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      Monocyte/macrophage regulation of vascular calcification in vitro.


      Enzyme Activation, Alkaline Phosphatase, metabolism, Animals, Aorta, cytology, Arteriosclerosis, complications, Calcinosis, Cattle, Cell Communication, drug effects, Cell Count, Cell Differentiation, Cells, Cultured, Coculture Techniques, Culture Media, Conditioned, pharmacology, Humans, Lipopolysaccharides, Lipoproteins, LDL, Macrophages, Monocytes, Osteoblasts, enzymology, Stem Cells, Tumor Necrosis Factor-alpha, biosynthesis

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          Calcification is a common complication of atherosclerosis and other chronic inflammatory processes that involves infiltration of monocytes and accumulation of macrophages. To determine whether these cells modulate vascular calcification in vitro, calcifying vascular cells (CVCs), a subpopulation of osteoblast-like cells derived from the artery wall, were cocultured with human peripheral blood monocytes for 5 days. Results showed that alkaline phosphatase (ALP) activity, a marker of osteoblastic differentiation, was significantly greater in cocultures than in cultures of CVCs or monocytes alone. Both ALP activity and matrix mineralization increased in proportion to the number of monocytes added. Activation of monocyte/macrophages (M/Ms) by oxidized LDL further increased ALP activity in cocultures. However, neither conditioned medium from oxidized-LDL-activated M/Ms or transwell coculture had this effect on CVCs, which suggests a need for cell-to-cell contact. In contrast, conditioned medium from lipopolysaccharide-activated M/Ms increased ALP activity of CVCs. ELISA showed that lipopolysaccharide-activated M/Ms secreted tumor necrosis factor-alpha, and neutralizing antibody to tumor necrosis factor-alpha attenuated the induction of ALP activity by the conditioned media. These results suggest that M/Ms enhance in vitro vascular calcification via 2 independent mechanisms: cell-cell interaction and production of soluble factors such as tumor necrosis factor-alpha.

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