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A Sabokbar

Publications and source records attributed to A Sabokbar.

43 records · Page 3Linked to original sources

Cells of the mononuclear phagocyte series differentiate into osteoclastic lacunar bone resorbing cells.

Although the osteoclast shares several features with other cells of the mononuclear phagocyte system (MPS), its precise cellular ontogeny is unknown, and its membership of the MPS is controversial. This study examined whether various cells of the MPS can be induced to differentiate into cells capable of the highly specialized osteoclastic function of lacunar bone resorption. We isolated mouse and rat monocytes, mouse (liver, peritoneal, alveolar, brain) tissue macrophages, and spleen and marrow haemopoietic cells, as well as foreign body macrophages and macrophage polykaryons derived from subcutaneous granulomas formed by implantation of latex beads and coverslips in mice. When these cells were incubated with UMR106 osteoblast-like cells on glass coverslips and human cortical bone slices in the presence of 1,25-dihydroxy vitamin D3 [1,25(OH)2D3] for 7 and 14 days, numerous tartrate-resistant acid phosphatase-positive cells formed in these co-cultures and scanning electron microscopy revealed extensive lacunar resorption of the bone surface. Bone resorption was seen as early as 4 days after monocytes were co-cultured with UMR106 cells. With the exception of bone marrow-derived cells, lacunar resorption was not seen in the absence of UMR106 cells. These findings show that a bone-derived stromal cell element is necessary for differentiation of monocytes and tissue and inflammatory macrophages into osteoclast-like cells capable of extensive lacunar bone resorption, and would argue in favour of osteoclast membership of the MPS.

Animals↗

Human osteoclast formation and bone resorption by monocytes and synovial macrophages in rheumatoid arthritis.

OBJECTIVE: To determine whether synovial macrophages and monocytes isolated from patients with rheumatoid arthritis patients are capable of differentiating into osteoclastic bone resorbing cells; and the cellular and humoral conditions required for this to occur. METHODS: Macrophages isolated from the synovium and monocytes from the peripheral blood of rheumatoid arthritis patients were cultured on bone slices and coverslips, in the presence and absence of UMR 106 rat osteoblast-like cells, 1,25-dihydroxy vitamin D3 (1,25(OH)2D3) and macrophage colony stimulating factor (M-CSF), and assessed for cytochemical and functional evidence of osteoclast differentiation. RESULTS: Isolated calcitonin receptor (CTR), tartrate resistant acid phosphatase (TRAP), and vitronectin receptor (VNR) negative, CD11b and CD14 positive monocytes and macrophages differentiated into CTR, TRAP, and VNR positive multinucleated cells capable of extensive lacunar bone resorption when co-cultured for 14 d with UMR 106 cells in the presence 1,25(OH)2D3 and M-CSF. CONCLUSIONS: Mononuclear phagocytes (monocytes and macrophages) from rheumatoid arthritis patients are capable of differentiating into multinucleated cells showing all the cytochemical and functional criteria of mature osteoclasts. Synovial macrophage-osteoclast differentiation may represent an important cellular mechanism in the bone destruction associated with rheumatoid arthritis.

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The human osteoclast precursor circulates in the monocyte fraction.

The osteoclast is known to be formed by fusion of circulating mononuclear precursor cells of haematopoietic origin. The precise nature of these circulating cells and, in particular, their relation to monocytes is unknown. We have developed an in vitro system of human osteoclast formation whereby human monocytes [CD14, CD11a, CD11b and HLA-DR positive, and tartrate-resistant acid phosphatase (TRAP), calcitonin receptor (CTR), vitronectin receptor (VNR) negative] were isolated and cocultured for up to 21 days with UMR106 rat osteoblast-like cells or ST2 mouse preadipocytic bone marrow stromal cells in the presence of 1 alpha, 25 dihydroxyvitamin D3 (1,25(OH)2D3) and macrophage colony stimulating factor (M-CSF). Numerous TRAP, VNR and CTR positive multinucleated cells, capable of extensive lacunar bone resorption, formed in these cocultures; the absolute requirements for this to occur were contact with the above bone stromal cells, 1,25(OH)2D3, and M-CSF. These results show that the human mononuclear osteoclast precursor circulates in the monocyte fraction and exhibits a monocyte phenotype, acquiring osteoclast phenotypic features in the process of differentiation into mature functional osteoclasts.

Acid Phosphatase↗

Bisphosphonate inhibition of bone resorption induced by particulate biomaterial-associated macrophages.

Aseptic loosening of total joint replacements is associated with bone resorption. A heavy infiltrate of foreign body macrophages in response to biomaterial wear particles is commonly found in the fibrous membrane surrounding loose components. It has recently been shown that foreign body macrophages can differentiate into osteoclastic cells. To determine whether pharmacological inhibitors of bone resorption have a role to play in controlling the osteolysis of aseptic loosening, we analyzed the effect of a bisphosphonate, disodium ethane-1, 1-diphosphonate (EHDP) on this process. Murine monocytes and foreign body macrophages (derived from granulomas formed by subcutaneous implantation of particles of prosthetic biomaterials) were co-cultured with UMR106 osteoblast-like cells in the presence of 1,25 dihydroxyvitamin D3 for 14 days on glass coverslips and bone slices. EHDP significantly inhibited bone resorption in these co-cultures. There was little or no expression of the osteoclast-associated enzyme, tartrate-resistant acid phosphatase (TRAP) in EHDP-treated co-cultures. Addition of EHDP to monocyte-UMR106 co-cultures after the appearance of TRAP-positive cells did not abolish bone resorption, indicating that EHDP, in addition to its known inhibitory effect on osteoclast function, suppresses differentiation of osteoclast precursors. EHDP inhibition of the osteolysis induced by particulate biomaterial-associated macrophages shows that pharmacological inhibition of bone resorption might be used to control the osteolysis of aseptic loosening.

Animals↗

Increased osteoclastic differentiation by PMMA particle-associated macrophages. Inhibitory effect by interleukin 4 and leukemia inhibitory factor.

To determine the influence of polymethylmethacrylate (PMMA) wear particles on macrophage-osteoclast differentiation, PMMA particles were added to mouse monocytes which were cocultured with UMR 106 osteoblast-like cells in the presence of 1,25 dihydroxy vitamin D3[1,25(OH)2D3] for up to 7 days on glass coverslips and for up to 14 days on human cortical bone slices. An increase in osteoclast differentiation, as evidenced by the expression of the osteoclast-associated enzyme tartrate-resistant acid phosphatase (TRAP) and the extent of lacunar bone resorption, was observed in monocyte cultures to which PMMA had been added. Interleukin 4 (IL-4) and Leukemia Inhibitory Factor (LIF) added to these cocultures caused considerably less expression of TRAP and significant inhibition of lacunar bone resorption. This inhibitory effect was reversed by the addition of specific neutralizing antibodies to LIF and IL-4. These findings show that PMMA-wear particle-associated macrophages exhibit an enhanced capacity for differentiation to osteoclastic bone-resorbing cells.

Acid Phosphatase↗

Role of inflammatory mediators and adhesion molecules in the pathogenesis of aseptic loosening in total hip arthroplasties.

Concentrations of prostaglandin E2, interleukin-6, and interleukin-8 were determined in the hip joint synovial fluid of 20 patients undergoing primary (n = 12) and revision (n = 8) total hip arthroplasties. Levels of soluble adhesion molecules were also investigated in these patients. There was a significant and marked increase in levels of prostaglandin E2 (P < .001), interleukin-6 (P < .011), interleukin-8 (P < .0002), soluble intercellular adhesion molecule 1 (P < .07), soluble vascular adhesion molecule 1 (P < .0006), and soluble endothelial leukocyte adhesion molecule 1 (P < .0003) in joint fluid of patients undergoing revision. On the basis of these observations, it is suggested that synovial fluid and its inflammatory contents could play a significant role in the pathogenesis of aseptic loosening in total hip arthroplasties.

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A rapid, quantitative assay for measuring alkaline phosphatase activity in osteoblastic cells in vitro.

Alkaline phosphatase (ALP) is the most widely recognized biochemical marker for osteoblast activity. Although its precise function is poorly understood, it is believed to play a role in skeletal mineralization. The aim of this study was to develop an assay suitable for measuring the activity of this enzyme in microtiter plate format. Using the well-characterized osteoblast-like cell line Saos-2, this paper describes an optimized biochemical assay suitable for measuring ALP activity in tissue culture samples. We have determined that a p-nitrophenyl phosphate substrate concentration of 9 mM provides highest enzyme activities. We have found that cell concentration, and hence enzyme concentration, affects both the kinetics and precision of the assay. We also tested several methods of enzyme solubilization and found that freeze-thawing the membrane fractions twice at -70 degrees C/37 degrees C or freeze-thawing once with sonication yielded highest enzyme activities. The activity of the enzyme decreased by 10% after 7 days storage. This assay provides a sensitive and reproducible method that is ideally suited for measuring ALP activity in isolated osteoblastic cells, although sample preparation and storage can influence results.

Alkaline Phosphatase↗