Department of Pharmaceutics, Nanomedicine Research Centre, Indo-Soviet Friendship College of Pharmacy, Moga, Punjab, India.
Drug Deliv. 2013 Feb;20(2):57-64. doi: 10.3109/10717544.2012.752419.
Small interfering RNA (siRNA) has been widely used as potential therapeutic for treatment of various genetic disorders. However, rapid degradation, poor cellular uptake and limited stability in blood limit the effectiveness of the systemic delivery of siRNA. Therefore, an efficient delivery system is required to enhance its transfection and duration of therapeutics. In the present study, plasmid DNA (pEGFPN3) expressing green fluorescent protein (GFP) was used as a reporter gene. Chitosan nanoparticles/polyplexes and cationic liposomes/lipoplexes were developed and compared for their transfectivity and therapeutic activity in mammalian cell line (HEK 293). The nanoparticulates were first characterized by assessing the surface charge (zeta potential), size (dynamic light scattering) and morphology (transmission electron microscope) followed by evaluation for their DNA retardation ability, transfection efficiency and cytotoxicity on HEK 293 cell line. The chitosan nanoparticles/plasmid DNA (pDNA) complex and liposomes/pDNA complex were co-transfected with GFP-specific siRNA into HEK 293 cells and it was found that both are efficient delivery vehicles for siRNA transfection, resulting in ~57% and ~70% suppression of the targeted gene (GFP), respectively, as compared with the mock control (cells transfected with nanocarrier/pDNA complexes alone). This strong inhibition of GFP expression indicated that cationic liposomes are better than chitosan nanoparticles and can be used as an effective carrier of siRNA in mammalian cells.
小干扰 RNA（siRNA）已被广泛用作治疗各种遗传疾病的潜在治疗方法。然而，siRNA 的快速降解、细胞摄取能力差和血液稳定性有限限制了其系统递送的效果。因此，需要一种有效的递送系统来增强其转染效率和治疗持续时间。在本研究中，表达绿色荧光蛋白（GFP）的质粒 DNA（pEGFPN3）被用作报告基因。开发了壳聚糖纳米粒子/聚阳离子和阳离子脂质体/脂质体，并比较了它们在哺乳动物细胞系（HEK 293）中的转染效率和治疗活性。首先通过评估表面电荷（zeta 电位）、粒径（动态光散射）和形态（透射电子显微镜）来表征纳米颗粒，然后评估其 DNA 阻滞能力、转染效率和对 HEK 293 细胞系的细胞毒性。壳聚糖纳米粒子/质粒 DNA（pDNA）复合物和脂质体/pDNA 复合物与 GFP 特异性 siRNA 共转染 HEK 293 细胞，结果发现两者都是有效的 siRNA 转染载体，与模拟对照（仅用纳米载体/pDNA 复合物转染的细胞）相比，分别导致靶向基因（GFP）的抑制率约为 57%和 70%。GFP 表达的强烈抑制表明阳离子脂质体优于壳聚糖纳米粒子，可作为哺乳动物细胞中 siRNA 的有效载体。
