Showing papers on "Tartrate-resistant acid phosphatase published in 1989"
TL;DR: While TRAP and multinuclearity are reliable markers for osteoclastic phenotype in bone, they are unreliable markers in culture and will not only overstate, to an unknown and probably variable degree, the number of multinucleate osteoclasts that develop, but will also fail to even identify what may be a considerable and more substantial population of mononuclear cells that possess osteoclast characteristics.
Abstract: The osteoclast is the cell that resorbs bone. It is known to derive from hemopoietic precursors, and a series of recent experiments has used enumeration of the tartrate-resistant acid phosphatase (TRAP)-positive multinucleate cells that develop in cultures of hemopoietic tissue as a means to analyze the regulation of osteoclast generation. These multinucleate cells have never been definitively characterized as osteoclasts, however, and we elected to assess the relationship among bone resorption (the primary function of the osteoclast), TRAP, and multinuclearity in mouse bone marrow cultures. Mouse bone marrow cells and peritoneal macrophages were incubated on plastic coverslips or bone slices for up to 14 days in the presence or absence of 1 alpha, 25-dihydroxyvitamin D3 [1 alpha,25-(OH)2D3]. Osteoclast generation, as judged by bone resorption, occurred in marrow cell cultures only in the presence of 1 alpha,25-(OH)2D3. However, TRAP-positive multinuclear cells developed both with and without the hormone. The multinuclear cells bound F4/80, a marker for macrophages that does not bind to osteoclasts. Peritoneal macrophages became multinucleate and developed TRAP positivity in culture to levels similar to those in freshly isolated osteoclasts, especially with 1 alpha,25-(OH)2D3, but remained nonresorptive. In cultures of marrow cells incubated with 1 alpha,25-(OH)2D3 bone resorption was more extensive than could readily be accounted for by the number of multinucleate cells present, and the size of excavations and extent of resorption suggested a major contribution by mononuclear cells with osteoclastic function. Thus, while TRAP and multinuclearity are reliable markers for osteoclastic phenotype in bone, they are unreliable markers in culture. Experiments designed to evaluate the regulation of osteoclast generation through enumeration of TRAP-positive multinucleate cells formed in bone marrow cultures will not only overstate, to an unknown and probably variable degree, the number of multinucleate osteoclasts that develop, but will also fail to even identify what may be a considerable and more substantial population of mononuclear cells that possess osteoclastic characteristics.
124 citations
TL;DR: The results suggest that the mononuclear cells are either precursors of the osteoclasts or perhaps release cytokines that affect osteoclast formation or activity, and strongly suggest that ALP moves from the base of theEnamel organ to the enamel itself over a period of time ranging from pre- to post-eruption.
Abstract: It was the aim of this study to determine the cellular changes that occur in the enamel organ, dental follicle, and surrounding bony crypt of the rat molar prior to and during tooth eruption. By use of light microscope histochemistry to detect cells containing tartrate-resistant acid phosphatase (TRAP), it was seen that TRAP-positive mononuclear cells were present in the dental follicle prior to the onset of eruption (e.g., three days postnatal age) and then declined in number during eruption. Concurrently, TRAP-positive osteoclasts were initially present in large numbers on the surface of the bony crypt surrounding the molars (three days postnatal age) and then declined in number as eruption progressed. Electron microscopy confirmed that these were mononuclear cells and osteoclasts. The results suggest that the mononuclear cells are either precursors of the osteoclasts or perhaps release cytokines that affect osteoclast formation or activity.Staining for alkaline phosphatase (ALP) activity indicated that...
113 citations
TL;DR: The purification of small amounts of TR-AP from human placenta is described, revealing the presence of a 1.5-kilobase transcript in white cells from a patient with hairy cell leukemia, in human K562 erythroleukemic cells, and in Epstein-Barr virus-transformed human B-cells, but not in a human T-cell line.
111 citations
TL;DR: Using a variety of substrates and inhibitors, it is demonstrated that there are differences between the osteoclastoma acid phosphatase and the enzyme purified from other sources.
Abstract: Tartrate-resistant acid phosphatase is one of the major enzymes produced and secreted by osteoclasts. To obtain sufficient enzyme for biochemical characterization, we have purified this enzyme from human osteoclastomas by sequential chromatography on SP-Sephadex, CM-Sephadex, hydroxylapatite, Sephadex G-150 and concanavalin A-Sepharose. The purification over the original tumour extract was about 2000-fold, with a yield of 10%. The enzyme appeared to be homogeneous when assessed by SDS/polyacrylamide-gel electrophoresis. Both gel filtration and SDS/polyacrylamide-gel electrophoresis indicated an Mr of about 30,000. The reduced and alkylated enzyme consists of two subunits with Mrs of 15,000 and 17,500. The N-terminal amino acid sequence of both subunits indicates that there is a high degree of identity between the osteoclastoma enzyme and similar enzymes purified from spleen and uterus. Using 4-methylumbelliferyl phosphate as substrate, the specific activity of the purified enzyme was 387 units.mg-1, and the Km was 284 microns. The pH optimum was 5.7. Unlike similar enzymes purified from human and bovine bone, osteoclastoma acid phosphatase is not activated by reducing agents (2-mercaptoethanol or ascorbic acid). The enzyme contains 4.8 mol of Fe2+/3+, 0.3 mol of Mn2+ and 1.7 mol of Mg2+ per mol of enzyme. Although the enzyme loses 50% of its activity in the presence of EDTA, it is not inhibited by the iron chelator 1,10-phenanthroline. However, the enzyme is activated to a small extent by Mn2+ and Mg2+. Using a variety of substrates and inhibitors, we demonstrate that there are differences between the osteoclastoma acid phosphatase and the enzyme purified from other sources.
106 citations
TL;DR: Osteoclastic acid phosphatase is a member of a widely-distributed class of iron-containing proteins with acid-phatase activity as mentioned in this paper, and when antibodies to one such protein, porcine uteroferrin, are added to medium in which rat osteoclasts are incubated on devitalised cortical bone, both bone resorption and acid-pase activity are markedly inhibited.
100 citations
TL;DR: It is suggested that sTr-AcP could be a useful marker for bone loss and consequently, for the measure of bone resorption rate in postmenopausal women.
Abstract: Serum tartrate-resistant acid phosphatase (sTr-AcP) and bone mineral content (BMC) were measured in 29 women with postmenopausal osteoporosis and in 12 control women. Serum Tr-AcP was higher in osteoporotic patients than in controls and a negative linear correlation was found between sTr-AcP and BMC in osteoporotic women. These results suggest that sTr-AcP could be a useful marker for bone loss and consequently, for the measure of bone resorption rate in postmenopausal women.
55 citations
TL;DR: The results suggest that the PGE2-induced stimulation of osteoclastic activity represents an indirect effect mediated by stromal cells derived from bone marrow.
35 citations
TL;DR: It is suggested that acid p-nitrophenyl phosphatase is a useful cytochemical marker for identification of the osteoclast family at electron-microscopic levels of resolution.
Abstract: By differentiation of substrate specificity, pH optimum range, and sensitivity to various inhibitors, 2 isoenzymes of acid phosphatase in bone cells have been studied at the electron-microscopic level. When p-nitrophenyl phosphate was used for the substrate, the demonstrable enzyme activity was affected by neither tartrate nor sodium fluoride. The reaction product, when incubated at pH 5-6, was detected in all sites along the pathway for the biosynthesis of acid phosphatase in the osteoclast, including the perinuclear space, cisternae of the endoplasmic reticulum, Golgi complex, various vesicles, and vacuoles. In the osteoclasts attached to bone, the enzymatic activity was demonstrated at the extracellular ruffled border and on the eroded bone surface. Reaction products became confined to lysosomes and extracellular ruffled border when incubated at pH 6-7. Unattached osteoclasts showed a similar intracytoplasmic localization of enzyme as the attached ones, except for the absence of the extracellular enzyme activity. The mononuclear, immature type of osteoclast also resembled the mature osteoclast in terms of enzymatic localization. Except for the osteoclasts, the acid p-nitrophenyl phosphatase activity was restricted to lysosomal vesicles in various bone cells, monocytes, and macrophages. Such activity was inhibited by adding 50 mM tartrate to the p-nitrophenyl phosphate medium. When beta-glycerophosphate or p-nitrocatechol sulfate was the substrate, most of the reaction product was localized intracellularly. Unlike the acid p-nitrophenyl phosphatase, the acid beta-glycerophosphatase or arylsulfatase activity in osteoclasts and other bone cells was inhibited completely by 10 mM tartrate or 10 mM sodium fluoride.(ABSTRACT TRUNCATED AT 250 WORDS)
28 citations
TL;DR: The data suggest that fetal mouse bone tissue induces the differentiation of osteoclast precursors, and fetal bone rudiments but not embryonic fibroblasts seem to produce a factor which stimulate the formation of cells showing characteristics of osteOClast precursor cells.
18 citations
TL;DR: In this paper, a morphometric and histochemical study was performed on trephine biopsies of the bone marrow to elucidate osseous remodeling by assessment of trabecular bone area (planimetry) and number of osteoclasts.
Abstract: Summary In 165 patients with chronic myeloproliferative disorders (CMPD) a morphometric and histochemical study was performed on trephine biopsies of the bone marrow to elucidate osseous remodeling by assessment of trabecular bone area (planimetry) and number of osteoclasts. Osteoclastic elements were identified by the tartrate-resistant acid phosphatase method. In addition to control specimens (n = 20) subtypes of CMPD included chronic myeloid leukemia (CML, n = 65), primary (essential) thrombocythemia (PTH, n = 25), polycythemia vera rubra (P. vera, n = 25) and agnogenic myeloid metaplasia (AMM, n = 50). AMM was discriminated into a so-called early hyperplastic stage without gross myelofibrosis (n = 19) and an overt or advanced stage showing fibro-osteosclerotic changes (n = 31). Total area of trabecular bone and counts for osteoclasts (uni- and multi-nucleated cells as well as a-nuclear cytoplasmic fragments) were not significantly increased in CML, PTH, P. vera and in the initial hypercellular stages of AMM. In contrast to these results, in advanced stages of AMM there was a significant increase in total bone area associated with a high count for all osteoclastic elements and apparently also an increased number of osteoblasts. It is speculated that the marked increase in osteoclastic-osteoblastic elements in late stages of AMM possibly reflects an imbalance of calcitriol (1,25-dihydroxyvitamin D 3) on skeletal homeostasis. This abnormal osseous remodeling may be mediated by the atypical megakaryocytic proliferation in this disorder, which is always a conspicuous feature of bone marrow biopsies.
17 citations
TL;DR: An immunoassay for human band-5 acid phosphatase in which antibodies to porcine uteroferrin, immobilized on Sepharose particles, are used, which is similar in reproducibility and sensitivity to assays based on inhibition by d-tartrate.
Abstract: We describe an immunoassay for human band-5 acid phosphatase in which antibodies to porcine uteroferrin, immobilized on Sepharose particles, are used. Band-5 acid phosphatase is the tartrate-resistant isoenzyme normally expressed in tissue macrophages such as osteoclasts and alveolar macrophages. The immunoassay is similar in reproducibility and sensitivity to assays based on inhibition by d-tartrate. However, compared with the latter, the greater specificity of the immunoassay makes it markedly less susceptible to errors arising from the presence of non-band-5 acid phosphatases, e.g., from prostate.
Journal Article•
TL;DR: A cytomorphometric study of the osteoclastic population was made after TRAP staining on transiliac bone biopsies from 10 human volunteers, and the diameter of each TRAP positive profile was determined with an automatic image analyser.
Abstract: Tartrate resistant acid phosphatase (TRAP) is considered as an enzyme marker for osteoclasts and their precursors A cytomorphometric study of the osteoclastic population was made after TRAP staining on transiliac bone biopsies from 10 human volunteers, and the diameter of each TRAP positive profile was determined with an automatic image analyser The frequency distribution of the cell diameter followed a lognormal law TRAP positive cells along bone trabeculae belong to a homogeneous osteoclastic population
Journal Article•
TL;DR: These isoenzyme serum levels were well correlated with those of two breast cancer markers (CEA and CA15.3) and radiograph and calculated according to several criteria.
Abstract: Bone alkaline phosphatase (B-ALP) and tartrate resistant acid phosphatase (TR-ACP) are markers of osteoblastic and osteoclastic activities respectively. During a period of up to two years, these isoenzymes have been assayed in the sera of 191 breast cancer patients; 80 had bone metastases (BM). In BM bearing patients, B-ALP activity was 261 IU/l and 63 IU/l for patients without BM; TR-ACP was respectively 6.6 and 3.3 IU/l. Specificity and sensitivity were calculated according to several criteria. These isoenzyme serum levels were well correlated with those of two breast cancer markers (CEA and CA15.3) and radiograph.
TL;DR: Conclusive proof for de novo synthesis of acid phosphatase in GA 3 -treated half-seeds was achieved by the in vivo labelling of enzyme with [ 35 S]sulphate, indicating the requirement of free sulfhydryl groups for the catalytic efficiency of the enzyme.