Agirre Mireia, Zarate Jon, Ojeda Edilberto, Puras Gustavo, Rojas Luis A, Alemany Ramón, Pedraz José L
NanoBioCel Group, University of the Basque Country, Vitoria-Gasteiz, Spain; Networking Research Center of Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Vitoria-Gasteiz, Spain.
Traslational Research Laboratory, IDIBELL-Institut Catalá d'Oncologia, L'Hospitalet de LLobregat, Barcelona, Spain.
Int J Pharm. 2015 Feb 20;479(2):312-9. doi: 10.1016/j.ijpharm.2014.12.062. Epub 2014 Dec 27.
Ultrapure oligochitosans have been recently reported as efficient non-viral vectors for the delivery of pCMS-EGFP plasmid (5.5kbp) to the cornea and retina. However, the delivery of oncolytic adenoviral plasmids (40kbp) represents a unique challenge. In this work, we elaborated self assembled O15 and O25 UOC/pAdTLRGD polyplexes, and we studied the influence of the N/P ratio, the pH of the transfection medium and the salt concentration on the particle size and zeta potential by an orthogonal experimental design. All polyplexes showed a particle size lower than 200nm and a positive zeta potential. These parameters were influenced by the N/P ratio, salt concentration, and pH of the transfection medium. The selected polyplexes were able to bind, release, and protect the plasmid from DNase degradation. Transfection experiments in HEK293 and A549 cell lines demonstrated that UOC/pAdTLRGD polyplexes were able to deliver the plasmid and transfect both cell lines. These results suggest that O15 and O25 UOC based polyplexes are suitable for future in vivo applications.
最近有报道称，超纯壳寡糖是将pCMS - EGFP质粒（5.5kbp）递送至角膜和视网膜的高效非病毒载体。然而，溶瘤腺病毒质粒（40kbp）的递送是一项独特的挑战。在这项工作中，我们制备了自组装的O15和O25 UOC/pAdTLRGD多聚体，并通过正交实验设计研究了N/P比、转染介质的pH值和盐浓度对粒径和zeta电位的影响。所有多聚体的粒径均低于200nm，且zeta电位为正。这些参数受N/P比、盐浓度和转染介质pH值的影响。所选的多聚体能够结合、释放并保护质粒免受DNase降解。在HEK293和A549细胞系中的转染实验表明，UOC/pAdTLRGD多聚体能够递送质粒并转染这两种细胞系。这些结果表明，基于O15和O25 UOC的多聚体适用于未来的体内应用。
