脂质体复合物,多组分非病毒基因载体,通常表现出优异的胶体稳定性,降低细胞毒性,和高转染效率。在这项研究中,一个新概念,光化学反应诱导的转染,使用光敏剂(PS)负载的脂质体复合物,通过光敏剂的光激发导致增强的转染和细胞毒性作用。金丝桃素,疏水性光敏剂,被包封在脂质体的脂质双层中。预制的纳米金丝桃素脂质体包裹了线性聚乙烯亚胺(lPEI)/pDNA复合物,导致金丝桃素脂质体复合物(Hy-LPP)的形成。含有50nM金丝桃素和0.25μgpDNA的Hy-LPP的直径为185.6±7.74nm和230.2±4.60nm,分别,通过动态光散射(DLS)和原子力显微镜(AFM)测量。凝胶电泳证实了金丝桃素和pDNA在脂质体复合物中的包封。此外,评估了在200、600和1000mJ/cm2的辐射下细胞内Hy-LPP的体外辐射。它证明体外荧光素酶表达比未照射细胞高60至75倍。乳酸脱氢酶(LDH)测定支持减少的转染是光细胞毒性的结果。开发的载有光敏剂的脂质体复合物提高了外源基因的转染效率或诱导的光细胞毒性;然而,前沿在于应用的光化学剂量。细胞内金丝桃素的光触发光激发导致活性氧(ROS)的产生,导致HepG2细胞中的光选择性转染。结论是,两种共同递送的治疗剂通过调整施加的光化学剂量而导致增强的转染和光动力效应。
The lipopolyplex, a multicomponent nonviral gene carrier, generally demonstrates superior colloidal stability, reduced cytotoxicity, and high transfection efficiency. In this study, a new concept, photochemical reaction-induced transfection, using photosensitizer (PS)-loaded lipopolyplexes was applied, which led to enhanced transfection and cytotoxic effects by photoexcitation of the photosensitizer. Hypericin, a hydrophobic photosensitizer, was encapsulated in the lipid bilayer of liposomes. The preformed nanosized hypericin liposomes enclosed the linear polyethylenimine (lPEI)/pDNA polyplexes, resulting in the formation of hypericin lipopolyplexes (Hy-LPP). The diameters of Hy-LPP containing 50 nM hypericin and 0.25 μg of pDNA were 185.6 ± 7.74 nm and 230.2 ± 4.60 nm, respectively, measured by dynamic light scattering (DLS) and atomic force microscopy (AFM). Gel electrophoresis confirmed the encapsulation of hypericin and pDNA in lipopolyplexes. Furthermore, in vitro irradiation of intracellular Hy-LPP at radiant exposures of 200, 600, and 1000 mJ/cm2 was evaluated. It demonstrated 60- to 75-fold higher in vitro luciferase expression than that in nonirradiated cells. The lactate dehydrogenase (LDH) assay supported that reduced transfection was a consequence of photocytotoxicity. The developed photosensitizer-loaded lipopolyplexes improved the transfection efficiency of an exogenous gene or induced photocytotoxicity; however, the frontier lies in the applied photochemical dose. The light-triggered photoexcitation of intracellular hypericin resulted in the generation of reactive oxygen species (ROS), leading to photoselective transfection in HepG2 cells. It was concluded that the two codelivered therapeutics resulted in enhanced transfection and a photodynamic effect by tuning the applied photochemical dose.