Martens Thomas F, Peynshaert Karen, Nascimento Thaís Leite, Fattal Elias, Karlstetter Marcus, Langmann Thomas, Picaud Serge, Demeester Jo, De Smedt Stefaan C, Remaut Katrien, Braeckmans Kevin
Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; Center for Nano-and Biophotonics (NB-Photonics), Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
Eur J Pharm Sci. 2017 May 30;103:27-35. doi: 10.1016/j.ejps.2017.02.027. Epub 2017 Feb 20.
Intravitreal administration of nanomedicines could be valuable for retinal gene therapy, if their mobility in the vitreous and therapeutic efficacy in the target cells can be guaranteed. Hyaluronic acid (HA) as an electrostatic coating of polymeric gene nanomedicines has proven to be beneficial on both accounts. While electrostatic coating provides an easy way of coating cationic nanoparticles, the stability of electrostatic complexes in vivo is uncertain. In this study, therefore, we compare electrostatic with covalent coating of gene nanocarriers with HA for retinal gene therapy via intravitreal administration. Specifically, DOTAP:DOPE/plasmid DNA lipoplexes coated with HA are evaluated in terms of intravitreal mobility using a previously optimized ex vivo model. We find that both electrostatic and covalent HA coating considerably improve the mobility of the lipoplexes in the vitreous humor of excised bovine eyes. In addition we evaluate in vitro uptake and transfection efficiency in ARPE-19 cells. Contrary to PEGylated lipoplexes it is found that HA coated lipoplexes are efficiently internalized into ARPE-19 cells. Covalent HA-coated lipoplexes had an 8-fold increase of transgene expression compared to the uncoated lipoplexes. We conclude that covalent HA-coating of gene nanomedicines is a promising approach for retinal gene therapy by intravitreal administration.
如果能确保纳米药物在玻璃体内的流动性及其在靶细胞中的治疗效果，那么玻璃体内注射纳米药物对于视网膜基因治疗可能具有重要价值。透明质酸（HA）作为聚合物基因纳米药物的静电涂层，已被证明在这两方面均有益处。虽然静电涂层为阳离子纳米颗粒提供了一种简便的包被方式，但静电复合物在体内的稳定性尚不确定。因此，在本研究中，我们比较了通过玻璃体内注射将基因纳米载体与HA进行静电包被和共价包被用于视网膜基因治疗的效果。具体而言，使用先前优化的体外模型，对HA包被的DOTAP:DOPE/质粒DNA脂质体在玻璃体内的流动性进行了评估。我们发现，静电包被和共价包被HA均能显著提高脂质体在切除的牛眼玻璃体液中的流动性。此外，我们还评估了其在ARPE - 19细胞中的体外摄取和转染效率。与聚乙二醇化脂质体不同，我们发现HA包被的脂质体能够有效地内化到ARPE - 19细胞中。与未包被的脂质体相比，共价HA包被的脂质体转基因表达增加了8倍。我们得出结论，基因纳米药物的共价HA包被是一种通过玻璃体内注射进行视网膜基因治疗的有前景的方法。
