Ostwald Ripening of Triacylglycerol Droplets Embedded in Glass-Supported Phospholipid Bilayers

Lipid droplets are fat storage organelles that consist of a neutral lipid core surrounded by a phospholipid monolayer. Because of their important biological functions, reconstituting model lipid droplets in synthetic phospholipid membranes is of great interest. In the present study, we investigated the incorporation of triacylglycerol droplets into glass-supported phospholipid bilayers by using fluorescence microscopy. We adsorbed triolein emulsions onto a glass surface that was partially covered with planar bilayers. After adsorption, triolein droplets were found to be immobilized in the bilayer membrane. The volume of each bound droplet varied over time. Large droplets grew, whereas small droplets shrank. Additionally, data on fluorescence recovery after photobleaching obtained for a phospholipid probe indicate that phospholipids on and near triolein droplets were fully mobile. Furthermore, photobleaching data obtained for a triacylglycerol probe indicate that triolein molecules diffused between different droplets along the planar bilayer. These results demonstrate Ostwald ripening, where triolein molecules in a small droplet dissolved in the bilayer, diffused laterally, and eventually bound to the interfaces of larger droplets. We investigated the ripening rate by using the average of the cube root of the fluorescence emission obtained for individual droplets. The ripening slowed after the addition of trilinolein to the triolein phase. Finally, we investigated the time dependence of the size distributions of the triolein droplets. The distribution was initially nearly unimodal and subsequently became bimodal.