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Abstract：
Cationic liposomes composed of a novel lipid (N-{6-amino-1-[N-(9Z) -octadec9-enylamino] -1-oxohexan-(2S) -2-yl} -N\'- {2- [N, N-bis(2-aminoethyl) amino] ethyl} -2-hexadecylpropandiamide) (OO4) and dioleoylphosphatidylethanolamine (DOPE) possess high amounts of amino groups and are promising systems for lipofection. Moreover, these cationic liposomes can also be used as a polycationic entity in multilayer formation using layer-by-layer technique (LbL), which is a method to fabricate surface coatings by alternating adsorption of polyanions and polycations. Since liposomes are suitable for endocytosis by or fusion with cells, controlled release of their cargo on site is possible. Here, a polyelectrolyte multilayer (PEM) system was designed of chondroitin sulfate (CS) and collagen type I (Col I) by LbL technique with OO4/DOPE liposomes embedded in the terminal layers to create an osteogenic microenvironment. Both, the composition of PEM and cargo of the liposomes were used to promote osteogenic differentiation of C2C12 myoblasts as in vitro model. The internalization of cargo-loaded liposomes from the PEM into C2C12 cells was studied using lipophilic (Rhodamine-DOPE conjugate) and hydrophilic (Texas Red-labeled dextran) model compounds. Besides, the use of Col I and CS should mimic the extracellular matrix of bone for future applications such as bone replacement therapies. Physicochemical studies of PEM were done to characterize the layer growth, thickness, and topography. The adhesion of myoblast cells was also evaluated whereby the benefit of a cover layer of CS and finally Col I above the liposome layer was demonstrated. As proof of concept, OO4/DOPE liposomes were loaded with dexamethasone, a compound that can induce osteogenic differentiation. A successful induction of osteogenic differentiation of C2C12 cells with the novel designed liposome-loaded PEM system was shown. These findings indicate that designed OH4/DOPE loaded PEMs have a high potential to be used as drug delivery or transfection system for implant coating in the field of bone regeneration and other applications.
摘要：
由新型脂质（N-{6-氨基-1-[N-（9Z）-十八烷基氨基]-1-氧代己烷-（2S）-2-基}-N'-{2-[N，N-双（2-氨基乙基）氨基]乙基}-2-十六烷基丙二酰胺）（OO4）和二油酰基磷脂酰乙醇胺（DOPE）具有大量的氨基，是有前途的脂质体转染系统。此外，这些阳离子脂质体也可以在使用逐层技术（LbL）的多层形成中用作聚阳离子实体，这是一种通过聚阴离子和聚阳离子的交替吸附来制造表面涂层的方法。由于脂质体适用于细胞内吞作用或与细胞融合，控制释放他们的货物在现场是可能的。这里,通过LbL技术设计了硫酸软骨素（CS）和I型胶原蛋白（ColI）的聚电解质多层（PEM）系统，其中OO4/DOPE脂质体嵌入在末端层中，以创建成骨微环境。两者,用PEM的组成和脂质体的货物促进C2C12成肌细胞的成骨分化作为体外模型。使用亲脂性(罗丹明-DOPE缀合物)和亲水性(得克萨斯红标记的葡聚糖)模型化合物研究装载货物的脂质体从PEM内化到C2C12细胞中。此外,ColI和CS的使用应模拟骨的细胞外基质，以用于未来的应用，例如骨替代疗法。PEM的物理化学研究进行了表征层的生长，厚度,和地形。还评估了成肌细胞的粘附性,从而证明了脂质体层上方的CS覆盖层和最后的ColI的益处。作为概念的证明，OO4/DOPE脂质体负载地塞米松，一种能诱导成骨分化的化合物。显示了用新型设计的脂质体负载的PEM系统成功诱导C2C12细胞的成骨分化。这些发现表明，设计的OH4/DOPE负载的PEM具有很高的潜力，可以用作骨再生和其他应用领域中植入物涂层的药物递送或转染系统。
