Figure 4.

Warfarin enhanced the susceptibility of Pi-induced calcification. (a) Effect of warfarin on Pi-induced AC in cultured explants of aorta. Pieces of rat aorta were cultured in calcification medium for 7 days. The calcified lesions were examined by von Kossa and Alizarin Red S staining. Scale bar: 100 μm. (b, c) Passage 5 (P5) and passage 15 (P15) VSMC were treated with phosphate (2.0 mmol/L), vehicle (methanol), and warfarin (2.0 μmol). Representative Alizarin Red staining for VSMC. Calcification in day 3 (b) and day 7 (c). Treatment with warfarin enhanced the VSMC calcification in both the senescent and young VSMC. Of note, calcification was further deteriorated by the addition of warfarin in the young VSMC compared with vehicle control in day 3.