探讨构建载基因脂多糖胺纳米囊泡(NPs，简称pNPs)/透明质酸(HA)聚电解质多层膜(PEM)的组装行为和机理。

利用含骨形成蛋白2(pBMP-2)基因的pNPs作为阳离子聚电解质，HA作为阴离子聚电解质，通过层层自组装技术在钛或石英玻璃表面构建PEM，记为Ti/Quartz-pNPs-(HA/pNPs)_n，其中HA和pNPs依次组装1次为1个组装循环，n为组装循环数。采用原子力显微镜(AFM)观察膜组装过程中形貌和粗糙度值改变；膜表面Zeta电位(electric potential)表征聚电解质膜表面的电荷和吸附特性；利用耗散型石英晶体微天平(QCM-D)实时监测膜的组装过程，探索膜组装规律。

AFM观察发现，pNPs最初以单个离散的纳米粒子形式吸附在石英玻璃表面，随着组装进程，形成越来越粗壮、密集的"树枝"状三维立体纳米结构，膜表面粗糙度值逐渐增大。Zeta电位结果表明，石英玻璃表面经过处理后Zeta电位为-4.83 mV，首层pNPs的表面电位为正，至第3个组装循环后Zeta电位稳定在+18 mV；而HA的Zeta电位最初为负值，随组装层数增加，其表面电荷逐渐趋正；组装过程中Zeta电位呈锯齿状交替上升。石英晶体微天平测量结果显示，随着组装进行膜质量和厚度逐渐增加，且以指数型增长。

载基因pNPs/HA通过层层自组装构建具有独特三维纳米结构的聚电解质多层膜，其增长方式为指数型，具有纳米级粗糙度和非致密性的特点。

To explore the mechanism and behavior of polyelectrolyte multilayer films (PEM)of gene-loaded lipopolysaccharide-amine nanopolymersomes/hyaluronic acid self assembled on titanium or quartz surface.

Via layer-by-layer self assembly technology, PEM were constructed on titanium or quartz surface by using bone morphogenetic protein-2(BMP-2)plasmid-loaded lipopolysaccharide-amine nanopolymersomes(pNPs)as a polycation, and hyaluronic acid(HA)as a polyanion. The constructed PEM was defined as substrate-pNPs-(HA/pNPs)_n, where a successive deposition of HA and pNPs on substrate surface was defined as one assembly cycle, and n was the cycle number. The changes in topography and roughness of films during assembly were observed by atomic force microscope(AFM). The surface zeta potential was determined by a zeta potential and nanoparticle size analyzer. The assembly procedure was monitored in real time by a quartz crystal microbalance with dissipation(QCM-D), and their assembly patterns were explored.

AFM results showed that pNPs discretely and uniformly adhere to the substrate surface at first, and then with self assembly, a dense and strong tree-like three-dimensional nanostructure is gradually formed, followed by the progressive increase in their surface roughness. The zeta potential of films increases in a zigzag pattern with self assembly. For quartz surface, it is -4.83 mV, and after pNPs deposition, it increases to positive. For films with outmost layer of pNPs, after 3 assembly cycles, it was stabilized at 18 mV. For films with outmost layer of HA, their zeta potential gradually changes from negative to positive with assembly. The QCM-D results showed that with self assembly, the film mass and thickness increase in an exponential type.

Gene-loaded lipopolysaccharide-amine nanopolymersomes/hyaluronic acid can construct polyelectrolyte multilayer films with distinctive three-dimensional nanostructure via layer-by-layer self assembly, their growth mode is exponential, and the films have nano-scale roughness with non-dense texture.