Subpopulations of chondrocytes from different zones of pig articular cartilage. Isolation, growth and proteoglycan synthesis in culture

1990 ◽  
Vol 97 (2) ◽  
pp. 349-360
Author(s):  
M. Siczkowski ◽  
F.M. Watt
Keyword(s):  
Articular Cartilage ◽  
Articular Surface ◽  
Polyacrylamide Gel Electrophoresis ◽  
Proteoglycan Synthesis ◽  
Lower Zone ◽  
Keratan Sulphate ◽  
Sulphate Content ◽  
Plating Density ◽  
Morphological Differences ◽  
Two Populations

Articular cartilage varies in ultrastructure and composition with distance from the articular surface. We have cultured chondrocytes from different zones of pig articular cartilage and investigated whether there are intrinsic differences in their behaviour that might account for the variation observed in intact tissue. On isolation, cells from the upper third of the cartilage were smaller than those of the lower third, but this difference was not maintained in culture. Upper zone cells attached and spread more slowly than lower zone cells; morphological differences between the two populations could be seen for several weeks. The growth rates of the two populations were similar, but upper zone cells reached a lower confluent density. Levels of protein synthesis were similar for both populations, but upper zone cells deposited less proteoglycan in the cell layer. On isolation, the percentage of upper zone cells that stained positive with MZ15, a monoclonal antibody to keratan sulphate, was smaller than the percentage of lower zone cells, but this difference was lost after several days in culture. Nevertheless, the keratan sulphate content of proteoglycan synthesised by lower zone chondrocytes at high density was greater than of that synthesised by upper zone cells. The proportion of nonaggregating proteoglycan was greater in upper than lower zone cartilage and this difference was also observed in long-term cultures. proteoglycans were further characterised by composite and polyacrylamide gel electrophoresis and by immunoblotting; differences detected in cartilage extracts were not, however, maintained in culture; instead, the small proteoglycans synthesised by both upper and lower zone cells varied with plating density. Finally, alkaline phosphatase, a marker of hypertrophic, calcifying cartilage, was only expressed in lower zone cultures. We conclude that some of the observed heterogeneity of articular cartilage reflects intrinsic differences between the cells of different zones, whereas some may reflect the response of chondrocytes to different environmental conditions.

scholarly journals Variations in the composition of bovine hip articular cartilage with distance from the articular surface

1981 ◽  
Vol 195 (3) ◽  
pp. 535-543 ◽  
Author(s):  
A Franzén ◽  
S Inerot ◽  
S O Hejderup ◽  
D Heinegård
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Chondroitin Sulphate ◽  
Articular Surface ◽  
Full Thickness ◽  
Rich Region ◽  
Keratan Sulphate ◽  
Average Size ◽  
Almost All ◽  
Cartilage Proteoglycans

Punch biopsies of bovine hip articular cartilage was sectioned according to depth and the proteoglycans were isolated. The mid-sections of the cartilage contained more proteoglycans than did either the superficial or the deepest portions of the cartilage proteoglycans than did either the superficial or the deepest portions of the cartilage. The most superficial 40 micrometer of the cartilage contained relatively more glucosaminoglycans compared with the remainder of the cartilage. The proteoglycans recovered from the surface 200 micrometer layer contained less chondroitin sulphate, were smaller and almost all of these molecules were able to interact with hyaluronic acid to form aggregates. From about 200 micrometer and down to 1040 micrometer from the surface, the proteoglycans became gradually somewhat smaller, probably owing to decreasing size of the chondroitin sulphate-rich region. The proportion of molecules that were able to interact with the hyaluronic acid was about 90% and remained constant with depth. The proteoglycans from the deepest layer near the cartilage-bone junction contained a large proportion of non-aggregating molecules, and the average size of the proteoglycans was somewhat larger. The alterations of proteoglycan structure observed with increasing depth of the articular cartilage beneath the surface layer (to 200 micrometer) are of the same nature as those observed with increasing age in full-thickness articular cartilage. The articular-cartilage proteoglycans were smaller and had much higher keratan sulphate and protein contents that did molecules isolated from bovine nasal or tracheal cartilage.

scholarly journals Turnover of proteoglycans in articular-cartilage cultures. Characterization of proteoglycans released into the medium

1989 ◽  
Vol 259 (1) ◽  
pp. 21-25 ◽  
Author(s):  
M A Campbell ◽  
C J Handley ◽  
S E D'Souza
Keyword(s):  
Articular Cartilage ◽  
Core Protein ◽  
Chondroitin Sulphate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Definitive Evidence ◽  
Core Proteins ◽  
The Matrix

By using an e.l.i.s.a. method it was demonstrated that the majority of proteoglycans released into the medium of both control and retinoic acid-treated explant cultures of bovine articular cartilage did not contain a hyaluronate-binding region. This supports our previous findings [Campbell & Handley (1987) Arch. Biochem. Biophys. 258, 143-155] that proteoglycans released into the medium of both cultures were of smaller hydrodynamic size, more polydisperse and unable to form aggregates with hyaluronate. Analysis of 35S-labelled core proteins associated with proteoglycans released into the medium of both cultures by using SDS/polyacrylamide-gel electrophoresis and fluorography indicated the presence of a series of core-protein bands (Mr approx. 300,000, 230,000, 215,000, 200,000, 180,000, 140,000, 135,000, 105,000, 85,000 and 60,000) compared with three core proteins derived from the proteoglycans remaining in the matrix (Mr 300,000, 230,000 and 215,000). Further analysis of the core proteins released into the medium indicated that the larger core proteins associated with medium proteoglycans contain both chondroitin sulphate and keratan sulphate glycosaminoglycans whereas the smaller core proteins contain only chondroitin sulphate chains. These experiments provide definitive evidence that the loss of proteoglycans from the matrix involves proteolytic cleavage at various sites along the proteoglycan core protein.

scholarly journals Synthesis of hyaluronate in cultured bovine articular cartilage

1989 ◽  
Vol 263 (3) ◽  
pp. 761-767 ◽  
Author(s):  
C K Ng ◽  
C J Handley ◽  
R M Mason ◽  
H C Robinson
Keyword(s):  
Articular Cartilage ◽  
Deep Layer ◽  
Superficial Layer ◽  
Explant Culture ◽  
Proteoglycan Synthesis ◽  
Hydrodynamic Size ◽  
Short Term ◽  
Bovine Articular Cartilage ◽  
Sulphated Glycosaminoglycans ◽  
Two Populations

The synthesis and distribution of hyaluronate and proteoglycan were studied in bovine articular cartilage in short-term explant culture with [3H]acetate and H2(35)SO4 as precursors. The incorporation of [3H]acetate into hyaluronate and sulphated glycosaminoglycans was linear with time, except that hyaluronate synthesis showed a marked lag at the beginning of the incubation. [3H]Hyaluronate represented 4-7% of the total [3H]glycosaminoglycans synthesized over a 6 h period. However, the distributions of [3H]hyaluronate and 3H-labelled sulphated glycosaminoglycans were different: about 50% of the newly synthesized [3H]hyaluronate appeared in the medium, compared with less than 5% of the 3H-labelled sulphated proteoglycans. A pulse-chase experiment revealed that the release of newly synthesized [3H]hyaluronate from cartilage was rapid. No difference was observed in the distribution of [3H]hyaluronate between medium and tissue by cartilage from either the superficial layer or the deep layer of articular cartilage. When articular cartilage was incubated with 0.4 mM-cycloheximide, proteoglycan synthesis was markedly inhibited, whereas the synthesis of hyaluronate was only partially inhibited and resulted in more of the newly synthesized hyaluronate being released into the medium. Analysis of the hydrodynamic size of [3H]hyaluronate isolated from cartilage on Sephacryl-1000 revealed one population that was eluted as a broad peak (Kav. less than 0.7), compared with two populations (Kav. greater than 0.5 and less than 0.5) appearing in the medium of cultures. These data suggest that hyaluronate is synthesized in excess of proteoglycan synthesis and that the hyaluronate that is not complexed with proteoglycans is rapidly lost from the tissue.

Structural studies of two populations of keratan sulphate chains from mature bovine articular cartilage

1989 ◽  
Vol 6 (2) ◽  
pp. 209-218 ◽  
Author(s):  
David J Thornton ◽  
Haydn G Morris ◽  
Gordon H Cockin ◽  
Thomas N Huckerby ◽  
Ian A Nieduszynski
Keyword(s):  
Articular Cartilage ◽  
Structural Studies ◽  
Keratan Sulphate ◽  
Bovine Articular Cartilage ◽  
Two Populations

scholarly journals Structure of proteoglycans from different layers of human articular cartilage

1983 ◽  
Vol 209 (2) ◽  
pp. 387-400 ◽  
Author(s):  
M T Bayliss ◽  
M Venn ◽  
A Maroudas ◽  
S Y Ali
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Chondroitin Sulphate ◽  
Articular Surface ◽  
Current Model ◽  
Gel Chromatography ◽  
Slice Thickness ◽  
Human Articular Cartilage ◽  
Guanidinium Chloride ◽  
Keratan Sulphate

Full-depth plugs of adult human articular cartilage were cut into serial slices from the articular surface and analysed for their glycosaminoglycan content. The amount of chondroitin sulphate was highest in the mid-zone, whereas keratan sulphate increased progressively through the depth. Proteoglycans were isolated from each layer by extraction with 4M-guanidinium chloride followed by centrifugation in 0.4M-guanidinium chloride/CsCl at a starting density of 1.5 g/ml. The efficiency with which proteoglycans were extracted depended on slice thickness, and extraction was complete only when cartilage from each zone was sectioned at 20 microns or less. When thick sections (250 microns) were extracted, hyaluronic acid was retained in the tissue. Most of the proteoglycans, extracted from each layer under optimum conditions, could interact with hyaluronic acid to form aggregates, although the extent of aggregation was less in the deeper layers. Two pools of proteoglycan were identified in all layers by gel chromatography (Kav. 0.33 and 0.58). The smaller of these was rich in keratan sulphate and protein, and gradually increased in proportion through the cartilage depth. Chondroitin sulphate chain size was constant in all regions. The changes in composition and structure observed were consistent with the current model for hyaline-cartilage proteoglycans and were similar to those observed with increasing age in human articular cartilage.

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

1999 ◽  
Vol 12 (02) ◽  
pp. 56-63 ◽  
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.

scholarly journals Anatomy of the Intermetatarsal Facets of the Fourth and Fifth Metatarsals

2021 ◽  
Vol 6 (1) ◽  
pp. 247301142097570
Author(s):  
Mossub Qatu ◽  
George Borrelli ◽  
Christopher Traynor ◽  
Joseph Weistroffer ◽  
James Jastifer
Keyword(s):  
Articular Cartilage ◽  
Digital Imaging ◽  
Articular Surface ◽  
Implant Design ◽  
Joint Surface ◽  
Fracture Classification ◽  
Articular Damage ◽  
Metatarsal Fracture ◽  
Metatarsal Fractures ◽  
The Mean

Background: The intermetatarsal joint between the fourth and fifth metatarsals (4-5 IM) is important in defining fifth metatarsal fractures. The purpose of the current study was to quantify this joint in order to determine the mean cartilage area, the percentage of the articulation that is cartilage, and to give the clinician data to help understand the joint anatomy as it relates to fifth metatarsal fracture classification. Methods: Twenty cadaver 4-5 IM joints were dissected. Digital images were taken and the articular cartilage was quantified by calibrated digital imaging software. Results: For the lateral fourth proximal intermetatarsal articulation, the mean area of articulation was 188 ± 49 mm2, with 49% of the area composed of articular cartilage. The shape of the articular cartilage had 3 variations: triangular, oval, and square. A triangular variant was the most common (80%, 16 of 20 specimens). For the medial fifth proximal intermetatarsal articulation, the mean area of articulation was 143 ± 30 mm2, with 48% of the joint surface being composed of articular cartilage. The shape of the articular surface was oval or triangular. An oval variant was the most common (75%, 15 of 20 specimens). Conclusion: This study supports the notion that the 4-5 IM joint is not completely articular and has both fibrous and cartilaginous components. Clinical Relevance: The clinical significance of this study is that it quantifies the articular surface area and shape. This information may be useful in understanding fifth metatarsal fracture extension into the articular surface and to inform implant design and also help guide surgeons intraoperatively in order to minimize articular damage.

A Review of the Collagen Orientation in the Articular Cartilage

Cartilage ◽  
2021 ◽  
pp. 194760352098877
Author(s):  
Roy D. Bloebaum ◽  
Andrew S. Wilson ◽  
William N. Martin
Keyword(s):  
Surface Layer ◽  
Articular Cartilage ◽  
Fiber Orientation ◽  
Collagen Fiber ◽  
Articular Surface ◽  
Imaging Techniques ◽  
Collagen Fibers ◽  
Deep Zone ◽  
Collagen Fiber Orientation ◽  
Critical Point Drying

Objective There has been a debate as to the alignment of the collagen fibers. Using a hand lens, Sir William Hunter demonstrated that the collagen fibers ran perpendicular and later aspects were supported by Benninghoff. Despite these 2 historical studies, modern technology has conflicting data on the collagen alignment. Design Ten mature New Zealand rabbits were used to obtain 40 condyle specimens. The specimens were passed through ascending grades of alcohol, subjected to critical point drying (CPD), and viewed in the scanning electron microscope. Specimens revealed splits from the dehydration process. When observing the fibers exposed within the opening of the splits, parallel fibers were observed to run in a radial direction, normal to the surface of the articular cartilage, radiating from the deep zone and arcading as they approach the surface layer. After these observations, the same samples were mechanically fractured and damaged by scalpel. Results The splits in the articular surface created deep fissures, exposing parallel bundles of collagen fibers, radiating from the deep zone and arcading as they approach the surface layer. On higher magnification, individual fibers were observed to run parallel to one another, traversing radially toward the surface of the articular cartilage and arcading. Mechanical fracturing and scalpel damage induced on the same specimens with the splits showed randomly oriented fibers. Conclusion Collagen fiber orientation corroborates aspects of Hunter’s findings and compliments Benninghoff. Investigators must be aware of the limits of their processing and imaging techniques in order to interpret collagen fiber orientation in cartilage.

On the Natural Lubrication of Synovial Joints: Normal and Degenerate

1977 ◽  
Vol 99 (2) ◽  
pp. 163-172 ◽  
Author(s):  
Joseph M. Mansour ◽  
Van C. Mow
Keyword(s):  
Articular Cartilage ◽  
Solid Phase ◽  
Articular Surface ◽  
Moving Load ◽  
Frictional Resistance ◽  
Elastic Solid ◽  
Surface Load ◽  
Two Phase ◽  
Tissue Properties ◽  
Synovial Joints

Fluid flow and mass transport mechanisms associated with articular cartilage function are important biomechanical processes of normal and pathological synovial joints. A three-layer permeable, two-phase medium of an incompressible fluid and a linear elastic solid are used to model the flow and deformational behavior of articular cartilage. The frictional resistance of the relative motion of the fluid phase with respect to the solid phase is given by a linear diffusive dissipation term. The subchondral bony substrate is represented by an elastic solid. The three-layer model of articular cartilage is chosen because of the known histological, ultrastructural, and biomechanical variations of the tissue properties. The calculated flow field shows that for material properties of normal healthy articular cartilage the tissue creates a naturally lubricated surface. The movement of the interstitial fluid at the surface is circulatory in manner, being exuded in front and near the leading half of the moving surface load and imbibed behind and near the trailing half of the moving load. The flow fields of healthy tissues are capable of sustaining a film of fluid at the articular surface whereas pathological tissues cannot.

scholarly journals Reduction of interleukin‐17–induced inhibition of chondrocyte proteoglycan synthesis in intact murine articular cartilage by interleukin‐4

2000 ◽  
Vol 43 (6) ◽  
pp. 1300-1306 ◽  
Author(s):  
Erik Lubberts ◽  
Leo A. B. Joosten ◽  
Fons A. J. Van De Loo ◽  
Liduine A. M. Van Den Bersselaar ◽  
Wim B. Van Den Berg
Keyword(s):  
Articular Cartilage ◽  
Interleukin 4 ◽  
Proteoglycan Synthesis ◽  
Interleukin 17
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