scholarly journals Articular-cartilage matrix γ-carboxyglutamic acid-containing protein. Characterization and immunolocalization

1992 ◽  
Vol 282 (1) ◽  
pp. 1-6 ◽  
Author(s):  
R Loeser ◽  
C S Carlson ◽  
H Tulli ◽  
W G Jerome ◽  
L Miller ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Matrix Vesicles ◽  
Protein Characterization ◽  
Pericellular Matrix ◽  
Guanidinium Chloride ◽  
Carboxyglutamic Acid ◽  
The Matrix ◽  
Important Regulator ◽  
Immunohistochemical Studies ◽  
Bovine Cartilage

Matrix gamma-carboxyglutamic acid (Gla)-containing protein (MGP) was found to be present in articular cartilage by Western-blot analysis of guanidinium chloride extracts of human and bovine cartilage and was further localized by immunohistochemical studies on human and monkey specimens. In newborn articular cartilage MGP was present diffusely throughout the matrix, whereas in growth-plate cartilage it was seen mainly in late hypertrophic and calcifying-zone chondrocytes. In adult articular cartilage MGP was present primarily in chondrocytes and the pericellular matrix. Immunoelectron microscopy studies revealed an association between MGP and vesicular structures with an appearance consistent with matrix vesicles. MGP may be an important regulator of cartilage calcification because of its localization in cartilage and the known affinity of Gla-containing proteins for Ca2+ and hydroxyapatite.

scholarly journals Chondrocyte-mediated depletion of articular cartilage proteoglycans in vitro

1985 ◽  
Vol 225 (2) ◽  
pp. 493-507 ◽  
Author(s):  
J A Tyler
Keyword(s):  
Articular Cartilage ◽  
Core Protein ◽  
Fold Increase ◽  
Acid Proteinase ◽  
Binding Region ◽  
Guanidinium Chloride ◽  
The Core ◽  
The Matrix ◽  
Average Size

The degradation of proteoglycan was examined in cultured slices of pig articular cartilage. Pig leucocyte catabolin (10 ng/ml) was used to stimulate the chondrocytes and induce a 4-fold increase in the rate of proteoglycan loss from the matrix for 4 days. Material in the medium of both control and depleted cultures was mostly a degradation product of the aggregating proteoglycan. It was recovered as a very large molecule slightly smaller than the monomers extracted with 4M-guanidinium chloride and lacked a functional hyaluronate binding region. The size and charge were consistent with a very limited cleavage or conformational change of the core protein near the hyaluronate binding region releasing the C-terminal portion of the molecule intact from the aggregate. The ‘clipped’ monomer diffuses very rapidly through the matrix into the medium. The amount of proteoglycan extracted with 4M-guanidinium chloride decreased during culture from both the controls and depleted cartilage, and the average size of the molecules initially remained the same. However, the proportion of molecules with a smaller average size increased with time and was predominant in explants that had lost more than 70% of their proteoglycan. All of this material was able to form aggregates when mixed with hyaluronate, and glycosaminoglycans were the same size and charge as normal, indicating either that the core protein had been cleaved in many places or that larger molecules were preferentially released. A large proportion of the easily extracted and non-extractable proteoglycan remained in the partially depleted cartilage and the molecules were the same size and charge as those found in the controls. There was no evidence of detectable glycosidase activity and only very limited sulphatase activity. A similar rate of breakdown and final distribution pattern was found for newly synthesized proteoglycan. Increased amounts of latent neutral metalloproteinases and acid proteinase activities were present in the medium of depleted cartilage. These were not thought to be involved in the breakdown of proteoglycan. Increased release of proteoglycan ceased within 24h of removal of the catabolin, indicating that the effect was reversible and persisted only while the stimulus was present.

Hypo-Osmolarity Decreases the Ability of Cells in Articular Cartilage to Survive a Load-Induced Injury

2003 ◽  
Author(s):  
Adam Levin ◽  
Peter A. Torzilli ◽  
C. T. Christopher Chen
Keyword(s):  
Cell Death ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Cartilage Explants ◽  
Hypotonic Solution ◽  
Pericellular Matrix ◽  
Isotonic Solution ◽  
Confined Compression ◽  
Superficial Zone ◽  
Bovine Cartilage

A recent study showed that exposure to hypotonic conditions increased chondrocyte surface area up to 234% by stretching the folded membrane, which may reduce the chondrocyte’s ability to deform under load. The goal of this study was to determine the effect of hypo-osmolarity on chondrocyte survival from load-induced injury. Bovine cartilage explants were incubated in either isotonic or hypotonic pH-buffered solution for 20 minutes, loaded with cyclic confined compression at 5MPa for 1 hour, and then assessed for cell viability using cell vital dyes and for pericellular matrix (PCM) using an type VI collagen antibody. Cell death in loaded explants was significantly greater than that of non-loaded controls (p<0.001). However, explants loaded in the hypotonic solution showed significantly greater cell death than those loaded in the isotonic solution. An increase of dead cells with flatten PCM were located in the superficial zone. Our findings suggest that hypo-osmolarity decreases the ability of chondrocytes in articular cartilage to survive from load-induced injury.

scholarly journals Immunohistochemical localization of fibronectin and chondronectin in canine articular cartilage.

1988 ◽  
Vol 36 (6) ◽  
pp. 581-588 ◽  
Author(s):  
N Burton-Wurster ◽  
V J Horn ◽  
G Lust
Keyword(s):  
Articular Cartilage ◽  
External Medium ◽  
Articular Surface ◽  
Immunohistochemical Localization ◽  
Pericellular Matrix ◽  
Osteoarthritic Cartilage ◽  
Matrix Interactions ◽  
The Matrix ◽  
Cut Surface ◽  
Disease Free

We compared the distribution of fibronectin and chondronectin within the matrix of canine articular cartilage. Fibronectin was found throughout the matrix as well as pericellularly. In contrast, chondronectin was observed predominantly associated with the cell or pericellular matrix. Interactions of these molecules with matrix components in the pericellular matrix probably differs, however, since concentrations of hyaluronidase which prevented detection of pericellular fibronectin allowed detection of chondronectin. Chondronectin and fibronectin were detected in osteoarthritic cartilage as well as in disease-free cartilage. Penetration of biotinylated fibronectin into cartilage from the external medium occurred only in osteoarthritic cartilage and proceeded only from the articular surface. Disease-free cartilage appeared to maintain a barrier to fibronectin penetration from the articular surface which was sustained even after the proteoglycan content was markedly depleted by incubation of cartilage with catabolin or lipopolysaccharide. In cartilage that was proteoglycan-depleted, the only detectable penetration of external fibronectin was from the cut surface.

High Frequency Acoustic Parameters of Human and Bovine Articular Cartilage following Experimentally-Induced Matrix Degradation

2001 ◽  
Vol 23 (2) ◽  
pp. 106-116 ◽  
Author(s):  
G.A. Joiner ◽  
E.R. Bogoch ◽  
K.P. Pritzker ◽  
M.D. Buschmann ◽  
A. Chevrier ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Speed Of Sound ◽  
Chemical Degradation ◽  
Matrix Degradation ◽  
Frequency Dependent ◽  
Bovine Articular Cartilage ◽  
Paired Samples ◽  
The Matrix ◽  
Interleukin 1Α ◽  
Bovine Cartilage

Matrix degradation and proteoglycan loss in articular cartilag eare features of early osteoarthritis. To determine the effect of matrix degradation and proteoglycan loss on ultrasound propagation in cartilage, we used papain and interleukin-1α to degrade the matrix proteoglycans of human and bovine cartilage samples, respectively. There is also minor collagen alteration associated with these chemical degradation methods. We compared the speed of sound and frequency dependent attenuation (20–40 MHz) of control and experimental paired samples. We found that a loss of matrix proteoglycans and collagen disruption resulted in a 20–30% increase in the frequency dependent attenuation and a 2% decrease in the speed of sound in both human and bovine cartilage. We conclude that the frequency dependent attenuation and speed of sound in articular cartilage are sensitive to experimental modification of the matrix proteoglycans and collagen. These findings suggest that ultrasound can potentially be used to detect morphologic changes in articular cartilage associated with the progression of osteoarthritis.

Effects of EDTA, Hyaluronidase and Collagenase on Epiphyseal Cartilage Matrix

1968 ◽  
Vol 26 ◽  
pp. 56-57
Author(s):  
H. Clarke Anderson ◽  
Priscilla R. Coulter
Keyword(s):  
Light And Electron Microscopy ◽  
Matrix Vesicles ◽  
Cartilage Matrix ◽  
Collagen Fibrils ◽  
Epiphyseal Cartilage ◽  
Dense Matrix ◽  
Type Iv ◽  
Bovine Testicular Hyaluronidase ◽  
The Matrix ◽  
Testicular Hyaluronidase

Epiphyseal cartilage matrix contains fibrils and particles of at least 5 different types: 1. Banded collagen fibrils, present throughout the matrix, but not seen in the lacunae. 2. Non-periodic fine fibrils <100Å in diameter (Fig. 1), which are most notable in the lacunae, and may represent immature collagen. 3. Electron dense matrix granules (Fig. 1) which are often attached to fine fibrils and collagen fibrils, and probably contain protein-polysaccharide although the possibility of a mineral content has not been excluded. 4. Matrix vesicles (Fig. 2) which show a selective distribution throughout the epiphysis, and may play a role in calcification. 5. Needle-like apatite crystals (Fig. 2).Blocks of formalin-fixed epiphysis from weanling mice were digested with the following agents in 0.1M phosphate buffer: a) 5% ethylenediaminetetraacetate (EDTA) at pH 8.3, b) 0.015% bovine testicular hyaluronidase (Sigma, type IV, 750 units/mg) at pH 5.5, and c) 0.1% collagenase (Worthington, chromatograhically pure, 200 units/mg) at pH 7.4. All digestions were carried out at 37°C overnight. Following digestion tissues were examined by light and electron microscopy to determine changes in the various fibrils and particles of the matrix.

scholarly journals P144 A CHONDROPROTECTIVE ROLE FOR FGF-2 IN THE PERICELLULAR MATRIX OF ARTICULAR CARTILAGE

2006 ◽  
Vol 14 ◽  
pp. S88
Author(s):  
T.L. Vincent ◽  
Y. Sawaji ◽  
C. McLean ◽  
L. Full ◽  
J. Saklatvala
Keyword(s):  
Articular Cartilage ◽  
Pericellular Matrix

Adult human chondrocytes cultured in alginate form a matrix similar to native human articular cartilage

1996 ◽  
Vol 271 (3) ◽  
pp. C742-C752 ◽  
Author(s):  
H. J. Hauselmann ◽  
K. Masuda ◽  
E. B. Hunziker ◽  
M. Neidhart ◽  
S. S. Mok ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cell Sorting ◽  
Proteolytic Enzymes ◽  
Alginate Beads ◽  
Human Chondrocytes ◽  
Human Articular Cartilage ◽  
Adult Human ◽  
The Matrix ◽  
Matrix Compartment ◽  
Associated Matrix

The matrix formed by adult human chondrocytes in alginate beads is composed of two compartments: a thin rim of cell-associated matrix that corresponds to the pericellular and territorial matrix of articular cartilage and a more abundant further-removed matrix, the equivalent of the interterritorial matrix in the tissue. On day 30 of culture, the relative and absolute volumes occupied by the cells and each of the two matrix compartments in the beads were nearly identical to those in native articular cartilage. Furthermore, the concentration of aggrecan in the cell-associated matrix was similar to that in adult human articular cartilage and was approximately 40-fold higher than in the further removed matrix compartment. Fluorescence-activated cell sorting revealed that the cell-associated matrix was built on the cell membrane in part via interactions between hyaluronic acid and CD44-like receptors. Approximately 25% of the aggrecan molecules synthesized by the chondrocytes during a 4-h pulse in the presence of [35S]sulfate on day 9 of culture were retained in the cell-associated matrix where they turned over with a half-life (t1/2) = 29 days. Most [35S]aggrecan molecules reached the further removed matrix compartment where they turned over much more slowly (t1/2 > 100 days). These results add support to the contention that aggrecan molecules residing in the pericellular and territorial areas of the adult human articular cartilage matrix are more susceptible to degradation by proteolytic enzymes synthesized by the chondrocytes than those that inhabit the interterritorial areas further removed from the cells.

scholarly journals Intact Pericellular Matrix of Articular Cartilage Is Required for Unactivated Discoidin Domain Receptor 2 in the Mouse Model

2011 ◽  
Vol 179 (3) ◽  
pp. 1338-1346 ◽  
Author(s):  
Lin Xu ◽  
Ilona Polur ◽  
Jacqueline M. Servais ◽  
Sirena Hsieh ◽  
Peter L. Lee ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Mouse Model ◽  
Pericellular Matrix ◽  
Discoidin Domain Receptor ◽  
Discoidin Domain Receptor 2

scholarly journals Biosynthesis of proteoglycan in vitro by cartilage from human osteochondrophytic spurs

1982 ◽  
Vol 206 (2) ◽  
pp. 329-341 ◽  
Author(s):  
Charles J. Malemud ◽  
Victor M. Goldberg ◽  
Roland W. Moskowitz ◽  
Lee L. Getzy ◽  
Robert S. Papay ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Deae Cellulose ◽  
Culture Conditions ◽  
Human Articular Cartilage ◽  
Guanidinium Chloride ◽  
Tissue Slices ◽  
Keratan Sulphate ◽  
Defined Culture

Proteoglycan biosynthesis by human osteochondrophytic spurs (osteophytes) obtained from osteoarthritic femoral heads at the time of surgical joint replacement was studied under defined culture conditions in vitro. Osteophytes were primarily present in two anatomic locations, marginal and epi-articular. Minced tissue slices were incubated in the presence of [35S]sulphate or [14C]glucosamine. Osteophytes incorporated both labelled precursors into proteoglycan, which was subsequently characterized by CsCl-isopycnic-density-gradient ultracentrifugation and chromatography on Sepharose CL-2B. The material extracted with 0.5m-guanidinium chloride showed 78.1% of [35S]sulphate in the A1 fraction after centrifugation. Only 23.0% of the [35S]sulphate in this A1 fraction was eluted in the void volume of Sepharose CL-2B under associative conditions. About 60–80% of the [35S]sulphate in the tissue 4m-guanidinium chloride extract was associated with monomeric proteoglycan (fraction D1). The average partition coefficient (Kav.) of the proteoglycan monomer on Sepharose CL-2B was 0.28–0.33. Approx. 12.4% of this monomer formed stable aggregates with high-molecular-weight hyaluronic acid in vitro. Sepharose CL-2B chromatography of fractions with lower buoyant densities (fractions D2–D4) demonstrated elution profiles on Sepharose CL-2B substantially different than that of fraction D1, indicative of the polydisperse nature of the newly synthesized proteoglycan. Analysis of the composition and chain size of the glycosaminoglycans showed the following: (1) preferential elution of both [35S]sulphate and [14C]glucosamine in the 0.5m-LiCl fraction on DEAE-cellulose; (2) the predominant sulphated glycosaminoglycan was chondroitin 6-sulphate (60–70%), with 9–11% keratan sulphate in the monomer proteoglycan; (3) Kav. values of 0.38 on Sephadex G-200 and 0.48 on Sepharose CL-6B were obtained with papain-digested and NaBH4-treated D1 monomer respectively. A comparison of the synthetic with endogenous glycosaminoglycans indicated similar types. These studies indicated that human osteophytes synthesized in vitro sulphated proteoglycans with some characteristics similar to those of mature human articular cartilage, notably in the size of their proteoglycan monomer and predominance of chondroitin 6-sulphate. They differed from articular cartilage primarily in the lack of substantial quantities of keratan sulphate and aggregation properties associated with monomer interaction with hyaluronic acid.

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