Department of Pharmacy, School of Health Sciences, Section of Pharmaceutical Technology, Laboratory of Pharmaceutical Nanotechnology, National and Kapodistrian University of Athens , Athens , Greece.
J Liposome Res. 2019 Dec;29(4):313-321. doi: 10.1080/08982104.2018.1468774. Epub 2019 Jun 7.
The aim of the present study is to investigate the interactions between liposomes and proteins and to evaluate the role of liposomal lipid composition and concentration in the formation of protein corona. Liposomes composed of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) or hydrogenated soybean phosphatidylcholine (HSPC) with 1,2-dipalmitoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (sodium salt) (DPPG), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-3000] (DPPE-PEG 3000), cholesterol (CH) or mixtures of these lipids, were prepared at different concentrations by the thin-film hydration method. After liposomes were dispersed in HPLC-grade water and foetal bovine serum (FBS), their physicochemical characteristics, such as size, size distribution, and -potential, were determined using dynamic and electrophoretic light scattering. Aggregation of DPPC, HSPC, DPPC:CH (9:1 molar ratio), and HSPC:CH (9:1 molar ratio) in FBS was observed. On the contrary, liposomes incorporating DPPG lipids and CH both in a molar ratio of 11% were found to be stable over time, while their size did not alter dramatically in biological medium. Liposomes containing CH and PEGylated lipids retain their size in the presence of serum as well as their physical stability. In addition, our results indicate that the protein binding depends on the presence of polyethylene glycol (PEG), CH, concentration and surface charge. In this paper, we introduce a new parameter, fraction of stealthiness (), for investigating the extent of protein binding to liposomes. This parameter depends on the changes in size of liposomes after serum incubation, while liposomes have stealth properties when is close to 1. Thus, we conclude that lipid composition and concentration affect the adsorption of proteins and the liposomal stabilization.
本研究旨在探究脂质体与蛋白质之间的相互作用，并评估脂质体的脂质组成和浓度在蛋白质冠形成中的作用。采用薄膜水化法，分别以 1,2-二棕榈酰-sn-甘油-3-磷酸胆碱（DPPC）或氢化大豆磷脂酰胆碱（HSPC）与 1,2-二棕榈酰-sn-甘油-3-磷酸-1'-rac-甘油（钠盐）（DPPG）、1,2-二棕榈酰-sn-甘油-3-磷酸乙醇胺-N-[甲氧基（聚乙二醇）-3000]（DPPE-PEG 3000）、胆固醇（CH）或这些脂质的混合物，制备了不同浓度的脂质体。将脂质体分散在高效液相色谱级水和胎牛血清（FBS）中后，采用动态光散射和电泳光散射法测定其粒径、粒径分布和 Zeta 电位等理化特性。在 FBS 中观察到 DPPC、HSPC、DPPC:CH（9:1 摩尔比）和 HSPC:CH（9:1 摩尔比）的聚集。相反，在摩尔比为 11%的情况下，同时包含 DPPG 脂质和 CH 的脂质体随着时间的推移被发现是稳定的，而其粒径在生物介质中没有发生显著变化。含有 CH 和 PEG 化脂质的脂质体在存在血清的情况下保持其粒径和物理稳定性。此外，我们的结果表明，蛋白质结合取决于聚乙二醇（PEG）、CH、浓度和表面电荷的存在。在本文中，我们引入了一个新参数，即隐形分数（），用于研究蛋白质与脂质体结合的程度。该参数取决于血清孵育后脂质体粒径的变化，当接近 1 时，脂质体具有隐形特性。因此，我们得出结论，脂质组成和浓度会影响蛋白质的吸附和脂质体的稳定性。
