scholarly journals STUDIES ON ULTRASTRUCTURAL IDENTIFICATION AND DISTRIBUTION OF PROTEIN-POLYSACCHARIDE IN CARTILAGE MATRIX

1967 ◽  
Vol 32 (2) ◽  
pp. 365-377 ◽  
Author(s):  
Victor J. Matukas ◽  
Bernard J. Panner ◽  
J. Lowell Orbison
Keyword(s):  
Articular Cartilage ◽  
Cartilage Matrix ◽  
Epiphyseal Cartilage ◽  
Ground Substance ◽  
Dense Matrix ◽  
Chick Embryos ◽  
Chemical Analyses ◽  
Electron Microscopic ◽  
Colloidal Iron ◽  
The Matrix

Previous reports on the ultrastructure of cartilage matrix have described fibers, amorphous ground substance and, in some instances, dense matrix granules. The fibers are presumably collagen, but the nature of the granules is unknown. The primary purpose of this study has been to investigate the ultrastructure of cartilage matrix ih chick embryos with particular emphasis on the distribution and composition of these granules. In matrix of the zone of articular cartilage, mature collagen fibers can be seen but granules are not present. In matrix of all other zones of cartilage, fibers are smaller and granules are present. When the matrix of epiphyseal cartilage is compared to that of the zone of hypertrophic cells, fibers are similar but the granules in the latter zone are larger and more numerous. The granules in both zones were digested by hyaluronidase and positive to colloidal iron staining. Chemical analyses of cartilage from these zones indicate the hexosamine and radiosulfate content of the zone of hypertrophic cells to be higher than that of the zone of epiphyseal cartilage. The increased hexosamine was shown by column chromatography to be principally sulfated mucopolysaccharide, thereby indicating a direct correlation between size and number of granules and sulfated mucopolysaccharide content in the two zones. These data and the results of the electron microscopic histochemical studies are consistent with the concept that the granules in cartilage matrix contain acidic mucopolysaccharide.

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 INCREASED FIXATION OF SULFUR35 BY CARTILAGE IN VITRO FOLLOWING DEPLETION OF THE MATRIX BY INTRAVENOUS PAPAIN

1960 ◽  
Vol 112 (5) ◽  
pp. 743-750 ◽  
Author(s):  
T. F. McElligott ◽  
J. L. Potter
Keyword(s):  
Articular Cartilage ◽  
Chondroitin Sulfate ◽  
Intravenous Injection ◽  
Experimental Situation ◽  
Costal Cartilage ◽  
Primary Lesion ◽  
Cartilage Matrix ◽  
Osteoarthritic Cartilage ◽  
The Matrix

The uptake in vitro of sulfur-35 by costal cartilage obtained from nine rabbits 11 days after an intravenous injection of crude papain solution was compared with that in costal cartilage from eight normal untreated rabbits. An increased fixation of the isotope was found in treated animals compared with controls. The depletion of cartilage matrix by papain provided an experimental situation to test the hypothesis that the depletion of matrix which occurs in osteoarthritic cartilage can stimulate increased synthesis of chondroitin sulfate. The results give further support to the view that the primary lesion in osteoarthritis occurs in the matrix rather than in the chondrocyte of articular cartilage.

RADIOAUTOGRAPHIC STUDY OF S35O4 UPTAKE BY EPIPHYSEAL CARTILAGE IN EXPERIMENTAL LATHYRISM

1962 ◽  
Vol 40 (1) ◽  
pp. 565-570 ◽  
Author(s):  
Yurika K. Shintani ◽  
H. E. Taylor
Keyword(s):  
Normal Diet ◽  
Cartilage Matrix ◽  
Epiphyseal Cartilage ◽  
Acid Mucopolysaccharides ◽  
The Matrix ◽  
Endochondral Growth ◽  
Experimental Lathyrism

Disturbances in the uptake of radiosulphate were observed in radioautographs of the epiphysis of rats made lathyritic by giving either beta-aminopropionitrile or semicarbazide. There was a decreased uptake of sulphate which became more marked as the lesions advanced and an increased uptake was noted as the lesions regressed when the rats were returned to a normal diet. In normal rats, the radio-sulphate shifted to the zone of calcifying cartilage by the 4th day postinjection and, by the 7th day, it was concentrated over the ossifying trabeculae. In lathyrism this shift was delayed and the radiosulphate image was still concentrated in the cartilage matrix at days 4 and 7 postinjection. It is believed these findings reflect a disturbance in the matrix acid mucopolysaccharides and an interference with endochondral growth in lathyrism.

scholarly journals Localization of proteoglycan monomer and link protein in the matrix of bovine articular cartilage: An immunohistochemical study.

1980 ◽  
Vol 28 (7) ◽  
pp. 621-635 ◽  
Author(s):  
A R Poole ◽  
I Pidoux ◽  
A Reiner ◽  
L H Tang ◽  
H Choi ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Articular Surface ◽  
Guanidine Hydrochloride ◽  
Cartilage Matrix ◽  
Binding Studies ◽  
Superficial Zone ◽  
Surface Culture ◽  
Bovine Articular Cartilage ◽  
Link Protein ◽  
The Matrix

Using monospecific antisera and immunofluorescence microscopy, proteoglycan monomer (PG), and link proteins were demonstrated throughout the extracellular matrix of bovine articular cartilage. A narrow band of strong pericellular staining was usually observed for both molecules, indicating a pericellular concentration of proteoglycan monomer: this conclusion was supported by dye-binding studies. Whereas PG was evenly distributed throughout the remaining matrix, more link protein was detectable in interterritorial sites in middle and deep zones. Well-defined zones of weaker territorial staining for link protein stained strongest for chondroitin sulfate. Trypsin treatment of cartilage resulted in a loss of most of the PG staining, but some selective retention of link protein, particularly around chondrocytes in the superficial zone at and near the articular surface. This residual staining was largely removed if sections were fixed after chondroitinase treatment. After extraction of cartilage with 4M guanidine hydrochloride, only PG remained and this was concentrated in the superficial zone. These observations are shown to support the concept of aggregation of PG and link protein with hyaluronic acid (HA) in cartilage matrix, and the binding of PG and link protein to HA, which is attached to the chondrocyte surface. Culture of cartilage depleted of PG and link protein by trypsin demonstrated that individual chondrocytes can secrete both PG and link proteins and that the organization of cartilage matrix can be regenerated in part over a period of 4 days.

Morphometric Evaluation of Rhesus Articular Cartilage: Correlation of Hexosamine and 35SO4= Incorporation

1989 ◽  
Vol 26 (4) ◽  
pp. 309-313 ◽  
Author(s):  
E. D. Roberts ◽  
D. H. Zeman ◽  
T. Miwa ◽  
H. Shoje
Keyword(s):  
Articular Cartilage ◽  
Inverse Correlation ◽  
Area Ratio ◽  
Cartilage Matrix ◽  
Significant Difference ◽  
The Matrix ◽  
Morphometric Assessment ◽  
Hexosamine Content ◽  
Morphometric Evaluation

Morphometric evaluation of 20 rhesus articular cartilage samples were correlated with hexosamine and an 18–hour incorporation of 35SO4= as a measure of proteoglycan production on samples from the same joint. Reduced chondrocyte cellularity was the basis of the reduced maintenance of the matrix by the chondrocyte. In the more cellular cartilage, the matrix/lacunae area ratio was <25, and the less cellular group had a ratio of >40. An inverse correlation existed between morphometric cartilage matrix/lacunar area ratio and hexosamine content. A significant difference of 35SO4= incorporation was not seen between the three morphometric grades. Morphometric assessment reduces the subjectivity of articular cartilage evaluation.

Protective Abilities of Interleukin-10 in Blood-Induced Joint Damage.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 494-494
Author(s):  
Nathalie W.D. Jansen ◽  
Joel A.G. Van Roon ◽  
Goris Roosendaal ◽  
Matthias Theobald ◽  
Johannes W.J. Bijlsma ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Articular Cartilage ◽  
Blood Cells ◽  
Joint Damage ◽  
Interleukin 10 ◽  
Cartilage Matrix ◽  
Proteoglycan Synthesis ◽  
Matrix Synthesis ◽  
Irreversible Inhibition ◽  
The Matrix

Abstract PURPOSE: Joint bleeds occur frequently in patients suffering from haemophilia or can occur as a consequence of joint trauma. Even a limited number of bleeds has been demonstrated to lead to severe joint damage in time (1,2). With respect to the mechanisms of blood induced joint damage, we have demonstrated that monocytes/macrophages within the mononuclear cell population together with the red blood cells as present in blood are responsible for the irreversible inhibition of cartilage matrix synthesis. Small amounts of IL-1 that are produced by activated monocytes/macrophages increase the production of hydrogen peroxide (H2O2) by chondrocytes. The hydrogen peroxide reacts with haemoglobin-derived iron from damaged and phagocytosed red blood cells, which results in the formation of hydroxyl radicals in the vicinity of chondrocytes. This leads to chondrocyte apoptosis and with that the irreversible inhibition of cartilage matrix synthesis (2). In search of possible interventions to prevent or limit the cartilage damaging effects of joint bleeds, we tested interleukin-10 (IL-10) as an inflammation controlling cytokine on blood-induced cartilage damage. METHODS: Healthy human articular cartilage tissue explants were cultured in the presence or absence of 50% v/v blood for 4 days, followed by a recovery period of 12 days. IL-10 was added in 0.1, 1 or 10 ng/ml. Also cartilage and synovium in case of haemophilic arthropathy, obtained at joint replacement surgery, was cultured in the presence of IL-10 (10 ng/ml). The effect on cartilage matrix proteoglycan synthesis, -release, and -content were determined as well as the IL-1β and TNFα production by the synovium. RESULTS: IL-10 was able to prevent the decrease in proteoglycan synthesis and the increase in proteoglycan release of cartilage exposed to blood dose dependently (p<0.05). As a consequence, the decrease in proteoglycan content after blood exposure could be prevented dose dependently (p<0.05). With 10 ng/ml IL-10, which is still a low dose for local therapeutic treatment, at least 50% reduction in adverse effects was observed (p<0.05). The matrix turnover of haemophilic cartilage improved upon addition of IL-10 and also the production of IL-1β and TNFα by haemophilic synovium decreased in the presence of IL-10 (all p<0.05). CONCLUSIONS: The present results show that interleukin-10 prevents the direct harmful effects of blood on articular cartilage and has a beneficial effect on the matrix turnover of haemophilic cartilage and pro-inflammatory cytokine production by haemophilic synovium. Therefore, although treatment studies in addition to prevention studies have to be performed; IL-10 might be worthwhile to test in clinical practice for local treatment of joint haemorrhages, to prevent joint damage in time.

RADIOAUTOGRAPHIC STUDY OF S35O4 UPTAKE BY EPIPHYSEAL CARTILAGE IN EXPERIMENTAL LATHYRISM

1962 ◽  
Vol 40 (5) ◽  
pp. 565-570 ◽  
Author(s):  
Yurika K. Shintani ◽  
H. E. Taylor
Keyword(s):  
Normal Diet ◽  
Cartilage Matrix ◽  
Epiphyseal Cartilage ◽  
Acid Mucopolysaccharides ◽  
The Matrix ◽  
Endochondral Growth ◽  
Experimental Lathyrism

Disturbances in the uptake of radiosulphate were observed in radioautographs of the epiphysis of rats made lathyritic by giving either beta-aminopropionitrile or semicarbazide. There was a decreased uptake of sulphate which became more marked as the lesions advanced and an increased uptake was noted as the lesions regressed when the rats were returned to a normal diet. In normal rats, the radio-sulphate shifted to the zone of calcifying cartilage by the 4th day postinjection and, by the 7th day, it was concentrated over the ossifying trabeculae. In lathyrism this shift was delayed and the radiosulphate image was still concentrated in the cartilage matrix at days 4 and 7 postinjection. It is believed these findings reflect a disturbance in the matrix acid mucopolysaccharides and an interference with endochondral growth in lathyrism.

scholarly journals COMPARISON OF THE EFFECTS OF PAPAIN AND VITAMIN A ON CARTILAGE

1960 ◽  
Vol 111 (5) ◽  
pp. 705-718 ◽  
Author(s):  
Lewis Thomas ◽  
Robert T. McCluskey ◽  
Jacobus L. Potter ◽  
Gerald Weissmann
Keyword(s):  
Vitamin A ◽  
Chondroitin Sulfate ◽  
Cartilage Matrix ◽  
Epiphyseal Cartilage ◽  
Blue Staining ◽  
Daily Injection ◽  
Alcian Blue Staining ◽  
The Matrix ◽  
Cartilage Cells ◽  
Ear Cartilage

The administration of large amounts of vitamin A to rabbits has been shown to result in depletion of cartilage matrix. The normal basophilic, metachromatic, and Alcian blue staining properties of the matrix are lost, especially in articular and epiphyseal cartilage. The cartilage cells remain intact, but are reduced in size. These changes sometimes appeared as early as 48 hours after the initiation of daily injection of 1 million units of vitamin A, and were usually well established by 5 days. Some rabbits failed to show changes in cartilage, even after 5 daily injections. Increased amounts of material presumed to be chondroitin sulfate were present in the sera of vitamin A-treated rabbits, usually by 72 hours after the first injection. This was demonstrated by a turbidimetric procedure using hexamminecobaltic chloride. In rabbits given sulfur-35 (Na2S35O4) 5 days before the initiation of vitamin A treatment, it was shown that sulfur-35 was lost from articular and epiphyseal cartilage. This was associated with an increase in the non-dialyzable sulfur-35 in both serum and in the cobalt-precipitable material. These rabbits also excreted more sulfur-35 than rabbits not given vitamin A. There was a reduction in sulfur-35 activity in chondromucoprotein extracted from the ear cartilage of vitamin A-treated rabbits. The changes are interpreted as indicating that the administration of large amounts of vitamin A to rabbits results in removal of chondroitin sulfate from cartilage matrix. The administration of small amounts of crude papain causes histologic changes in cartilage that are remarkably similar to those seen in vitamin A-treated rabbits. The possibility is suggested that the changes in cartilage produced by administration of vitamin A to rabbits may be the result of activation of a proteolytic enzyme or enzymes, with properties similar to those of papain.

Intercellular Substances of the Uterine Cervix: Electron Microscopic Histochemical Studies of Hormone-Dependent Changes in the Mouse

1968 ◽  
Vol 26 ◽  
pp. 60-61
Author(s):  
T. J. Leppi ◽  
P. A. Kinnison ◽  
S. P. Gaffney
Keyword(s):  
Connective Tissue ◽  
Prussian Blue ◽  
Uterine Cervix ◽  
Histochemical Staining ◽  
Ground Substance ◽  
Extracellular Material ◽  
Electron Microscopic ◽  
Colloidal Iron ◽  
Cytochemical Study ◽  
Ovariectomized Mice

Earlier study of the connective tissue ground substance in uterine cervical walls of ovariectomized mice established the presence of a histochemically stainable extracellular material which varied in amount depending on hormonal conditions. The administration of estrogen singly or in combination with relaxin resulted in the appearance within the stroma of colloidal iron-positive material visible as Prussian blue. No iron staining was present in cervices from mice treated with relaxin alone or the hormone vehicles. The stainable material was tentatively identified as acid muco-polysaccharide(s) on the basis of its lability toward testicular hyaluroni-dase. The present ultrastructural histo- and cytochemical study of uterine cervices from mice in similar hormonal states was undertaken to: 1) identify clearly the connective tissue ground substance in this site; 2) establish a correlation with the previous light microscopic results; and 3) characterize further the histochemical staining properties of the intercellular material.

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