NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain.
NanoBioCel Group, Laboratory of Pharmaceutics, School of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain; Histology and Cell Biology Department, Faculty of Medicine, University of Alexandria, Alexandria, Egypt; Department of Basic Sciences, The American University of Antigua-College of Medicine, Coolidge, Antigua and Barbuda.
J Control Release. 2019 Jun 28;304:181-190. doi: 10.1016/j.jconrel.2019.05.010. Epub 2019 May 6.
The incorporation of chloroquine within nano formulations, rather than as a co-treatment of the cells, could open a new avenue for in vivo retinal gene delivery. In this manuscript, we evaluated the incorporation of chloroquine diphosphate into the cationic niosome formulation composed of poloxamer 188, polysorbate 80 non-ionic surfactants, and 2,3-di (tetradecyloxy) propan-1-amine (hydrochloride salt) cationic lipid, to transfect rat retina. Niosome formulations without and with chloroquine diphosphate (DPP80, and DPP80-CQ, respectively) were prepared by the reverse phase evaporation technique and characterized in terms of size, PDI, zeta potential, and morphology. After the incorporation of the pCMS-EGFP plasmid, the resultant nioplexes -at different cationic lipid/DNA mass ratios- were further evaluated to compact, liberate, and secure the DNA against enzymatic digestion. In vitro procedures were achieved in ARPE-19 cells to assess transfection efficacy and intracellular transportation. Both nioplexes formulations transfected efficiently ARPE-19 cells, although the cell viability was clearly better in the case of DPP80-CQ nioplexes. After subretinal and intravitreal injections, DPP80 nioplexes were not able to transfect the rat retina. However, chloroquine containing vector showed protein expression in many retinal cells, depending on the administration route. These data provide new insights for retinal gene delivery based on chloroquine-containing niosome non-viral vectors.
将氯喹纳入纳米制剂中，而不是作为细胞的共同处理，可能为体内视网膜基因传递开辟新途径。在本手稿中，我们评估了二磷酸氯喹（chloroquine diphosphate）掺入由泊洛沙姆 188、聚山梨酯 80 非离子表面活性剂和 2,3-二（十四烷氧基）丙-1-胺（盐酸盐）阳离子脂质组成的阳离子脂质体配方中，以转染大鼠视网膜。通过反相蒸发技术制备不含和含有二磷酸氯喹（DPP80 和 DPP80-CQ，分别）的脂质体配方，并根据粒径、PDI、zeta 电位和形态进行表征。在掺入 pCMS-EGFP 质粒后，进一步评估所得的 nioplexes-在不同阳离子脂质/DNA 质量比下-以使其紧凑、释放并保护 DNA 免受酶消化。体外程序在 ARPE-19 细胞中进行，以评估转染效率和细胞内运输。两种 nioplexes 配方均有效地转染了 ARPE-19 细胞，尽管 DPP80-CQ nioplexes 的细胞活力明显更好。经视网膜下和玻璃体内注射后，DPP80 nioplexes 无法转染大鼠视网膜。然而，含有氯喹的载体显示出在许多视网膜细胞中表达蛋白，这取决于给药途径。这些数据为基于含有氯喹的脂质体非病毒载体的视网膜基因传递提供了新的见解。
