Modelling cartilage mechanobiology

The growth, maintenance and ossification of cartilage are fundamental to skeletal development and are regulated throughout life by the mechanical cues that are imposed by physical activities. Finite element computer analyses have been used to study the role of local tissue mechanics on endochondral ossification patterns, skeletal morphology and articular cartilage thickness distributions. Using single-phase continuum material representations of cartilage, the results have indicated that local intermittent hydrostatic pressure promotes cartilage maintenance. Cyclic tensile strains (or shear), however, promote cartilage growth and ossification. Because single-phase material models cannot capture fluid exudation in articular cartilage, poroelastic (or biphasic) solid/fluid models are often implemented to study joint mechanics. In the middle and deep layers of articular cartilage where poroelastic analyses predict little fluid exudation, the cartilage phenotype is maintained by cyclic fluid pressure (consistent with the single-phase theory). In superficial articular layers the chondrocytes are exposed to tangential tensile strain in addition to the high fluid pressure. Furthermore, there is fluid exudation and matrix consolidation, leading to cell 'flattening'. As a result, the superficial layer assumes an altered, more fibrous phenotype. These computer model predictions of cartilage mechanobiology are consistent with results of in vitro cell and tissue and molecular biology experiments.     

Changes in the distribution of proteinpolysaccharide complexes within the articular cartilage in experimental osteoarthrosis

The quantity and type of proteinpolysaccharide complexes in the matrix determine up to a great extent the mechanical properties of articular cartilage. It is the purpose of this study to evaluate the changes in the mentioned matrix components against the background of experimentally induced osteoarthrosis. As shown by electron microscopic and morphometric studies, the changes in the superficial layer are promptly occurring and clearcut, whereas those in the deep layers are recorded in late observation terms only. A reduction of proteoglycan quantity is noted with a simultaneous differentiation of their fine structure in the various stages of osteoarthrosis development. Initially the alteration in the cell organization of chondroblasts is associated with occurrence of differences in proteoglycan content, and subsequently--in the collagen structures of the matrix too.     

The early development of Jacobson's organ in the nasal cavity of the rat before the inception of the secondary palatal development

The early development of Jacobson's organ was studied by means of a series of embryos of the rat which were of various ages and exactly dated. Already at the youngest stage of those rats, the nasal cavity is just an open groove, the organ is a thickened epithelial layer at the medial nasal process. Only 15 h later, while the nasal grooves start to close from caudal to rostral, Jacobson's organ has acquired the shape of a deep, long cleft, situated within the broad nasal opening. On the 13th d of fetal life, a complete, caudally closed nasal cavity appears. By the means of fundamental growth changes, the already well developed organ has become shifted to a more caudal position and lies now above the primary palate. A shorter caudal part of the still cleft-like organ just starts to close itself thus forming its typical tube-like structure. Moreover strong nerve bundles running from Jacobson's organ to the brain indicate that in the meantime a sensory epithelium can be distinguished. Up to the 15th d of development, the tube-forming process of Jacobson's organ is completed. Parallel to this procedure, the surrounding nasal cavity acquires a caudal apertura nasalis interna by the rupture of the membrana bucconasalis while Jacobson's organ still lies above the rostral primary palate. Primary in the medial, somewhat later in the lateral part of the nasal cavity, first outlines of cartilage appear, visible as dense cell formations. Together with this, the paraseptal cartilage, in these stages closely connected to the septal cartilage, develops quite early. Between the 14th and 15th d of its fetal life, the flat, tube-formed Jacobson's organ of the rat gets turned from a primary horizontal into a vertical position, which brings its sensory epithelium to the medial side. It is assumed that this happens for functional reasons. Because of the obviously early and progressive development of Jacobson's organ within that of the nasal cavity, it seems to be probable that already the origin of the nose, the olfactory placodes, are determined in the directions both of the nasal cavity and of Jacobson's organ. Furthermore the results demonstrate an early preferential development of Jacobson's organ in comparison to that of the surrounding nasal cavity.     

Two subpopulations of differentiated chondrocytes identified with a monoclonal antibody to keratan sulfate

We have prepared a monoclonal antibody, named MZ15, that specifically binds keratan sulfate. Immunofluorescence studies showed that the distribution of keratan sulfate in articular cartilage was not uniform: the amount of keratan sulfate increased with distance from the articular surface. Two subpopulations of chondrocytes could be distinguished after isolation from cartilage by the presence or absence of cell surface keratan sulfate. Keratan sulfate-negative chondrocytes were shown to come from the upper cartilage layers. There was therefore a direct correlation between biochemical heterogeneity of cartilage matrix and heterogeneity within the chondrocyte population. During growth in monolayer culture, superficial chondrocytes began to synthesize keratan sulfate, but the cells could still be distinguished from cultures of deep or unfractionated chondrocytes by their reduced substrate adhesiveness and tendency to remain rounded.     

An immunohistochemical study of localization of type I and type II collagens in mandibular condylar cartilage compared with tibial growth plate

Immunohistochemical localization of type I and type II collagens was examined in the rat mandibular condylar cartilage (as the secondary cartilage) and compared with that in the tibial growth plate (as the primary cartilage) using plastic embedded tissues. In the condylar cartilage, type I collagen was present not only in the extracellular matrix (ECM) of the fibrous, proliferative, and transitional cell layers, but also in the ECM of the maturative and hypertrophic cell layers. Type II collagen was present in the ECM of the maturative and hypertrophic cell layers. In the growth plate, type II collagen was present in the ECM of whole cartilaginous layers; type I collagen was not present in the cartilage but in the perichondrium and the bone matrices. These results indicate that differences exist in the components of the ECM between the primary and secondary cartilages. It is suggested that these two tissues differ in the developmental processes and/or in the reactions to their own local functional needs.     

应力作用下软骨细胞信号传导的研究进展

研究显示应力刺激对软骨细胞生长及基质代谢具重要作用。软骨正常结构形态以及应力下的软骨细胞形态和基质代谢的变化是力-生物信号转化的基础,信号分子及信号通路则是应力信号传导的核心,二者是对软骨细胞应力下信号传导过程深入了解不可或缺的信息组成,了解应力对软骨细胞的作用方式及作用机制有助于软骨相关疾病诊治、组织工程等领域的研究,本文就这两个方面研究进展做一综述。     

An immunohistochemical study of localization of type I and type II collagens in mandibular condylar cartilage compared with tibial growth plate

Summary Immunohistochemical localization of type I and type II collagens was examined in the rat mandibular condylar cartilage (as the secondary cartilage) and compared with that in the tibial growth plate (as the primary cartilage) using plastic embedded tissues. In the condylar cartilage, type I collagen was present not only in the extracellular matrix (ECM) of the fibrous, proliferative, and transitional cell layers, but also in the ECM of the maturative and hypertrophic cell layers. Type II collagen was present in the ECM of the maturative and hypertrophic cell layers. In the growth plate, type II collagen was present in the ECM of whole cartilaginous layers; type I collagen was not present in the cartilage but in the perichondrium and the bone matrices. These results indicate that differences exist in the components of the ECM between the primary and secondary cartilages. It is suggested that these two tissues differ in the developmental processes and/or in the reactions to their own local functional needs.     

Effect of various periods of social isolation of juvenile rats on the level of their intraspecies aggression at a sexually mature age

Isolation of male outbred white rats from the same age rats at 12-30 and 15-30 days of postnatal life increases their aggressiveness in pubertal age and causes a change of aggressive reactions spectrum towards the decrease of attacks and increase of threats in relation to intruders. These changes in the ontogenetic formation of rats aggressive behaviour are not compensated by the experience of rats social intercourse in the period 30-70th days. A hypothesis is made about the existence of sensitive period of rats aggressive behaviour formation with the approximate limits 15th-30th days of postnatal life during which under the influence of contacts with rats of the same age prerequisites are created for consolidation of various agonistic reactions in resultant act of mature aggression.     

Immunohistochemical localization of type II and type I collagens in articular cartilage of the femoral head of dexamethasone-treated rats

The immunohistochemical localization of type II and type I collagens was examined in the articular cartilage of the femoral head of growing rats injected systemically with 5 mg kg−1 dexamethasone for 2 weeks every other day. The intensities of immunostaining for type II collagen, measured by microphotometry, was highest in the flattened cell layer and high in the hypertrophic cell layer, moderate in the proliferative cell and transitional cell layers and low in the superficial layer. After dexamethasone administration, the intensities decreased markedly in the flattened cell layer and slightly in the hypertrophic cell layer, although the decreases in other layers were negligible. The staining intensities for type I collagen were highest in the flattened cell layer, low in the superficial and transitional cell layers and very low in the proliferative and hypertrophic cell layers. After dexamethasone administration, the intensities increased markedly in the flattened cell layer and slightly in the superficial and proliferative cell layers, but did not change in the transitional and hypertrophic cell layers. Thus, dexamethasone administration caused a decrease in type II collagen and an increase in type I collagen in the matrix of the surface portion of articular cartilage. The composition of isoforms of collagen in the matrix changed after the steroid administration. The results strongly suggest that the shift in collagen composition from type II to type I predominance is a cause of the degeneration of the articular cartilage after glucocorticoid administration. This revised version was published online in November 2006 with corrections to the Cover Date.     

Physiology and pathophysiology of the growth plate

Longitudinal growth of the skeleton is a result of endochondral ossification that occurs at the growth plate. Through a sequential process of cell proliferation, extracellular matrix synthesis, cellular hypertrophy, matrix mineralization, vascular invasion, and eventually apoptosis, the cartilage model is continually replaced by bone as length increases. The regulation of longitudinal growth at the growth plate occurs generally through the intimate interaction of circulating systemic hormones and locally produced peptide growth factors, the net result of which is to trigger changes in gene expression by growth plate chondrocytes. This review highlights recent advances in genetics and cell biology that are illuminating the important regulatory mechanisms governing the structure and biology of the growth plate, and provides selected examples of how studies of human mutations have yielded a wealth of new knowledge regarding the normal biology and pathophysiology of growth plate cartilage.     

Weaning and the histology of the mandibular condyle in the rat.

Eighty-eight Long Evans/Turku rats were used in the study. The effect of the articulatory function on the mandibular condyle was observed histologically during normal growth, when the rat is changing its diet from milk to whole pellets as a part of weaning. Six animals each were killed at the age of 10, 15, 20, 25, 30, 35, 40 and 50 days for histological tissue processing. For further information, 30 animals were fed a soft diet (6 animals each were killed at the age of 25, 30, 35, 40 and 50 days), and 10 animals were fed hardened pellets (2 animals each were killed at the ages of 25, 30, 35, 40 and 50 days). An even and regular transition from mesenchymal cells via immature chondroblasts into mature chondroblasts and hypertrophied chondrocytes was found at 10, 15 and 20 days during normal growth and also at 25, 30, 35, 40 and 50 days when animals were fed a soft diet. This maturing process appeared to be disturbed at the age of 25, 30, 35 and 40 days in the superior aspect of the condyle in animals fed ordinary pellets. The density of the mesenchymal cell layer was decreased, and the amount of intercellular matrix seemed to be evaluated in mesenchymal and intermediate cell layers. These features were later manifest deeper in the cartilage as acellular regions and as cell clusters. The changes were similar but more severe when the animals were fed hardened pellets.(ABSTRACT TRUNCATED AT 250 WORDS)     

Impact of sex hormones, insulin, growth factors and peptides on cartilage health and disease

Sex hormones contribute to the pathogenesis of osteoarthritis (OA) in both sexes. OA is normally not seen in pre-menopausal women, whereas men may develop the disease as early as the 30th year of life. OA also shows increased incidence in association with diseases such as diabetes mellitus. Recent years have seen characterization of essential components of a functional endocrinal network in the articular cartilage comprising not only sex hormones but apparently insulin, growth factors and various peptides as well. In this review, we summarize the latest information regarding the influence of sex hormones, insulin, growth factors and some peptides on healthy cartilage and their involvement in osteoarthritis. Both animal and human research data were considered. The results are presented in an information matrix that identifies what is known, with supporting references, and identifies areas for further investigation.     

FORMATION,STRUCTURE AND FUNCTION OF CARTILAGE

Improved investigative techniques including electron microscopy, isotope tracings and improved histochemistry have greatly increased knowledge of the function of cartilage as a body tissue. Highly complex and delicate enzyme systems contained in the cartilage cell are involved in cartilage matrix formation and in the processes of calcification and cartilage repair. Heat, various drugs, freezing, and changes in the chemical environment damage or destroy these enzyme systems and interfere with the growth and function of cartilage. Hyaline cartilage to be transplanted must be handled with great care to preserve the cellular enzyme systems—otherwise the graft will be resorbed and clinical failure will result.     

New aspects of the histology of the mandibular condyle in the rat

The function of the multipotential mesenchymal cells in the mandibular condyle was studied histochemically and histologically in 27 Long Evans/Turku rats. Sagittal sections from the temporomandibular joint were stained with haematoxylin and eosin, toluidine blue, or van Gieson's stain. A weakly orthochromatically stained fibrous layer was followed in the upper region by a weakly metachromatically stained mesenchymal cell layer. Deep within this was a strongly metachromatically stained layer of immature chondroblasts. The metachromasia of the matrix of these layers disappeared abruptly in an anterior direction and gradually in a posterior direction. The changes in the staining reactions are explained by the fact that mesenchymal cells can differentiate into chondrogenic or osteogenic cells depending on the environmental conditions. A new hypothesis is presented according to which regulation of the direction of condylar growth is achieved by choosing the cells for chondrogenesis more posteriorly or anteriorly from among the mesenchymal cells covering the whole condylar cartilage.     

Compression-induced changes in the shape and volume of the chondrocyte nucleus

Changes in cell shape and volume are believed to play a role in the process of mechanical signal transduction by chondrocytes in articular cartilage. One proposed pathway through which chondrocyte deformation may be transduced to an intracellular signal is through cytoskeletally mediated deformation of intracellular organelles, and more specifically, of the cell nucleus. In this study, confocal scanning laser microscopy was used to perform in situ three-dimensional morphometric analyses of the nuclei of viable condrocytes during controlled compression of articular cartilage explants from the canine patellofemoral groove. Unconfined compression of the tissue to a 15% surface-to-surface strain resulted in a significant decrease of chondrocyte height and volume by 14.7 ± 6.4 and 11.4 ± 8.4%, respectively, and of nuclear height and volume by 8.8 ± 6.2% and 9.8 ± 8.8%, respectively. Disruption of the actin cytoskeleton using cytochalasin D altered the relationship between matrix deformation and changes in nuclear height and shape, but not volume. The morphology and deformation behavior of the chondrocytes were not affected by cytochalasin treatment. These results suggest that the actin cytoskeleton plays an important role in the link between compression of the extracellular matrix and deformation of the chondrocyte nuclei and imply that chondrocytes and their nuclei undergo significant changes in shape and volume in vivo.     

Type X collagen alterations in rachitic chick epiphyseal growth cartilage

We examined collagens of both normal and vitamin D-deficient chick epiphyseal growth cartilage. Special emphasis was placed on the study of Type X collagen, a recently described product of hypertrophic chondrocytes. Scanning electron microscopy of the epiphyseal growth cartilage of vitamin D-deficient chickens showed an enlarged growth cartilage with a disorganized extracellular matrix. The cartilage collagens were solubilized by proteolytic digestion and disulfide bond reduction of both normal and rachitic growth tissues. Sequential extraction with neutral salt and acetic acid buffers followed by pepsin digestion at 4 degrees C solubilized about 12% of normal tissues and about 7% of collagen from rachitic growth cartilage. Treatment of the pepsin-resistant collagens with neutral salt-dithiothreitol buffer under nondenaturing conditions and a subsequent pepsin digestion increased the yield of solubilized collagen to greater than 95% of the total tissue collagen. Results of the biochemical studies showed a marked increase in the relative proportion of Type X collagen (from 5.6 to 27.9%), a corresponding decrease in the proportions of Types II and IX collagens, and a moderate increase in Type XI collagen in rachitic cartilage. Amino acid analysis indicated that there were no differences in the Types II and X collagens of normal and rachitic cartilage. However, an abnormality in the relative proportions of the CNBr peptides of Type X collagen was detected in the rachitic cartilage. We suggest that the increase in collagen in the rachitic state may reflect increased levels of Type X collagen synthesis by cells in the hypertrophic region. It is likely that in rickets the overproduction of Type X collagen may be a compensatory mechanism by which the hypertrophic chondrocyte attempts to provide a maximum area of calcifiable matrix for the calcium-depleted serum.     

Immunolocation analysis of glycosaminoglycans in the human growth plate.

Monoclonal antibodies were used in this study to immunolocate glycosaminoglycans throughout the human growth plate. Chondroitin-4-sulfate, chondroitin-6-sulfate, and keratan sulfate were observed in the extracellular matrix of all zones of the growth plate and persisted into the cartilage trabeculae of newly formed metaphyseal bone. Also present in the extracellular matrix was an oversulfated chondroitin/dermatan sulfate glycosaminoglycan which appeared to be specific to the proliferative and hypertrophic zones of the growth plate. As with the other extracellular matrix molecules, this epitope persisted into the cartilage trabeculae of the metaphyseal bone. Zonal differences between the extracellular and pericellular or lacunae matrix were also observed. The hypertrophic chondrocytes appeared to synthesize chondroitin sulfate chains containing a non-reducing terminal 6-sulfated disaccharide, which were located in areas immediately adjacent to the cells. This epitope was not found to any significant extent in the other zones. The pericellular region around hypertrophic chondrocytes also contained a keratan sulfate epitope which was also observed in the resting zone but not in the proliferative zone. These cell-associated glycosaminoglycans were not found in the cartilage trabeculae of metaphyseal bone, indicating their removal as the terminal hypertrophic chondrocytes and their lacunae are removed by invading blood vessels. These changes in matrix glycosaminoglycan content, both in the different zones and within zones, indicate constant subtle alterations in chondrocyte metabolic products as they proceed through their life cycle of proliferation, maturation, and hypertrophy.     

Localization of CD44 (hyaluronan receptor) and hyaluronan in rat mandibular condyle.

CD44 is a multifunctional adhesion molecule that binds to hyaluronan (HA), type I collagen, and fibronectin. We investigated localization of CD44 and HA in mandibular condylar cartilage compared with the growth plate and the articular cartilage, to clarify the characteristics of chondrocytes. We also performed Western blotting using a lysate of mandibular condyle. In mandibular condyle, CD44-positive cells were seen in the surface region of the fibrous cell layer and in the proliferative cell layer. Western blotting revealed that the molecular weight of CD44 in condyle was 78 to 86 kD. Intense reactivity for HA was detected on the surface of the condyle and the lacunae of the hypertrophic cell layer. Moderate labeling was seen in cartilage matrix of the proliferative and maturative layer. Weak labeling was also seen in the fibrous cell layer. In growth plate and articular cartilage, HA was detected in all cell layers. However, chondrocytes of these cartilages did not exhibit reactivity for CD44. These results suggest that chondrocytes in the mandibular condylar cartilage differ in expression of CD44 from those in tibial growth plate and articular cartilage. Cell-matrix interaction between CD44 and HA may play an important role in the proliferation of chondrocytes in the mandibular condyle.