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PubMed1994

[Magnetic resonance of the cartilages of the large joints].

MRI of the articular cartilage requires a careful technical approach since this structure is very thin, with a peculiar internal architecture between the supporting matrix and the cell component. On MR images the normal articular cartilage has a zonal appearance. To optimize the variables for best visualization of the internal architecture of the hyaline articular cartilage, an ex vivo and in vivo study was carried out. Accurate T1 and T2 relaxation times of the articular cartilage were obtained with a particular mixed sequence and then used to create isocontrast intensity graphs. The latter allowed, in all pulse sequences (SE and GRE), the best combination of TR, TE and FA to optimize signal differences between cartilage areas. A trilaminar pattern was demonstrated, with a superficial and a deep hypointense areas, in all sequences, together with an intermediate area which was moderately hyperintense on SE images and markedly hyperintense on GRE images. In the current literature, MRI appears to have been widely used to investigate hyaline cartilage conditions. In many series, the technique proved its efficacy in assessing both acute (traumatic cartilage fractures, osteochondritis dissecans, arthritis) and chronic (arthrosis, chondromalacia patellae, Hoffa's disease, synovial plica syndrome) conditions of the articular cartilage. T2-weighted sequences (both SE and GRE) are widely known as the most accurate in assessing cartilage conditions, which depends mainly on the arthrographic effect of synovial fluid on T2-weighted images. Of late, also MR arthrography, especially MR arthrography after the i.v. administration of Gadolinium, has emerged as an outstanding technique to investigate articular cartilage conditions. On the basis of MR arthrography findings, some authors suggested a classification of osteochondritis dissecans, arthrosis and chondromalacia patellae on MR images. MR stages seem to be closely correlated with the histologic classification suggested for these conditions.

Cartilage Diseases↗

PubMedDate not supplied

Analysis of the physical properties of costal cartilage in a porcine model.

OBJECTIVE: To determine the impact of interventions on the degree of warping of costal cartilage. METHODS: The project was conducted at a large university animal research laboratory. The costal cartilage of eight 30-kg domestic pigs was harvested. The cartilage was cut into central and peripheral segments with a standard cutting die. Two sizes of rectangular cubes were compared. The central portions of costal cartilage were segmented and glued with octyl-2-cyanoacrylate. The shape of the cartilage was documented with both digital and film photography. The cartilage was placed into subdermal pockets on the dorsum of the pigs. The animals were killed at 4 weeks, and the cartilage was photographed. Adobe Photoshop software was used to measure the degree of warping. Statistical analysis was calculated by t test analysis. RESULTS: A total of 115 rectangular costal cartilage blocks were treated. Large blocks warped less than small blocks (P<.02). Centrally cut blocks warped less than peripherally cut blocks (P<.03). The octyl-2-cyanoacrylate incited a significant sterile inflammatory response such that the blocks could not be accurately assessed for warpage. CONCLUSION: Costal cartilage can be effectively used for grafting in rhinoplasty with minimal warping if large grafts from the central portion of cartilage are used.

Animals↗

PubMed2000

Comparison of thyroid, auricular, and costal cartilage donor sites for laryngotracheal reconstruction in an animal model.

OBJECTIVE: To evaluate and compare the use of autogenous thyroid cartilage with that of auricular and costal cartilage in laryngotracheoplasty (LTP). DESIGN: A blinded comparison of LTP techniques using anterior thyroid, auricular, or costal cartilage as graft material in a rabbit model. Histological and anatomical analyses were performed on the laryngeal specimens 1, 4, and 6 weeks after surgery. The following factors were analyzed in each specimen: graft viability, cartilage proliferation, perichondrial viability, degree of necrosis, inflammatory response, and degree of epithelialization. SUBJECTS: Fifty-seven New Zealand adult male rabbits, aged 6 months, were divided into 3 study groups (19 animals in each group) initially and equally into the 3 time periods. RESULTS: No episodes of respiratory compromise occurred in any of the animals in the 3 study groups. Gross inspection of the laryngotracheal complex in the thyroid cartilage group revealed no evidence of laryngeal structural compromise. There was no statistical difference between the 3 types of cartilage used for reconstruction for the variables of graft or perichondrial viability, degree of necrosis, or inflammatory response at 1, 4, or 6 weeks. Cartilage proliferation in the thyroid cartilage group was decreased compared with that in the other 2 groups at 1 week. The amount of proliferation increased in this group and was equal to the amount present in the other 2 groups 4 and 6 weeks after surgery. Complete epithelialization of the graft material was present in all 3 groups at 4 and 6 weeks after reconstruction. CONCLUSIONS: The use of autogenous thyroid cartilage for LTP compares favorably with that of other methods of reconstruction that use either auricular or costal cartilage in the rabbit model. This technique is a viable alternative for single-stage LTP and has the added advantage of using a single incision.

Animals↗

PubMed2003

Possible impedance of luminal reepithelialization by tracheal cartilage metalloproteinases.

BACKGROUND: Rapid reepithelialization of respiratory epithelium after injury to the large conducting airway (eg, trachea and bronchus) is poor. Our laboratory has developed an in vitro model of the trachea that allows us to examine reepithelialization in a complex culture system. We previously described how the presence of cartilage inhibited respiratory epithelial cell (REC) migration/proliferation. In the present study, we examined the effect of cartilage-conditioned medium (CCM) on REC proliferation. We hypothesized that a potential cause of delayed reepithelialization of the large conducting airway after injury could be excessive or aberrant secretion of matrix metalloproteinases (MMPs) by cartilage. DESIGN: We assessed cartilage-derived MMP production and effects on REC proliferation by adding CCM to primary cultures of porcine RECs on type I collagen and determining the cell number and viability. Cartilage-conditioned medium-derived MMP activity was determined by means of gelatin zymography in pooled samples from different times during in vitro cartilage culture. RESULTS: We detected MMP-2 and a small amount of MMP-9 in CCM. Enzyme activity was abolished by EDTA, confirming MMP identity. Cartilage-conditioned medium inhibited REC attachment and proliferation. Addition of the MMP inhibitor GM6001 to cartilage cultures yielded CCM that did not inhibit REC growth, indicating a role for cartilage-derived MMPs in modulating REC proliferation. CONCLUSION: Cartilage production and activity of MMP after injury to the large conducting airway may be a factor in the failure of luminal reepithelialization, resulting in aberrant repair.

Animals↗

PubMed1997

The interaction of the zone of calcified cartilage and subchondral bone in osteoarthritis.

The zone of calcified cartilage (ZCC) forms an important interface between cartilage and bone for transmitting force, attaching cartilage to bone, and limiting diffusion from bone to the deeper layers of cartilage. The height of the ZCC is a relatively constant percent of articular cartilage and the height is maintained by a balance between progression of the tidemark into the unmineralized cartilage and changing into bone by vascular invasion and bony remodeling. During its formation, the cells that form the ZCC have properties similar to the cells of the growth plate. In the adult, the ZCC becomes quiescent but not inactive. The ZCC may be reactivated in osteoarthritis and may progressively calcify the unmineralized cartilage. This might contribute to cartilage thinning which would increase the concentration of forces across the uncalcified cartilage leading to more damage. Although the subchondral bony plate remodels extensively in osteoarthritis, there is little evidence that a change in the biomechanics of the plate directly initiates the osteoarthritic process in cartilage. However, increased repair by endochondral ossification of vertical cracks in the ZCC that penetrate into the marrow space could contribute to progression via changes in the ZCC.

Animals↗

PubMed1999

Morphology and mechanical function of long-term in vitro engineered cartilage.

Cartilage tissue can be engineered in vitro with articular chondrocytes and poly(glycolic acid) nonwoven scaffolds as previously shown over 12 weeks in vitro. This study addressed whether engineered cartilage would further evolve and approach natural cartilage in extracellular matrix organization and biomechanical properties, especially aggregate modulus through longer term in vitro cultivation. Cellularity, cell size, compressive modulus, and permeability of the in vitro engineered cartilage stabilized within the 12-week cultivation time and remained at the same levels as those of natural cartilage thereafter. The linear range of the stress-strain curve was from 0 to a strain value between 5 and 10% for all the engineered cartilage tissues that were in vitro cultured for longer than 2 weeks, which was the same linear range for natural cartilage. The aggregate modulus further increased from week 12 to week 20 and remained approximately the same value thereafter during a 25-week in vitro cultivation. The aggregate modulus of the engineered cartilage reached 179+/-9 kPa after 20 weeks of in vitro cultivation, which was 40% that of natural articular cartilage. To our knowledge this is the highest aggregate modulus value yet reported of any in vitro engineered cartilage tissue.

Animals↗

PubMed2000

Mechanical quality of tissue engineered cartilage: results after 6 and 12 weeks in vivo.

Traumatic events are a primary cause for local lesions of articular cartilage. If treated early, restoration of the initial joint geometry and integrity may be achieved. In large defects, sufficient material is not available to bridge the affected area. Heterologeous transplantation is not well accepted due to the risk of infection and immune response. Alternatives are cartilage-like structures, which may be cultured in vitro and transplanted into the defect site. Critical to the success of these new tissues are their mechanical properties. Goals of this study were to generate a hyaline-like cartilage structure, to evaluate its performance in vivo and to verify that its cellular and material properties meet those of native cartilage. Hyaline-like cartilage specimens were generated in vitro and implanted in the backs of nude mice. Specimens were explanted after 6 and 12 weeks, mechanically tested using an indentation test and histologically examined. In mechanical testing, stiffness and failure load significantly increased between weeks 6 and 12. At 12 weeks, mechanical properties of the hyaline-like cartilage were comparable to those of native nasal septal cartilage. Compared to native articular cartilage, the engineered tissue achieved up to 30-50% in strength and mechanical stiffness. In histological examination, specimens showed neocartilage formation. The mechanical testing procedure proved to be sufficiently sensitive to identify differences in properties between cartilage specimens of different origin and at different stages of healing. As an adjunct to histological analysis, mechanical testing may be a valuable tool for judging the utility of engineered cartilage prior to a broad clinical usage.

Adult↗

PubMed2000

Heterogeneity of cartilage laminae in MR imaging.

The purpose of this study was to investigate the discrepancy in the number of laminae observed in magnetic resonance (MR) images of articular cartilage (the magic angle effect in MRI of cartilage). Microscopic MR imaging (muMRI) experiments were carried out at 14-micrometer pixel resolution on full-depth cartilage-bone plugs from several locations (central, intermediate, and peripheral) on the humeral heads of two young healthy beagles. When the articular surface of the plug was perpendicular to the direction of the magnetic field, the cartilage appeared to have two layers in the plugs from the central locations of the humeral head, three layers in the plugs from the greater tubercle side of the humeral head, and three or five layers in the plugs from the lesser tubercle side. This heterogeneity of cartilage laminae was observed within a single humeral head and was symmetrical about the median plane of the animal. This result suggests that some structural variations related to cartilage structure in various regions of load bearing may cause some unique laminar patterns seen in MRI of cartilage. This novel and new observation may resolve the controversy about whether cartilage appears as two or three layers in MR images. A comprehensive model for the collagen structure over a curved two-dimensional surface of a joint is suggested as a replacement of the classic three-zone model of fiber orientation in collagen. This heterogeneity of cartilage laminae is speculated to be related to the load-bearing status of the tissue in the joint. The ability to visualize such structural heterogeneity is important because of the direct connection between collagen structure and the mechanical characteristics of cartilage.

Animals↗

PubMed1998

Progressive polyarthritis induced in BALB/c mice by aggrecan from normal and osteoarthritic human cartilage.

OBJECTIVE: To find an "unlimited" source of antigenic material (aggrecan) for arthritis induction in BALB/c mice; to analyze the specificities of immune reactions to aggrecan and type II collagen in 2 arthritis-susceptible murine strains, BALB/c mice for proteoglycan (aggrecan)-induced arthritis and DBA/1j mice for collagen-induced arthritis; to compare the histopathologic features of arthritis induced by purified aggrecans or total extracts of osteoarthritic (OA) cartilage; and to determine arthritis susceptibility in various BALB/c colonies. METHODS: Aggrecans from total extracts of human fetal, normal adult, OA, and rheumatoid cartilage samples and from osteophytes were isolated, purified by gradient centrifugation, deglycosylated, characterized, and tested for arthritis induction. Purified type II collagen and salt-soluble collagens from OA cartilage were denatured, stromelysin treated, and used for immunization and arthritis induction in arthritis-susceptible (DBA/1j and BALB/c) murine strains. RESULTS: Chondrocytes from OA cartilage synthesize predominantly fetal-type aggrecan, which is the most efficient antigenic material for arthritis induction in BALB/c mice. The critical autoimmune/arthritogenic T cell epitopes of aggrecan are located in the G1 domain. Although most of the aggrecan molecules are heavily degraded and lost from OA cartilage, the G1 domain-containing fragments accumulate in OA cartilage. The amount of G1-containing fragments is approximately twice as much in OA than in normal adult articular cartilage, and the arthritogenic epitope(s) remains intact in G1-containing fragments retained in cartilage. Thus, total extracts of OA cartilage (without additional purification), if deglycosylated appropriately, can be used as arthritogenic material in BALB/c mice. CONCLUSION: Predominantly G1 domain-containing fragments of aggrecan accumulate in OA cartilage, and these are the fragments which induce arthritis in BALB/c mice. Arthritis induction is highly specific for aggrecan epitopes and dictated by the genetic background of the BALB/c strain.

Adult↗

PubMed1998

Exercise protects against articular cartilage degeneration in the hamster.

OBJECTIVE: It has been reported that osteoarthritis can occur in hamsters. The present study was undertaken to determine the effects of exercise on the composition of articular cartilage and synovial fluid and on the development of cartilage degeneration in these animals. METHODS: Young (2.5-month-old) group-housed hamsters were compared with 5.5-month-old hamsters that had undergone 3 months of daily wheel running exercise (6-12 km/day) or 3 months of sedentary, individually housed living. The condition of the femoral condyles was determined by scanning electron microscopy in 12 exercising hamsters, 12 sedentary hamsters, and 6 of the young controls. The content of proteoglycan, hyaluronic acid, hydroxyproline, and proline in synovial fluid and patellar cartilage was measured. RESULTS: By scanning electron microscopy, the femoral articular cartilage was smooth and undulating in young controls and older exercising hamsters. In contrast, the femoral condyles were fibrillated in all 12 of the sedentary hamsters. There was no difference in the patellar cartilage collagen content between the 3 groups, but proteoglycan content and synthesis were lower in the patellar cartilage of the sedentary group. Synovial fluid volume was also decreased in the sedentary group compared with the young controls or the older exercising hamsters. CONCLUSION: A sedentary lifestyle in the hamster leads to a lower proteoglycan content in the cartilage and a lower synovial fluid volume. These changes are associated with cartilage fibrillation, pitting, and fissuring. Daily exercise prevents early cartilage degeneration and maintains normal articular cartilage.

Animals↗

PubMed1999

Degradation of cartilage type II collagen precedes the onset of osteoarthritis following anterior cruciate ligament rupture.

OBJECTIVE: To determine if degradation of cartilage matrix in primary osteoarthritis (OA) or in OA secondary to rupture of the anterior cruciate ligament (ACL) is a gradual response to excessive loading or an early, initiating event in the disease process. METHODS: Biopsy samples were obtained from the low-weight-bearing articular cartilage of the intercondylar notch, in patients undergoing knee arthroscopy (ACL injury) or arthroplasty (late-stage primary OA) or in controls. In some cases, biopsy samples were also removed from the high-weight-bearing articular cartilage of the femoral condyles. Biopsy specimens were extracted and assayed for total and denatured type II collagen (CII) by inhibition enzyme-linked immunosorbent assay and for proteoglycan using a colorimetric method. All patients were assessed radiographically for cartilage erosion. In addition, the cartilage of patients with ACL injury was assessed at arthroscopy, and the knee function of patients with primary OA was assessed using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). RESULTS: Increased CII degradation was detected in the low- as well as the high-weight-bearing cartilage of patients with late-stage OA, and there was a positive correlation between the percentage denatured collagen and the WOMAC score. Most of the patients with ACL injury had no clinical signs of OA or macroscopic cartilage erosion. However, the low-weight-bearing articular cartilage from these patients showed a significant increase in CII degradation, similar to that observed in late-stage OA. The proteoglycan content of articular cartilage did not change significantly in patients with OA or ACL injury compared with controls. CONCLUSION: CII degradation is an early event following ACL injury and is unlikely to be a direct result of mechanical loading, since it was observed in cartilage obtained from a low-weight-bearing site.

Adolescent↗

PubMed2000

Cartilage degradation and invasion by rheumatoid synovial fibroblasts is inhibited by gene transfer of a cell surface-targeted plasmin inhibitor.

OBJECTIVE: Joint destruction in rheumatoid arthritis (RA) is a result of degradation and invasion of the articular cartilage by the pannus tissue. The present study was undertaken to examine the role of the plasminogen activation system in cartilage degradation and invasion by synovial fibroblasts and investigate a novel gene therapeutic approach using a cell surface-targeted plasmin inhibitor (ATF.BPTI). METHODS: Adenoviral vectors were used for gene transfer. The effects of ATF.BPTI gene transfer on RA synovial fibroblast-dependent cartilage degradation were studied in vitro, and cartilage invasion was studied in vivo in the SCID mouse coimplantation model. RESULTS: The results indicate that cartilage matrix degradation by rheumatoid synovial fibroblasts is plasmin mediated and depends on urokinase-type plasminogen activator for activation. Targeting plasmin inhibition to the cell surface of the fibroblasts by gene transfer of a cell surface-binding plasmin inhibitor resulted in a significant reduction of cartilage matrix degradation in vitro and of cartilage invasion in vivo. Compared with uninfected rheumatoid synovial fibroblasts, the mean +/-SEM cartilage degradation in vitro was reduced to 87.9+/-0.9% after LacZ gene transfer versus a reduction to 24.0+/-1.6% after ATF.BPTI gene transfer (P<0.0001). The mean +/- SEM in vivo cartilage invasion score was 3.1+/-0.4 in the control-transduced fibroblasts and 1.8+/-0.4 in the ATF.BPTI-transduced fibroblasts (P<0.05). CONCLUSION: These results indicate a role of the plasminogen activation system in synovial fibroblast-dependent cartilage degradation and invasion in RA, and demonstrate an effective way to inhibit this by gene transfer of a cell surface-targeted plasmin inhibitor.

Activating Transcription Factors↗

PubMed2001

Osteopontin: an intrinsic inhibitor of inflammation in cartilage.

OBJECTIVE: To identify extracellular and intraarticular matrix components that are differentially expressed in normal and osteoarthritis (OA)-affected cartilage and to investigate their functions with respect to regulation of mediators of inflammation. METHODS: Differential-display reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of a pool of messenger RNA (mRNA) from 10 human OA cartilage samples and 5 normal cartilage samples was performed using arbitrary primers. Confirmatory analysis of the up-regulated transcripts of fibronectin (FN) and osteopontin (OPN) was performed by RT-PCR of individual RNA samples from a separate set of donors. The effect of recombinant OPN (or anti-OPN antiserum) on chondrocyte function was examined by analyzing the spontaneous or interleukin-1 (IL-1)-induced release of nitric oxide (NO) and prostaglandin E2 (PGE2) from human OA-affected cartilage under ex vivo conditions. RESULTS: Up-regulation (300-700%) of FN and OPN mRNA was observed in human OA-affected cartilage as compared with normal cartilage. Functional analysis of the role of OPN in OA cartilage showed that 1) Addition of 1 microg/ml (20 nM) of recombinant OPN to human OA-affected cartilage under ex vivo conditions inhibited spontaneous and IL-1beta-induced NO and PGE2 production, and 2) neutralization of intraarticular OPN with anti-OPN antiserum augmented NO production. CONCLUSION: The data indicate that one of the functions of intraarticular OPN, which is overexpressed in OA cartilage, is to act as an innate inhibitor of IL-1, NO, and PGE2 production. These findings suggest that the production of pleiotropic mediators of inflammation that influence cartilage homeostasis, such as NO and PGE2, is regulated by the interaction of chondrocytes with differentially expressed proteins within the extracellular matrix.

Aged↗

PubMed2001

The splice variants VEGF121 and VEGF189 of the angiogenic peptide vascular endothelial growth factor are expressed in osteoarthritic cartilage.

OBJECTIVE: Vascular endothelial growth factor (VEGF) has recently been shown to play an important role during endochondral bone formation in hypertrophic cartilage remodeling, ossification, and angiogenesis, but it is not expressed in normal adult cartilage. Since genes expressed during development often reappear in the disease state, we investigated whether VEGF and its receptors (VEGFRs) are expressed in osteoarthritic (OA) cartilage. METHODS: VEGF production in OA cartilage from the tibial plateau was measured by enzyme-linked immunosorbent assay. Deposition of VEGF and VEGFR was determined by immunohistochemistry. Expression of messenger RNA for the different VEGF splice forms and for VEGFR was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Increased VEGF concentrations were measured in OA cartilage from the tibial plateau, while VEGF was almost undetectable in normal cartilage but could be immunostained within the intracellular and pericellular matrices of OA chondrocytes. In analyses of cartilage samples from all 10 OA patients evaluated, VEGF121 and VEGF189 were identified as the only VEGF splice forms expressed. RT-PCR and immunohistochemistry for VEGF in normal hyaline cartilage yielded negative findings. In addition to VEGF, VEGFR-2 (kinase domain region/fetal liver kinase 1), but not VEGFR-1 (fms-like tyrosine kinase 1), could be detected by RT-PCR in OA cartilage and immunostained on OA chondrocytes. CONCLUSION: Apart from its production in hypertrophic chondrocytes, VEGF is also produced in chondrocytes of OA cartilage. While the splice variant VEGF189 binds to extracellular matrix proteoglycans, VEGF121 is diffused freely. Both proteins should contribute to the inflammatory process by autocrine/paracrine stimulation of chondrocytes, chemotaxis of macrophages, and promotion of angiogenesis.

Alternative Splicing↗

PubMed2001

Activation of procollagenases is a key control point in cartilage collagen degradation: interaction of serine and metalloproteinase pathways.

OBJECTIVE: Bovine and human cartilages in explant culture respond to proinflammatory cytokines with the up-regulation of procollagenases. In stimulated bovine nasal cartilage (BNC), >90% of collagen is released by day 14 of culture, but collagen release is rarely seen before day 7. The aim of this study was to investigate if activation of procollagenases is a rate-limiting step in cartilage collagen breakdown. METHODS: BNC and human articular cartilage explants were cultured with interleukin-1alpha (IL-1alpha) and/or oncostatin M (OSM) with or without test reagents. Collagen levels were determined by assay of hydroxyproline. Collagenase activity was measured using the diffuse fibril assay. RESULTS: The addition of procollagenase activators, matrix metalloproteinase 3 (MMP-3), and APMA to IL-1alpha/OSM-stimulated BNC resulted in early release of collagen. The release with APMA was completely blocked by the addition of tissue inhibitor of metalloproteinases 1. This shows that procollagenases are present early in the culture period, but cartilage collagen breakdown does not happen until activation occurs. The addition of plasminogen to IL-1alpha/OSM-stimulated cartilage produced early collagen release in bovine and a significant increase in human cartilage. Thus, plasminogen activators (PAs) are present and convert plasminogen to plasmin, a known activator of several MMPs, including collagenases. Addition of alpha1-proteinase inhibitor or a urokinase-type PA inhibitor, 7-amino-4-chloro-3-(3-isothiureidopropoxy) isocoumarin, partially blocked the breakdown of collagen from IL-1alpha/OSM-treated bovine cartilage. This suggests that serine proteinases are involved in the activation cascades of procollagenases that result in cartilage collagen breakdown. CONCLUSION: The activation of procollagenases is a key control point in cartilage collagen breakdown, and serine proteinase pathways activate MMPs.

Animals↗

PubMed1980

A light and electron microscopic study of the region of cartilage resorption in the embryonic chick femur.

It has long been known that uncalcified cartilage of embryonic chick long bones is removed to make way for invading marrow. However, no one has clearly established which cells are responsible for this erosion. Using the light and electron microscopes, we have studied the cartilage-marrow interface, which we presume to be the region of resorption. Here, we found two types of mononuclear cells in intimate contact with cartilage matrix. 1. The predominate cell type had a euchromatic nucleus with a nucleolus and a cytoplasm containing extensive profiles of rough endoplasmic reticulum; also, processes extended from these cells into the adjoining cartilage matrix. 2. Macrophages containing many lysosomal vesicles, which often became swollen, were found on or near the surface of cartilage. In addition, a few cells with an intermediate appearance were present. A decrease in the amount of sulfated material in a 25-30 micrometer zone of cartilage in advance of the interface and an alteration in the orientation, and in some cases the integrity, of collagenous fibers were associated with the presence of the above mentioned cells. These alterations in cartilage were not due to the synthesis of sulfated or of collagenous material. The above evidence, although not conclusive, suggests that these mononuclear cells are responsible for cartilage resorption. In this respect, the removal of avian uncalcified cartilage is similar to the resorption of uncalcified articular cartilage which occurs in rheumatoid arthritis.

Animals↗

PubMed1982

The cellular organization of fibroblastic cells and macrophages at regions of uncalcified cartilage resorption in the embryonic chick femur as revealed by alkaline and acid phosphatase histochemistry.

Resorption of uncalcified cartilage in the embryonic chick femur appears to be mediated by two types of mononuclear cells. One cell type lies flattened and adherent along the surface of the cartilage matrix into which it extends cellular processes. Cytological characteristics of a large, euchromatic nucleus containing a nucleolus, and cytoplasm containing moderate to extensive amounts of rough endoplasmic reticulum indicate that these are protein synthetic cells. Macrophages, characterized by a pleomorphic shape and cytoplasm containing numerous mitochondria and vesicles, comprise the second cell type. These may be seen lying in contact with cartilage matrix, but are more likely located in the nonhematopoietic marrow adjacent to resorbing cartilage, where they establish close cellular associations with protein synthetic cells. Alkaline and acid phosphatase histochemical studies differentiate these two cellular types. Marrow alkaline phosphatase activity is restricted to the cartilage-marrow interface from which it diffuses a short distance into cartilage matrix, but does not diffuse into nearby marrow. Intracellular alkaline phosphatase is present only in protein synthetic cells that line the surface of cartilage, and thus appears to be produced by these cells. Acid phosphatase positive macrophages are scattered throughout the marrow, but are found in greatest concentrations in the region of cartilage resorption. They are rarely in direct contact with cartilage, and there is no evidence that acid phosphatase is released from these cells. The relative localizations and the presence of cellular interactions of these two cell types suggests that protein synthetic cells may be of fibroblastic origin, and may play a primary role in cartilage degradation, while macrophages, in keeping with biochemical evidence, play an adjunct or possibly a regulative role.

Acid Phosphatase↗

PubMed1984

Ultrastructural cytochemistry of proteoglycans associated with calcification of shark cartilage.

Proteoglycans (PGs) as well as sulfated glycosaminoglycans (GAGs) are closely associated with cartilage calcification. An inner zone of endoskeletal tesserae of sharks is composed of a unique calcified hyaline cartilage. Initial calcification can be seen in the cartilage close to the inner zone. We have ultrastructurally examined shark, Triakis scyllia, noncalcifying, calcifying, and calcified cartilage using the tannic acid-ferric chloride (TA-Fe), the high iron diamine (HID), and the HID-thiocarbohydrazide-silver proteinate (HID-TCH-SP) methods for localization of sulfated complex carbohydrates. In noncalcifying cartilage, TA-Fe and HID strongly stained matrix granules which were round, ovoid, elongated, or irregularly shaped and presumably represented PG monomers. The size and staining intensity of the reactive matrix granules progressively decreased in calcifying cartilage toward the calcification front of the calcified cartilage. Similarly, a progressive decrease in the size of the HID-TCH-SP stain deposits in the matrix granules was observed in the calcifying cartilage close to the calcification front and was interpreted as a decrease in length of sulfate containing GAG chains. In the calcified cartilage, the highly calcified areas were often localized in the calcification front and contained few or no small HID-TCH-SP stain deposits, whereas the weakly calcified regions contained more stain deposits. These results indicate that partial and complete degradation of sulfated GAGs and/or PGs may be a requisite for calcification of shark cartilage.

Animals↗