Brito Barrera Y A, Hause G, Menzel M, Schmelzer C E H, Lehner E, Mäder K, Wölk C, Groth T
Department Biomedical Materials, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Heinrich Damerow Strasse 4, 06120, Halle (Saale), Germany.
Martin Luther University Halle-Wittenberg, Biocenter, Weinbergweg 22, 06120, Halle (Saale), Germany.
Mater Today Bio. 2020 Jul 31;7:100071. doi: 10.1016/j.mtbio.2020.100071. eCollection 2020 Jun.
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.
由新型脂质（N-{6-氨基-1-[N-(9Z)-十八碳-9-烯基氨基]-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）和二油酰磷脂酰乙醇胺（DOPE）组成的阳离子脂质体含有大量氨基，是有前景的脂质转染系统。此外，这些阳离子脂质体还可以作为聚阳离子实体用于采用层层技术（LbL）的多层形成，层层技术是一种通过聚阴离子和聚阳离子交替吸附来制备表面涂层的方法。由于脂质体适合细胞的内吞作用或与细胞融合，因此可以在现场实现其负载物的控释。在此，通过层层技术设计了一种由硫酸软骨素（CS）和I型胶原（Col I）组成的聚电解质多层（PEM）系统，在终端层嵌入OO4/DOPE脂质体以创建成骨微环境。PEM的组成和脂质体的负载物都被用于促进C2C12成肌细胞的成骨分化，以此作为体外模型。使用亲脂性（罗丹明-DOPE共轭物）和亲水性（德克萨斯红标记的葡聚糖）模型化合物研究了负载有货物的脂质体从PEM内化到C2C12细胞中的情况。此外，使用Col I和CS应该能够模拟骨的细胞外基质，以用于未来诸如骨替代疗法等应用。对PEM进行了物理化学研究以表征层生长、厚度和形貌。还评估了成肌细胞的黏附情况，结果表明在脂质体层上方存在CS层以及最终的Col I层的益处。作为概念验证，OO4/DOPE脂质体负载了地塞米松，一种能够诱导成骨分化的化合物。结果显示，新型设计的负载脂质体的PEM系统成功诱导了C2C12细胞的成骨分化。这些发现表明，设计的负载OH4/DOPE的PEMs在骨再生领域及其他应用中作为植入物涂层的药物递送或转染系统具有很高的潜力。
