CARTILAGE MATRIX DEPLETION BY RHEUMATOID SYNOVIAL CELLS IN TISSUE CULTURE

Articular cartilage fragments were added to monolayer cultures of synovial membrane cells. After 3 wk of incubation, the cartilage fragments were examined histologically for metachromasia and basophilia, and for fluorescent staining using a rabbit antiserum to cartilage protein-polysaccharide. Cartilage incubated with cells derived from rheumatoid synovial membranes showed striking loss of metachromasia and basophilia as well as diminished to absent fluorescent staining. Cartilage fragments incubated with cells from normal synovia, or with cells from the synovial membrane of a patient with Reiter's syndrome, did not show these changes and resembled control cartilage incubated in tissue culture medium alone. It appears, therefore, that rheumatoid synovial cells in tissue culture are able to deplete the matrix of articular cartilage.

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Protease activated receptor 2 and matriptase expression in the joints of cats with and without osteoarthritis

Journal of Feline Medicine and Surgery ◽

10.1177/1098612x20977796 ◽

2020 ◽

pp. 1098612X2097779

Author(s):

Siti M Zainal Ariffin ◽

David Bennett ◽

William R Ferrell ◽

John C Lockhart ◽

Lynette Dunning ◽

...

Keyword(s):

Articular Cartilage ◽

Mrna Expression ◽

Synovial Membrane ◽

Serine Proteases ◽

Cartilage Degradation ◽

Significant Difference ◽

Adult Cats ◽

Novel Therapeutic Strategies ◽

Protease Activated Receptor 2 ◽

Synovial Membranes

Objectives The aim of this study was to determine the presence of protease-activated receptor 2 (PAR2) and matriptase proteins and quantify PAR2 and matriptase mRNA expression in the articular cartilage and synovial membrane of cats with and without osteoarthritis (OA). Methods A total of 28 articular cartilage samples from adult cats (14 OA and 14 normal), 10 synovial membranes from adult cats (five OA and five normal) and three cartilage samples from 9-week-old fetal cats were used. The presence of PAR2 and matriptase in the cartilage and synovial membrane of the adult samples was detected by immunohistochemical (IHC) staining, while real-time PCR was used for mRNA expression analyses in all samples. Results PAR2 was detected in all OA and normal articular cartilage and synovial membrane samples but confined to only a few superficial chondrocytes in the normal samples. Matriptase was only detected in OA articular cartilage and synovial membrane samples. PAR2 and matriptase mRNA expression were, however, detected in all cartilage and synovial membrane samples. PAR2 and matriptase mRNA expression levels in OA articular cartilage were five ( P <0.001) and 3.3 ( P <0.001) times higher than that of the healthy group, respectively. There was no significant difference ( P = 0.05) in the OA synovial membrane PAR2 and matriptase mRNA expression compared with the normal samples. Conclusions and relevance Detection of PAR2 and matriptase proteins and gene expression in feline articular tissues is a novel and important finding, and supports the hypothesis that serine proteases are involved in the pathogenesis of feline OA. The consistent presence of PAR2 and matriptase protein in the cytoplasm of OA chondrocytes suggests a possible involvement of proteases in cartilage degradation. Further investigations into the PAR2 and matriptase pathobiology could enhance our understanding of the proteolytic cascades in feline OA, which might lead to the development of novel therapeutic strategies.

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Production of Mucopolysaccharides by Synovial Cells in a Simplified Tissue Culture Medium

Experimental Biology and Medicine ◽

10.3181/00379727-94-22853 ◽

1957 ◽

Vol 94 (1) ◽

pp. 51-56 ◽

Cited By ~ 24

Author(s):

C. W. Castor

Keyword(s):

Tissue Culture ◽

Culture Medium ◽

Tissue Culture Medium ◽

Synovial Cells

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Biosynthesis and secretion of procollagenase by rabbit synovial fibroblasts. Inhibition of procollagenase secretion by monensin and evidence for glycosylation of procollagenase

Biochemical Journal ◽

10.1042/bj2140281 ◽

1983 ◽

Vol 214 (2) ◽

pp. 281-288 ◽

Cited By ~ 69

Author(s):

H Nagase ◽

C E Brinckerhoff ◽

C A Vater ◽

E D Harris

Keyword(s):

Gel Electrophoresis ◽

Culture Medium ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Synovial Fibroblasts ◽

Synovial Cells ◽

Intracellular Accumulation ◽

Myristate Acetate ◽

Monolayer Cultures ◽

Continuous Labelling

Monolayer cultures of rabbit synovial fibroblasts stimulated with phorbol myristate acetate to produce large amounts of collagenase (EC 3.4.24.7) were used to study the biosynthesis and secretion of this enzyme. [3H]Leucine was added to cell cultures for pulse-chase and continuous-labelling experiments. The labelled procollagenase synthesized was identified by immunoprecipitation followed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and fluorography. The amounts of intracellular and extracellular proenzyme were quantified by measuring radioactivity incorporated into the proteins. procollagenase was synthesized as doublet proteins of Mr 57 000 and Mr 61 000. Immunoprecipitable proenzyme proteins were first detected in culture medium 35 min after [3H]leucine was added to the cells. Monensin treatment of the cells inhibited procollagenase secretion and led to intracellular accumulation of the proenzyme. Cells treated with tunicamycin produced only the 57 000-Mr form, indicating that in rabbit synovial cells the 61 000-Mr form was post-translationally modified by addition of oligosaccharides to asparagine residues. The ratios of glycosylated to unglycosylated forms in cell lysates and in culture medium were 0.22:1 and 0.07:1 respectively.

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Topographic localization of a 116,000-dalton protein in cartilage.

Journal of Histochemistry & Cytochemistry ◽

10.1177/33.2.3881518 ◽

1985 ◽

Vol 33 (2) ◽

pp. 127-133 ◽

Cited By ~ 11

Author(s):

R S Fife ◽

G L Hook ◽

K D Brandt

Keyword(s):

Articular Cartilage ◽

Matrix Protein ◽

Articular Surface ◽

Intense Staining ◽

Normal Cartilage ◽

Subunit Protein ◽

Monolayer Cultures ◽

Noncollagenous Protein ◽

The Matrix ◽

Surface Staining

A disulfide-bonded greater than 400,000-dalton (greater than 400-kD) protein with 116-kD subunits in hyaline cartilage from several species has recently been described. It constitutes 2-4% of the total noncollagenous protein in 4 M guanidinium chloride extracts of normal articular cartilage and accounts for most of the total noncollagen, nonproteoglycan protein synthesized in short-term organ cultures of canine articular cartilage. In the present study, immunofluorescence techniques were used to examine the topographic distribution of the 116-kD subunit protein in normal cartilage. In specimens of normal adult articular cartilage from several species, the protein was located throughout the matrix. More intense staining was observed at the articular surface than in the remainder of the uncalcified cartilage. In contrast, in fetal cartilage, the protein was uniformly distributed throughout the matrix without a marked increase in surface staining. Normal canine menisci and annulus fibrosus also demonstrated moderate fluorescence after incubation with the antiserum to the 116-kD subunit protein. Normal canine nucleus pulposus, synovium, aorta, and monolayer cultures of canine synovial cells exhibited only weak immunofluorescence after incubation with the antiserum. Therefore, the 116-kD subunit protein appears to be a ubiquitous matrix protein in cartilage.

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THE OCCURRENCE OF INTRACELLULAR CHONDROITIN SULFATE

The Journal of Cell Biology ◽

10.1083/jcb.18.1.13 ◽

1963 ◽

Vol 18 (1) ◽

pp. 13-17 ◽

Cited By ~ 24

Author(s):

Frank K. Thorp ◽

Albert Dorfman

Keyword(s):

Tissue Culture ◽

Infrared Spectroscopy ◽

Chemical Analysis ◽

Chondroitin Sulfate ◽

Culture Medium ◽

Extracellular Polysaccharide ◽

Tissue Culture Medium ◽

Extracellular Polysaccharides ◽

Chick Embryos ◽

The Matrix

Suspensions of chondrocytes were prepared by treatment with trypsin of the epiphyses of tibias and femurs of 13-day-old chick embryos. After washing to remove the matrix, such suspensions readily incorporate radioactive sulfate into both intracellular and extracellular chondroitin sulfate. Following disruption of the cells, the cell constituents were fractionated by centrifugation. Fractions obtained from cells incubated for 10 minutes showed a concentration of radioactivity in the material which sediments at 10,000 to 20,000 g. At this time the radioactivity of the extracellular chondroitin sulfate is low, but at 1 hour the radioactivity of the intracellular material is relatively unchanged, while that of the extracellular polysaccharide is markedly increased. Following incubation of the chondrocyte suspensions in a tissue culture medium, the intracellular chondroitin sulfate was isolated. This was compared with chondroitin sulfate isolated from the cartilage matrix. Chemical analysis and infrared spectroscopy indicated that both the intracellular and extracellular polysaccharides consist of a mixture of chondroitin sulfuric acids A and C. A portion of the chondroitin sulfate is not sulfated.

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Mycoplasmas in cell cultures from rheumatoid synovial membranes

Journal of Hygiene ◽

10.1017/s0022172400021203 ◽

1971 ◽

Vol 69 (1) ◽

pp. 17-25 ◽

Cited By ~ 10

Author(s):

K. B. Fraser ◽

P. V. Shirodaria ◽

Margaret Haire ◽

D. Middleton

Keyword(s):

Rheumatoid Arthritis ◽

Tissue Culture ◽

Fluorescent Antibody ◽

Fluorescent Staining ◽

Antibody Staining ◽

Synovial Fluids ◽

Direct Fluorescent Antibody ◽

The Common ◽

Synovial Membranes ◽

Fluorescent Antibody Staining

SUMMARYTen strains of myocoplasmas were recovered from cultures of synovium or cultures inoculated with synovial fragments from rheumatoid arthritis and one from osteo-arthritis. The source of the organisms is not known. Patients with rheumatoid arthritis had no complement-fixing antibody and no fluorescent staining antibody against the mycoplasmas isolated and no mycoplasma antigen was detected by immunofluorescence in sections of synovia and in synovial fluids.The strains isolated were of two main serological types and could be distinguished by direct fluorescent antibody staining from standard types of human commensals and the common tissue-culture contaminants. One may beMycoplasma laidlawii.

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Glycoconjugate markers of joint diseases

Biochemical Society Transactions ◽

10.1042/bst0390331 ◽

2011 ◽

Vol 39 (1) ◽

pp. 331-335 ◽

Cited By ~ 1

Author(s):

Janusz Popko ◽

Sławomir Olszewski ◽

Tomasz Guszczyn ◽

Krzysztof Zwierz ◽

Sławomir Pancewicz

Keyword(s):

Articular Cartilage ◽

Synovial Fluid ◽

Human Blood ◽

Synovial Membrane ◽

Lyme Arthritis ◽

Synovial Cells ◽

Joint Diseases ◽

Rheumatoid Disease ◽

Joint Tissue ◽

Different Types

A number of different types of glycoconjugate are found associated with joint tissue and fluids, comprising glycoproteins, glycolipids and glycosaminoglycans. Oligosaccharide chains of glycoconjugates are degraded by exoglycosidases, and the dominant exoglycosidase found in human blood, synovial fluid, the synovial membrane and chondrocytes of articular cartilage is HEX (N-acetyl-β-hexosaminidase). HEX is localized mostly intracellularly in synovial cells. Serum activity of HEX may be used to monitor the course and efficiency of treatment of Lyme arthritis, and activity of HEX, above 10 μkat/kg of protein in the synovial fluid, suggests rheumatoid disease. There is a shortage of HEX inhibitors able to penetrate synoviocytes, so the development of drugs which inhibit synthesis and/or the activity of HEX will be a promising field for future investigations.

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The Effect of Total Meniscectomy versus Caudal Pole Hemimeniscectomy on the Stifle Joint of the Sheep

Veterinary and Comparative Orthopaedics and Traumatology ◽

10.1055/s-0038-1632463 ◽

1999 ◽

Vol 12 (02) ◽

pp. 56-63 ◽

Cited By ~ 5

Author(s):

C. R. Bellenger ◽

P. Ghosh ◽

Y. Numata ◽

C. Little ◽

D. S. Simpson

Keyword(s):

Tissue Culture ◽

Articular Cartilage ◽

Fibrous Tissue ◽

Cartilage Degeneration ◽

Medial Compartment ◽

Proteoglycan Synthesis ◽

Stifle Joint ◽

Medial Meniscectomy ◽

Tibial Condyle ◽

Articular Cartilage Degeneration

SummaryTotal medial meniscectomy and caudal pole hemimeniscectomy were performed on the stifle joints of twelve sheep. The two forms of meniscectomy produced a comparable degree of postoperative lameness that resolved within two weeks of the operations. After six months the sheep were euthanatised and the stifle joints examined. Fibrous tissue that replaced the excised meniscus in the total meniscectomy group did not cover as much of the medial tibial condyle as the residual cranial pole and caudal fibrous tissue observed following hemimeniscectomy. The articular cartilage from different regions within the joints was examined for gross and histological evidence of degeneration. Analyses of the articular cartilage for water content, glycosaminoglycan composition and DNA content were performed. The proteoglycan synthesis and release from explanted articular cartilage samples in tissue culture were also measured. There were significant pathological changes in the medial compartment of all meniscectomised joints. The degree of articular cartilage degeneration that was observed following total meniscectomy and caudal pole meniscectomy was similar. Caudal pole hemimeniscectomy, involving transection of the meniscus, causes the same degree of degeneration of the stifle joint that occurs following total meniscectomy.The effect of total medial meniscectomy versus caudal pole hemimeniscectomy on the stifle joint of sheep was studied experimentally. Six months after the operations gross pathology, histopathology, cartilage biochemical analysis and the rate of proteoglycan synthesis in tissue culture were used to compare the articular cartilage harvested from the meniscectomised joints. Degeneration of the articular cartilage from the medial compartment of the joints was present in both of the groups. Caudal pole hemimeniscectomy induces a comparable degree of articular cartilage degeneration to total medial meniscectomy in the sheep stifle joint.

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