CAJ | 학술논문

目的：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。方法利用含骨形成蛋白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。结果 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。结论载基因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。
목적：탐토구건재기인지다당알납미낭포(NPs,간칭pNPs)/투명질산(HA)취전해질다층막(PEM)적조장행위화궤리。방법이용함골형성단백2(pBMP-2)기인적pNPs작위양리자취전해질,HA작위음리자취전해질,통과층층자조장기술재태혹석영파리표면구건PEM,기위Ti/Quartz-pNPs-(HA/pNPs)n,기중HA화pNPs의차조장1차위1개조장순배,n위조장순배수。채용원자력현미경(AFM)관찰막조장과정중형모화조조도치개변；막표면Zeta전위(electric potential)표정취전해질막표면적전하화흡부특성；이용모산형석영정체미천평(QCM-D)실시감측막적조장과정,탐색막조장규률。결과 AFM관찰발현,pNPs최초이단개리산적납미입자형식흡부재석영파리표면,수착조장진정,형성월래월조장、밀집적“수지”상삼유입체납미결구,막표면조조도치축점증대。 Zeta전위결과표명,석영파리표면경과처리후Zeta전위위-4.83 mV,수층pNPs적표면전위위정,지제3개조장순배후Zeta전위은정재+18 mV；이HA적Zeta전위최초위부치,수조장층수증가,기표면전하축점추정；조장과정중Zeta전위정거치상교체상승。석영정체미천평측량결과현시,수착조장진행막질량화후도축점증가,차이지수형증장。결론재기인pNPs/HA통과층층자조장구건구유독특삼유납미결구적취전해질다층막,기증장방식위지수형,구유납미급조조도화비치밀성적특점。
Objective 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. Methods 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. Results 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. Conclusion 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.