State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, P. R. China.
J Mater Chem B. 2021 Dec 8;9(47):9784-9793. doi: 10.1039/d1tb01930a.
Liquid coacervate microdroplets have been widely explored as membrane-free compartment protocells for cargo delivery in therapeutic applications. In this study, coacervate protocells were developed as gene carriers for transfection of nitric oxide synthase (NOS) and overproduction of nitric oxide (NO) for killing of cancer cells. The coacervate microdroplet protocells were formed the liquid-liquid phase separation of oppositely charged diethylaminoethyl-dextran/polyacrylic acids. The coacervate microdroplet protocells were found to facilitate gene transfection, which was demonstrated by cell imaging of the internalized coacervate microdroplets containing plasmids of enhanced green fluorescent protein. Due to their high transfection capability, the coacervate protocells were subsequently utilized for the delivery of NOS plasmids (pNOS). The cellular internalization of pNOS-containing coacervate carriers was found to result in high NOS expression coupled with NO overproduction, which then induced cell apoptosis and decreased cell viability. The cell apoptosis is associated with NO-mediated mitochondrial damage. The enhanced gene transfection was attributed to coacervate microdroplets' unique high sequestration capability and liquid-like fluidity. Overall, the incorporation of genes in coacervate microdroplets was demonstrated as a viable and novel strategy for the development of cargo biocarriers for biomedical applications.
液相聚集体微滴已被广泛探索为无膜隔室原代细胞，用于治疗应用中的货物输送。在这项研究中，聚集体原代细胞被开发为基因载体，用于转染一氧化氮合酶（NOS）和过量产生一氧化氮（NO）以杀死癌细胞。聚集体微滴原代细胞是通过带相反电荷的二乙氨基乙基-葡聚糖/聚丙烯酸的液-液相分离形成的。研究发现，聚集体微滴原代细胞有利于基因转染，这可以通过含有增强型绿色荧光蛋白质粒的聚集体微滴内化的细胞成像来证明。由于其高转染能力，随后将聚集体原代细胞用于NOS 质粒（pNOS）的递送。发现含有 pNOS 的聚集体载体的细胞内吞导致高 NOS 表达与过量产生的 NO 相结合，从而诱导细胞凋亡和降低细胞活力。细胞凋亡与 NO 介导的线粒体损伤有关。增强的基因转染归因于聚集体微滴独特的高隔离能力和类似液体的流动性。总体而言，将基因纳入聚集体微滴被证明是开发用于生物医学应用的货物生物载体的一种可行且新颖的策略。
