Aied Ahmed, Zheng Yu, Newland Ben, Wang Wenxin
Charles Institute of Dermatology, University College Dublin , Dublin 4, Ireland.
Biomacromolecules. 2014 Dec 8;15(12):4520-7. doi: 10.1021/bm5013162. Epub 2014 Nov 20.
Polymer-based transfection vectors are increasingly becoming the preferred alternative to viral vectors thanks to their safety and ease of production, but low transfection potency has limited their application. Many polycationic vectors show high efficiency in vitro, but their excessive charge density makes them toxic for in vivo applications. Herein, we demonstrate the synthesis of new and unique disulfide-reducible polymeric gene nanocarriers that exhibit significantly enhanced transfection potency and low cytotoxicity, particularly in skin cells, surpassing the efficiency of the well-known transfection reagents polyethylenimine (PEI) and Lipofectamine2000. The unique three-dimensional (3D) "multiknot" vectors were synthesized from in situ deactivation enhanced atom transfer radical (co)polymerization (DE-ATRP) of multivinyl monomers (MVMs). The high transfection levels and low toxicity of this multiknot structured polymer in vitro, combined with its ability to mediate collagen VII expression in 3D skin equivalents made from cells of recessive dystrophic epidermolysis bullosa patients, demonstrates its use as a platform nanotechnology which should be investigated further for dermatological disease therapies. Our findings suggest that the marked improvements stem from the dense multiknot architecture and degradable property, which facilitate both the binding and releasing process of the plasmid DNA.
基于聚合物的转染载体因其安全性和易于生产，正日益成为病毒载体的首选替代品，但转染效力低限制了它们的应用。许多聚阳离子载体在体外显示出高效率，但其过高的电荷密度使其在体内应用时具有毒性。在此，我们展示了新型独特的可二硫键还原的聚合物基因纳米载体的合成，该载体表现出显著增强的转染效力和低细胞毒性，尤其是在皮肤细胞中，超过了著名的转染试剂聚乙烯亚胺（PEI）和脂质体2000的效率。独特的三维（3D）“多结”载体是通过多乙烯基单体（MVMs）的原位失活增强原子转移自由基（共）聚合（DE-ATRP）合成的。这种多结结构聚合物在体外具有高转染水平和低毒性，再加上它能够在由隐性营养不良性大疱性表皮松解症患者细胞制成的3D皮肤等效物中介导VII型胶原蛋白的表达，证明了它作为一种平台纳米技术的用途，应进一步研究其在皮肤病治疗中的应用。我们的研究结果表明，显著的改进源于密集的多结结构和可降解特性，这有利于质粒DNA的结合和释放过程。
