The influence of formulation variables on in vitro transfection efficiency and physicochemical properties of chitosan-based polyplexes
The influence of formulation variables on in vitro transfection efficiency and physicochemical properties of chitosan-based polyplexes
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DOI：
10.1016/s0378-5173(03)00301-6
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发表时间：
2003-08-11
影响因子：
5.8
通讯作者：
Thu, BJ
中科院分区：
文献类型：
--
作者：
Romoren, K;Pedersen, S;Thu, BJ
本研究的目的是研究选择的配方变量如何影响基于壳聚糖的聚合物的体外转染效率和物理化学性质（粒径、zetapotential和壳聚糖-质粒结合）。应用实验设计结合多变量数据分析来揭示制剂变量对响应的影响。以下配方变量进行了研究：壳聚糖的分子量和乙酰化度，pH值和离子强度的缓冲液中，壳聚糖被溶解，电荷比的聚合物，质粒浓度和包含在质粒溶液中的凝聚剂。复合物的电荷比、复合物中DNA的浓度以及壳聚糖的分子量和乙酰化程度均影响壳聚糖在Epithelioma papulosum cyprini（EPC）细胞中的体外转染效率。在更彻底的调查中确定了两种有利的配方。这些制剂由SC 113（理论电荷比10）和SC 214（理论电荷比3）制成。复合物的大小受乙酰化程度、DNA浓度、pH、凝聚剂的加入和电荷比的影响。电荷比、pH和离子强度决定了粒子的zeta电位，而电荷比对质粒与壳聚糖之间的缔合是重要的。(C)2003 Elsevier B. V.保留所有权利。
The aim of this study was to investigate how a selection of formulation variables affects the in vitro transfection efficiency and physicochemical properties (particle size, zetapotential and chitosan-plasmid association) of chitosan-based polyplexes. Experimental designs in combination with multivariate data analysis were applied to reveal the effects of the formulation variables on the responses. The following formulation variables were studied: molecular weight and degree of acetylation of chitosan, pH and ionic strength of the buffer in which chitosan was dissolved, charge ratio of polyplexes, plasmid concentration and inclusion of a coacervation agent in the plasmid solution. The in vitro transfection efficiency in Epithelioma papulosum cyprini (EPC) cells was affected by the polyplex charge ratio, the DNA concentration in the complexes as well as the molecular weight and degree of acetylation of the chitosans. Two favourable formulations were identified in a more thorough investigation. These formulations were made of SC 113 (theoretical charge ratio 10) and SC214 (theoretical charge ratio 3). The size of the complexes was affected by the degree of acetylation, concentration of DNA, pH, inclusion of a coacervation agent and the charge ratio. The charge ratio, pH and ionic strength determined the zetapotential of the particles, while the charge ratio was important for the association between the plasmid and chitosan. (C) 2003 Elsevier B.V. All rights reserved.