acid-phosphatase and Osteoarthritis

The levels of six lysosomal enzymes (acid phosphatase, beta-acetylglucosaminidase, cathepsin D, beta-galactosidase, arylsulfatase A, and beta-glucuronidase) and four neutral and alkaline hydrolases (esterase, inorganic phyrophosphatase, alkaline phosphatase, and 5'-nucleotidase) were measured in osteoarthritic, rheumatoid and control synovia. All enzyme levels in diseased synovium except esterase values in osteoarthritis were significantly elevated compared with controls. The mean values of the group of acid hydrolases and the group of neutral and alkaline hydrolases in osteoarthritic synovia were 1.9- and 2.0-fold greater than those of control specimens. In rheumatoid synovia, the values were 4.2- and 4.5 fold greater than control for the same enzymes. Levels in rheumatoid synovia were significantly higher than those in osteoarthritic synovia with the exception of 5'-nucleotidase. Only a limited correlation between the extents of inflammation present in the synovia and the levels of a lysosomal marker enzyme (cathepsin D) was observed. These results demonstrate that whatever the mechanism, increased levels of acid hydrolases as well as certain neutral and alkaline hydrolases are present in osteoarthritic and rheumatoid synovia, and these enzymes are probably contained in the synovial lining cells.

Topics: Acetylglucosaminidase; Acid Phosphatase; Alkaline Phosphatase; Arthritis, Rheumatoid; Cathepsins; Cerebroside-Sulfatase; Esterases; Galactosidases; Glucuronidase; Humans; Hydrolases; Nucleotidases; Osteoarthritis; Peroxidases; Polyethylene Glycols; Pyrophosphatases; Synovial Membrane

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Cathepsins; Chondroitin; Collagen; DNA; Glycosaminoglycans; Hexosamines; Histocytochemistry; Humans; Keratins; Mitosis; Osteoarthritis; Proteoglycans; Rabbits; Sulfates; Water

Osteoarthritis (OA) is a most commonly multifactorial degenerative joint disease along with the aging population, particularly in postmenopausal women. During the onset of OA, articular cartilage and subchondral bone act in concert as a functional unit. This present study is to investigate the effects of early or late treatment with recombinant lubricin on the onset of osteoarthritis (OA) in ovariectomized (OVX) rats. We found that both early and late recombinant lubricin treatments attenuated the onset of OA by positive feedback loop between articular cartilage and subchondral bone, although late treatment contributed to a lesser effect compared with early treatment. Specifically, treatment with recombinant lubricin protected articular cartilage from degeneration, demonstrated by lower proteoglycan loss, lower OARSI scores, less calcification cartilage zone and reduced immunostaining for collagen X (Col X) and matrix metalloproteinase (MMP-13) but increased the expression of lubricin, in comparison with vehicle-treated OVX rat group. Further, chondroprotective effects of lubricin normalized bone remodeling in subchondral bone underneath. It's suggested that treatment with recombinant lubricin inhibited the elevation of TRAP and Osterix positive cells in OVX rats and led to the normalization of subchondral bone microarchitectures with the suppression of subsidence of bone volume ratio (BV/TV) and trabecular thickness (Tb.Th) and the increase of trabecular separation (Tb.Sp) in vehicle-treated OVX rats. What's more, the normalization of subchondral bone in turn attenuated the articular cartilage erosion by inhibiting vascular invasion from subchondral bone to calcified cartilage zone, exemplified by inhibiting the elevation of CD31 positive cells in calcified cartilage and angiography in subchondral bone. Together, these results shed light that both early and late recombinant lubricin treatments attenuate the onset of OA by balancing the interplay between articular cartilage and subchondral bone in OVX rats, while also providing a further rationale for its therapeutic targeting to postmenopausal OA and suggesting that treatment timing is a pivotal factor for better effect acquisition.

Topics: Acid Phosphatase; Alendronate; Animals; Cartilage, Articular; Collagen Type I; Collagen Type X; Feedback, Physiological; Female; Glycoproteins; Humans; Isoenzymes; Matrix Metalloproteinase 13; Osteoarthritis; Ovariectomy; Peptides; Platelet Endothelial Cell Adhesion Molecule-1; Protective Agents; Rats, Sprague-Dawley; Recombinant Proteins; Tartrate-Resistant Acid Phosphatase; Tibia; Transcription Factors; X-Ray Microtomography

Subchondral bone cyst (SBC) growth, caused by osteoclast activity during early knee osteoarthritis (OA) pathogenesis, should be treated to prevent further progressions of OA. In the present study, we evaluated the effects of gentle treadmill walking on subchondral bone and cartilage changes in an experimental rat model of destabilized medial meniscus (DMM).. Twelve-week-old Wistar rats underwent DMM surgery in their right knee and sham surgery in their left knee and were assigned to either the sedentary group or walking group (n = 42/group). Animals in the walking group were subjected to treadmill exercise 2 days after surgery, which included walking for 12 m/min, 30 min/day, 5 days/week for 1, 2, and 4 week(s). Subchondral bone and cartilage changes were evaluated by micro-CT analysis, histological analysis, and biomechanical analysis.. Treadmill walking had a tendency to suppress SBC growth, which was confirmed by micro-CT (P = 0.06) and positive staining for tartrate-resistant acid phosphatase (TRAP) activity for the osteoclast number per bone surface (P = 0.09) 4 weeks after surgery. These changes coincide with the prevention of cartilage degeneration as evaluated by the Osteoarthritis Research Society International (OARSI) score (P < 0.05) and biomechanically softening (P < 0.05). Furthermore, treadmill walking could suppressed increasing osteocyte deaths (P < 0.01), which was positively correlated with the OARSI score (r = 0.77; P < 0.01).. These results indicate biomechanical and biological links exist between cartilage and subchondral bone; preventive effects of treadmill walking on subchondral bone deterioration might be partly explained by the chondroprotective effects.

Topics: Acid Phosphatase; Animals; Apoptosis; Cartilage, Articular; Cell Death; Disease Models, Animal; Exercise Test; Immunohistochemistry; In Situ Nick-End Labeling; Male; Menisci, Tibial; Osteoarthritis; Osteoarthritis, Knee; Osteocytes; Osteophyte; Rats; Rats, Wistar; Tibia; Walking; X-Ray Microtomography

Multinucleated giant cells have been noticed in diverse arthritic conditions since their first description in rheumatoid synovium. However, their role in the pathogenesis of osteoarthritis (OA) or rheumatoid arthritis (RA) still remains broadly unknown. We aimed to study the presence and characteristics of multinucleated giant cells (MGC) both in synovium and in subchondral bone tissues of patients with OA or RA.. Knee synovial and subchondral bone samples were from age-matched patients undergoing total joint replacement for OA or RA, or non-arthritic post mortem (PM) controls. OA synovium was stratified by histological inflammation grade using index tissue sections. Synovitis was assessed by Krenn score. Histological studies employed specific antibodies against macrophage markers or cathepsin K, or TRAP enzymatic assay.. Inflamed OA and RA synovia displayed more multinucleated giant cells than did non-inflamed OA and PM synovia. There was a significant association between MGC numbers and synovitis severity. A TRAP negative/cathepsin K negative Langhans-like subtype was predominant in OA, whereas both Langhans-like and TRAP-positive/cathepsin K-negative foreign-body-like subtypes were most commonly detected in RA. Plasma-like and foam-like subtypes also were observed in OA and RA synovia, and the latter was found surrounding adipocytes. TRAP positive/cathepsin K positive osteoclasts were only identified adjacent to subchondral bone surfaces. TRAP positive osteoclasts were significantly increased in subchondral bone in OA and RA compared to PM controls.. Multinucleated giant cells are associated with synovitis severity, and subchondral osteoclast numbers are increased in OA, as well as in RA. Further research targeting multinucleated giant cells is warranted to elucidate their contributions to the symptoms and joint damage associated with arthritis.

Topics: Acid Phosphatase; Aged; Anti-Inflammatory Agents, Non-Steroidal; Antirheumatic Agents; Arthritis, Rheumatoid; Biomarkers; Calcium; Cathepsin K; Cross-Sectional Studies; Diphosphonates; Female; Giant Cells; Giant Cells, Langhans; Glucocorticoids; Humans; Isoenzymes; Knee Joint; Macrophages; Male; Middle Aged; Osteoarthritis; Osteoclasts; Research Design; Single-Blind Method; Synovial Membrane; Tartrate-Resistant Acid Phosphatase; Tibia; Vitamin D

The analysis of indicators of mineral metabolism in patients with degenerative dystrophic affections of joints demonstrated that under development of osteoarthrosis process the alteration of indicators of concentration of electrolytes in blood serum, urine and synovial fluid occurs. The stage II of process is characterized by maximal alterations of indicators. The indicator of relationship between concentration of phosphate-ion and index of phosphatases of blood serum turned out the significant coefficient of correlation.

Topics: Acid Phosphatase; Adult; Aged; Alkaline Phosphatase; Biomarkers; Calcium; Disease Progression; Female; Humans; Isoenzymes; Knee Joint; Male; Middle Aged; Osteoarthritis; Phosphates; Synovial Fluid; Tartrate-Resistant Acid Phosphatase

This study was carried out to investigate the effect of Spatholobus suberectus Dunn (SS) on the protection of chondral defect and inhibition of osteoclastogenesis. To examine these effects, we measured the matrix metalloproteinase (MMP) levels in SW1353 chondrosarcoma cells and performed tartrate-resistant acid phosphatase (TRAP) staining in bone marrow macrophage (BMM)-derived osteoclasts. To investigate the anti-osteoarthritis (OA) effects, we assessed TNF-α-induced MMP-1, -3, -9 and tissue inhibitors of matrix metalloproteinase (TIMP) expression levels in SW1353 cells. We observed that SS extract significantly inhibited MMP and TIMP expression in SW1353 cells. Also, SS extract inhibited the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. These results suggest that SS extract may have a potential in the treatment of bone loss and chondral defect by suppressing osteoclast differentiation and decreasing the expression of OA factors. Therefore, clarification of the mechanism of the action of SS extract and its active components is needed.

Topics: Acid Phosphatase; Animals; Cell Differentiation; Chondrogenesis; Chondrosarcoma; Depression, Chemical; Fabaceae; Isoenzymes; Macrophages; Male; Matrix Metalloproteinases; Mice; Mice, Inbred ICR; Osteoarthritis; Osteoclasts; Phytotherapy; Plant Extracts; Plant Roots; RANK Ligand; Stimulation, Chemical; Tartrate-Resistant Acid Phosphatase; Tissue Inhibitor of Metalloproteinases; Tumor Cells, Cultured

Bone destruction is a common feature of inflammatory arthritis and is mediated by osteoclasts, the only specialized cells to carry out bone resorption. Aberrant expression of receptor activator of nuclear factor kappa β ligand (RANKL), an inducer of osteoclast differentiation has been linked with bone pathology and the synovial fibroblast in rheumatoid arthritis (RA). In this manuscript, we challenge the current concept that an increase in RANKL expression governs osteoclastogenesis and bone destruction in autoimmune arthritis. We isolated human fibroblasts from RA, pyrophosphate arthropathy (PPA) and osteoarthritis (OA) patients and analyzed their RANKL/OPG expression profile and the capacity of their secreted factors to induce osteoclastogenesis. We determined a 10-fold increase of RANKL mRNA and protein in fibroblasts isolated from RA relative to PPA and OA patients. Peripheral blood mononuclear cells (PBMC) from healthy volunteers were cultured in the presence of RA, PPA and OA synovial fibroblast conditioned medium. Osteoclast differentiation was assessed by expression of tartrate-resistant acid phosphatase (TRAP), vitronectin receptor (VNR), F-actin ring formation and bone resorption assays. The formation of TRAP(+), VNR(+) multinucleated cells, capable of F-actin ring formation and lacunar resorption in synovial fibroblast conditioned medium cultures occured in the presence of osteoprotegerin (OPG) a RANKL antagonist. Osteoclasts did not form in these cultures in the absence of macrophage colony stimulating factor (M-CSF). Our data suggest that the conditioned medium of pure synovial fibroblast cultures contain inflammatory mediators that can induce osteoclast formation in human PBMC independently of RANKL. Moreover inhibition of the TNF or IL-6 pathway was not sufficient to abolish osteoclastogenic signals derived from arthritic synovial fibroblasts. Collectively, our data clearly show that alternate osteoclastogenic pathways exist in inflammatory arthritis and place the synovial fibroblast as a key regulatory cell in bone and joint destruction, which is a hallmark of autoimmune arthritis.

Topics: Acid Phosphatase; Actins; Arthritis, Rheumatoid; Bone Resorption; Cell Differentiation; Cells, Cultured; Chondrocalcinosis; Culture Media, Conditioned; Fibroblasts; Gene Expression Regulation; Humans; Integrin alphaVbeta3; Interleukin-6; Isoenzymes; Leukocytes, Mononuclear; Osteoarthritis; Osteoclasts; Osteoprotegerin; RANK Ligand; Signal Transduction; Synovial Fluid; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

The levels of tartrate resistant acid phosphatase (TRAP), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in synovial fluid (SF) and serum in cases of canine osteoarthritis (OA) were measured. OA was induced by a surgically-created medial patellar luxation in the left stifle of 24 dogs. SF and blood samples were collected at 1.5- and 3-month intervals, respectively. Every 3 months, one dog was euthanatized to collect tissue samples from both stifles. TRAP levels in SF and serum were measured using a spectrophotometer, and TRAP-positive cells in joint tissues were identified by enzyme histochemistry. MMP-2 and TIMP-2 in SF and serum were detected by Western blotting and ELISA, respectively. TRAP in SF from the stifles and serum was significantly increased (p < 0.05) after 3 months. TIMP-2 in SF and serum was significantly decreased (p < 0.05), whereas MMP-2 in SF was significantly increased (p < 0.05) during the progression of OA. Histochemistry revealed an increased number of TRAP-positive cells in tissues from OA-affected joints. Assays measuring TRAP, MMP-2, and TIMP-2 in SF and serum, and methods that detect increased numbers of TRAP-positive cells in the joint tissues can play an important role in identifying the early phases of degenerative changes in canine joint components.

Topics: Acid Phosphatase; Animals; Arthritis, Experimental; Biomarkers; Blotting, Western; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Isoenzymes; Joint Dislocations; Male; Matrix Metalloproteinase 2; Osteoarthritis; Spectrophotometry; Stifle; Synovial Fluid; Tartrate-Resistant Acid Phosphatase; Tissue Inhibitor of Metalloproteinase-2

This study was undertaken to determine the effect of toll-like receptor-3 (TLR3) on the regulation of osteoclastogenic activity in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS). The expression of receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and protein in RA-FLS after TLR3 activation was determined using RT-PCR, real-time PCR, western blot analysis, and immunohistochemistry. Human monocytes were cocultured with RA-FLS that had been prestimulated by the TLR3 ligand polyriboinosinic-polyribocytidylic acid and then stained for tartrate-resistant acid phosphatase (TRAP) activity. Other markers of osteoclasts were measured using RT-PCR and real-time PCR. The expression of TLR3 and RANKL was much higher in the RA synovium than in the osteoarthritis (OA) synovium. TLR3 activation induced RANKL expression in RA-FLS, but not in OA-FLS or in normal skin fibroblasts. TLR3 activation also induced the production of IL-1beta but had no effect on IL-17 or TNF-alpha production in RA-FLS. Inhibition of IL-1beta reversed the TLR3-induced upregulation of RANKL expression. Coculture of human monocytes with TLR3-activated RA-FLS or TLR3 ligand-stimulated human monocytes increased the expression of TRAP, RANK, cathepsin K, calcitonin receptor, and MMP-9, reflecting the differentiation of monocytes into osteoclasts. Our results suggest that TLR3 promotes osteoclastogenesis in the RA synovium both directly and indirectly. TLR3 stimulates human monocytes directly to promote osteoclast differentiation. TLR3 induces RANKL expression indirectly in RA-FLS, and the expression of RANKL promotes the differentiation of osteoclasts in the RA synovium. Targeting the TLR3 pathway may be a promising approach to preventing inflammatory bone destruction in RA.

Topics: Acid Phosphatase; Adult; Aged; Arthritis, Rheumatoid; Coculture Techniques; Cytokines; Female; Fibroblasts; Humans; Immunohistochemistry; Isoenzymes; Male; Middle Aged; Monocytes; Osteoarthritis; Osteoclasts; Osteogenesis; RANK Ligand; Synovial Fluid; Tartrate-Resistant Acid Phosphatase; Toll-Like Receptor 3; Up-Regulation

Tartrate-resistant acid phosphatase (TRAP) is highly expressed in osteoclasts and chondroclasts. The present study investigated changes in TRAP activity after chondrocyte death and cartilage damage, and also evaluated the possible use of TRAP as a diagnostic factor in a model of osteoarthritis. We induced experimental osteoarthritis in beagle dogs and separated chondrocytes from articular cartilage using an enzyme probe. Chondrocyte death was induced by proteasome inhibition and TRAIL treatment, and levels of lactate dehydrogenase, reactive oxygen species (ROS), caspase activation and TRAP activity were measured in the chondrocytes and synovial fluid. Proteasome inhibition and TRAIL treatment significantly enhanced chondrocyte death via caspase-8 activation and ROS generation in the primary cultured canine chondrocytes. TRAP activity was highly increased in damaged chondrocytes, but was decreased by blocking chondrocyte death using caspase inhibition or an ROS scavenger. In the synovial fluid of osteoarthritic dogs, TRAP activity as well as caspase activation and ROS levels were higher than those in the normal joint. Our study demonstrated that TRAP is activated by apoptosis and oxidative stress in primary cultured chondrocytes and osteoarthritic joints and also suggests that TRAP may be used as a diagnostic biomarker for detection of cartilage-related diseases, including osteoarthritis.

Topics: Acid Phosphatase; Animals; Arthritis, Experimental; Cartilage, Articular; Caspase 8; Cell Death; Cells, Cultured; Chondrocytes; Dogs; Female; Isoenzymes; Osteoarthritis; Reactive Oxygen Species; Synovial Fluid; Tartrate-Resistant Acid Phosphatase; TNF-Related Apoptosis-Inducing Ligand

Human serum contains two related isoforms of TRACP: TRACP 5a and TRACP 5b. Serum TRACP 5a protein is increased in about one third of rheumatoid arthritis (RA) sera. This study was undertaken to examine the significance of serum TRACP isoforms 5a and 5b as disease markers of inflammation and bone destruction in RA. One hundred eighteen patients were recruited including 50 with RA (25 with nodules), 26 with osteoarthritis (OA), and 42 with other rheumatic diseases. Twenty-six healthy adults served as controls. Serum TRACP 5a activity, TRACP 5a protein, and TRACP 5b activity were determined by in-house immunoassays. C-reactive protein (CRP) was determined by in-house immunoassay using commercial antibodies and CRP. Other commercial markers included bone-specific alkaline phosphatase (BALP), C-telopeptides of type-I collagen (ICTP), cartilage glycoprotein-39 (YKL-40), and IgM rheumatoid factors (IgM-RF). Mean TRACP 5a protein was significantly elevated only in RA compared with healthy controls and other disease groups. TRACP 5a protein correlated significantly only with IgM-RF in RA. Among RA patients, mean TRACP 5a protein and IgM RF were significantly higher in nodule formers. In contrast, TRACP 5b activity was slightly elevated in RA and correlated with BALP, ICTP, and YKL-40 but not with IgM-RF or CRP. Mean TRACP 5b activity was no different in RA patients with or without nodules. TRACP isoforms could be useful disease markers in RA; TRACP 5a protein may be a measure of systemic inflammatory macrophage burden and disease severity. TRACP 5b activity is a marker for osteoclast number and perhaps local or systemic bone destruction.

Topics: Acid Phosphatase; Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Biomarkers; Female; Humans; Immunohistochemistry; Isoenzymes; Male; Middle Aged; Multivariate Analysis; Osteoarthritis; Regression Analysis; Tartrate-Resistant Acid Phosphatase

Cathepsin K and MMP-9 are considered to be the most abundant proteases in osteoclasts. TRAP is a marker for osteoclasts, and there is increasing evidence of its proteolytic role in bone resorption. RANKL is a recently discovered regulator of osteoclast maturation and activity and induces expression of many genes. This study compared cathepsin K, MMP-9, TRAP, RANKL, OPG, and osteocalcin gene expression in the proximal femur of patients with osteoarthritis with that of patients with femoral neck fracture. Fifty-six patients undergoing arthroplasty because of osteoarthritis or femoral neck fracture were included in the study. Total mRNA was extracted from the bone samples obtained from the intertrochanteric region of the proximal femur. Real-time RT-PCR was used to quantify CTSK (cathepsin K), MMP-9 (matrix metalloproteinase 9), ACP5 (TRAP), TNFSF11 (RANKL), TNFRSF11B (OPG), and BGLAP (osteocalcin) mRNAs. The levels of mRNAs coding for MMP-9 and osteocalcin indicated higher expression in the osteoarthritic group (P = 0.011, P = 0.001, respectively), whereas RANKL expression and the ratio RANKL/OPG were both significantly lower in the osteoarthritic group than in the fracture group. Expression of cathepsin K, MMP-9, and TRAP relative to RANKL was significantly higher in the osteoarthritic group. Ratios of all three proteolytic enzymes relative to formation marker osteocalcin were higher in the fracture group. Gene expression of cathepsin K, MMP-9, TRAP, RANKL, OPG, and osteocalcin and the association between their mRNA levels pointed to higher bone resorption and bone formation in osteoarthritis, differences in balance between them, and differences in regulation of bone resorption in osteoarthritic and osteoporotic bone.

Topics: Acid Phosphatase; Aged; Aged, 80 and over; Bone Resorption; Cathepsin K; Cathepsins; Female; Gene Expression Regulation; Humans; Isoenzymes; Male; Matrix Metalloproteinase 9; Osteoarthritis; Osteocalcin; Osteoporosis; Osteoprotegerin; RANK Ligand; Tartrate-Resistant Acid Phosphatase

Vascular endothelial growth factor (VEGF) plays an essential role in the angiogenesis of growing cartilage. Although VEGF expression in cartilage vanishes in normal adults, VEGF is known to be expressed in chondrocytes of osteoarthritic (OA) cartilage. As little information is available on the VEGF expression in the cartilage of OA-like lesions of the temporomandibular joint (TMJ), VEGF expression in the condylar cartilage of TMJs of rats affected with OA was examined. To evoke OA, mechanical stress was applied by forced jaw opening for 10 or 20 days. After 20 days, marked OA-like lesions were observed in the condyle. VEGF was expressed in the chondrocytes of the mature and hypertrophic cell layers of the intermediate and posterior region of the condyle. The percentage of VEGF immunopositive chondrocytes significantly increased with the period of applied mechanical stress. Furthermore, tartrate-resistant acid phosphatase (TRAP) staining of the condylar cartilage showed significant increment of osteoclasts in the mineralized layer subjacent to the hypertrophic layer where high VEGF expression could be detected. The results suggest that VEGF plays an important role in the progression of OA.

Topics: Acid Phosphatase; Animals; Autocrine Communication; Chondrocytes; Immunohistochemistry; Isoenzymes; Male; Osteoarthritis; Osteoclasts; Paracrine Communication; Rats; Rats, Wistar; Staining and Labeling; Tartrate-Resistant Acid Phosphatase; Temporomandibular Joint; Vascular Endothelial Growth Factors

Our purpose was to develop a specific immunoassay for tartrate-resistant acid phosphatase (TRACP) 5b using naphthol ASBI phosphate (N-ASBI-P) as a selective substrate for isoform 5b and heparin as a selective inhibitor of isoform 5a.. Serum TRACP 5a and 5b and recombinant TRACP 5a were used to optimize and standardize the immunoassay for specificity, linearity, analytical recovery, sensitivity and reproducibility. Serum N-telopeptide cross-links (NTX) and bone alkaline phosphatase (bone ALP) were also measured. The clinical sensitivity and specificity were assessed in healthy control subjects and patients with osteoarthritis (OA), rheumatoid arthritis (RA) and endstage renal disease (ESRD).. TRACP 5b specificity was achieved at pH 6.3 with 2.5 mmol/l substrate and 25 U/ml heparin. Isoform 5b specificity was increased over our original immunoassay using 4-nitrophenyl phosphate (4-NPP) without heparin. The alternative immunoassay was linear with 110% analytical recovery and no serum matrix effects. The average intra-assay error was 10.65%; the average inter-assay error was 10.11% for values of 1-3 U/l and 6.5% for values of 7-11 U/l. Mean serum TRACP 5b in OA and RA were not significantly different from control using either immunoassay. Mean serum TRACP 5b was significantly increased in ESRD with both immunoassays. Serum TRACP 5b levels correlated significantly with NTX and bone ALP in the disease groups, but not always in the control group.. This alternative immunoassay for TRACP 5b activity is highly specific. It should have applications in evaluating patients with bone disease and will improve our understanding of the biological significance of TRACP 5b expression in health and disease.

Topics: Acid Phosphatase; Adult; Aged; Aged, 80 and over; Anticoagulants; Arthritis, Rheumatoid; Biomarkers; Bone and Bones; Chromatography, Ion Exchange; Enzyme Inhibitors; Female; Fluorescent Dyes; Heparin; Humans; Hydrolysis; Immunoassay; Indicators and Reagents; Isoenzymes; Kidney Failure, Chronic; Male; Middle Aged; Neuraminidase; Organophosphorus Compounds; Osteoarthritis; Recombinant Proteins; Tartrate-Resistant Acid Phosphatase; Trypsin

To investigate the expression of extracellular matrix metalloproteinase inducer (EMMPRIN) at sites of joint destruction in rheumatoid arthritis (RA) and to correlate it with the production of matrix metalloproteinases (MMPs).. Reverse transcription-polymerase chain reaction was performed to study the existence of EMMPRIN in synovial tissue derived from RA and osteoarthritis (OA) patients. In situ hybridization with a human complementary DNA specific for EMMPRIN and immunohistochemistry were performed to characterize the EMMPRIN-expressing cells at sites of joint destruction, including bone. Northern blot analysis was performed to detect the level of expression of EMMPRIN messenger RNA (mRNA) in synovial tissue. The production of MMP-1 and MMP-3 by synovial tissue from RA patients was examined by enzyme-linked immunosorbent assay.. Expression of EMMPRIN mRNA was detected in synovium from 9 of 11 patients with RA and 1 of 5 patients with OA. The presence of mRNA encoding EMMPRIN was recognized in the invasive synovium at sites of joint destruction in RA but not OA. Fibroblast-like synovial cells and granulocytes were demonstrated to express EMMPRIN mRNA. MMP-1 and MMP-3 production by synovial tissue was correlated with levels of expression of EMMPRIN mRNA, as detected by Northern blotting.. The expression of EMMPRIN stimulates the production of MMP-1 and MMP-3 in the synovial tissue of affected joints in RA. The results of this study suggest that EMMPRIN may be one of the important factors in progressive joint destruction in RA.

Topics: Acid Phosphatase; Antigens, CD; Antigens, Neoplasm; Arthritis, Rheumatoid; Basigin; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gene Expression; Humans; In Situ Hybridization; Isoenzymes; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Membrane Glycoproteins; Osteoarthritis; RNA, Messenger; Synovial Membrane; Tartrate-Resistant Acid Phosphatase

Our objective was to evaluate the significance and source of serum tartrate-resistant acid phosphatase (TRACP) in patients with rheumatoid arthritis (RA).. Thirty-five RA, 32 osteoarthritis (OA) and 16 control subjects were studied. Serum TRACP-5b activity and total TRACP protein were determined by immunoassay. TRACP isoforms were analyzed by non-denaturing polyacrylamide gel electrophoresis (PAGE). Serum bone alkaline phosphatase (BAP), cross-linked N-terminal telopeptides (NTx), and C-terminal telopeptides (ICTP) of type I collagen were estimated as markers of bone turnover. C-reactive protein (CRP) was measured as a marker of chronic inflammation. Macrophages and dendritic cells (DC) were developed from peripheral blood monocytes. Cell lysates and culture supernatants were analyzed for TRACP isoforms by immunoassay and PAGE.. In RA, mean TRACP-5b activity was normal, but median total TRACP protein was increased twofold (p<0.001). In OA, TRACP-5b activity and protein were normal. In RA, TRACP-5b activity correlated weakly with ICTP (r=0.56) while TRACP protein levels correlated weakly with NTx (r=0.43). Additionally, TRACP protein, but not TRACP-5b activity correlated significantly with CRP (r=0.42). Macrophage and DC lysates contained TRACP-5b, while tissue culture supernatants contained TRACP-5a.. Increased total TRACP protein in RA sera was probably due to TRACP-5a and not derived from osteoclasts. Rather, it could be a secreted product of inflammatory macrophages and DC.

Topics: Acid Phosphatase; Adolescent; Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Biomarkers; C-Reactive Protein; Cells, Cultured; Dendritic Cells; Electrophoresis, Polyacrylamide Gel; Humans; Immunoassay; Inflammation; Isoenzymes; Macrophages; Middle Aged; Osteoarthritis; Sensitivity and Specificity; Tartrate-Resistant Acid Phosphatase

A macrophage infiltrate is commonly found in enlarging subchondral cysts in osteoarthrosis (OA) and the surrounding bone. To determine whether osteoclast differentiation by these cells contributes to the increase in the number of osteoclasts and bone resorption that accompanies OA cyst enlargement, we isolated macrophages from the wall of OA cysts and co-cultured them with osteoblast-like UMR106 cells in the presence or absence of 1,25(OH)2D3 and M-CSE After 14 days of incubation, co-cultures of UMR106 cells and cyst-derived macrophages showed evidence of osteoclast differentiation by expression of TRAP, VNR and formation of numerous lacunar pits. We found that, unlike osteoclast precursors in monocyte and other tissue macrophage populations, the addition of M-CSF to medium is not required for osteoclast differentiation. Our findings suggest that macrophage-osteoclast differentiation is one means whereby the osteolysis associated with the enlargement of OA cysts could be effected.

Topics: Acetabulum; Acid Phosphatase; Aged; Bone Cysts; Bone Resorption; Calcitriol; Cell Differentiation; Coculture Techniques; Female; Femur Head; Humans; Isoenzymes; Macrophage Colony-Stimulating Factor; Macrophage-1 Antigen; Macrophages; Male; Middle Aged; Osteoarthritis; Osteoclasts; Receptors, Vitronectin; Tartrate-Resistant Acid Phosphatase

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Electromagnetic Phenomena; Glucuronidase; Humans; Lysosomes; Osteoarthritis; Rabbits

Chronic immune responses and inflammatory reactions in rheumatoid arthritis (RA) often cause severe destruction of cartilage and bone, but its mechanism is still a matter of controversy. We reported that interleukin-6 (IL-6) alone does not induce osteoclast formation, but soluble interleukin-6 receptors (sIL-6R) triggered the formation in the presence of IL-6 in cocultures of murine osteoblastic cells and bone marrow cells. In this study, we examined the involvement of sIL-6R and IL-6 in joint destruction in patients with RA. Although the frequency of patients having osteoclast-like multinucleated cells in synovium derived from the knee joint was not significantly different between RA (65%) and osteoarthritis (OA) patients (43%), the number of osteoclast-like cells found in the synovium was greater in the former than in the latter. Multinucleated cells obtained from RA synovium expressed the osteoclast-specific phenotype such as tartrate-resistant acid phosphatase, carbonic anhydrase II, vacuolar proton-ATPase and vitronectin receptors at similar levels to those from a human giant cell tumor of bone. The concentration of both IL-6 and sIL-6R was significantly higher in the synovial fluids from patients with RA than with OA. The concentration of IL-6 and sIL-6R correlated well with the roentgenologic grades of joint destruction. Dose-response curves for human IL-6 and human sIL-6R in inducing osteoclast-like cell formation in cocultures indicated that the RA synovial fluids contained sufficient IL-6 and sIL-6R to induce osteoclastogenesis. When synovial fluids from RA and OA patients were added to the cocultures, some of the RA synovial fluids containing high levels of IL-6 and sIL-6R stimulated osteoclast-like cell formation, which was strikingly inhibited by adding anti-IL-6R antibody simultaneously. These results suggest that IL-6 in the RA synovial fluids is at least in part responsible for joint destruction in the presence of sIL-6R through osteoclastogenesis.

Topics: Acid Phosphatase; Adult; Aged; Aged, 80 and over; Animals; Antigens, CD; Arthritis, Rheumatoid; Cells, Cultured; Female; Humans; Interleukin-6; Male; Mice; Middle Aged; Osteoarthritis; Osteoblasts; Osteoclasts; Phenotype; Receptors, Interleukin; Receptors, Interleukin-6; Solubility; Synovial Fluid; Synovial Membrane

Animal model and in vitro cultures suggest that osteoclasts and cells of the mononuclear phagocyte system share a common precursor. However, the human osteoclast precursor has not been positively identified. We attempted to identify the precursor in situ by using a number of osteoclast- and macrophage-selective markers, together with the expression of osteopontin mRNA, previously shown to be abundant in human osteoclasts. Sections of osteophytic bone and a panel of inflammatory connective tissues were processed for in situ hybridization; serial sections were analyzed for tartrate-resistant acid phosphatase (TRAP) and nonspecific esterase (NSE) activity, selective cytochemical markers for the osteoclast and cells of the macrophage/monocyte lineage, respectively. The murine anti-human osteoclast monoclonal antibodies 23C6 (vitronectin receptor) and C35 (osteoclast-selective) were used to further identify the osteoclast phenotype. We compared osteoclasts, giant cells, and their respective putative mononuclear precursors. At resorption sites within osteophytic bone, osteopontin mRNA was expressed in osteoclasts and a distinct population of TRAP+, NSE- mononuclear cells. Adjacent clusters of mononuclear cells were TRAP- and NSE+ or were active for both enzymes; these cells demonstrated variable expression of osteopontin mRNA. In the inflammatory connective tissues, abundant macrophage-like cells (NSE+/TRAP-) did not express osteopontin mRNA. However, TRAP+ mononuclear cells observed among clusters of NSE+ cells did express osteopontin mRNA. At these sites, clusters of putative macrophage polykaryons removing fragments of bone debris were observed. These giant cells and associated mononuclear cells were NSE- and distinctly TRAP+, and expressed osteopontin mRNA, C35, and 23C6 (human osteoclast) reactivity. Therefore, cells involved in the remodeling (resorption) of bone or the removal of bone debris, together with their immediate precursors, switch from being NSE+/TRAP- to NSE-/TRAP+ cells that express osteopontin mRNA. We propose that the clusters of NSE+/TRAP- mononuclear cells represent the immature osteoclast precursor. In support of this, TRAP+/NSE+ cells were occasionally observed in both tissues, representing an intermediate stage in differentiation. These results further suggest that cells of the mononuclear phagocyte lineage within bone and inflammatory connective tissue have the potential to differentiate into osteoclasts.

Topics: Acid Phosphatase; Bone and Bones; Cell Differentiation; Connective Tissue; Esterases; Giant Cells; Humans; Immunohistochemistry; Isoenzymes; Osteoarthritis; Osteoclasts; Osteopontin; Phenotype; RNA, Messenger; Sialoglycoproteins; Synovial Membrane; Tartrate-Resistant Acid Phosphatase

Osteopontin is a phosphorylated glycoprotein believed to be secreted by osteoblasts and deposited into the bone matrix to facilitate osteoclasts adhesion or to initiate osteoid mineralization. Previously we have presented contradictory evidence that osteoclasts express osteopontin mRNA in human remodeling bone. The aim of this study was to ascertain whether osteoclasts synthesize and deposit osteopontin in resorption lucunae. We characterized expression of osteopontin mRNA and protein expression in both intramembranous and endochondral ossification, as well as remodeling bone, in the human osteophyte. Osteopontin mRNA was expressed in osteoclast with tartrate-resistant acid phosphatase (TRAP) positivity within resorption lacunae. The osteoclasts and immediate resorption surfaces also expressed osteopontin. However, osteopontin mRNA and protein were weak (transient) or undetectable in osteoblasts at adjacent bone formation sites; no osteopontin expression was observed in the osteoid, although occasional reactivity was observed in osteocytes and the mineral-osteoid interface. In contrast, osteopontin was highly expressed in the osteoblasts and matrix of woven bone during intramembranous and endochondral ossification. The matrix expression correlated with mineralization; however, in some instances osteopontin deposition was observed prior to mineralization. Similarly, osteopontin expression was evident in cartilage matrix, solely at foci of mineralization. Chondroclasts expressed osteopontin mRNA and protein: the surfaces of resorbed calcified cartilage also expressed osteopontin. Abnormal, unmineralized matrices apparently lacked deposited osteopontin, but were nevertheless resorbed by osteoclasts; the osteoclasts and resorbed surfaces expressed no osteopontin protein. That osteoclasts are responsible for the deposition of osteopontin was confirmed in vitro, whereby resorption pits in whale dentine and bovine bone slices, produced by isolated human osteoclasts, contained deposited osteopontin. Osteopontin may facilitate the adhesion (or detachment) of the osteoclast to the bone surface. Alternatively, the possibility that osteopontin may act as a postresorptive signal to recruit osteoblasts, or to polarize and direct the mineralization of the formed osteoid, is discussed.

Topics: Acid Phosphatase; Bone Remodeling; Bone Resorption; Calcification, Physiologic; Cartilage; Cell Adhesion; DNA, Complementary; Femur Head; Histocytochemistry; Humans; Immunohistochemistry; In Situ Hybridization; Osteoarthritis; Osteoclasts; Osteopontin; Promoter Regions, Genetic; RNA, Messenger; Sialoglycoproteins; Tartrates

The influx of cells into the synovial intima in rheumatoid joints may include osteoclasts and their precursors. The distribution of osteoclast markers--namely, tartrate resistant acid phosphatase activity and the expression of vitronectin receptor (shown with monoclonal antibodies 13C2 and 23C6)--was therefore examined in synovium obtained from patients with rheumatoid (RA) or degenerative (OA) arthritis. Tartrate resistant acid phosphatase positive cells were found in frozen sections of 60% (n = 30) of RA and 69% (n = 29) of OA synovial membranes. Whereas all synovia tested (four RA, four OA) showed diffuse staining of the lining cells with 13C2, 55% (n = 11) of RA and 57% (n = 14) of OA synovial membranes contained isolated cells stained with 23C6 scattered throughout the tissue. In cultures of synovial cells, tartrate resistant acid phosphatase positive, multinuclear, and 23C6 positive cells were found; these cells did not, however, form resorption pits on bone slices. The results show that fully differentiated osteoclasts are uncommon in synovium from patients with either degenerative or inflammatory arthropathies.

Topics: Acid Phosphatase; Arthritis, Rheumatoid; Bone Resorption; Cells, Cultured; Humans; Immunochemistry; Integrins; Microscopy, Electron, Scanning; Osteoarthritis; Receptors, Cytoadhesin; Receptors, Vitronectin; Synovial Membrane

Giant cells are commonly present in inflamed synovium, often in close association with the intimal layer. The nature of these multinucleate cells has been reassessed using new cytochemical and immunochemical techniques.. Cryostat sections of non-inflamed, rheumatoid arthritic and osteoarthritic synovia were analysed for the presence of CD68 and non-specific esterase, markers associated with macrophages; activity of uridine diphosphoglucose dehydrogenase, associated with fibroblast-like synoviocytes; and tartrate resistant acid phosphatase and the vitronectin receptor subunit CD51, associated with osteoclasts.. Giant cells were not seen in non-inflamed tissue. In diseased tissue giant cells in the intimal layer fell into two major groups: CD68 negative or dull cells with high uridine diphosphoglucose dehydrogenase (UDPGD) activity suggestive of true synoviocyte polykaryons; and CD68 positive cells with low UDPGD activity suggestive of macrophage polykaryons. The two groups were seen in samples from patients with rheumatoid arthritis (RA) and patients with osteoarthritis (OA), but the former were more prominent in OA and the latter in RA. Most CD68 positive giant cells also showed tartrate resistant acid phosphatase activity and prominent expression of CD51. As such they were histochemically indistinguishable from osteoclasts, but their bone resorbing capacity remains unknown.. Giant cells in arthritic synovium appear to be of two types, one related to true synoviocytes and one to macrophages.

Topics: Acid Phosphatase; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Arthritis, Rheumatoid; Giant Cells; Humans; Immunohistochemistry; Osteoarthritis; Receptors, Cytoadhesin; Receptors, Vitronectin; Rheumatic Diseases; Synovial Membrane; Uridine Diphosphate Glucose Dehydrogenase

This study evaluates the progression of osteoarthritis (OA) in the adult New Zealand white rabbit temporomandibular joint following unilateral disc perforation. Thirty-seven animals were divided into five groups: control (n = 8), 6-week sham (n = 5), and experimental 6-, 12-, and 24-weeks (n = 8 each). Quantitative data was examined with two-way analysis of variance, and followed by Scheffe pair-wise comparisons. Transmission electron microscopy, acid phosphatase [AcP] activity, uronic acid content, and gross morphologic analysis indicated that disc perforation induced remodeling activity and degenerative changes in the condylar cartilage and bone as early as 6 weeks postoperatively. AcP activity of homogenized cartilage samples was significantly increased in experimental joints versus the side that did not undergo surgery at 6 and 12 weeks (P < .05). Uronic acid content was significantly greater in experimental joints versus the side that did not undergo surgery at 6 weeks (P < .05). Heightened cellular activity was present in the deep zone of osteoarthritic fibrocartilage of the 6- and 12-week experimental groups. Degenerating chondrocytes appeared to contain greater proportions of intracytoplasmic filaments and lysosome-like bodies. Disc perforation provided the impetus for degenerative or remodeling changes in the condylar cartilage of experimental joints, and is consistent with secondary OA. These dynamic events were most significant in the deep zone of articular fibrocartilage.

Topics: Acid Phosphatase; Animals; Body Water; Bone Remodeling; Calcinosis; Cartilage, Articular; Collagen; Cytoplasm; Elastic Tissue; Endoplasmic Reticulum; Glycosaminoglycans; Golgi Apparatus; Mandibular Condyle; Microscopy, Electron; Osteoarthritis; Rabbits; Temporomandibular Joint; Time Factors; Tissue Adhesions; Uronic Acids

Leukemia inhibitory factor/differentiation-stimulating factor (LIF/D-factor), expression of its mRNA, and possible roles in bone metabolism were studied in murine primary and clonal osteoblast-like cells. Local bone-resorbing factors such as IL-1, TNF alpha, and LPS strongly induced expression of LIF/D-factor mRNA in both clonal MC3T3-E1 cells and primary osteoblast-like cells. Neither parathyroid hormone nor 1 alpha,25-dihydroxyvitamin D3 stimulated expression of LIF/D-factor mRNA. LIF/D-factor per se did not stimulate expression of its own mRNA. Appreciable amounts of LIF/D-factor were detected in synovial fluids from rheumatoid arthritis (RA) patients but not in those with osteoarthritis (OA). Simultaneous treatment with LIF/D-factor, IL-1, and IL-6 at the concentrations found in synovial fluids from RA patients greatly enhanced bone resorption, though these cytokines did not stimulate bone resorption when separately applied. This suggests that LIF/D-factor produced by osteoblasts is in concert with other bone-resorbing cytokines such as IL-1 and IL-6 involved in the bone resorption seen in the joints of RA patients. LIF/D-factor specifically bound to MC3T3-E1 cells with an apparent dissociation constant of 161 pM and 1,100 binding sites/cell. LIF/D-factor dose-dependently suppressed incorporation of [3H]thymidine into MC3T3-E1 cells. In addition, it potentiated the alkaline phosphatase activity induced by retinoic acid, though LIF/D-factor alone had no effect on enzyme activity. These results suggest that LIF/D-factor is involved in not only osteoclastic bone resorption but also osteoblast differentiation in conjugation with other osteotropic factors.

Topics: Acid Phosphatase; Animals; Arthritis, Rheumatoid; Bone and Bones; Bone Resorption; Cells, Cultured; Dose-Response Relationship, Drug; Female; Growth Inhibitors; Interleukin-1; Interleukin-6; Leukemia Inhibitory Factor; Lipopolysaccharides; Lymphokines; Mice; Mice, Inbred C57BL; Osteoarthritis; Osteoblasts; Pregnancy; RNA, Messenger; Synovial Fluid; Tretinoin; Tumor Necrosis Factor-alpha

The purpose of this study was to correlate histologic findings in temporomandibular joint (TMJ) condyles and discs with their macroscopic appearance at surgery. The 24 patients with internal derangement of the joint included 20 women and 4 men (mean age, 37 years; range, 18 to 61 years). The tissue lesions varied in degree from mild soft-tissue fraying and bone remodeling to extensive resorption and new cartilage and bone formation with high phosphatase enzyme activities, and even to loss of articular soft tissue and breakdown of cortical bone. Reactions may arise in the hard tissues before they occur in the articular surface layers.

Topics: Acid Phosphatase; Adolescent; Adult; Alkaline Phosphatase; Cartilage, Articular; Female; Glycosaminoglycans; Humans; Male; Mandibular Condyle; Middle Aged; Osteoarthritis; Proteoglycans; Staining and Labeling; Synovial Membrane; Temporomandibular Joint Disorders

Alkaline phosphatase (AP), acid phosphatase and myeloperoxidase (MP) activity and the level of cation protein (CP) in blood and synovial fluid (SF) neutrophils were studied and compared in 54 patients with rheumatoid arthritis (RA) and in 22 patients suffering from primary osteoarthrosis deformans (OAD) combined with reactive synovitis. As compared to the patients with OAD, the patients with RA manifested a significant rise of AP, acid phosphatase and MP activity together with a decrease of the level of CP in blood neutrophils. Meanwhile in SF neutrophils from the patients with RA, all the parameters appeared higher than in OAD and were lower that in blood neutrophils in both the groups. As compared to the routine biochemical and cytological tests, the diagnostic information content of the cytochemical parameters of blood neutrophils (AP, acid phosphatase) and SF neutrophils (AP, acid phosphatase, MP) from the patients with RA (against the patients suffering from OAD) was noticeably higher.

Topics: Acid Phosphatase; Adult; Aged; Alkaline Phosphatase; Arthritis, Rheumatoid; Blood Proteins; Clinical Enzyme Tests; Diagnosis, Differential; Female; Histocytochemistry; Humans; Male; Middle Aged; Neutrophils; Osteoarthritis; Peroxidase; Synovial Fluid

Topics: Acid Phosphatase; Adult; Aged; Arthritis, Rheumatoid; Endodeoxyribonucleases; Female; Humans; Osteoarthritis; Synovial Fluid

In order to investigate the relationship between the synovial inflammatory response and lysosomal enzyme activity in osteoarthritis, synovial specimens obtained from 19 osteoarthritic patients and control specimens from 10 normal joints were analysed for cathepsin D and acid phosphatase enzyme levels. In estimating enzyme activities methods previously developed for quantitative enzyme determination in cartilage were modified and applied to synovial tissues for the first time. In addition, samples of osteoarthritic synovium were histologically graded according to their degree of inflammation. It was found that in osteoarthritic synovium cathepsin D and acid phosphatase, which is a general marker for lysosomal enzyme activity, were significantly increased compared with normal control synovium. No significant relationship was found between the degree of synovial tissue inflammation and lysosomal enzyme activity.

Topics: Acid Phosphatase; Cathepsin D; Humans; Lysosomes; Osteoarthritis; Synovial Membrane; Synovitis

In osteoarthritis, despite increased matrix synthesis, there is a reduction of both major matrix components, proteoglycan and collagen. This study suggests that this is the result of enhanced degradative activity intrinsic to the cartilage. Because osteoarthritis is a focal disease, histologic controls were used to measure the severity in different areas of the cartilage and in different specimens. Neutral proteoglycanase and collagenase were both described in human cartilage, and their levels matched the severity of the disease as did acid phosphatase, a marker of lysosomal enzymes. Articular cartilage collagenase has an inhibitor in the cartilage and has negligible activity in normal cartilage. This was found not to be a lysosomal enzyme. A model of osteoarthritis was studied and found to have the same biochemical pattern as human disease. Using this method, inhibitors of degradative enzymes were used as a treatment. The chelator of metallic cations EDTA was found to have a significant effect on reduction of degradative enzyme activity and altered the arthritic process.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Cytidine; Disease Models, Animal; Edetic Acid; Egtazic Acid; Endopeptidases; Glycosaminoglycans; Humans; Hydrogen-Ion Concentration; Metalloendopeptidases; Microbial Collagenase; Osteoarthritis; Rabbits; Thymidine

We studied the light microscopic, ultrastructural, and cytochemical characteristics of the temporomandibular joints of male ICR mice, from early neonatal life until they reached senescence, when spontaneous osteoarthritis is a common phenomenon. Aging of mandibular condylar cartilage was accompanied by decreasing total proteoglycan content and by an unmasking of collagen fibers, with no shift in collagen type. Fibronectin was also commonly present on the articular surface of specimens from old animals. Chondrocytes of aged mice contained an increased number of lysosomes, and their adjacent matrix vesicles reacted positively for acid phosphatase and arylsulfatase, but not for alkaline phosphatase. Such vesicles were also found to be devoid of calcium complexes and, thus, did not appear to be involved in the mineralization process. Similar age-related changes have been described in human mandibular condyles; hence, the male ICR mouse could serve as a useful model for studies of spontaneous osteoarthritis in the human mandibular joint.

Topics: Acid Phosphatase; Aging; Animals; Arylsulfatases; Cartilage, Articular; Collagen; Fibronectins; Histocytochemistry; Male; Mice; Mice, Inbred ICR; Osteoarthritis; Proteoglycans; Temporomandibular Joint

A partial lateral meniscectomy procedure has been developed for the induction of a predictable and reproducible degenerative joint disease in knees of rabbits. The procedure adopted involves section of the fibular collateral and sesamoid ligaments and removal of 4-5 mm of the anterior lateral meniscus. In most experiments the animals are killed and tissues obtained for histologic examination at 6 weeks. Section of the ligaments alone (with or without penetration of the joint space) did not result in significant pathologic change. Significant degeneration was observed in tibial and femoral cartilage when the meniscus as well as the ligaments were cut, but the most extensive lesions were seen when a piece of the anterolateral meniscus was actually removed. These lesions included fibrillation, ulceration and erosion, "clone" and osteophyte formation, loss of chondrocytes, and loss of safraninophilic staining in the articular cartilage. The incidence and distribution of lesions with time following surgery were also investigated. Lesions were observed as early as 1-2 weeks post-surgery and increased in number and severity up to 12 weeks. A global scoring system has been devised to permit statistical comparisons of lesion incidence and severity in different groups of rabbits. This scoring system has enabled us to test drug efficacy in the rabbit lateral meniscectomy model of osteoarthritis.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Histocytochemistry; Knee Joint; Ligaments, Articular; Male; Menisci, Tibial; Osteoarthritis; Rabbits

A combination of immunochemical staining for HLA-DR antigens and the histochemical demonstration of enzyme activity has been used to identify specific cell populations in the normal and arthritic synovial lining layers. Such combined staining has revealed that the normal synovial lining contains a proportion of HLA-DR + ve cells, all of which show strong lysosomal enzyme activity. This population is greatly expanded in biopsies from patients with osteoarthritis and these positive cells also express strong ATPase activity. In the rheumatoid synovium five distinct cell types can be identified; all of which are HLA-DR + ve but differ in their morphology and pattern of enzyme activity. Of special interest was the discovery that a small but significant proportion of these cells have the characteristics of the interdigitating cells of the lymph node paracortex. The relationship between the emergence of these heterogeneous populations and the immunological basis of this inflammatory response is discussed.

Topics: Acid Phosphatase; Adenosine Triphosphatases; Arthritis, Rheumatoid; Carboxylesterase; Carboxylic Ester Hydrolases; Fluorescent Antibody Technique; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Osteoarthritis; Synovial Membrane

Topics: Acid Phosphatase; Aged; Female; Hip Joint; Humans; Methods; Osteoarthritis; Time Factors

Topics: Acid Phosphatase; Adolescent; Adult; Aged; Arthritis, Rheumatoid; Autoimmune Diseases; Female; Humans; Lymphocyte Activation; Male; Middle Aged; Osteoarthritis; Rheumatic Diseases

Topics: Acid Phosphatase; Alkaline Phosphatase; Bone Marrow; Cartilage, Articular; Glycosaminoglycans; Hip Joint; Humans; Osteoarthritis; Phosphates; Pressure; Radiography; Radionuclide Imaging; Synovial Fluid; Synovial Membrane

In order to determine the localization and activity of alkaline and acid phosphatase in the synovial membrane of osteoarthritic hip joints, enzymo-histochemical analyses were performed using Burstone's and Barka & Anderson's methods. Frozen sections of synovial biopsy material from 12 osteoarthritic and 6 control hip joints were studied. Alkaline phosphatase was found located in fibroblasts below the lining cells and in capillaries and precapillary arterioles. Acid phosphatase was seen in the lysosomes in the lining cells. Semiquantitative evaluation by means of initial time determination showed significantly greater activity in osteoarthritic synovia than in the control group. Whilst the increased activity of lysosomal enzymes is presumably implicated in the joint cartilage damage seen in osteoarthritis, the significance of elevated alkaline phosphatase levels is not yet clear.

Topics: Acid Phosphatase; Aged; Alkaline Phosphatase; Female; Hip Joint; Histocytochemistry; Humans; Male; Middle Aged; Osteoarthritis; Synovial Membrane

In order to analyze the presence of alkaline and acid phosphatase activity in osteoarthrotic subchondral bone frozen sections from 24 femoral heads were prepared for semiquantitative analysis, using the enzyme-histochemical methods described by Burstone and by Barka and Anderson. Different areas of the subchondral bone, viz. weight-bearing, nonweight-bearing and osteophytes as well as central regions were analyzed. Furthermore, the cartilagenous changes were determined by histological-histochemical grading. There were wide variations within the same femoral head with significantly greater activity of both alkaline and acid phosphatase in weight-bearing than in nonweight-bearing areas and in subchondral bone than in central regions. Osteophytes excluded, the enzyme activities correlated directly with the severity of the cartilage lesions. These enzyme activities presumably reflect the degree of bone regeneration and bone resorption respectively.

Topics: Acid Phosphatase; Aged; Alkaline Phosphatase; Female; Hip Joint; Humans; Joint Diseases; Male; Osteoarthritis

Topics: Acid Phosphatase; Adult; Aged; Calcium; Connective Tissue; Female; Hexosamines; Humans; Hydroxyproline; Male; Middle Aged; Orosomucoid; Osteoarthritis; Phosphorus

A correlation between increased arylsulfatase activities and decreased sulfated proteoglycan content in human osteoarthritic articular cartilage suggested a possible interrelationship between these parameters. Since we had previously shown that ascorbate caused a decrease in levels of arylsulfatase A and B activities in normal chondrocyte cultures, the validity of the above relationship was examined by measuring the effect of vitamin C on the biosynthesis and distribution of 35S-labeled proteoglycans and arylsulfatase A and B activities in cell extracts of chondrocytes derived from normal and osteoarthritic tissue. Arylsulfatase A and B activities were found to be reduced in the presence of ascorbic acid in all normal and osteoarthritic cell lines examined when measured 3, 6, 10, and 13 days after the introduction of the vitamin in the culture medium. Acid phosphatase activity, on the other hand, was found to be elevated in the presence of ascorbate. The inhibitory effect by ascorbic acid on arylsulfatase activities could be reversed by withdrawing the vitamin from the nutrient medium. Addition of EDTA to the cell extracts before assay also reversed the inhibiton. Sulfated proteoglycan biosynthesis as reflected in 35S-sulfate uptake per milligram of DNA was significantly increased in the presence of ascorbic acid. The distribution of the newly synthesized molecules between the cell layer and medium fractions was altered. In the presence of ascorbate, more deposition into the cell layer of newly synthesized macromolecules occurred. These data suggest an inverse relationship between arylsulfatase activities and the stability of the newly synthesized sulfated proteoglycans in the extracellular matrix.

Topics: Acid Phosphatase; Arylsulfatases; Ascorbic Acid; Cartilage, Articular; Cell Line; Cerebroside-Sulfatase; Chondro-4-Sulfatase; Chondroitin Sulfate Proteoglycans; Edetic Acid; Humans; Osteoarthritis; Proteoglycans; Sulfatases

Sera and synovial fluid were investigated in 45 patients with rheumatoid Arthritis and 50 patients with osteoarthritis in inflammatory exacerbation (control group). The following tests were performed: IgG, IgM, IgA determinations, complement components C3, C3, C4, C3-proactivator, ceruloplasmin, electrophoresis, LDH and total acid phosphatase. 1. Serum levels of the ceruloplasmin, alpha 1, alpha 2 and gamma fractions of electrophoresis are significantly higher in patients with rheumatoid arthritis than in patients with osteoarthritis. 2. Synovial fluid: a) There is a significantly higher concentration of IgG, IgM, IgA, C3-proactivator and total acid phosphatase in the synovial fluid of patients with rheumatoid arthritis. b) C4 is significantly lower in patients with rheumatoid arthritis. c) Both groups were also compared with the help of a point system. Every patient received a plus point when the following criteria were seen: IgM greater than 150 mg/100 ml, C3 greater than 50 mg/100 ml, ceruloplasmin greater than 35 mg/100 ml, alpha 1 greater than 0.21 g%, alpha 2 greater than 0.44 g%, beta greater than 0.60 g% and gamma fraction on electrophoresis greater than 0.90 g%. Another point was added if the criteria ceruloplasmin greater than 22 mg/100 ml and C4 less than 17 mg/100 ml were simultaneously seen. With the help of this points system 48 out of the 50 osteoarthritis patients (96%) received zero points, one received 1 point and one 2 points, as opposed to the patients with rheumatoid arthritis where 35 out of 45 (78%) received one or more points. d) The differentation is not improved through additional testing of the rheumatic factors.

Topics: Acid Phosphatase; Arthritis, Rheumatoid; Blood; Ceruloplasmin; Complement System Proteins; Humans; Immunoglobulins; L-Lactate Dehydrogenase; Osteoarthritis; Rheumatoid Factor; Synovial Fluid

In a system consisting of synovial samples taken surgically and maintained in culture, the authors studied the regulation of enzyme synthesis by taking acid phosphatase as the test enzyme. By means of double labelling and the use of protein synthesis inhibitors, they were able to demonstrate that this synthesis was stimulated by the addition of rheumatoid factor to the culture medium and that it depended on the transcription activity of the genome and on the translation of the information molecules.

Topics: Acid Phosphatase; Animals; Carbon Radioisotopes; Cattle; Chromatography, Gel; Culture Techniques; Cycloheximide; Dactinomycin; Humans; Isoenzymes; Lysosomes; Osteoarthritis; Protein Biosynthesis; Synovial Membrane

Synovial cells from patients with rheumatoid arthritis (RA) when grown in vitro in media supplemented with 20% fetal calf serum failed to show any difference in growth rate, life span, uptake of tritiated thymidine or cellular and nuclear characteristics when compared with synovial cells from patients with osteoarthritis or other joint diseases grown similarly in 20% serum enriched medium. There was also no evidence that lymphocytes and/or sera from RA patients were more cytotoxic to autologous synovial cells than sera and/or lymphocytes from OA patients. It is unlikely that antisynovial antibodies or lymphocytes from RA patients act as triggers for synovial damage.

Topics: Acid Phosphatase; Arthritis, Rheumatoid; Autoradiography; Cells, Cultured; Cytoplasm; Cytotoxicity Tests, Immunologic; Fibroblasts; Fluorescent Antibody Technique; Humans; Lymphocytes; Microscopy, Electron; Osteoarthritis; Photomicrography; Synovial Fluid; Thymidine

Section of the medial collateral and both cruciate ligaments combined with resection of the medial meniscus in rabbit knees caused instability and during the ensuing six months these knees showed progressive histological changes similar to those of human osteoarthritis. Biochemical analysis of the cartilage from such knee joints demonstrated a decrease in proteoglycan, an increase in acid phosphatase, and increases in the rates of synthesis of protein and glycosaminoglycan. These findings, which are quite consistent with those in human osteoarthritis, suggest that this animal model may be of value in the study of the pathogenesis and treatment of human disease.

Topics: Acid Phosphatase; Animals; Cartilage, Articular; Disease Models, Animal; Femur; Glycine; Glycosaminoglycans; Hexosamines; Hydroxyproline; Knee Joint; Osteoarthritis; Protein Biosynthesis; Proteoglycans; Rabbits; Tritium

Topics: Acid Phosphatase; Aminosalicylic Acids; Arthritis, Rheumatoid; Chondrocalcinosis; Enzymes; Esterases; Exudates and Transudates; Glucuronidase; Histocytochemistry; Humans; Knee Joint; Lymphocyte Activation; Lymphocytes; Monocytes; Naphthalenes; Osteoarthritis; Osteochondritis; Punctures; Rheumatic Diseases; Synovial Fluid

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Cartilage, Articular; Cathepsins; Centrifugation; Chickens; DNA; Ear, External; Glycosaminoglycans; Haplorhini; Hip Joint; Humans; Hyaluronoglucosaminidase; Hydrogen-Ion Concentration; In Vitro Techniques; Microbial Collagenase; Osteoarthritis; Rabbits

Topics: Acid Phosphatase; Adult; Aged; Alanine Transaminase; Alkaline Phosphatase; Arthritis, Rheumatoid; Blood Sedimentation; Female; Hemoglobinometry; Histocytochemistry; Humans; Inflammation; Male; Middle Aged; Neutrophils; Nucleotidases; Osteoarthritis; Serum Albumin; Synovial Fluid; Synovial Membrane; Synovitis

Topics: Acid Phosphatase; Adolescent; Aged; Cartilage, Articular; Child; Histocytochemistry; Humans; Hydrogen-Ion Concentration; Middle Aged; Osteoarthritis; Surface-Active Agents

Topics: Acid Phosphatase; Adult; Aged; Arthritis, Rheumatoid; Female; Humans; Immunoglobulin A; Immunoglobulin D; Immunoglobulin G; Immunoglobulin M; Leukocyte Count; Lymphocytes; Male; Middle Aged; Monocytes; Nucleotidases; Osteoarthritis; Spondylitis, Ankylosing; Synovial Fluid

Topics: Acid Phosphatase; Arginase; Arthritis, Rheumatoid; Glycoside Hydrolases; Gout; Humans; Joint Diseases; Osteoarthritis; Peptide Hydrolases; Rheumatic Diseases; Synovial Fluid; Transaminases

Topics: Acid Phosphatase; Alanine Transaminase; Alkaline Phosphatase; Arthritis, Rheumatoid; Aspartate Aminotransferases; Glucose; Humans; Hydrarthrosis; Hydrogen-Ion Concentration; Isoenzymes; L-Lactate Dehydrogenase; Osteoarthritis; Proteins; Recurrence; Synovial Fluid

Topics: Acid Phosphatase; Aminopeptidases; Arthritis, Rheumatoid; Blood Sedimentation; Centrifugation; Hemoglobins; Humans; Hydrogen-Ion Concentration; Hydrolysis; Osteoarthritis; Synovial Fluid

Topics: Acid Phosphatase; Alkaline Phosphatase; Arthritis, Rheumatoid; Bursitis; Fructose-Bisphosphate Aldolase; Gout; Humans; Joint Diseases; Osteoarthritis; Phosphoric Monoester Hydrolases; Synovial Fluid; Transaminases

Topics: Acid Phosphatase; Adult; Arthritis, Infectious; Arthritis, Rheumatoid; Blood Cell Count; Electrophoresis; Humans; Inflammation; Isoenzymes; Leukocyte Count; Lymphocytes; Osteoarthritis; Synovial Fluid; Synovitis

Topics: Acid Phosphatase; Arthritis, Rheumatoid; Bradykinin; Humans; Kinins; Knee Joint; L-Lactate Dehydrogenase; Leukocyte Count; Osteoarthritis; Pain; Proteins; Synovial Fluid

Topics: Acid Phosphatase; Arthritis; Arthritis, Reactive; Aspartate Aminotransferases; Bradykinin; Female; Glucuronidase; Humans; Hydrogen-Ion Concentration; Kinins; L-Lactate Dehydrogenase; Male; Muscle, Smooth; Osteoarthritis; Synovial Fluid; Uterus

Topics: Acid Phosphatase; Animals; Anti-Inflammatory Agents; Arthritis; Arthritis, Rheumatoid; Aspirin; Cathepsins; Depression, Chemical; Glucuronidase; Gold; Humans; Hydrocortisone; Hydrogen-Ion Concentration; Hydrolases; In Vitro Techniques; Indomethacin; Knee Joint; Liver; Lysosomes; Osteoarthritis; Phenylbutazone; Phosphoric Monoester Hydrolases; Rabbits; Sulfhydryl Compounds; Synovial Fluid

Topics: Acid Phosphatase; Adult; Arthritis, Rheumatoid; Culture Techniques; Female; Humans; Lysosomes; Male; Middle Aged; Osteoarthritis; Proteins; Synovial Membrane

Topics: Acid Phosphatase; Arthritis; Arthritis, Rheumatoid; Histocytochemistry; Humans; Knee Joint; Microscopy, Electron; Osteoarthritis; Synovial Fluid

Topics: Acid Phosphatase; Adolescent; Alkaline Phosphatase; Arthritis; Arthritis, Infectious; Arthritis, Rheumatoid; Chemistry Techniques, Analytical; Diagnosis; Geriatrics; Gout; Humans; Knee Injuries; Osteoarthritis; Phosphoric Monoester Hydrolases; Synovial Fluid; Synovitis; Tuberculosis; Tuberculosis, Osteoarticular