Showing papers on "Tartrate-resistant acid phosphatase published in 1997"
TL;DR: It is demonstrated here that expression of a wild-type transgene in only a limited number of tissues can fully rescue the src-/- phenotype, and biochemical examination of osteoclasts from these mice suggest that Src may function in part by recruiting or activating other tyrosine kinases.
Abstract: The Src tyrosine kinase has been implicated in a wide variety of signal transduction pathways, yet despite the nearly ubiquitous expression of c-src, src-/- mice show only one major phenotype-osteopetrosis caused by an intrinsic defect in osteoclasts, the cells responsible for resorbing bone. To explore further the role of Src both in osteoclasts and other cell types, we have generated transgenic mice that express the wild-type and mutated versions of the chicken c-src proto-oncogene from the promoter of tartrate resistant acid phosphatase (TRAP), a gene that is expressed highly in osteoclasts. We demonstrate here that expression of a wild-type transgene in only a limited number of tissues can fully rescue the src-/- phenotype. Surprisingly, expression of kinase-defective alleles of c-src also reduces osteopetrosis in src-/- animals and partially rescues a defect in cytoskeletal organization observed in src-/- osteoclasts. These results suggest that there are essential kinase-independent functions for Src in vivo. Biochemical examination of osteoclasts from these mice suggest that Src may function in part by recruiting or activating other tyrosine kinases.
279 citations
TL;DR: It is shown that osteoclast-like cells are formed in co-culture of peripheral blood mononuclear cells and rheumatoidsynovial fibroblasts obtained by continued sub-cultures, implicating that osteoclasts generated within the synovial membrane are probably involved in bone destruction in rhearatoid arthritis.
182 citations
TL;DR: Serum TRAP activity is confirmed as a valid cytochemical marker for identification of osteoclasts, but is an osteoclastic marker of weak sensitivity due to known factors, such as synthesis of the enzyme not being unique to osteoclast, enzyme instability, and the presence of inhibitors in serum.
Abstract: Tartrate-resistant acid phosphatase (TRAP) activity is regarded as an important cytochemical marker of osteoclasts; its concentration in serum is utilized as a biochemical marker of osteoclast function and degree of bone resorption. This study was carried out to assess the sensitivity of TRAP activity both as a cytochemical marker in histological sections and as a biochemical marker in serum in comparison with the standardized histomorphometric variables of osteoclasts. To this end we investigated 24 patients (21 women, 3 men; 60 +/- 17 years of age) affected with various metabolic bone diseases. Osteoclast surface (OcS/BS) and osteoclast number (OcN/BS) were evaluated by standardized histomorphometry in iliac crest biopsies. On the basis of TRAP cytochemical activity, TRAP-positive osteoclast surface (TRAP + OcS/BS) and number (TRAP + OcN/BS) were measured. TRAP-positive cells adjacent to bone and showing one nucleus or no nuclei at all in the plane of section were included in the counts as osteoclasts. Serum TRAP activity was determined by spectrophotometric assay. Values of OcS/BS and OcN/BS were much lower than those of TRAP + OcS/BS (-50%) and TRAP + OcN/BS (-60%), respectively. Correlations between OcS/BS and TRAP + OcS/BS, and between OcN/BS and TRAP + OcN/BS, were highly significant. Serum TRAP was significantly correlated with OcS/BS, OcN/BS, and TRAP + OcN/BS. These correlations, however, were rather low. Moreover, serum TRAP did not correlate with TRAP + OcS/BS. From these results, the conclusion can be drawn that while TRAP activity is confirmed as a valid cytochemical marker for identification of osteoclasts, serum TRAP activity is an osteoclastic marker of weak sensitivity. This may be due to known factors, such as synthesis of the enzyme not being unique to osteoclasts, enzyme instability, and the presence of inhibitors in serum. Mononucleated osteoclasts do not significantly influence the serum enzyme levels.
128 citations
TL;DR: Bone resorption by mononucleated cells was studied in the acellular bone of a teleost fish (Oreochromis niloticus) by histological and enzyme histochemical observations and by transmission electron microscopy.
Abstract: Bone resorption by mononucleated cells was studied in the acellular bone of a teleost fish (Oreochromis niloticus) by histological and enzyme histochemical observations and by transmission electron microscopy. Bone resorbing cells (osteoclasts) were identified by their location at the sites of bone resorption, their frequent association with a band of concentrated activity of tartrate-resistant acid phosphatase at the bone surface and by the presence or lack of certain enzymes. Tartrate-resistant acid phosphatase was used as a marker for osteoclasts, and alkaline phosphatase as a marker for osteoblasts. Osteoclasts in O. niloticus are not multinucleated; however, during intense bone resorption, they form cell aggregations that resemble multinucleated giant cells in mammals. Conversely, during less intense bone degradation, osteoclasts are flat, have long narrow cytoplasmic processes and resemble the bone-lining cells of mammals. All bone-resorbing cells in O. niloticus are mononucleated and lack a ruffled border. Similarities to and differences from bone resorption by mononucleated cells in mammals are discussed.
103 citations
TL;DR: Findings indicate that osteopontin is expressed during the early stages of the differentiation of osteoclast and osteoblast progenitors in the bone marrow and that its cell adhesion properties are required for osteooclastogenesis.
Abstract: Osteoclast development requires cell-to-cell contact between hematopoietic osteoclast progenitors and bone marrow stromal/osteoblastic support cells. Based on this, we hypothesized that osteopontin, an adhesion protein produced by osteoclasts and osteoblasts, plays a role in osteoclastogenesis. Using in situ hybridization, we demonstrate that cells expressing the osteopontin messenger RNA (mRNA) appear after 3 days of culturing murine bone marrow cells. The number of these cells increases thereafter, reaching a peak on day 5. In the same cultures, cells expressing alkaline phosphatase (AP) or tartrate resistant acid phosphatase (TRAP), phenotypic markers for osteoblastic and osteoclast-like cells, respectively, appeared subsequent to the appearance of the osteopontin-positive cells. By means of a combination of in situ hybridization and histostaining, it was shown that the osteopontin mRNA was localized in 30–50% of the AP-positive or the TRAP-positive, as well as in nonspecific esterase (NSE)-positive, c...
88 citations
TL;DR: The recombinant PAP should be useful in future studies on the properties and regulation of the mammalian PAP enzyme, and was shown to be substituted with N-linked oligosaccharides.
Abstract: The tartrate-resistant acid phosphatase (TRAP) of rat osteoclasts has been shown to exhibit high (85-94%) identity at the amino acid sequence level with the purple acid phosphatase (PAP) from bovine spleen and with pig uteroferrin. These iron-containing purple enzymes contain a binuclear iron centre, with a tyrosinate-to-Fe(III) charge-transfer transition responsible for the purple colour. In the present study, production of rat osteoclast TRAP could be achieved at a level of 4.3 mg/litre of medium using a baculovirus expression system. The enzyme was purified to apparent homogeneity using a combination of cation-exchange, hydrophobic-interaction, lectin-affinity and gel-permeation chromatography steps. The protein as isolated had a purple colour, a specific activity of 428 units/mg of protein and consisted of the single-chain form of molecular mass 34 kDa, with only trace amounts of proteolytically derived subunits. The recombinant enzyme had the ability to dephosphorylate bone matrix phosphoproteins, as previously shown for bone TRAP. Light absorption spectroscopy of the isolated purple enzyme showed a lambda max at 544 nm, which upon reduction with ascorbic acid changed to 515 nm, concomitant with the transition to a pink colour. EPR spectroscopic analysis of the reduced enzyme at 3.6 K revealed a typical mu-hydr(oxo)-bridged mixed-valent Fe(II)Fe(III) signal with g-values at 1.96, 1.74 and 1.60, proving that recombinant rat TRAP belongs to the family of PAPs. To validate the use of recombinant PAP in substituting for the rat bone counterpart in functional studies, various comparative studies were carried out. The enzyme isolated from bone exhibited a lower K(m) for p-nitrophenyl phosphate and was slightly more sensitive to PAP inhibitors such as molybdate, tungstate, arsenate and phosphate. In contrast with the recombinant enzyme, TRAP from bone was isolated predominantly as the proteolytically cleaved, two-subunit, form. Both the recombinant enzyme and rat bone TRAP were shown to be substituted with N-linked oligosaccharides. A slightly higher apparent molecular mass of the monomeric form and N-terminal chain of bone TRAP compared with the recombinant enzyme could not be accounted for by differential N-glycosylation. Despite differences in specific post-translational modifications, the recombinant PAP should be useful in future studies on the properties and regulation of the mammalian PAP enzyme.
85 citations
TL;DR: Two other markers of bone resorption, hydroxylysyl pyridinoline and lysyl pyrIDinoline, were found in peptide linkage in the culture medium but not in free form; indicating that the osteoclasts had degraded the bone collagen to peptides but not to the free cross-linking amino acids.
76 citations
TL;DR: The changes in IL-6 production induced by the hormones could not be the sole determinant for the extent of TRAP+ MNC formation, however, the stimulatory effect of 1α,25(OH)2D3 on osteoclastogenesis nevert...
Abstract: In mouse bone marrow primary cultures, the formation of osteoclast-like, i.e. tartrate-resistant acid phosphatase (TRAP)- and calcitonin receptor-positive multinucleated cells (MNC), when induced by 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3), can be suppressed by 17β-estradiol (17β-E2), whereas 17α-E2 is without any effect. 17β-E2, above 10−11 m, significantly reduced 1α,25(OH)2D3-mediated TRAP+ MNC formation in cultured bone marrow cells from both female and male mice. The estrogen at 10−8 m suppressed the peak response to the vitamin D sterol by 50%. 17β-E2 significantly suppressed basal and 1α,25(OH)2D3-stimulated cellular production of interleukin (IL)-6. IL-6 alone, although bone marrow cells in hormone-free culture produced appreciable amounts of the cytokine, did not induce any TRAP+ MNC. Therefore, the changes in IL-6 production induced by the hormones could not be the sole determinant for the extent of TRAP+ MNC formation. However, the stimulatory effect of 1α,25(OH)2D3 on osteoclastogenesis nevert...
55 citations
TL;DR: The data suggest that orthodontic tooth movement after appliance activation requires the recruitment of osteoclasts to sites of compression and that this is indomethacin-sensitive, and that indomethire enhances root resorption at compression sites 10 days after appliance reactivation.
48 citations
TL;DR: In the search for a new class of bone‐sparing agents for treating osteopenic disorders, it was hypothesized that tartronic acid derivatives, sharing the chemical characteristics both of bisphosphonates and of Gla residues contained in matrix proteins such as osteocalcin, could positively affect bone metabolism.
Abstract: In the search for a new class of bone-sparing agents for treating osteopenic disorders, we hypothesized that tartronic acid derivatives, sharing the chemical characteristics both of bisphosphonates and of Gla residues contained in matrix proteins such as osteocalcin, could positively affect bone metabolism. A series of tartronates was therefore tested for their ability to affect bone metabolism. In vitro resorption tests were performed examining pit formation by freshly isolated rat and rabbit osteoclasts plated onto bone slices and exposed to the drugs for 48 h. Tartronates bearing a linear side-chain (DF 1222 and DF 1363A) were the most effective in inhibiting pit excavation in the pM-nM range. Tartronates did not affect osteoclast viability, number, adhesion, or tartrate resistant acid phosphatase activity. Transient cell retraction was observed in osteoclasts plated onto glass and exposed to DF 1222. The maximal effect was seen in cells treated for 4 h at a concentration of 1 pM. DF 1222 accelerated mineralization in cultures of periosteal cells without affecting other osteoblast-like functions. This product was therefore tested in vivo in ovariectomized mice. Bone mass in femur was evaluated, by ash gravimetry, 21 days after ovariectomy. Unfortunately, DF 1222, the most active of tartronates in vitro, was inactive in this test because of its high hydrophilicity and the subsequent too short residence time. On the contrary, its tetrahydropyranyl ether derivative, DF 1363A, endowed with a significantly higher lipophilicity, showed a dose-dependent bone-sparing effect when administered subcutaneously at 10, 30, and 100 mg/kg/die, thus confirming the activity seen in in vitro tests. Because of their feasible parallel effect on both bone resorption and formation, tartronate derivatives may be tested to candidate this class of products for clinical studies.
41 citations
Journal Article•
TL;DR: Increased bone resorption and enhanced osteoclastogenesis were specifically observed in the iliac bone marrow of patients with RA, especially those with severe RA, and these phenomena can be considered to accompany generalized osteoporosis in RA.
Abstract: Objective. To investigate osteoclastogenesis in bone marrow cells from patients with various pathogenic backgrounds known to induce osteoporosis, to identify specific factors that may cause generalized osteoporosis in patients with rheumatoid arthritis (RA). Methods. Bone marrow blood was obtained from 59 women, 36 with RA and 23 without RA. Patients with RA were classified as severe (26) and mild RA (10: 5 patients with and 5 without corticosteroid therapy). The non-RA subjects were divided into 3 groups, premenopausal (7), menopausal (8), and elderly (8). As a marker of bone resorption, the pyridinoline crosslinked telopeptide domain of type I collagen (ICTP) concentration in the bone marrow supernatant was measured by radioimmunoassay. The bone marrow cells were cultured 14 days in the presence or absence of autologous bone marrow supernatant; then the number of tartrate resistant acid phosphatase positive multinucleated cells (TRAP positive MNC) was counted as an indicator of osteoclastogenesis. Results. ICTP concentration of the bone marrow supernatant and the number of TRAP positive MNC showed remarkable enhancement in some patients with severe RA, but these features were not observed in the 3 control groups. Conclusion. Increased bone resorption and enhanced osteoclastogenesis were specifically observed in the iliac bone marrow of patients with RA, especially those with severe RA. These phenomena can be considered to accompany generalized osteoporosis in RA.
TL;DR: R reverse transcriptase‐polymerase chain reaction (RT‐PCR) analysis showed that osteoblasts express SCF mRNA and that parathyroid hormone increases expression levels about fourfold.
Abstract: Stem cell factor (SCF) is a polypeptide growth factor active on multiple cell types, mainly of hematopoietic origin. We studied the effects of avian SCF on the differentiation of chicken osteoclasts from their putative progenitors as well as on the bone-resorbing activity of terminally differentiated osteoclasts. Osteoclast formation was analyzed in long-term cocultures of osteoblasts and nonadherent, osteoclast-depleted bone marrow cells. Osteoclast activity was studied in short-term (48 h) cultures of bone marrow cell populations enriched for osteoclasts, on dentine slices. SCF strongly enhanced osteoclast differentiation. The IL-6-related chicken myelomonocytic growth factor (cMGF) had a similar effect, and the effects of SCF and cMGF were additive. SCF, but not cMGF, also stimulated the bone-resorbing activity of existing osteoclasts. As osteoblasts have been found to regulate osteoclast activity and formation, chicken osteoblasts were tested for their ability to express and secrete SCF. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that osteoblasts express SCF mRNA and that parathyroid hormone increases expression levels about fourfold. SCF did not accumulate in the culture medium, but remained cell (osteoblasts) surface associated.
TL;DR: The giant cells in this case of PVNS express all the phenotypical features of osteoclasts including the ability to carry out lacunar resorption, which may account for the bone destruction associated with this aggressive synovial lesion.
Abstract: AIM: To determine the cytochemical and functional phenotype of multinucleated giant cells in pigmented villo nodular synovitis (PVNS). METHODS: Giant cells isolated from a patient with PVNS of the knee were assessed for a number of markers used to distinguish osteoclasts from macrophages/ macrophage polykaryons: evidence of tartrate resistant acid phosphatase (TRAP) activity; expression of CD11b, CD14, CD51, and calcitonin receptors; and the ability of the giant cells to carry out lacunar resorption. RESULTS: Isolated giant cells expressed an osteoclast antigenic phenotype (positive for CD51, negative for CD11b and CD14) and were TRAP and calcitonin receptor positive. They also showed functional evidence of osteoclast differentiation, producing numerous lacunar bone resorption pits on bone slices in short term culture. CONCLUSIONS: The giant cells in this case of PVNS express all the phenotypical features of osteoclasts including the ability to carry out lacunar resorption. This may account for the bone destruction associated with this aggressive synovial lesion.
TL;DR: The enhanced TRAP activity in individual osteoclasts supports the concept that osteoclast are more active following estrogen withdrawal in agreement with theoretical arguments advanced previously.
Abstract: The effects of estrogen suppression on osteonal remodeling in young women was investigated using transiliac biopsies (eight paired biopsies + four single pre; three single post biopsies) taken before and after treatment for endometriosis (6 months) with analogs of gonadotrophin releasing hormone (GnRH) Estrogen withdrawal increased the proportion of Haversian canals with an eroded surface (106%, p = 0047), a double label (238%, p = 0004), osteoid (71%, p = 0002), and alkaline phosphatase (ALP) 116%, p = 0043) but not those showing tartrate-resistant acid phosphatase (TRAP) activity (p = 025) or a single label (p = 030) Estrogen withdrawal increased TRAP activity in individual osteoclasts in canals with diameters greater than 50 microns (p = 00089) and also the number of osteons with diameters over 250 microns (p = 0049) ALP activity in individual osteoblasts was increased but not significantly following treatment (p = 0051) Wall thickness was significantly correlated with osteon diameter (p < 0001) In a separate group of patients (four pairs + one post biopsy) on concurrent treatment with tibolone, there was no significant increase in the osteon density, cortical porosity, median canal diameter, or the markers of bone formation and resorption Enzyme activities and numbers of active canals were also not increased with the concurrent treatment, but there was still an increase in the osteon diameter As previously shown for cancellous bone, estrogen withdrawal increased cortical bone turnover We have now shown that resorption depth within Haversian systems was also increased with treatment The enhanced TRAP activity in individual osteoclasts supports the concept that osteoclasts are more active following estrogen withdrawal in agreement with theoretical arguments advanced previously Understanding the cellular and biochemical mechanisms responsible for increased depth of osteoclast resorption when estrogen is withdrawn may allow the development of new strategies for preventing postmenopausal bone loss
TL;DR: It is concluded that mononuclear phagocytes are capable of differentiating into mature functional OCs when cultured under specific cellular and hormonal conditions and should prove useful in further analysing factors controlling OC generation in bone remodelling and repair.
TL;DR: This report is the first documentation that human osteoclast-like FLG 29.1 cells express, in a developmentally regulated fashion, a homolog of the specific 150 kD avian osteoclasts surface antigen that is related to superoxide dismutase, a protective free radical scavenging enzyme and is essential for osteoclastic bone resorption.
Abstract: Osteoclast development from hematopoietic bone marrow precursors is associated with the expression of various enzymes, receptors, adhesion molecules, and other specialized components. Among these is a novel 150 kD superoxide dismutase-related membrane glycoprotein, originally identified by its reaction with the anti-osteoclast monoclonal antibody 121F. This antigen is uniquely restricted to osteoclasts in bone, universally present on osteoclasts from multiple species, induced during osteoclast differentiation in vitro and in ovo, and required at high levels for avian osteoclastic bone pit resorption. Expression of a comparable human antigen was investigated using human leukemic FLG 29.1 cells capable of differentiating towards an osteoclast-like phenotype. Phorbol ester, 1,25 (OH)2 vitamin D3, and osteoblast-derived soluble factors elicited dose and time-dependent inductions of this antigen as measured by enzyme-linked immunosorbent assay (ELISA) and immunocytochemical staining, coincident with their display of multiple other osteoclastic features. Synergistic interactions of these modulators led to further elevations in the ultimate expression levels of this antigen, although not to the full extent associated with in vivo-formed avian osteoclasts. The potent antiresorptive hormone 17beta-estradiol, but not its inactive alpha isomer, partially suppressed the phorbol ester-induced elevation of the 121F antibody-reactive antigen in FLG 29.1 cells as it does in avian osteoclast-like cells. Characterization of the human antigen isolated from FLG 29.1 cells by 121F immunoaffinity purification demonstrated that this regulated membrane component was synthesized by these human cells, more abundant following their differentiation into osteoclast-like cells, and similar biochemically and immunologically to the 150 kD integral membrane glycoprotein previously described from avian osteoclasts. Therefore, this report is the first documentation that human osteoclast-like FLG 29.1 cells express, in a developmentally regulated fashion, a homolog of the specific 150 kD avian osteoclast surface antigen that is related to superoxide dismutase, a protective free radical scavenging enzyme and is essential for osteoclastic bone resorption.
TL;DR: Results showed that OPN can be an important mechanism for regulating differentiation and bone resorption.
Abstract: Osteopontin (OPN) is an acidic phosphoprotein synthesized by osteoblasts and osteoclastic cells that are localized in the mineralized phase of bone matrix. OPN is thought to bind to the vitronection receptor on the osteoclast membrane and regulates bone resorption by the osteoclast. In this study, we investigated whether or not OPN can relate to osteoclast differentiation and bone resorption in a co-culture system. When C57Black/6N mouse bone marrow cells suspended on ivory slices coated with collagen were inoculated onto a MC3T3-G2/PA6 cell layer, colonies containing TRAP(+) mononuclear and multinuclear cells were formed in the presence of 1 alpha, 25-dihydroxyvitamin D3 and dexamethasone. At the end of culture period the number of TRAP(+) osteoclast-like cells were counted and the resorption pits were evaluated by reflected light microscopy. The mRNA of OPN was detected by in situ hybridization. Osteoclast-like cells expressed OPN mRNA. The addition of an OPN antisense oligomer (5' AAT CAC TGC CAA TCT CAT 3') at the start of the co-culture period decreased the number of TRAP(+) cells present after 7 d (30.3 +/- 3.4 vs 56.9 +/- 12.4), and the ratio of mononuclear and multinucleated cells was changed (77.6:23.2 vs 60.8:39.3). The total area of pits per ivory slice was also decreased by adding the OPN antisense oligomer (246813 vs 303139 microns2). These results showed that OPN can be an important mechanism for regulating differentiation and bone resorption.
TL;DR: After 10 d of culture a small but consistent number of early multinucleated osteoclasts were identified with a frequency of one cell per 700 granulocytic/monocytic cells, as revealed by cytologic examination.
Abstract: Circulating CD34 + cells were isolated from leukapheresis products collected from patients with ovarian cancer. CD34 - contaminating cells, identified immediately after immunoselection, ranged from 5% to 25% in five different experiments and were predominantly CD3 + T-lymphocytes (range 2-12%); CD3 - /CD16 + /CD56 + natural killer cells (range 2-11%) and rare mature CD15 + / CDllb + granulocytes (range 1-2%). CD34 + cells were cultured in liquid medium in the presence of interleukin-3, granulocyte-macrophage colony stimulating factor, stem cell factor. granulocyte colony stimulating factor and a powerful proliferation with prevalent differentiation along the granulocytic/monocytic lineage was obtained. After 10 d of culture a small but consistent number of early multinucleated osteoclasts were identified with a frequency of one cell per 700 granulocytic/monocytic cells, as revealed by cytologic examination. This observation was confirmed by staining for tartrate-resistant acid phosphatase activity which revealed red multinucleated elements with a frequency comparable to that reported above. Conversely. no osteoclasts were observed in those cultures in which macrophage overgrowth was obtained by culturing CD 34 + cells until day 35. These observations suggest that circulating progenitors have a multilineage potential in vitro and contribute to the clarification of osteoclast development in humans; additionally, they provide the basis for the future development of optimized osteoclast culture techniques in liquid medium and the basic culture system, to test the distinct activity of 1,25(OH) 2 D 3 , parathyroid hormone, interleukin-11 and of other cytokines on osteoclast development in humans..
TL;DR: Having characterized several properties of these anti-TRAP antibodies, 9C5 and 14G6 may be useful for development of TRAP-specific immunoassays in bone pathology and hematology and will certainly be of use for the study of biosynthesis, regulation, expression, and function ofTRAP.
Abstract: A major product of osteoclasts, tartrate-resistant acid phosphatase (TRAP) is an essential but insufficient enzyme for bone resorption. TRAP is an excellent cell marker for osteoclasts and macropha...
TL;DR: It is suggested that bone resorption in response to continuous mechanical deformation is regulated by cells of the osteoblast lineage such as preosteoblasts, osteoblast, bone lining cells and osteocytes in vivo, and that prostaglandins, but not prostag landins E2, are involved in the stretch-enhanced, osteoclast-like cell formation.
Journal Article•
TL;DR: Investigating the role of a nitric oxide donor, 3-morpholinosydnonimine hydrochloride, on the FLG 29.1 cells confirmed the bidirectional effect ofNitric oxide whose basal production is necessary in promoting osteoclast differentiation, while at high levels it is effective in inhibiting osteOClast activity.
Abstract: Nitric oxide is a short-lived free radical produced by different isoforms of the enzyme nitric oxide synthase. It regulates a whole range of functions in the body but little is known about its effects on bone. Rat osteoclasts and a human preosteoclast cell line (FLG 29.1) have been shown to produce nitric oxide and to express nitric oxide synthases. In the present study we investigated the role of a nitric oxide donor, 3-morpholinosydnonimine hydrochloride, on the FLG 29. 1 cells. 3-Morpholinosydnonimine hydrochloride has been shown to significantly increase IL6 production and tartrate resistant acid phosphatase activity in FLG 29.1 cells, indicating a positive modulation of osteoclast differentiation. However FLG 29.1 cell adhesion on osteoblast-like cells was significantly inhibited, suggesting an inhibition of osteoclast motility All these results confirm the bidirectional effect of nitric oxide whose basal production is necessary in promoting osteoclast differentiation, while at high levels it is effective in inhibiting osteoclast activity.
Journal Article•
TL;DR: Deoxypyridinoline can be a more sensitive marker than hydroxyproline, with some advantages, such as its quantitation in a urine specimen and its high bone specificity, and tartrate resistant acid phosphatase was the only biochemical marker of bone resorption with increased levels in patients with renal failure.
Abstract: Collagen type 1 represents more than 90% of bone matrix. Therefore, quantitation of collagen crosslinks, such as deoxypyridinoline, can provide information on bone resorption degree. An evaluation was made of deoxypyridinoline as well as other bone markets, such as alkaline phosphatase, tartrate resistant acid phosphatase, and hydroxyproline in patients with the diagnosis of osteoporosis. Paget's disease, hyperthyroidism, and chronic renal failure on haemodialysis or not. Deoxypyridinoline levels were significantly increased in patients with osteoporosis, Paget's disease, and hyperthyroidism. Hydroxyproline levels were increased in patients with Paget's disease, and tartrate resistant acid phosphatase was increased in all the entities studied. Deoxypyridinoline can be a more sensitive marker than hydroxyproline, with some advantages, such as its quantitation in a urine specimen and its high bone specificity. In patients with renal failure, tartrate resistant acid phosphatase was the only biochemical marker of bone resorption with increased levels.
30 Dec 1997
TL;DR: A simple and inexpensive household vegetable steamer provided equivalent or superior HIER for demonstration of TRAP with 9C5 in a variety of tissues and found no significant differences in immunoreactivity or nuclear detail between formalin and B-5 fixed bone marrow biopsy sections.
Abstract: Tartrate-resistant acid phosphatase (TRAP) is an important cell marker in both hematology and bone pathology. Cytochemical stains for TRAP activity are used to identify leukemic cells of hairy cell leukemia, inflammatory or activated macrophages, and bone resorbing osteoclasts. A monoclonal antibody to TRAP (9C5) was developed for immunohistochemical identification of these and other TRAP-expressing cells in paraffin sections. In our continuing efforts to maximize the sensitivity and specificity of 9C5 for diagnostic and investigational purposes, we conducted studies to compare steam to microwave irradiation for heat induced epitope retrieval (HIER) and to compare formalin fixation to B-5 fixation. We found that a simple and inexpensive household vegetable steamer provided equivalent or superior HIER for demonstration of TRAP with 9C5 in a variety of tissues. We also found no significant differences in immunoreactivity or nuclear detail between formalin and B-5 fixed bone marrow biopsy sections. ...