Department of Pharmaceutics and Biopharmaceutics, University of Marburg, Robert-Koch-Str. 4, 35037 Marburg, Germany; Punjab University College of Pharmacy, University of the Punjab, 54000 Lahore, Pakistan.
Department of Pharmaceutics and Biopharmaceutics, University of Marburg, Robert-Koch-Str. 4, 35037 Marburg, Germany.
Eur J Pharm Biopharm. 2019 Feb;135:72-82. doi: 10.1016/j.ejpb.2018.12.013. Epub 2018 Dec 24.
Non-viral vectors are a safe, efficient and non-toxic alternative to viral vectors for gene therapy against many diseases ranging from genetic disorders to cancers. Polyamidoamine (PAMAM), a positively charged dendrimer has a tendency to complex with nucleic acids (to form dendriplexes) like plasmid DNA (pDNA) and small interfering RNA (siRNA) and can shield them from enzymatic degradation, thereby facilitating endocytosis and endosomal release. In this study, we developed an advanced variant of the dendriplexes by encapsulating them within liposomes to enhance their gene delivery efficiency. This liposome encapsulated dendriplex system can further reduce unwanted cytotoxicity and enhance cellular uptake of nucleic acids. A broad range of lipid combinations were used to optimize the lipodendriplexes in terms of their physicochemical characteristics including size, shape and zeta potential. The optimized lipodendriplexes were tested for pDNA transfection, in vitro cell viability, cellular uptake, siRNA mediated knockdown, hemocompatibility, metastatic progression and in ovo in chorioallantoic membrane model (CAM). The optimized system has shown significant improvement in pDNA transfection (p < 0.01) with higher GFP expression and gene silencing and has shown improved cell viability (p < 0.05) compared to the parent dendriplex system. The hemocompatibility and CAM analysis, revealed an efficient yet biocompatible gene delivery system in the form of lipodendriplexes.
非病毒载体是基因治疗许多疾病（从遗传疾病到癌症）的安全、高效、无毒的病毒载体替代物。多聚酰胺-胺（PAMAM）是一种带正电荷的树枝状聚合物，有与核酸（如质粒 DNA（pDNA）和小干扰 RNA（siRNA）形成树枝状聚合物的趋势）并可以保护它们免受酶降解，从而促进内吞作用和内涵体释放。在这项研究中，我们通过将其包裹在脂质体中来开发树枝状聚合物的高级变体，以提高其基因传递效率。这种脂质体包裹的树枝状聚合物系统可以进一步降低不必要的细胞毒性并增强核酸的细胞摄取。使用广泛的脂质组合来优化脂质体的物理化学特性，包括大小、形状和 zeta 电位。优化的脂质体用于测试 pDNA 转染、体外细胞活力、细胞摄取、siRNA 介导的基因沉默、血液相容性、转移进展以及鸡胚尿囊膜模型（CAM）中的基因沉默。优化的系统在 pDNA 转染方面表现出显著改善（p<0.01），具有更高的 GFP 表达和基因沉默，并显示出与母体树枝状聚合物系统相比，细胞活力提高（p<0.05）。血液相容性和 CAM 分析显示，脂质体形式的树枝状聚合物是一种高效且生物相容的基因传递系统。
