Abstract

Background: Liposomes are utilized as a drug delivery carrier in various fields of biomedicine. They are synthesized in the nanometer-size range and are becoming a viable drug delivery carrier for the treatment of different diseases. MicroRNAs as regulatory elements could be transferred to cells for changing their morphology or physiology. The study’s major aim is to find the optimized formula of liposomes for transfection of microRNA to human mesenchymal stem cell line S1939 (HMSCs). Materials and Methods: Various ratios of soybean phosphatidylcholine (SPC), cholesterol, 1, 2 dioleoyloxy-3- (trimethylammonium) propane (DOTAP), and polyethylene glycol (PEG) were combined. The mean diameter of all formulations and their surface properties were determined by a zeta sizer device and scanning electron microscope, respectively. The cytotoxicity of formulations was assessed using MTT (3,4,5-dimethyl thiazol-2-yl) (2,5-diphenyltetrazolium bromide) assay. The transfection effectiveness of liposomal miRNA vs empty liposomes was determined using agarose gel electrophoresis. Results: The optimized liposome vesicles were prepared using 45:30:27.5:5 molar ratios of SPC:DOTAP:cholesterol: DSPE-PEG. The liposome formulations F10 and F18 were the best in terms of biocompatibility because of the higher viabilities of treated cells. The best formulation (F18, containing 0.7 µg of miRNA and 10 µg of liposome) was nearly 100% efficient in sequestering and fixing miRNA. Phase-contrast and fluorescent microscopic examinations showed intra-nuclear as well as intracytoplasmic localization of the particles. Conclusion: Some easily prepared liposomal formulation vehicles are quite efficient in the transfection of miRNA into the HMSCs and could be used for in vitro applications in regenerative medicine.

摘要

背景：脂质体被用作生物医学各个领域的药物递送载体。它们在纳米尺寸范围内合成，正在成为治疗不同疾病的可行药物输送载体。作为调控元件的 MicroRNA 可以转移到细胞中以改变它们的形态或生理学。该研究的主要目的是找到用于将 microRNA 转染到人间充质干细胞系 S1939 (HMSCs) 的优化脂质体配方。材料和方法：将各种比例的大豆磷脂酰胆碱 (SPC)、胆固醇、1, 2 二油酰氧基-3-（三甲基铵）丙烷 (DOTAP) 和聚乙二醇 (PEG) 组合在一起。所有配方的平均直径及其表面特性分别由 zeta sizer 装置和扫描电子显微镜测定。使用 MTT（3,4,5-二甲基噻唑-2-基）（2,5-二苯基四唑溴化物）测定法评估制剂的细胞毒性。使用琼脂糖凝胶电泳测定脂质体 miRNA 与空脂质体的转染效果。结果：使用 45:30:27.5:5 摩尔比的 SPC:DOTAP:胆固醇:DSPE-PEG 制备优化的脂质体囊泡。脂质体制剂 F10 和 F18 在生物相容性方面是最好的，因为处理过的细胞具有更高的活力。最佳配方（F18，含有 0.7 µg miRNA 和 10 µg 脂质体）在隔离和固定 miRNA 方面几乎 100% 有效。相衬和荧光显微镜检查显示颗粒位于核内和胞质内。结论：一些易于制备的脂质体制剂载体在将 miRNA 转染到 HMSCs 中非常有效，可用于再生医学的体外应用。