The ultrastructure and biomechanical significance of the tidemark of articular cartilage.

Thirty specimens of human articular cartilage obtained at surgery were examined by scanning electron microscopy to determine the ultrastructure of the tidemark, the junction of the non-calcified and calcified portions of mature articular cartilage. Three distinct variations of the collagen framework of the tidemark were observed: (1) A band of randomly oriented compacted fibrils that appeared to be continuous with those of the non-calcified and calcified zones. (2) A band of flattened fibrils paralleling the undulating surface of the calcified cartilage. (3) A band of perpendicularly oriented fibrils having a distinct continuous transition between the non-calcified and calcified zones, the amount of calcified material applied about the fibrils rapidly increasing as the fibrils entered the calcified zone. The tidemark may serve to provide a tethering mechanism for the relatively flexible and perpendicularly oriented collagen fibrils of the deepest portion of the non-calcified articular cartilage and may prevent them from being sheared at their point of anchorage to the calcified zone. The undulating pattern of the tidemark affords a strong geometric pattern in providing resistance to the shearing action of articulation. Small gaps present in the tidemark may provide pathways for the passage of nutrients into the deep non-calcified zone of articular cartilage from the subchondral bone.     

正常膝关节软骨钙化层形态结构研究

目的探索关节软骨钙化层的结构形态及其与软骨非钙化层和软骨下骨之间的界面连接方式。方法组织库获取自愿捐献的人体正常股骨髁新鲜标本20个,男10个,女10个;年龄17～45岁。常规制备石蜡横、纵切片,番红O／固绿和冯库萨染色观察钙化层形态结构;扫描电镜观察软骨各层之间的界面连接方式;连续切片结合建模技术建立骨软骨三维模型。结果关节骨软骨复合组织番红O／固绿染色结果示软骨红染,软骨下骨蓝染,钙化层位于潮线与黏合线之间;冯库萨染色结果示钙化层黑染,结构及边界清晰,上界面以波浪状潮线结构与非钙化层紧密连接,下界面以凹凸不平的梳齿状结构与软骨下骨相互锚合：关节骨软骨剥离面及断面扫描电镜示钙化层与非钙化层以沟壑镶嵌方式相互嵌合;关节骨软骨复合组织纵切面观察,钙化层与软骨下骨间的黏合线呈凹凸不平的梳齿状结构;骨软骨三维模型观察结果与关节软骨自然剥离横断面扫描电镜观察结果基本一致。结论钙化层是关节软骨的重要结构,它通过特有界面连接方式将软骨牢牢固定在软骨下骨上。     

Variations in articular calcified cartilage by site and exercise in the 18-month-old equine distal metacarpal condyle

OBJECTIVES: To interrelate articular calcified cartilage thickness, mineralisation density, tidemark count and tidemark linear accretion rate by site in the equine third metacarpal distal condyle. To determine the effects of exercise during early life on articular calcified cartilage. METHOD: Six of 12 pasture-raised Thoroughbred horses were exercised from 10 days old. Calcein labels were given 19 and 8 days prior to euthanasia at 18 months old. Osteochondral specimens were cut from the distal third metacarpal condyle and imaged using confocal scanning light microscopy (CSLM) and quantitative backscattered electron scanning electron microscopy (qBSE). Articular calcified cartilage thickness and total thickness mineralisation density were measured on montaged qBSE image sets, and inter-label mineralisation density, tidemark count and linear accretion rate measured on registered CSLM-qBSE image pairs. RESULTS: Calcified cartilage thickness, mineralisation density, tidemark count and linear accretion rate varied significantly between sites. Regions with thinner calcified cartilage had greater linear accretion rates, hence rapid chondroclastic resorption. Mineralisation density was positively correlated with linear accretion rate. Fewer multiple tidemarks were counted in regions with greater linear accretion rates. Lag time between the tidemark and cement line was estimated (180 days; in the range of 0-648 days). Exercise had little effect on measured parameters. CONCLUSION: The major determinant of articular calcified cartilage thickness is the rate of chondroclastic resorption, not tidemark linear accretion rate. Our evidence supports coupled, mechanosensitive regulation of chondroclastic resorption and linear accretion rate in articular calcified cartilage. Exercising pasture-reared foals causes little additional adaptation in distal third metacarpal articular calcified cartilage.     

The cartilage-bone interface

In the knee joint, the purpose of the cartilage-bone interface is to maintain structural integrity of the osteochondral unit during walking, kneeling, pivoting, and jumping--during which tensile, compressive, and shear forces are transmitted from the viscoelastic articular cartilage layer to the much stiffer mineralized end of the long bone. Mature articular cartilage is integrated with subchondral bone through a approximately 20 to approximately 250 microm thick layer of calcified cartilage. Inside the calcified cartilage layer, perpendicular chondrocyte-derived collagen type II fibers become structurally cemented to collagen type I osteoid deposited by osteoblasts. The mature mineralization front is delineated by a thin approximately 5 microm undulating tidemark structure that forms at the base of articular cartilage. Growth plate cartilage is anchored to epiphyseal bone, sometimes via a thin layer of calcified cartilage and tidemark, while the hypertrophic edge does not form a tidemark and undergoes continual vascular invasion and endochondral ossification (EO) until skeletal maturity upon which the growth plates are fully resorbed and replaced by bone. In this review, the formation of the cartilage-bone interface during skeletal development and cartilage repair, and its structure and composition are presented. Animal models and human anatomical studies show that the tidemark is a dynamic structure that forms within a purely collagen type II-positive and collagen type I-negative hyaline cartilage matrix. Cartilage repair strategies that elicit fibrocartilage, a mixture of collagen type I and type II, are predicted to show little tidemark/calcified cartilage regeneration and to develop a less stable repair tissue-bone interface. The tidemark can be regenerated through a bone marrow-driven growth process of EO near the articular surface.     

Interaction between zonal populations of articular chondrocytes suppresses chondrocyte mineralization and this process is mediated by PTHrP

OBJECTIVE: Articular cartilage is separated from subchondral bone by the tidemark and a calcified cartilage zone. Advancement of the calcified region and tidemark duplication are both hallmarks of osteoarthritis (OA). Currently the mechanisms controlling post-natal articular cartilage mineralization are poorly understood. The objective of this study is to test the hypothesis that cellular communication between different cartilage layers regulates articular chondrocyte mineralization. DESIGN: Co-culture models were established to evaluate the interaction of chondrocytes derived from the surface, middle and deep zones of articular cartilage. The cultures were stimulated with triiodothyronine (T3) to promote chondrocyte hypertrophy. The effects of zonal chondrocyte interactions on chondrocyte mineralization were examined over time. RESULTS: Co-culture of deep zone chondrocytes (DZCs) with surface zone chondrocytes (SZCs) suppressed the T3-induced increase in alkaline phosphatase (ALP) activity and related mineralization. Moreover, SZC-DZC co-culture was associated with a significantly higher parathyroid hormone-related peptide (PTHrP) expression when compared to controls. When PTHrP(1-40) was added to the DZC-only culture, it suppressed DZC ALP activity similar to the inhibition observed in co-culture with SZC. In addition, treatment with PTHrP reversed the effect of T3 stimulation on the expression of hypertrophic markers (Indian hedgehog, ALP, matrix metalloproteinases-13, Type X collagen) in the DZC cultures. Moreover, blocking the action of PTHrP significantly increased ALP activity in SZC+DZC co-culture. CONCLUSION: Our findings demonstrate the role of zonal chondrocyte interactions in regulating cell mineralization and provide a plausible mechanism for the post-natal regulation of articular cartilage matrix organization. These findings also have significant implications in understanding the pathology of articular cartilage as well as devising strategies for functional cartilage repair.     

Site-specific immunostaining for type X collagen in noncalcified articular cartilage of canine stifle knee joint

Type X collagen is a short-chain collagen that is strongly expressed in hypertrophic chondrocytes. In this study, we used an immunohistochemical technique exploiting a prolonged hyaluronidase unmasking of type X collagen epitopes to show that type X collagen is not restricted to calcified cartilage, but is also present in normal canine noncalcified articular cartilage. A 30° valgus angulation procedure of the right tibia was performed in 15 dogs at the age of 3 months, whereas their nonoperated sister dogs served as controls. Samples were collected 7 and 18 months after the surgery and immunostained for type X collagen. The deposition of type X collagen increased during maturation from age 43 weeks to 91 weeks. In the patella, most of the noncalcified cartilage stained for type X collagen, whereas, in the patellar surface of the femur, it was present mainly in the femoral groove close to cartilage surface. In femoral condyles, the staining localized mostly in the superficial cartilage on the lateral and medial sides, but not in the central weight-bearing area. In tibial condyles, type X collagen was often observed close to the cartilage surface in medial parts of the condyles, although staining could also be seen in the deep zone of the cartilage. Staining for type X collagen appeared strongest at sites where the birefringence of polarized light was lowest, suggesting a colocalization of type X collagen with the collagen fibril arcades in the intermediate zone. No significant difference in type X collagen immunostaining was observed in lesion-free articular cartilage between controls and dogs that underwent a 30° valgus osteotomy. In osteoarthritic lesions, however, there was strong immunostaining for both type X collagen and collagenase-induced collagen cleavage products. The presence of type X collagen in the transitional zone of cartilage in the patella, femoropatellar groove, and in tibial cartilage uncovered by menisci suggests that it may involve a modification of collagen fibril arrangement at the site of collagen fibril arcades, perhaps providing additional support to the collagen network.     

The structure of the human subchondral plate

To study the anatomy of subarticular bone and cartilage, fresh specimens of cartilage on bone from the human shoulder, hip and knee were treated with bleach or papain, or were fixed and decalcified. All were compared using scanning electron microscopy. Papain digestion selectively removed cartilage to the tidemark. The tidemark contour was highly variable; irregularities were indirectly related to degenerative lesions and were most prominent in peripheral non-weight-bearing areas of joints with central fibrillation. Decalcification exposed the interface between the bone and calcified cartilage. Collagen fibrils in articular cartilage did not interdigitate with those of bone. The subchondral bone was appositional, avascular, smooth and very thin in most areas of human joints. Perforations through subchondral bone or calcified cartilage were rare. Bleach maceration destroyed important details.     

Cyclical articular joint loading leads to cartilage thinning and osteopontin production in a novel in vivo rabbit model of repetitive finger flexion

OBJECTIVE: An in vivo rabbit model of repetitive joint flexion and loading was used to characterize the morphological effects of cyclical loading on articular cartilage. DESIGN: The forepaw digits of eight anesthetized New Zealand White adult female rabbits were repetitively flexed at 1 Hz with a mean peak digit load of 0.42 N for 2 h per day for 60 cumulative hours. Metacarpophalangeal joints were collected from loaded and contra-lateral control limbs, fixed, decalcified, embedded, and thin-sectioned. Serial sections were stained for histology or for immunohistochemistry. Morphometric data including the mean thicknesses of the uncalcified cartilage and of the calcified cartilage were collected from digital photomicrographs of safranin O-stained sections. The number of cells stained with anti-osteopontin antibody was counted. RESULTS: We observed a decrease in uncalcified cartilage mean thickness with no significant change in calcified cartilage thickness. We also observed a significant increase in the number of cells positive for osteopontin (OPN) in the uncalcified cartilage. These changes occurred without overt cartilage surface degeneration. CONCLUSIONS: Cyclical loading leads to changes at the tissue and cellular levels in articular cartilage. These changes are suggestive of tidemark advancement and may indicate a reactivation of cartilage mineralization steps analogous to endochondral ossification. This novel in vivo rabbit model of repetitive flexion and loading can be used to investigate the effects of cyclical loading on articular joints.     

Variations in site and levels of expression of chondrocyte nucleotide pyrophosphohydrolase with aging.

The aim of this study was to identify changes in cartilage intermediate layer protein/nucleotide pyrophosphohydrolase (CILP/NTPPH) expression in articular cartilage during aging. Adult (3-4 years old) and young (7-10 days old) porcine articular hyaline cartilage and fibrocartilage were studied by Northern blot analysis, in situ hybridization, and immunohistochemistry using a complementary DNA (cDNA) probe encoding porcine CILP/NTPPH and antibody to a synthetic peptide corresponding to a CILP/NTPPH sequence. Northern blot analysis of chondrocytes showed lower expression of CILP/NTPPH messenger RNA (mRNA) in young cartilage than in adult cartilage. In adult cartilage, extracellular matrix from the surface to the middeep zone was immunoreactive for CILP/NTPPH, especially in the pericellular matrix surrounding the middeep zone chondrocytes. In young cartilage, chondrocytes were moderately immunoreactive for CILP/NTPPH throughout all zones except the calcified zone. The matrix of young cartilage was negative except in the superficial zone. In young cartilage, CILP/NTPPH mRNA expression was undetectable. In adult cartilage, chondrocytes showed strong mRNA expression for CILP/NTPPH throughout middeep zones. Protein and mRNA signals were not detectable below the tidemark. CILP/NTPPH secretion into matrix around chondrocytes increases with aging. In this extracellular site it may generate inorganic pyrophosphate and contribute to age-related calcium pyrophosphate dihydrate crystal deposition disease.     

Localization of type X collagen in canine growth plate and adult canine articular cartilage

Type X collagen was extracted from ends of canine growth plates by pepsin digestion after 4 M guanidine hydrochloride extraction, purified by stepwise salt precipitation (2.0 M NaCl in 0.5 M acetic acid), and chromatographed on a Bio-Gel A1.5 M column in 1.0 M CaCl2. Without reduction on sodium dodecyl sulfate (SDS) polyacrylamide gels, the preparation yielded a single, high-molecular-weight (mol wt) band; after reduction, a single band of relative mol wt 5.0 x 10(4) was found. Polyclonal sera were raised against the purified collagen and used in the immunolocalization of canine type X collagen. As expected, indirect immunoperoxidase (IP) or indirect immunofluorescent staining with the polyclonal sera demonstrated that most of the immunoreactivity was localized in the zone of provisional calcification of the growth plate and in cartilage remnants in the metaphyseal region of the physis. A progressive decrease in staining toward the diaphysis of the fetal canine long bone was apparent as the trabecular structures were remodeled to bone. Unexpectedly, type X collagen was also detected in the zone of calcified, mature articular cartilage. It was concentrated in the pericellular matrix of the chondrocytes, appeared at or just above the tidemark, and was expressed immediately before mineralization. Identification of type X collagen in both the canine growth plate and the zone of calcified articular cartilage suggests that cells in the deep layer of cartilage and in the zone of calcified cartilage in the adult animal retain some characteristics of a growth plate and may be involved in regulation of mineralization at this critical interface. The expression of growth plate-like properties would allow the deep chondrocytes of mature articular cartilage to play a role in remodeling of the joint with age and in the pathogenesis of osteoarthritis.     

Lead accumulation in tidemark of articular cartilage

OBJECTIVE: Determination of the spatial distribution of the toxic element lead (Pb) and other trace elements in normal articular cartilage and subchondral bone from adult humans with no history of work-related exposure to Pb. METHODS: Four macroscopically normal femoral heads and three patellas were harvested from randomly selected forensic autopsies. All subjects died of acute illnesses, had no history of work-related exposure to Pb and had no metabolic bone disease. The elemental distribution of lead (Pb) together with zinc (Zn), strontium (Sr) and calcium (Ca) in the chondral and subchondral region was detected using high resolution synchrotron radiation induced micro X-ray fluorescence (SR mu-XRF) analysis. SR mu-XRF line scans in conventional and SR mu-XRF area scans in confocal geometry were correlated to backscattered electron (BE) images visualizing the mineralized tissue. RESULTS: In all samples, we found a highly specific accumulation of Pb in the tidemark, the transition zone between calcified and non-calcified articular cartilage. Pb fluorescence intensities in the tidemark, which is thought to be a metabolically active mineralization front, were 13-fold higher when compared to subchondral bone. Pb intensities in the subchondral region were strongly correlated with Zn, but were distinctly different from Ca and Sr. CONCLUSIONS: The finding of the highly specific accumulation of lead in the tidemark of human articular cartilage is novel. However at this point, the exact mechanisms of the local Pb accumulation as well as its clinical implications are unknown.     

Articular cartilage preservation and storage. III. Quantitative zonal analysis of cytoplasmic components of stored versus in vivo articular cartilage chondrocytes

The cytoplasmic components of chondrocytes in the various zones of articular cartilage of the medial femoral condyle of six-week-old male New Zealand white rabbits stored in tissue culture medium at 37 degrees in 5% CO2 and air were quantitated from electron micrographs, and the results were compared statistically with the cytoplasmic components of chondrocytes in the corresponding zones of in vivo articular cartilage. The major changes that occurred during storage were: (1) an increase in the amount of lipid bodies in chondrocytes in the tangential, transitional, and calcified zones; (2) a decrease in the number of holes in the cytoplasm of chondrocytes in the radial and calcified zones; (3) a decrease in the amount of endoplasmic reticulum in the radial and calcified zones; and (4) an increase in cell size and cytosol area in the tangential and transitional zones but a significant decrease in cell size and cytosol area in the calcified zone. Stored articular cartilage chondrocytes demonstrated cellular changes associated with aging, whereas in vivo articular cartilage chondrocytes demonstrated changes associated with degeneration. The matrix of stored articular cartilage in the tangential, transitional, and upper part of the radial zones showed a decrease in opacity due to a decrease in the number of collagen fibers per unit area of matrix, a condition termed "chondroporosis." This study demonstrates that articular cartilage stored in standard tissue culture medium under ideal physiological conditions is morphologically abnormal. Based on these findings, one would not expect such stored cartilage to remain functionally intact when transplanted to replace articular cartilage loss.     

颈椎软骨终板钙化与颈椎间盘退变和椎体骨赘形成的关系

目的：研究颈椎软骨终板钙化与颈椎间盘退变和颈椎椎体骨赘形成的关系。方法：应用组织学方法观察颈前路环锯手术切下的18例脊髓型颈椎病和4例颈椎过伸性损伤致颈椎间盘突出患者的颈椎间盘及相邻的上下椎体标本,研究不同退变程度颈椎间盘软骨终板和椎间盘的形态学变化及椎体骨赘形成过程。结果：退变程度较轻或基本正常的颈椎间盘软骨终板结构良好,潮标清晰,退变程度较重的颈椎间盘软骨终板发生明显纤维化,潮标前移,钙化软骨和软骨下骨板增厚,退变颈椎间盘周边软骨终板潮标明显前移,钙化和骨化层增厚,形成突向外侧的椎体边缘的骨赘。结论：颈椎软骨终板的不断钙化和骨化导致颈椎间盘营养发生障碍可能是启动颈椎间盘退变的关键因素,退变椎体周边软骨终板的不断钙化和骨化是椎体骨赘形成的根本原因。     

Healing of defects in canine articular cartilage: Distribution of nonvascular α-smooth muscle actin-containing cells

The objective of this study was to evaluate the types of tissue resulting from spontaneous healing of surgically created defects in adult canine articular cartilage up to 29 weeks postoperatively, with specific attention directed toward the presence and distribution of cells containing the contractile actin isoform, alpha-smooth muscle actin. Two 4-mm diameter defects were made in the trochlear groove to the depth of the tidemark in 20 adult mongrel dogs. The areal percentage of specific tissue types in the reparative material was determined histomorphometrically. Immunohistochemistry was employed to evaluate the percentage of alpha-smooth muscle actin-containing cells. The results showed that approximately 50% of the chondrocytes in the superficial zone of the uninvolved articular cartilage expressed alpha-smooth muscle actin. A significantly lower percentage of alpha-smooth muscle actin-positive chondrocytes appeared in the uninvolved deep zone. Notably, the deep zone adjacent to the defect contained a greater percentage of such cells than in the uninvolved deep zone. Also of interest was that a greater percentage of nonvascular cells in the hyaline cartilage and fibrocartilage of the reparative tissue contained alpha-smooth muscle actin-positive cells, compared to the fibrous tissue in the defects. The findings of this study revealed that canine articular cartilage has some potential for spontaneous regeneration, including integration with the calcified cartilage zone. By 29 weeks, up to 40% of an areal cross section of an untreated full-thickness chondral defect was found to fill with hyaline cartilage, with up to 19% judged histologically similar to articular cartilage. The results warrant further consideration of the role of alpha-smooth muscle actin in chondrocytes in normal articular cartilage and in reparative tissue.     

Characterization of lesions of the talus and description of tram-track lesions

BACKGROUND: The ankle joint, although not prone to primary osteoarthritis (OA), is known to be susceptible to secondary OA as a result of sports injuries and other trauma. Unlike the knee joint, a thorough investigation of talar cartilage lesions has not been previously reported. METHODS: One hundred and five human tali from 67 donors were used to determine the type and location of the most common lesions through gross examination, radiography, diffraction enhanced imaging, and histology. "tram-track lesions" also are described. RESULTS: The most anterior and posterior regions of the talar dome, along with the medial and lateral borders of the dome were most affected by cartilage degeneration. These are regions that appear to be most subjected to frictional forces from their articulating counterparts during high stress activities that move articulating surfaces slightly out of congruence with each other. One particularly striking cartilage degeneration pattern that, to our knowledge, has not been described histologically, is the tram-track lesion. These lesions displayed a longitudinally oriented groove in the cartilage, running from anterior to posterior, in which the deepest portion of the groove was located in the middle of the anterior-posterior axis of the talus. Several of these cartilage grooves had a bony ridge beneath, mirroring the cartilage groove whereas others did not. This suggests that the cartilage groove develops before the interruption of the tidemark (border between the calcified and uncalcified cartilage). In specimens for which the articulating tibial articular surface was available, (17) it was found that the tibia displayed small osteophytes on the anterior articular margin that exactly corresponded to their articulation with the talar cartilage grooves as the ankle articulated through plantar flexion and dorsiflexion. CONCLUSION: This study provides an in-depth histologic and gross anatomic look at the most common lesions of the talus of the ankle joint. In particular, the "tram-track" lesion was shown to be a consequence of its tibial articulation and to include both subchondral bone and articular cartilage changes.     

Role of endochondral ossification of articular cartilage and functional adaptation of the subchondral plate in the development of fatigue microcracking of joints

The mechanisms that regulate functional adaptation of the articular ends of long bones are poorly understood. However, endochondral ossification of articular cartilage and modeling/remodeling of the subchondral plate and epiphyseal trabeculae are important components of the adaptive response. We performed a histologic study of the distal end of the third metacarpal/metatarsal bone of Thoroughbreds after bones were bulk-stained in basic fuchsin and calcified sections were prepared. The Thoroughbred racehorse is a model of an extreme athlete which experiences particularly high cyclic strains in distal limb bones. The following variables were quantified: microcrack boundary density in calcified cartilage (N.Cr/B.Bd); blood vessel boundary density in calcified cartilage (N.Ve/B.Bd); calcified cartilage width (Cl.Cg.Wi); duplication of the tidemark; and bone volume fraction of the subchondral plate (B.Ar/T.Ar). Measurements were made in five joint regions (lateral condyle and condylar groove; sagittal ridge; medial condylar and condylar groove). N.Cr/B.Bd was site-specific and was increased in the condylar groove region; this is the joint region from which parasagittal articular fatigue (condylar) fractures are typically propagated. Formation of resorption spaces in the subchondral plate was co-localized with microcracking. N.Ve/B.Bd was also site-specific. In the sagittal ridge region, N.Ve/B.Bd was increased, Cl.Cg.Wi was decreased, and B.Ar/T.Ar was decreased, when compared with the other joint regions. Multiple tidemarks were seen in all joint regions. Cumulative athletic activity was associated with a significant decrease in B.Ar/T.Ar in the condylar groove regions. N.Cr/B.Bd was positively correlated with B.Ar/T.Ar (P < 0.05, r(s) = 0.29) and N.Ve/B.Bd was negatively correlated with B.Ar/T.Ar (P < 0.005, r2 = 0.14) and Cl.Cg.Wi (P < 0.05, r2 = 0.07). We conclude that endochondral ossification of articular cartilage and modeling/remodeling of the subchondral plate promote initiation and propagation of site-specific fatigue microcracking of the joint surface, respectively, in this model. Microcracking of articular calcified cartilage likely represents mechanical failure of the joint surface. Propagation of microcracks into the subchondral plate is a critical factor in the pathogenesis of articular condylar fatigue (stress) fracture. Functional adaptation of the joint likely protects hyaline cartilage from injury in the short-term but may promote joint degeneration and osteoarthritis with ongoing athleticism.     

Age-related histological changes in rat mandibular condyle

The effect of aging on rat mandibular condyle was histologically assessed using 1-, 4-, 9-, and 16-month-old male Fischer rats (Eight rats of each group). The medial and anterior portions of the condyle protruded with age. The hypertrophic cell layer of the condylar cartilage occurring up to 4 months was no longer observable at 9 months, after which calcification of the cartilage layer with a tidemark was featured. The chondrocytes in the calcified cartilage layer were reduced in size and intensely stained by toluidine blue. A mineralizing front was formed parallel to, and migrated toward, the condyle surface. At 9 months, osteon-like osteogenesis occurred around blood vessels on the border between the calcified cartilage and the subchondral bone. Condyles of 16-month-old rats exhibited osteosclerotic changes. The calcified cartilage of the middle portion of the condyle formed a remarkably thick layer. The increased volume of the trabecular bone appears to lead to enhanced osteosclerotic changes of the condyle. The aging-related protrusion of the anterior and medial portions of the condyle could be the result of a combination of mechanical forces, e.g., occlusion and mastication. Further, our study revealed that the turnover from cartilage to bone occurring in rat condyle through the process of aging involved an osteon-like bone formation around the blood vessels on the border between the calcified cartilage layer and the bone. This process differs from the endochondral ossification process observed in younger rats.     

Solute transport in the deep and calcified zones of articular cartilage

OBJECTIVES: (1) To establish whether the tidemark and calcified cartilage are permeable to low molecular weight solutes, thereby providing a potential pathway for nutrition of cells in the deep cartilage. (2) To investigate transport from the subchondral microcirculation into calcified cartilage in an intact perfused joint and the effects on transport of static loading. METHODS: The permeability of the tidemark and calcified cartilage was investigated in plugs of cartilage and subchondral bone which formed the membrane of a diffusion cell. Transport from the subchondral microcirculation and the effects of load were studied in an intact perfused joint. Both preparations used the metacarpophalangeal joints of mature horses and fluorescein and rhodamine (m.w. approximately 400 Da) were employed as tracers, assayed by quantitative fluorescence microscopy on histological sections. RESULTS: Calcified cartilage was permeable to both solutes, both from the superficial and the subchondral sides. The effective diffusivity of both solutes was of the order of 9 x 10(-9) cm(2) s(-1), fivefold less than in the uncalcified cartilage. The calcified zone was heterogeneous, with high uptake of both tracers in the vicinity of the tidemark. The distribution volume of rhodamine B was higher than for fluorescein, consistent with a significant anionic charge in the calcified matrix. Static loading of the intact joint did not affect the transport of rhodamine B but caused a significant decrease in concentration of fluorescein both in the surface and deep zones of the tissue. CONCLUSIONS: Calcified cartilage is permeable to small solutes and the subchondral circulation may make a significant contribution to the nutrition of deep cartilage in the mature horse. Static loading reduces the uptake of small anionic solutes in the intact joint.     

Anatomy, histologic features, and vascularity of the adult acetabular labrum

Acetabular labrum tears have been implicated as a cause of hip pain in adult patients. Few studies describe the anatomy, histologic features, and microvasculature of the acetabular labrum and labral tears. Fifty-five embalmed and 12 fresh-frozen adult hips with a mean age of 78 years (range, 61-98 years) were studied. Of these, 96 % (53 of 55) of the hips had labral tears, with 74% of the tears located in the anterosuperior quadrant. Histologically, the fibrocartilaginous labrum was contiguous with the acetabular articular cartilage through a 1- to 2-mm zone of transition. A consistent projection of bone extends from the bony acetabulum into the substance of the labrum that is attached via a zone of calcified cartilage with a well-defined tidemark. Two distinct types of tears of the labrum were identified histologically. The first consisted of a detachment of the fibrocartilaginous labrum from the articular hyaline cartilage at the transition zone. The second consisted of one or more cleavage planes of variable depth within the substance of the labrum. Both types of labral tears were associated with increased microvessel formation seen within the tear. The acetabular labrum tear appears to be an acquired condition that is highly prevalent in aging adult hips. Labral tears occur early in the arthritic process of the hip and may be one of the causes of degenerative hip disease.     

膝关节原发性骨关节炎软骨和软骨下骨病理改变的定量研究

目的观察膝关节原发性骨关节炎(osteoarthritis,OA)胫骨平台软骨和软骨下骨病理改变特点,对比内、外侧平台软骨和软骨下骨结构参数,探讨钙化层和软骨下骨在OA发病机制中的作用。方法取2009年10月-2011年5月行人工全膝关节置换术治疗的30例30膝原发性OA患者自愿捐赠的新鲜胫骨平台标本进行实验。其中男11例,女19例;年龄55～78岁,平均65.1岁。病程10～25年,平均16.6年;患膝内翻畸形1～23°,平均9.3°。大体观察胫骨平台后在内、外侧中央负重区取材,常规制备脱钙石蜡切片,行HE和番红O/固绿染色,观察关节软骨退变特点,参照Mankin评分标准评分并分期;观察钙化层及软骨下骨病理改变。应用Image Pro Plus 6.0图像分析软件测量软骨和软骨下骨结构参数,包括软骨全层(total articular cartilage,TAC)厚度、钙化层(articular calcified cartilage,ACC)厚度、ACC/TAC比值、软骨下骨板(subchondral bone plate,SCP)厚度以及骨小梁体积分数(trabecular bone volume,BV/TV)。结果大体观察内侧平台软骨退变较外侧严重,内侧平台软骨Mankin评分为(12.4±1.1)分,显著高于外侧平台的(8.3±1.6)分(t=12.173,P=0.000)。根据Mankin评分结果在60个标本中,14个为OA早期,可见软骨浅表层裂隙、潮线复制和软骨下骨增厚;19个为OA中期,可见软骨深层裂隙、多发软骨下骨吸收陷窝和明显增厚的软骨下骨;27个为OA晚期,可见软骨全层缺失、软骨内化骨和"象牙化"软骨下骨。软骨和软骨下骨结构参数测定示:内侧平台TAC厚度显著低于外侧平台,ACC/TAC比值、BV/TV及SCP厚度显著高于外侧平台,差异均有统计学意义(P<0.05)。内、外侧平台ACC厚度比较,差异无统计学意义(P>0.05)。结论钙化层和软骨下骨可能在OA发生与进展中发挥了重要作用。