Howard HsuAntonio ArtiguesMaria Villar2009-05-052009-05-052006-10-16Howard Hsu;Antonio Artigues;Maria Villar: Induction of calcification by serum depletion in cell culture: a model for focal calcification in aortas related to atherosclerosis. Lipids Health Dis 2008, 7(1):2.https://hdl.handle.net/2271/600BACKGROUND:Since aortic calcification has been shown to initiate in the lower zone of well-thickened plaques (LZP) adjacent to the aortic media of rabbits fed supplemental cholesterol diets, a restricted supply of serum to vascular cells could play a role in vascular calcification. This study was designed to use a cell culture model to support this hypothesis.RESULTS:Rabbit aortic smooth muscle cells were grown to confluence in a culture media containing 10 % fetal bovine serum (FBS). The confluent cells were then exposed to the media for 2 hrs with or without serum at a Ca × P ion product range of 4.5 9.4 mM2. In contrast to the cells cultured in the presence of FBS, confluent cells in its absence displayed marked mineral-positive alizarin red staining and infrared absorption of mineral phosphate. A kinetic parameter C1/2 was used to designate the concentration of serum or its protein constituents needed to reduce the deposition of Ca and P by half. The C1/2 for FBS and rabbit serum was 0.04 0.07 % The C1/2 value for rabbit serum proteins was 13.5 µg/ml corresponding to the protein concentration in 0.06 % of serum. This C1/2 was markedly smaller than 86.2 µg/ml for bovine serum albumin present in 0.37 % serum (p < 0.05). Serum depletion also caused marked membrane translocation as evidenced through a specific apoptosis dye uptake by cells. The proteomic analysis of calcifying vesicles, which can be released by serum depletion, revealed several calcification-related proteins.CONCLUSION:The aortic smooth muscle cell culture model suggests that serum depletion may play a role in the initiation of aortic calcification. The serum exhibits remarkable ability to inhibit cell-mediated calcification.en-USThis is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.http://creativecommons.org/licenses/by/2.0AnimalsAorta, Thoracic/metabolism/ pathologyAortic Diseases/etiology/metabolism/pathologyBiological Markers/metabolismCalcinosis/metabolism/ pathologyCholesterol, Dietary/adverse effects/ metabolismCholesterol, LDL/ metabolismCoronary Artery Disease/etiology/metabolism/pathologyCytoplasmic Vesicles/metabolism/ pathologyDiet, AtherogenicDietary Supplements/adverse effectsHypercholesterolemia/bloodMaleMicrosomes/metabolism/pathologyRabbitsTunica Intima/metabolism/ pathologyArticle10.1186/1476-511X-7-2PMC17042957openAccess