Lyso-phosphatidylcholine induces osteogenic gene expression and phenotype in vascular smooth muscle cells.

Objective: Calcifying vascular cells in human atherosclerotic plaques actively contribute to ectopic vascular mineralization. Lyso-phosphatidylcholine (LPC), a product of oxidized phosphatidylcholine hydrolysis, is found at concentrations of 1-12 microg/g tissue throughout the atheroma. The objective of this study was to determine if LPC induces an osteogenic phenotype in vascular smooth muscle cells.

Methods And Results: Proliferating human aortic smooth muscle cells were treated with a wide-range of LPC concentrations (0.1 nM to 100 microM) over 14 days. Von Kossa, Alizarin Red S, and alkaline phosphatase staining were used to identify mineralizations. RT-PCR, ELISA, alkaline phosphatase activity, and 45Ca incorporation assays were used to evaluate the osteo-inductive effect of LPC on smooth muscle phenotype. Histology and morphometry revealed that cells treated with as little as 10 nM LPC produced calcium phosphate deposits in culture. LPC-treated vascular smooth muscle cells showed a significant increase in 45Ca incorporation and alkaline phosphatase activity. Furthermore, LPC treatment induced a significant loss of Schnurri 3 protein, a key repressor of Runt-related transcription factor 2 stability. Genomic studies revealed that osteogenic gene expression was significantly up-regulated in LPC-treated cells, which is attributed to increased Runt-related transcription factor 2 expression and transcriptional activity.

Conclusion: LPC induces osteogenic morphology, physiology, gene expression, and phenotype in vascular smooth muscle cells. The present study suggests that localized concentrations of LPC in human atherosclerotic plaques may be a contributing factor to the generation of calcifying vascular cells.