Open AccessCellular Component of Vascular Calcification
Author(s) -
Yuko Watanabe,
Miyuki Suzuki,
Yuhta Oyama,
E Kusano,
Kaichiro Tamba,
Osamu Ito,
Chihiro Ito,
Muneaki Imai,
Yoshihide Asano
Publication year - 2002
Publication title -
the nephron journals/nephron journals
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.951
H-Index - 72
eISSN - 2235-3186
pISSN - 1660-8151
DOI - 10.1159/000065461
Subject(s) - calcium , von kossa stain , calcification , phosphate , staining , deposition (geology) , medicine , biochemistry , endocrinology , chemistry , biology , pathology , enzyme , alkaline phosphatase , paleontology , sediment
It has been reported that calcium deposition and calcium content in cultured human aortic smooth muscle cells (SMC) increased when the cells are incubated in a medium with a high phosphate concentration (2 mM). To determine the cellular components or soluble factors contributing to the deposition, we cultured commercially available SMC, fibroblasts (Fb) and endothelial cells (Ed). These cells and their mixtures were incubated for 10 days in normal or high-phosphate media. Calcium crystals were stained by the von-Kossa staining and counted in the defined area. Calcium content was measured by a colorimetric assay. SMC were incubated in high-phosphate media (up to 2 mM) or beta-glycerophosphate (beta-GP) media, resulting in no obvious deposition of calcium crystals, irrespective of the coating of type I collagen on the dish. Next, various combinations of cells were cultured, and a significant number of depositions were observed only when Fb were included in the combination. The calcium content was significantly higher in cultures of SMC and Fb. The calcium deposition on single or mixture of the cells did not increase compared with control when cells were incubated in a high concentration of phosphate, cultured in the existence of beta-GP or uremic serum. We therefore conclude that Fb, rather than SMC or Ed, are essential for calcium deposition and calcium accumulation in culture. Phosphate concentration in the medium and uremic serum did not influence the deposition of calcium.