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.

scholarly journals Keratan Sulfate Epitopes Exhibit a Conserved Distribution During Joint Development That Remains Undisclosed on the Basis of Glycosaminoglycan Charge Density

2002 ◽  
Vol 50 (8) ◽  
pp. 1039-1047 ◽  
Author(s):  
Emma Kavanagh ◽  
Anne C. Osborne ◽  
Doreen E. Ashhurst ◽  
Andrew A. Pitsillides
Keyword(s):  
Chondroitin Sulfate ◽  
Alcian Blue ◽  
Functional Role ◽  
Keratan Sulfate ◽  
Epiphyseal Cartilage ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Joint Development ◽  
Joint Cavity ◽  
And Cartilage

Changes in glycosaminoglycan (GAG) content and distribution are vital for joint development. However, their precise character has not been established. We have used immunohistochemistry (IHC) and “critical electrolyte” Alcian blue staining to assess such changes in developing chick and rabbit joints. IHC showed chondroitin sulfate labeling in chick epiphyseal cartilage but not in interzones. In contrast, prominent labeling for keratan sulfate (KS) was restricted to chick cartilage–interzone interfaces. In rabbit knees, KS labeling was also prominent at presumptive cavity borders, but weak in interzone and cartilage. Selective pre-digestion produced appropriate loss of label and undersulfated KS was undetectable. Quantification of Alcian blue staining by scanning and integrating microdensitometry showed prominent hyaluronan-like (HA-like) interzone staining, with chondroitin sulfate and weaker KS staining restricted to epiphyseal cartilage. Hyaluronidase decreased HA-like staining in the interzone. Surprisingly, keratanases also reduced HA-like but not sulfated GAG (sGAG-like) staining in the interzone. Chondroitinase ABC had little effect on HA-like staining but decreased sGAG staining in all regions. Rabbit joints also showed HA-like but not KS staining in the interzone and strong chondroitin sulfate-like staining in epiphyseal cartilage. Our findings show restricted KS distribution in the region close to the presumptive joint cavity of developing chick and rabbit joints. Alcian blue staining does not detect this moiety. Therefore, it appears that although histochemistry allows relatively insensitive quantitative assessment of GAGs, IHC increases these detection limits. This is particularly evident for KS, which exhibits immunolabeling patterns in joints from different species that is consistent with a conserved functional role in chondrogenesis.

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.

Spatiotemporal patterns of fibronectin distribution during embryonic development

Development ◽  
1981 ◽  
Vol 63 (1) ◽  
pp. 193-206
Author(s):  
Michael Melnick ◽  
Tina Jaskoll ◽  
Anna G. Brownell ◽  
Mary Macdougall ◽  
Conny Bessem ◽  
...  
Keyword(s):  
Limb Development ◽  
Spatial Organization ◽  
Cartilage Matrix ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Major Constituent ◽  
Alcian Blue Staining ◽  
In Ovo ◽  
Tissue Organization ◽  
Testicular Hyaluronidase

It has been suggested that an extracellular matrix - and cell surface - associated glycoprotein, fibronectin, plays a role in the positioning of cells in morphogenesis and in the maintenance of orderly tissue organization. In the present study the appearance and distribution of fibronectin during in ovo chick limb development has been investigated by indirect immunofluorescence techniques in H.H. stages 20–30. Fibronectin is not detectable until just prior to the transition from the morphogenetic to the cytodifferentiation phase of development. Beginning at H.H. stage 25, successive nonrandom patterns of fibronectin detection and distribution, which resemble the subsequent cartilaginous elements, precede overt chondrogenesis as detected by Alcian blue staining. This corresponds to the onset of the cytodifferentiation phase of limb development. As the accumulation of acidic proteoglycan increases in the cartilage matrix and the mesenchymal cells become more round in appearance, the presence of detectable fibronectin decreases and is ultimately seen only in the perichondria and basement membrane. However, predigestion of developed cartilage tissue with testicular hyaluronidase, prior to fibronectin staining, indicated that fibronectin remains a major constituent of cartilage matrix and is apparently masked by cartilagespecific proteoglycans. This study of chick limb development is consistent with the hypothesis that fibronectin may be a molecule that facilitates the spatial organization of cartilaginous primordia cytodifferentiation.

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 COMPARISON OF THE EFFECTS OF PAPAIN AND VITAMIN A ON CARTILAGE

1960 ◽  
Vol 111 (5) ◽  
pp. 719-744 ◽  
Author(s):  
Honor B. Fell ◽  
Lewis Thomas
Keyword(s):  
Chick Embryo ◽  
Vitamin A ◽  
Culture Medium ◽  
Cartilage Matrix ◽  
Chick Embryos ◽  
Hypervitaminosis A ◽  
Normal Medium ◽  
Fetal Mouse ◽  
Fetal Mice ◽  
Cartilage Cells

The effects of papain protease and of vitamin A on explanted limb bone rudiments from 7- and 13-day chick embryos and fetal mice have been studied and compared. The incubation of cartilaginous rudiments from 7-day chick embryos in a solution containing papain and cysteine resulted in complete loss of the metachromasia of the cartilage matrix within 1 hour; explants treated in this fashion recovered normal metachromatic staining properties when grown in normal medium for 4 days. The incubation of 7-day chick cartilage rudiments in a solution containing papain without cysteine resulted in partial loss of metachromasia from cartilage within 1 hour; the addition of vitamin A to the solution did not enhance the effect of papain during this period. The addition of papain to the culture medium in which 7-day chick embryo cartilage rudiments were grown resulted in uniform loss of the metachromasia of the cartilage matrix; similar explants grown in the presence of excess vitamin A also showed loss of the metachromasia of cartilage, but certain regions of the cartilage were affected earlier and more severely than others. Changes in cartilage cells, including loss of glycogen, occurred when the rudiment was grown in medium containing excess vitamin A, but not when it was grown in the presence of papain. Bone rudiments from 13-day chick embryos showed changes in cartilage similar to those seen in 7-day chick embryo rudiments when grown in the presence of papain or of excess vitamin A; the existing bone was not affected under these conditions. When grown in the presence of papain or excess vitamin A, the cartilage of late fetal mouse bone underwent changes similar to those already described in chick embryo rudiments. In contrast to the chick embryo rudiments, those from the fetal mouse showed rapid resorption of bone when grown in the presence of excess vitamin A. Papain had no effect on bone from either source. The changes seen in cartilage of explants grown in the presence of vitamin A and papain together were greater than those seen with either agent alone. The changes seen in fetal mouse bone grown in the presence of vitamin A were not enhanced by the additional presence of papain. On the basis of these observations, it is suggested that the changes in cartilage seen in experimental hypervitaminosis A may be the result of activation of a proteolytic enzyme or enzymes with properties similar to papain.

scholarly journals VITAMIN A AND ENDOCHONDRAL OSSIFICATION IN THE RAT AS INDICATED BY THE USE OF SULFUR-35 AND PHOSPHORUS-32

1954 ◽  
Vol 100 (1) ◽  
pp. 11-24 ◽  
Author(s):  
Dominic D. Dziewiatkowski
Keyword(s):  
Vitamin A ◽  
Chondroitin Sulfate ◽  
Endochondral Ossification ◽  
Specific Activity ◽  
Epiphyseal Cartilage ◽  
Sulfate Sulfur ◽  
A Value ◽  
Sulfur Containing ◽  
Inorganic Sulfate

The administration of vitamin A to vitamin A-deficient rats resulted in a decreased concentration of inorganic sulfate-sulfur in the serum from a value of 2.5 mg. per cent to 1.8 mg. per cent, the latter being close to the value of 2.0 mg. per cent found in normal rats of the same age. The uptake of sulfate and phosphate by femurs and tibiae of vitamin A-deficient rats was less than that in normal rats of the same age. An increased uptake followed the administration of vitamin A: radioautography indicated that in the case of sulfate, its uptake was particularly increased in the epiphyseal cartilage; an increased uptake of phosphate was particularly evident in the diaphysis immediately adjacent to the epiphyseal cartilage plate. The specific activity of the sulfate-sulfur in the chondroitin sulfate samples isolated from the skeletons of vitamin A-deficient rats fell progressively as the deficiency continued. Following administration of vitamin A, the specific activity approached and exceeded the value given by the sample from the skeletons of normal rats of the same age. A substantial increase was found in the value of the specific activity of the sulfate-sulfur of sulfomucopolysaccharides isolated from skins of vitamin A-deficient rats that had been given vitamin A. Following administration of vitamin A to rats deficient in this vitamin, an increased accumulation of some sulfur-containing material was found in regions of active calcification.

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.

scholarly journals Hybrid Hydrogel Composed of Hyaluronic Acid, Gelatin, and Extracellular Cartilage Matrix for Perforated TM Repair

2021 ◽  
Vol 9 ◽  
Author(s):  
Yili Wang ◽  
Feng Wen ◽  
Xueting Yao ◽  
Lulu Zeng ◽  
Jiaming Wu ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Cartilage Matrix ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Hybrid Hydrogel ◽  
Composite Hydrogels ◽  
Further Development ◽  
Perforated Tympanic Membrane

A novel series of composite hydrogels, built from the three components 1), hyaluronic acid methacryloyl (HAMA); 2), gelatin methacryloyl (GelMA), and 3), extracellular cartilage matrix (ECM), was prepared and studied regarding the possible utility in the surgical repair of damaged (perforated) tympanic membrane (TM). Noteworthy is component 3), which was harvested from the ribs of α-1,3-galactosidyltransferase-knockout (α-1,3 GalT-KO) pigs. The absence of α-1,3-galactosyl glycoprotein is hypothesized to prevent rejection due to foreign-body immunogenicity. The composite hydrogels were characterized by various aspects, using a variety of physicochemical techniques: aqueous swelling, structural degradation, behavior under compression, and morphology, e.g., in vitro biocompatibility was assessed by the CCK-8 and live–dead assays and through cytoskeleton staining/microscopy. Alcian blue staining and real-time PCR (RT-PCR) were performed to examine the chondrogenic induction potential of the hydrogels. Moreover, a rat TM defect model was used to evaluate the in vivo performance of the hydrogels in this particular application. Taken together, the results from this study are surprising and promising. Much further development work will be required to make the material ready for surgical use.

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.

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