Heterogeneity of proteoglycans extracted before and after collagenase treatment of human articular cartilage: I. Physical properties related to age

Stadt, R J van de; Kuijer, Roel; Kampen, G. P. J. Van; Koning, M H M T de; Voorde‐Vissers, Els Van De; Korst, J. K. van der

doi:10.1002/art.1780291009

Cited by 11 publications

(2 citation statements)

References 24 publications

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“…Washed cartilage was then treated for 24 hours with 4M guanidine hydrochloride in the presence of enzyme inhibitors, including 100 mM 6-amino-n-hexanoic acid, 10 mM disodium EDTA, 10 mM N-ethylmaleimide, 5 mM benzamidine, and 1 mM phenylmethylsulfonyl fluoride (all from Sigma). Residual cartilage was digested with collagenase (type I1 clostridial; Sigma) for 24 hours at 37°C (17). The guanidine hydrochloride extract and the collagenase extract were treated in the same way as the medium compartment, except that PD-10 columns were equilibrated with 4M guanidine hydrochloride before the extracts were added.…”

Section: Methodsmentioning

confidence: 99%

Cartilage inorganic pyrophosphate elaboration is independent of sulfated glycosaminoglycan synthesis

Ryan

Kurup

McCarty

et al. 1990

Arthritis & Rheumatism

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Inorganic pyrophosphate (PPi), a product of glycosaminoglycan synthesis, may be cosecreted with matrix proteoglycan to reach the extracellular site where calcium pyrophosphate dihydrate crystals form. To test this hypothesis, sulfated glycosaminoglycan synthesis by articular cartilage in culture was stimulated or inhibited while the effect on extracellular PP, was measured. When stimulated by 0.8 mM xyloside to increase 35S0, incorporation (mean f SEM % of control 183 f 16, n = 5), PP, accumulation changed little (from 54 2 6 pmoleshg to 63 -+ 8 pmoleshg of cartilage wet weight). Inhibition of sulfation with monensin or diethylcarbamazine disproportionately lowered 35S0, incorporation compared with PP, elaboration. Using 60 mM diethylcarbamazine, PP, production was preserved (105 -+ 8% mean -t SEM) compared with control cultures, while sulfation was markedly inhibited (7 f 1 %). This dissociation of sulfate incorporation and PP, secretion indicates that it is not likely that glycosaminoglycan sulfation is the source of the PP, that escapes from chondrocytes to participate in the formation of extracellular crystals.

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Section: Methodsmentioning

confidence: 99%

Cartilage inorganic pyrophosphate elaboration is independent of sulfated glycosaminoglycan synthesis

Ryan

Kurup

McCarty

et al. 1990

Arthritis & Rheumatism

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“…Cells and media were processed as described above. The proteoglycans were extracted (4 M GuHCI) from the cultured tissues and corresponding culture media and purified (CsC1 density centrifugation) as described previously [3,4]. By sequential digestion with proteinase-K and chondroitinase-AC disaccharides from chondroitin sulphate were obtained.…”

Section: High Density and Monolayer Culturesmentioning

confidence: 99%

Effects of tissue disorganization on proteoglycan synthesis by articular chondrocytesin vitro

Vankampen¹,

Vandestadt²,

Kiljan³

et al. 1988

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Heterogeneity of proteoglycans extracted before and after collagenase treatment of human articular cartilage: II. Variations in composition with age and tissue source

Kuijer¹,

Vandestadt²,

Vankampen³

et al. 1986

Arthritis & Rheumatism

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Proteoglycans (A1 fractions) were extracted with 4M guanidine hydrochloride (GuHC1) from human articular cartilage samples of a wide age range. Distinctions were made between hip and knee, and upper and lower layers. The residues of these extractions were digested with purified collagenase, and a second extraction with 4M GuHCl was performed, which yielded appreciable amounts of proteoglycans. When proteoglycans from second extractions were compared with those from first extractions, the following changes were observed: an increase in chondroitin sulfate; a relative decrease in keratan sulfate; a decrease in protein content; and a decrease in the ratio of chondroitin 6-sulfate to chondroitin 4-sulfate. The same changes were found when nonaggregating proteoglycans were compared with proteoglycan aggregates, when proteoglycans from young cartilage were compared with those from mature carailage, when proteoglycans from knee cartilage were compared with those from hip cartilage, and when proteoglycans from upper layers of cartilage were compared with those from deeper layers. It is suggested that the differences found between first and second extrac- tions of cartilage, between upper and lower layers of cartilage, and between knee and hip cartilage are caused by variations in the relative amount of nonaggregating proteoglycans and/or variations in proteoglycan size.The most obvious biochemical alterations in aging cartilage are found in the composition and structure of the proteoglycan population (1-5). Proteoglycan size decreases, chondroitin sulfate chain length decreases, and the length of the core protein shortens (2). The content of keratan sulfate increases relatively, as well as absolutely (2,6). These changes were found to be caused by shifts in the relative amounts of the several proteoglycan subclasses (4,7).The generally used proteoglycan extraction procedure liberates, depending on tissue geometry (8), about 5040% of the proteoglycans from mature articular cartilage. In this study, proteoglycans were isolated from human articular cartilage by 2 consecutive extractions with 4M guanidine hydrochloride (GuHCl) intercalated by a collagenase digestion of the residue of the first extraction (5). The chemical properties of the high buoyant density proteoglycan preparations obtained are described in relation to age and to layer of the cartilage (upper or lower), because of reported differences in the composition of directly extracted proteoglycans that occur with depth of the cartilage (8).Proteoglycans isolated from hip and knee cartilage were compared because the knee joint seems to be prone to developing osteoarthritis at an earlier age than does the hip joint (9).

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Heterogeneity of proteoglycans extracted before and after collagenase treatment of human articular cartilage: I. Physical properties related to age

Cited by 11 publications

References 24 publications

Cartilage inorganic pyrophosphate elaboration is independent of sulfated glycosaminoglycan synthesis

Cartilage inorganic pyrophosphate elaboration is independent of sulfated glycosaminoglycan synthesis

Effects of tissue disorganization on proteoglycan synthesis by articular chondrocytesin vitro

Heterogeneity of proteoglycans extracted before and after collagenase treatment of human articular cartilage: II. Variations in composition with age and tissue source

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