acid-phosphatase and Cartilage-Diseases

Initial studies indicated that bone and cartilage, when treated with a hypertonic glutaraldehyde fixative for a short period, retained significant enzyme activity for histochemistry and also maintained excellent fine structure. We used 6% glutaraldehyde in 0.1 M cacodylate buffer, pH = 7.2, 4 degrees C to fix small pieces of bone or cartilage for three hours while the tissues were being constantly agitated. These samples were demineralized in 10% ethylene diamine tetraacetic acid, buffered to pH = 7.2 with 0.1 M Tris HC1, at 4 degrees C. The demineralized tissue was frozen and cryostat sections 32 microns thick were taken for incubation at 37 degrees C in various media for histochemistry. For electron microscopic localization of enzymes a heavy metal capturing method had to be used. For light microscopy, the azo dye methods were frequently used, but these were not usable for electron microscopy. Alkaline phosphatase was found on the outer surface of osteoblast and hypertrophic cartilage cell membranes. The only intracellular enzyme activity was found on the mitochondrial membranes of the osteoclast and only when the pH of the media was lowered from the optimum 9.5 to 8.5. Alkaline phosphatase was not found along the osteocyte or young cartilage cell membranes...

Topics: Acid Phosphatase; Alkaline Phosphatase; Bone and Bones; Bone Neoplasms; Cartilage; Cartilage Diseases; Cell Membrane; Giant Cell Tumors; Golgi Apparatus; Histocytochemistry; Humans; Lysosomes; Mucolipidoses; Organoids; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis Imperfecta; Osteosarcoma; Phosphoric Monoester Hydrolases

To examine the role of tartrate resistant acid phosphatase (TRAP) positive mononuclear and multinucleated cells in the destruction of articular cartilage in patients with rheumatoid arthritis (RA).. The presence of TRAP positive cells in the synovial tissue of patients with RA was examined by enzyme histochemistry and immunohistochemistry. Expression of mRNAs for matrix metalloproteinases (MMPs) was assessed by the reverse transcriptase-polymerase chain reaction (RT-PCR) and northern blot analysis. Production of MMPs by mononuclear and multinucleated TRAP positive cells was examined by immunocytochemistry, enzyme linked immunosorbent assay (ELISA) of conditioned medium, and immunohistochemistry of human RA synovial tissue. In addition, a cartilage degradation assay was performed by incubation of (35)S prelabelled cartilage discs with TRAP positive cells.. TRAP positive mononuclear cells and multinucleated cells were found in proliferating synovial tissue adjacent to the bone-cartilage interface in patients with RA. Expression of MMP-2 (gelatinase A), MMP-9 (gelatinase B), MMP-12 (macrophage metalloelastase), and MMP-14 (MT1-MMP) mRNA was detected in TRAP positive mononuclear and multinucleated cells by both RT-PCR and northern blot analysis. Immunocytochemistry for these MMPs showed that MMP-2 and MMP-9 were produced by both TRAP positive mononuclear and multinucleated cells, whereas MMP-12 and MMP-14 were produced by TRAP positive multinucleated cells. MMP-2 and MMP-9 were detected in the conditioned medium of TRAP positive mononuclear cells. TRAP positive mononuclear cells also induced the release of (35)S from prelabelled cartilage discs.. This study suggests that TRAP positive mononuclear and multinucleated cells located in the synovium at the cartilage-synovial interface produce MMP-2 and MMP-9, and may have an important role in articular cartilage destruction in patients with RA.

Topics: Acid Phosphatase; Aged; Animals; Arthritis, Rheumatoid; Blotting, Northern; Cartilage Diseases; Cartilage, Articular; Cattle; Cells, Cultured; Female; Humans; Immunohistochemistry; Isoenzymes; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Middle Aged; Monocytes; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Synovial Membrane; Tartrate-Resistant Acid Phosphatase

The mechanism of laryngotracheitis virus-induced dissolution of chick nasal turbinate cartilage was studied by lysosomal enzyme histochemistry. Five-day-old chicks were infected by intranasal instillation, and changes in lysosomal enzyme distribution were followed at daily intervals through the tissue regeneration stage, Day 28. In the mucosa the lysosomes were activated beginning on Day 1, and glycerol acid phosphatase and a diffuse form of beta-glucuronidase were released concomitant with tissue cell destruction. In the chondrocytes (where glycerol acid phosphatase was absent), beginning on Day 2, particulate (lysosomal) beta-glucuronidase decreased as diffuse beta-glucuronidase increased and extended out into the matrix. The cartilage lost its metachromatic staining properties and became soft and pliable. Regeneration of the mucosa started on Day 6 and gradual reappearance of metachromatic staining of the cartilage began on Day 8 with considerable recovery of original turbinate structure by Day 12. A lysosomal membrane labilizer, vitamin A, exacerbated the cartilage pathology, whereas a stabilizer, cortisone, retarded it.

Topics: Acid Phosphatase; Animals; Cartilage; Cartilage Diseases; Chickens; Cortisone; Glucuronidase; Herpesviridae Infections; Herpesvirus 1, Gallid; Lysosomes; Nose Diseases; Poultry Diseases; Turbinates; Vitamin A

In recent years, histochemistry and electron microscopy have been applied more and more to the investigation of bone tumors. The contributions and limitations of these methods in differential diagnosis are discussed. The levels of glycosaminoglycans in cartilaginous tumors display distinct differences between slow- and fast-growing types. All cartilaginous tumors are poor in phosphatase activity. Demonstration of these enzymes at acid and alkaline pH in bone-forming conditions reveals differences between benign and malignant tumors. Osteosarcomas display a rich activity of both phosphatases in bone-forming and in bone-free regions. Acid phosphatase may play a rôle in the breakdown of the host tissue infiltrated by the tumor. Electron microscopy of bone tumors has brought out some interesting findings. In fibrous dysplasia a particular kind of very fine fibrillar structures was observed besides the regular collagen fibrils. This may indicate retardation of collagen maturation. Cell organelles in benign and malignant bone tumors usually differ quantitatively. They resemble active fibroblasts. In bone- and in cartilage-forming tumors we observed large quantities of microfilaments in the cytoplasm. Nuclear indentations and invaginations probably indicate increased nuclear activity. The intense acid phosphatase activity demonstrated histochemically seems inconsistent with the low number of lysosomes in the cytoplasm of osteosarcoma cells, but other organelles (Golgi apparatus and vesicles) may also contain the enzyme. Virus-like particles have not been observed in human osteosarcomas up to now. Other authors have observed a correlation between the number of cell organelles and the grade of differentiation, but this was not detected in our sample of benign and malignant cartilaginous tumors. Histochemistry and electron microscopy of bone tumors are still in the early stage of material gathering. Some histochemical findings, however, can already be used as diagnostic tools.

Topics: Acid Phosphatase; Alkaline Phosphatase; Bone Neoplasms; Cartilage Diseases; Cell Nucleus; Chondrosarcoma; Collagen; Cytoplasm; Cytoskeleton; Fibrous Dysplasia of Bone; Humans; Osteosarcoma

An experimental model of degenerative joint disease on chondromalacia consists of a surgically-scarified articular surface of the adult dog knee joint. In 52 dogs, evaluated by histologic and enzymatic assays over a period of 1 to 110 weeks post-surgery, the levels of acid hydrolase activity varied on various areas of articular cartilage within the same joint. There was a transient rise in most of the acid hydrolases in the synovium as a response to arthrotomy of the knee joint. All of the acid hydrolases studied did not respond uniformly to surgically created trauma. There was evidence of repair of the cartilage lacerations even when the subchondral zone was not breached. Lacerations in the central portion of the patella rarely showed healing in contrast to those placed more to the periphery of the articular surface. There was no gross or histologic evidence of progressive degenerative joint disease up to 2 years post-surgery. Thus an injury inflicted to the surface of the articular cartilage may be in itself insufficient in severity to produce destructive changes in the joint. This should not be too surprising, since, clinically, all joint surface injury does not lead to degenerative arthritis. The joint seems to have an injury threshold whereby chondrocytes are capable of repairing surface injury if the damage is not massive or repetitive. Insofar as lacerations in the center of the patella rarely healed, while the peripheral ones showed consistent signs of healing, the site of injury, as well as the magnitude of injury, may be critical.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Arylsulfatases; Cartilage Diseases; Cartilage, Articular; Cathepsins; Deoxyribonucleases; Disease Models, Animal; Dogs; Glucuronidase; Joint Diseases; Knee Joint; Patella; Synovial Membrane; Time Factors

Topics: Acid Phosphatase; Animals; Cartilage; Cartilage Diseases; Female; Glucuronidase; Histocytochemistry; Hydrolases; Lysosomes; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Connective Tissue; Neoplasms, Experimental; Papain; Rats; Staining and Labeling; Sternum; Transplantation, Homologous; Vitamin A