Unveiling growth and dynamics of liposomes by graphene liquid cell-transmission electron microscopy

Liposome is a model system for biotechnological and biomedical purposes spanning from targeted drug delivery to modern vaccine research. Yet, the growth mechanism of liposomes is largely unknown. In this work, the formation and evolution of phosphatidylcholine-based liposomes are studied in real-time by graphene liquid cell-transmission electron microscopy (GLC-TEM). We reveal important steps in the growth, fusion and denaturation of phosphatidylcholine (PC) liposomes. We show that initially complex lipid aggregates resembling micelles start to form. These aggregates randomly merge while capturing water and forming small proto-liposomes. The nanoscopic containers continue sucking water until their membrane becomes convex and free of redundant phospholipids, giving stabilized PC liposomes of different sizes. In the initial stage, proto-liposomes grow at a rate of 10-15 nm s(-1), which is followed by their growth rate of 2-5 nm s(-1), limited by the lipid availability in the solution. Molecular dynamics (MD) simulations are used to understand the structure of micellar clusters, their evolution, and merging. The liposomes are also found to fuse through lipid bilayers docking followed by the formation of a hemifusion diaphragm and fusion pore opening. The liposomes denaturation can be described by initial structural destabilization and deformation of the membrane followed by the leakage of the encapsulated liquid. This study offers new insights on the formation and growth of lipid-based molecular assemblies which is applicable to a wide range of amphiphilic molecules.

Automated summary

In this study, the formation and evolution of phosphatidylcholine-based liposomes were investigated in real-time using graphene liquid cell-transmission electron microscopy (GLC-TEM). The growth, fusion, and denaturation steps of liposomes were revealed. The study showed that complex lipid aggregates resembling micelles formed initially, which randomly merged and captured water to form small proto-liposomes. The liposomes grew in size until their membrane became convex and free of redundant phospholipids. Liposomes were found to fuse through lipid bilayers docking and the formation of a fusion pore. Denaturation of liposomes involved structural destabilization and membrane deformation followed by the leakage of encapsulated liquid.