Degradable copolymer based on amphiphilic N-octyl-N-quatenary chitosan and low-molecular weight polyethylenimine for gene delivery.
Degradable copolymer based on amphiphilic N-octyl-N-quatenary chitosan and low-molecular weight polyethylenimine for gene delivery.
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DOI：
10.2147/ijn.s36179
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发表时间：
2012
影响因子：
8
通讯作者：
Liu K
中科院分区：
文献类型：
--
作者：
Liu C;Zhu Q;Wu W;Xu X;Wang X;Gao S;Liu K
壳聚糖具有较高的生物相容性和较低的细胞毒性。然而，壳聚糖在生理ph下不溶，且缺乏电荷，转染效果不佳。为了开发一种高转染效率、低细胞毒性的新型基因载体，合成了两亲性壳聚糖，并与低分子量聚乙烯亚胺（PEI）连接。首先合成了两亲性n -辛基- n -四烷基壳聚糖（OTMCS），然后将低分子量PEI与两亲性壳聚糖交联制备了可降解的PEI衍生物，得到了一种新的聚合基因载体（OTMCS - PEI）。利用多种物理化学方法对新基因载体进行了表征。在体外和体内测定了其细胞毒性和基因转染效率。载体表现出可控的退化。它非常稳定，并表现出优异的缓冲能力。OTMCS-PEI /DNA复合物的粒径在150 ~ 200 nm之间，zeta电位在10 ~ 30 mV之间。当DNA酶I/μ DNA浓度为2.25 U时，该聚合物可以保护质粒DNA不被DNA酶I消化。此外，50%胎牛血清和1100 μg/mL肝素钠诱导的解离作用对其具有抗性。即使在高剂量下，OTMCS-PEI也显示出较低的细胞毒性。与PEI 25 KDa相比，OTMCS-PEI /DNA复合物在体外和体内也表现出更高的转染效率。OTMCS-PEI是一种安全、高效的基因载体。
Chitosan shows particularly high biocompatibility and fairly low cytotoxicity. However, chitosan is insoluble at physiological pH. Moreover, it lacks charge, so shows poor transfection. In order to develop a new type of gene vector with high transfection efficiency and low cytotoxicity, amphiphilic chitosan was synthesized and linked with low-molecular weight polyethylenimine (PEI). We first synthesized amphiphilic chitosan – N-octyl-N-quatenary chitosan (OTMCS), then prepared degradable PEI derivates by cross-linking low-molecular weight PEI with amphiphilic chitosan to produce a new polymeric gene vector (OTMCS–PEI). The new gene vector was characterized by various physicochemical methods. We also determined its cytotoxicity and gene transfecton efficiency in vitro and in vivo. The vector showed controlled degradation. It was very stable and showed excellent buffering capacity. The particle sizes of the OTMCS–PEI/DNA complexes were around 150–200 nm with proper zeta potentials from 10 mV to 30 mV. The polymer could protect plasmid DNA from being digested by DNase I at a concentration of 2.25 U DNase I/μg DNA. Furthermore, they were resistant to dissociation induced by 50% fetal bovine serum and 1100 μg/mL sodium heparin. OTMCS–PEI revealed lower cytotoxicity, even at higher doses. Compared with PEI 25 KDa, the OTMCS–PEI/DNA complexes also showed higher transfection efficiency in vitro and in vivo. OTMCS–PEI was a potential candidate as a safe and efficient gene vector for gene therapy.