Advanced glycation end-products (AGEs), senescent macroprotein derivatives formed at an accelerated rate in diabetes, are closely associated with vascular calcification in humans. In this study, the hypothesis that AGEs enhance calcification in cultured vascular smooth muscle cells (VSMCs) was tested. Using real-time polymerase chain reaction (PCR) and specific protein assays, it was demonstrated that rat aortic VSMCs incubated with AGEs exhibited an increased expression of the AGE receptor (RAGE) and typical bone proteins, such as osteopontin and alkaline phosphatase. Incubation with AGEs also enhanced calcium accumulation in VSMCs in time- and dose-dependent manners. These AGEs-mediated changes in VSMCs were partially attenuated by a neutralizing antibody to RAGE. The results suggest that AGEs that accumulate in diabetes could elicit the osteoblastic differentiation of VSMCs, thereby contributing to vascular calcification via the RAGE pathway. Interruption of the AGE-RAGE interaction might be a promising target for therapeutic intervention to prevent diabetic vascular calcification.