. Cells derived from fibrous and adipose synovium had higher proliferative potential and colonyforming efficiency compared with subcutaneous fatderived cells, both in mixed-population and in singlecell-derived cultures. In chondrogenic assays, pellets from fibrous synovium-and adipose synovium-derived cells produced more cartilage matrix than did cell pellets from subcutaneous fat. Osteogenic ability was also higher in fibrous synovium-and adipose synovium-derived cells, whereas adipogenic potential was nearly indistinguishable among the 3 populations. Differentiation potential of the cells was similar between young and elderly donors.Conclusion. Cells derived from the fibrous synovium and from the adipose synovium demonstrate comparable chondrogenic potential. Adipose synoviumderived cells are more similar to fibrous synoviumderived cells than to subcutaneous fat-derived cells.

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Quantitation of chondroitin 4—sulfate and chondroitin 6—sulfate in pathologic joint fluid

Cited by 149 publications

References 12 publications

Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

Inhibition of interleukin‐1β‐stimulated production of matrix metalloproteinases by hyaluronan via CD44 in human articular cartilage

Higher chondrogenic potential of fibrous synovium– and adipose synovium–derived cells compared with subcutaneous fat–derived cells: Distinguishing properties of mesenchymal stem cells in humans

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