Cholesterol loading affects osteoblastic differentiation in mouse mesenchymal stem cells

[Display omitted] ► We separate the effects of cholesterol itself from other components of modified lipoproteins. ► Accumulation of cholesterol positively affects MSC osteoblastic differentiation. ► The osteogenic potency of cholesterol was mostly due to CE levels. ► BMP2 and Runx2 are involved in the effects of cholesterol on MSC osteoblastic differentiation. Effects of cholesterol on osteoblastic differentiation was evaluated in mouse bone marrow derived mesenchymal stem cells (MSCs). Cholesterol-treated MSCs showed a stimulated differentiation process with induced mRNA and protein levels of osteogenic lineage markers, increased alkaline phosphatase (AKP) activity and more mineralized nodules. However, the stimulation extent was reduced when incubating the cells with cholesterol plus the ACAT (acyl-CoA:cholesterol acyltransferase) inhibitor Sandoz58035 or SiRNA-ACAT1 [which blocks the esterification of free cholesterol (FC) to cholesteryl ester (CE)], indicating the osteogenic potency of cholesterol was mostly due to CE levels. The key role of BMP2 and Runx2 in the effects of cholesterol on MSC osteogenesis was elucidated. These results point to cholesterol as a modulator of osteoblastic differentiation, which separate cholesterol itself from other components of modified lipoproteins.