Growth kinetics of lipid-based nanodiscs to unilamellar vesicles-a time-resolved small angle neutron scattering (SANS) study.

Mixtures of dimyristoyl-phosphatidylcholine (DMPC), dimyristoyl-phosphatidylglycerol (DMPG) and dihexanoyl-phosphatidylcholine (DHPC) in aqueous solutions spontaneously form monodisperse, bilayered nanodiscs (also known as "bicelles") at or below the melting transition temperature of DMPC (T(M) ~23°C). In dilute systems above the main transition temperature T(M) of DMPC, bicelles coalesce (increasing their diameter) and eventually self-fold into unilamellar vesicles (ULVs). Time-resolved small angle neutron scattering was used to study the growth kinetics of nanodiscs below and equal to T(M) over a period of hours as a function of temperature at two lipid concentrations in presence or absence of NaCl salt. Bicelles seem to undergo a sudden initial growth phase with increased temperature, which is then followed by a slower reaction-limited growth phase that depends on ionic strength, lipid concentration and temperature. The bicelle interaction energy was derived from the colloidal theory of Derjaguin and Landau, and Verwey and Overbeek (DLVO). While the calculated total energy between discs is attractive and proportional to their growth rate, a more detailed mechanism is proposed to describe the mechanism of disc coalescence. After annealing at low temperature (low-T), samples were heated to 50°C in order to promote the formation of ULVs. Although the low-T annealing of samples has only a marginal effect on the mean size of end-state ULVs, it does affect their polydispersity, which increases with increased T, presumably driven by the entropy of the system.

Effects of charge density and thermal history on the morphologies of spontaneously formed unilamellar vesicles.

A phospholipid mixture consisting of dimyristoyl phosphatidylcholine (DMPC), dihexanoyl phosphatidylcholine (DHPC), and the negatively charged dimyristoyl phosphatidylglycerol (DMPG) lipid is found to spontaneously form uniform-size unilamellar vesicles (ULVs). Small angle neutron scattering (SANS) is used to examine ULV size as a function of net charge, dilution, and thermal history. It shows that ULVs only exist within a narrow window of charge densities, where larger size ULVs can be obtained at a lower charge density through slow temperature annealing. There is also a 6-fold change in the size of low polydispersity ULVs, confirming a previously proposed model for spontaneously forming ULVs [Nieh, M.-P. et al. Langmuir 2005, 21, 6656]. Finally, the stability of these ULVs was confirmed through a series of high temperature dilution experiments, further making the case that these nanoparticles can be used as carriers for drugs and contrast imaging agents.