Showing papers on "Cartilage published in 1971"

Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data.

Abstract: For thirty-two areas of cartilage from nine osteo-arthritic and four "normal" femoral heads a histologic-histochemical grade was assigned as an index of severity of the osteo-arthritic process. The DNA and hexosamine concentrations were determined as indicators of cell density and polysaccharide con

Ruthenium red and violet. II. Fine structural localization in animal tissues.

Abstract: The inorganic dye, ruthenium red, stains extracellular materials in animal tissues which probably are acidic mucopolysaccharides. It complements other techniques, its advantages being fine grain, high resolution and good contrast. Localization is shown in mouse and rat muscle, heart, lung and intestine, frog cartilage and cells scraped from oral epithelium of human beings. Attention is paid to collagen bundles, the cell/collagen interface and particularly the myotendinal junction, cartilage matrix and agar gel, desmosomes, intestinal microvilli, erythrocytes and vascular endothelium, nerve fibers and the T-system of striated muscle. Although ruthenium red generally is excluded by plasma membranes, it penetrates giving intracellular density, if the membrane is broken. Even when the cell membrane is intact, exceptions occur with selective staining of the T-tubules or the sarcoplasmic sacs depending upon the state of contraction of the muscle cell, and with intracellular staining of certain nuclei and epithelial cells. Ruthenium red stains intracellular lipid droplets revealing lamellae, and stains myelin forms grown from crude egg lecithin but cannot penetrate deeply. It is localized in extracellular materials which have an important mechanical function. Its exclusion by cell membranes permits tracing tortuous cellular invaginations and those exceptions to its exclusion invite a comparison of the localization of the dye with the function of the cell.

The Glycosaminoglycans of Normal and Arthritic Cartilage

Abstract: The cartilages from the hip joints of 13 normal and 15 osteoarthritic humans were analyzed for glycosaminoglycan content and distribution. The GAGs were separated by elution with CPC on a short cellulose column by the technique of Svejcar and Robertson after digestion of the tissue with pronase and papain. The eluates were identified by a variety of methods including determination of molar ratios, N-acetyl-hexosamine determinations after hyaluronidase treatment and thin-layer chromatography of unhydrolyzed and hydrolyzed GAGs. From the data obtained, it was demonstrated that cartilage from arthritic patients showed a significant increase in the concentration of chondroitin 4-sulfate and a significant decrease in keratan sulfate, with only slight changes in the total amount of GAG present. Calculations of the molar ratios showed variation in the sulfation with chondroitin 4-sulfate appearing in the "supersulfated" state in the arthritic cartilage. The data lead to speculation regarding the process of osteoarthritis, and it is concluded that the changes seen are more likely to represent an altered pattern of synthesis rather than selective degradation. Since the changes suggest a younger cartilage, a theory is advanced that the chondrocyte responds to the chronic stress of osteoarthritis by modulation to a chondroblastic phase.

A NEW CHONDRODYSTROPHIC MUTANT IN MICE : Electron Microscopy of Normal and Abnormal Chondrogenesis

Abstract: The occurrence of a new mutation affecting cartilage and bone in mice is reported. The gene is lethal, shows autosomal recessive inheritance, and has high penetrance. It is not allelic to shorthead and probably not to phocomelia or achondroplasia. It results in a foreshortened face, cleft palate, defective trachea, and shortened long bones with flared metaphyses. Chondrocytes of epiphyseal cartilage from the mutant are not aligned in columns, and there is a decrease in the usual staining of the cartilage matrix. Electron microscope observations show large, wide collagen fibrils with "native" banding in the matrix of mutant cartilage, which are not present in normal cartilage. Possible explanations for the expression of this genetic disorder of cartilage development are put forward.

The fine structure of bovine nasal cartilage. Extraction as a technique to study proteoglycans and collagen in cartilage matrix.

Abstract: Bovine nasal cartilage was studied by electron microscopy before and after extraction with 4 M guanidinium chloride or 1.9 M CaCl 2 . These solvents removed matrix granules, basophilia, and 85% of the proteoglycan complex, measured as hexuronate. Simultaneously, many collagen fibrils were disaggregated into component microfibrils (approximately 40 A thick). In contrast to the above solvents, exhaustive extraction with 0.5 M guanidinium chloride removed 20% of the proteoglycan complex, and matrix granules were reduced in size but not in number. Extraction with 4 M CaCl 2 removed only 10% of the proteoglycan complex, did not remove matrix granules, and caused the normal banding pattern of collagen to disappear. The banding was restored by further treatment with trypsin. Trypsin, before or after 4 M CaCl 2 , removed matrix granules and 90% of the proteoglycan complex. We conclude that matrix granules are an electron microscopic representation of the proteoglycan complex, and consist of more than one proteoglycan macromolecule. It would appear that 4 M guanidinium chloride and 1.9 M CaCl 2 , in addition to removing most of the proteoglycan complex, also disaggregate some of the collagen fibrils into their component microfibrils.

Papain-induced degenerative arthritis of the hip in rabbits

Abstract: 1. Degenerative arthritis has been produced consistently in adult rabbits by the injection of the proteolytic plant enzyme papain into the hip joint. Arthritic changes were recognisable radiographically after six weeks. 2. A progression of changes occurred, from loss of acid mucopolysaccharide staining in the matrix, fibrillation, fissuring and erosion of articular cartilage with death of chondrocytes in the weight-bearing areas, to secondary bony changes of subchondral sclerosis, occasional cysts and osteophyte formation. 3. Synovial inflammation occurred with accumulation of cartilage and bone debris in the inferior capsule and later capsular thickening. 4. It is suggested that this arthritis is sufficiently similar to human osteoarthritis to be useful as a model for further studies of the pathogenesis of the disease and the effects of different methods of treatment.

Contribution of Meckel's Cartilage to Ossification of the Mandible in Mice:

Abstract: The role of Meckel's cartilage in prenatal and neonatal osteogenesis of the mandible was studied in mice. Three ossification processes, two intramembranous and one endochondral, occurred concurrently in the anterior part of the mandible. The contribution of Meckel's cartilage was a transient one, but related to possible congenital defects.

The Nutritional Pathways of Articular Cartilage: An Autoradiographic Study In Rabbits Using 35s Injected Intravenously

Abstract: An autoradiographic study of the nutrition of articular cartilage in rabbits using intravenous 35S described. In immature rabbits (open epiphyses) the isotope appeared to enter the cartilage from both the synovial fluid and the subchondral bone, while in the mature animals (closed epiphyses) the isotope appeared to enter articular cartilage only from the synovial fluid. The experiment provides additional evidence that the route of nutrients for the articular cartilage is affected by skeletal maturity.

Collagen synthesis in mature articular cartilage of the rabbit

Abstract: 1. The utilisation of labelled proline in normal and injured mature rabbit articular cartilage has been studied and compared simultaneously in one phase of the study with radiosulphate utilisation. The morphologic features of lacerative injury paralleled those reported previously. 2. Labelled proline is actively utilised by mature articular cartilage and can be recovered in time from the matrix as labelled hydroxyproline. This is taken as evidence of collagen synthesis. 3. Evidence is presented to suggest that the rate of formation of labelled hydroxyproline may be augmented after lacerative trauma. 4. Parallel utilisation of radiosulphate and labelled proline suggests that the synthesis of chondromucoprotein and collagen are closely related and that the continual synthesis of both moieties is necessary for the maintenance of normal matrix. 5. Despite evidence of increased chondromucoprotein and collagen synthesis no significant contribution is made to the healing of lacerative defects in mature rabbit articular cartilage.

Histological, autoradiographic and microchemical studies of spontaneously healing osteochondral articular defects in adult rabbits.

Abstract: A limited, standardized osteochondral defect was created in adult rabbits and the regenerated tissue was examined histologically and autoradiographically after labellingin vitro with35S-sulphate, and microchemically for its content and composition of glycosaminoglycans. The principal findings were: 1. Between 1 week and 4 to 8 weeks, non-metachromatic connective tissue differentiated to metachromatically stained cartilage, and the proportion of the chondroitin sulphate increased significantly at the expense of the glycoproteins. 2. Up to 4 weeks, the major part of the defect area was filled with newly formed bone; after this time, the area lay above the level of the “tidemark”, towards the articular surface. 3. Hyaline cartilage with morphological, autoradiographic and chemical resemblance to the articular cartilage in terms of the distribution of glycosaminoglycans constituted the articular surface of the defect area in one-third of the cases at observation times after 8 weeks. Hyaline cartilage was observed mainly in areas where newly formed bone had closed the medullary cavity. 4. In two-thirds of the cases, after 8 weeks, parts of the articular surface of the defect consisted not only of cartilage but also of fibrous tissue, occurring mainly in the lateral parts of the defect and in areas where the medullary cavity facing the articular surface had not been sealed by bone tissue. The glycoprotein fraction increased relative to the chondroitin sulphate fraction. 5. In most cases after 20 weeks, newly-formed cartilage underwent degenerative changes, which were reflected in some reduction of the chondroitin sulphate.

Scale effects in animal joints. II. Thickness and elasticity in the deformability of articular cartilage

Abstract: The deformability of articular cartilage was studied in vitro in 64 bovine, canine and human joints with an indenting compressometer. Two factors contributed to the depth of the indentation: the thickness of the cartilage and its intrinsic elasticity. Within a given species, there was in general a consistent curvilinear relationship between the magnitude of the deformation and the thickness of the articular cartilage under a given level of static loading (usually 106 pounds per square inch). Although it was not possible to establish a satisfactory Young's modulus, differences in intrinsic elasticity were demonstrated by variations in the indentations produced in cartilages of comparable thickness, when subjected to the same compressive stress. These species differences corresponded roughly to variations in the water content of the articular cartilages (bovine, 87; canine, 80; human, 79%). The data are consistent with the hypothesis that deformability of articular cartilage serves to increase the congruence of joints and thereby reduce the stresses on their surfaces. A frequently postulated senescent thinning of articular cartilage received no support from measurements of 28 intact human patellas through the fifth decade of life.

LYSOZYME IN EPIPHYSEAL CARTILAGE : IV. Embryonic Chick Cartilage Lysozyme-Its Localization and Partial Characterization.

Abstract: The localization of chick embryonic lysozyme was determined by two techniques: by studying the rate of release from the tissue during sequential enzymatic digestion and by immunocytochemistry. Both techniques indicate that, in this tissue, lysozyme is primarily extra-cellular. Cartilage lysozyme was isolated and partially characterized and found to be identical with egg white lysozyme in its immunologic and enzymatic behavior. In addition, a method for the isolation of large numbers of viable chondrocytes is described.

The effects of maternal hypercortisonism and hypervitaminosis D2 on fetal osteogenesis and ossification in rats.

Abstract: Pregnant rats were treated daily with intramuscular injections of 5 or 10 mg cortisone acetate dissolved in 0.5 ml normal saline, or orally with 20,000 IU vitamin D2 dissolved in 0.5 ml olive oil, or both. Controls received olive oil and normal saline, or vitamin D2. Treatment was from day 8 or 10 of gestation until the animals were killed on day 18, 20, or 22. The higher dose of cortisone shortened the diaphyses and epiphyses of long bones. Concomitant administration of vitamin D2 caused further bone shortening and hypomineralization. The main microscopic alterations in the fetal bones of cortisone-treated rats were changes in cartilaginous intercellular substance, narrowing of the hypertrophic and calcifying cartilage cell layer, and inhibition of endochondral ossification. Bone trabeculae were very wide and irregular. The bone matrix had abnormal staining properties. Bone marrow spaces were obliterated and contained a very large number of fibroblasts, preosteoblasts, and osteoclasts. The number of osteoclasts was further increased by treatment with vitamin D2. It is presumed that cortisone passed through the placenta and affected fetal bones, causing the microscopic changes. Concomitant administration of vitamin D2 potentiated the effects of cortisone on bone, and also produced certain specific changes.

Immature joint cartilage and the homograft reaction

Abstract: 1. Grafts of joint cartilage from immature lambs were used to repair articular cartilage defects in other lambs and in adult sheep. 2. Stability of these grafts in a functional state was found in most for periods up to fourteen months. Although a limited homograft reaction occurred this did not lead to destruction of the cartilage, even though parts of it were well vascularised. 3. The results suggest that the process of endochondral ossification is associated with the liberation of antigenic material leading to sensitisation of the host. Destruction ofthe cartilage is prevented by an inhibitory action which the matrix appears to exert on the destructive elements themselves and which is itself dependent on the vitality of the chondrocytes. 4. The avascularity of cartilage is not a sufficient explanation for its privileged position in relation to the homograft reaction.

Distribution of hexamethonium and other quaternary ammonium compounds in cartilage

Abstract: N-methyl-C14- and H3-labeled hexamethonium was administered to rats and mice in a dose of 10 to 30 mg/kg. Tissue sections for whole body and cellular autoradiography were cut from animals sacrificed at various intervals after injection. In contrast to the distribution pattern of most drugs hexamethonium was found to he preferentially accumulated in certain avascular cartilaginous tissues, including those of intervertebral discs. epiphyseal and articular cartilages of hip. knee and other joints. costal and articular cartilage of the ribs, circular cartilage of trachea and nasal and ear cartilages. On the other hand, the compact bone itself or the bone marrow of femur and other bones showed little radiodensity in the autoradiograms. A similar pattern of distribution is observed after C14-decamethonium. Cellular autoradiograms and histochemical studies showed localization of the labeled hexamethonium in areas rich in acid mucopolvsaccharides. Hexamethonium was also found, in vitro , to bind strongly to chondroitin sulfate. It is hypothesized that the diquaternary ammonium compounds, under physiologic conditions, are bound to the polyanionic mucopolysaccharide components of cartilaginous tissue.

Cartilage compositions for dental use

Abstract: CARTILAGE POWDER IS DESCRIBED HEREIN FOR USE IN DENTAL APPLICATIONS INCLUDING THE TREATMENT AND PREVENTION OF DRY SOCKETS, GINGIVAL TISSUES AND MANDIBULAR CYSTECTOMIES, THE CARTILAGE POWDER BEING USED IN THE FORM OF A PASTE OF LOW MOBILITY.

Organ-culture studies of achondroplastic rabbit cartilage: evidence for a metabolic defect in glucose utilization.

Abstract: Organ-culture studies were made using cartilage from achondroplastic (dwarf) rabbits ( ac / ac ) and their phenotypically normal litter-mates. A significantly higher incorporation of 14C from glucose and galactose was measured in the dwarf; incorporation of 35S from sulfate and 3H from thymidine was equal in the two types of explant. Also, 14CO2 and [14C]lactate production from glucose by the dwarf cartilage was increased. No difference in the ratio of 14CO2 evolution from [1-14C]- and [6-14C]glucose was found between the two cartilage types. Explants of dwarf cartilage utilized more glucose from the medium than did the controls. Radioautographs of tissue sections from the explants showed an increased number of grains from [14C]glucose overlying the dwarf cartilage, and this difference was particularly great over the central portions of cartilage. The proportion of 14C grains from glucose was greater over the dwarf nuclei, less over the dwarf matrix and equal over the cytoplasm of the two tissues. Grain counts of sulfate and of thymidine did not differ in the two types of cartilage.

Zonal analysis of electrolytes in epiphyseal cartilage and bone of normal and rachitic chickens and pigs

Abstract: Electrolytes were analyzed in serum, and in mineralizing tissues at varying stages of calcification, in normal and vitamin D-deficient chickens and pigs. Moisture, ash, and organic matter were also measured. Results revealed that in addition to Ca, other serum electrolytes were altered in rickets. Mg and inorganic P were diversely affected by the vitamin deficiency. Larger changes in mineral level were seen in the tissues in the early stages of calcification than were seen at corresponding times in the serum, suggesting a direct effect of the vitamin deficiency on the calcifying tissue itself. Relative to the serum level, Ca was deposited in the tissues of rachitic animals to a greater extent than in normal animals. This was not true for Mg or inorganic P, indicating a preferential affinity for Ca in the rachitic tissue. Gravimetric analyses of organic and dry matter in epiphyseal zones revealed that the amounts of proliferating and hypertrophic cartilage were increased, and calcified cartilage decreased, in rickets, in accord with previous histological observations. Unexpectedly, the proportion of resting cartilage was also increased.

Lactic-malic dehydrogenase quotients during transformation of fibroblasts into cartilage and bone.

Abstract: SummaryThe concurrent assay of lactic and malic dehydrogenase in the cytosol of homogenized tissues is a simple and useful device in studying the biochemistry of transformation of fibroblasts from fascia into cartilage, followed by bone. The quotient, LDH/MDH, in cartilage exceeds 1 whereas in fascia and bone it is less than 1.

ENZYME ACTIVITIES IN CHICK EMBRYONIC CARTILAGE Their Subcellular Distribution in Isolated Chondrocytes

Abstract: Some characteristic enzymatic activities were determined in chick embryonic cartilage and compared with the analogous activities in bone and liver. Chondrocytes were isolated, broken by sonication, and subjected to subcellular fractionation to yield a nuclear pellet, the mitochondrial, lysosomal, and microsomal fractions, and the high speed supernatant solution. It was established that these fractions are characterized by enzymatic activities usually associated with similar fractions in other tissues, but with some quantitative differences. Lysozyme, a particulate-associated enzyme in other tissues, was not detected in any subcellular fraction even by the sensitive technique of microzone electrophoresis and is therefore considered to be primarily extracellular in cartilage.