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.

阳离子脂质体，由新型脂质（N- {6-氨基-1- [N-（9Z）-octadec9-烯基氨基] -1-氧己酮-（2S）-2-基} –N'- {2- [N ，N-双（2-氨基乙基）氨基]乙基} -2-十六烷基丙酰胺（OO4）和油酰磷脂酰乙醇胺（DOPE）具有大量的氨基，是有希望的脂质转染系统。此外，这些阳离子脂质体还可以在使用逐层技术（LbL）的多层形成中用作聚阳离子实体，该方法是通过交替吸附聚阴离子和聚阳离子来制造表面涂层的方法。由于脂质体适用于细胞内吞或与细胞融合，因此有可能在现场控制其货物的释放。这里，通过LbL技术设计了硫酸软骨素（CS）和I型胶原（Col I）的聚电解质多层（PEM）系统，并在末端层中嵌入了OO4 / DOPE脂质体，从而形成了成骨微环境。作为体外模型，PEM的组成和脂质体的载量均用于促进C2C12成肌细胞的成骨分化。使用亲脂性（若丹明-DOPE共轭物）和亲水性（得克萨斯红标记的葡聚糖）模型化合物研究了从PEM到C2C12细胞的载有脂质体的内在化。此外，Col I和CS的使用应模仿骨骼的细胞外基质，以用于将来的应用，例如骨替代疗法。对PEM进行了物理化学研究，以表征层的生长，厚度和形貌。还评估了成肌细胞的粘附，从而证明了CS的覆盖层以及最终脂质体层上方的Col I的益处。作为概念的证明，OO4 / DOPE脂质体装有地塞米松，地塞米松是一种可以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，设计的负载OH4 / DOPE的PEM具有很高的潜力，可用作骨再生和其他应用领域中植入物涂层的药物输送或转染系统。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，设计的负载OH4 / DOPE的PEM具有很高的潜力，可用作骨再生和其他应用领域中植入物涂层的药物输送或转染系统。显示了用新颖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，设计的负载OH4 / DOPE的PEM具有很高的潜力，可用作骨再生和其他应用领域中植入物涂层的药物输送或转染系统。