Evaluating Vascular Calcification Induced by High Dietary Calcium

In both studies, feed was withdrawn 4 h prior to sacrifice. All mice were euthanized by decapitation under anesthesia with diethyl ether. Blood was collected into micro tubes (Serum Z, Sarstedt, Nümbrecht, Germany) to gain serum. Serum concentrations of ionized calcium and inorganic phosphate were measured spectrophotometrically using Fluitest® CA CPC and Fluitest® PHOS kits (Analyticon Biotechnologies AG, Lichtenfels, Germany). Serum PTH was analyzed using the mouse PTH 1-84 ELISA Kit (#60-2305) and serum iFGF23 was quantified by using the mouse/rat FGF-23 (intact) ELISA Kit (#60-6800) both purchased from Immunotopics, Inc. (San Clemente, CA, USA). To identify whether high dietary calcium concentrations may induce vascular calcification, aortic root sections were prepared and quantified via von Kossa staining (Schmidt et al., 2012 (link)). The area of calcification was related to the aortic valve tissue.