Tailoring of Chitosans for Gene Delivery: Novel Self-Branched Glycosylated Chitosan Oligomers with Improved Functional Properties
Tailoring of Chitosans for Gene Delivery: Novel Self-Branched Glycosylated Chitosan Oligomers with Improved Functional Properties
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DOI：
10.1021/bm800832u
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发表时间：
2008-11-01
影响因子：
6.2
通讯作者：
Artursson, Per
中科院分区：
文献类型：
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作者：
Strand, Sabina P.;Issa, Mohamed M.;Artursson, Per
壳聚糖是一种很有前途的生物材料，具有诱人的安全性，但其在生理pH值下的相容性较差，阻碍了其在基因传递方面的应用潜力。为了改善壳聚糖低聚物的功能性质和基因转移效率，我们定制了壳聚糖的分子结构，并开发了含有N-乙酰氨基葡萄糖的三糖取代的自支化糖化壳聚糖低聚物(SB-TCO)。以壳聚糖为原料，通过控制解聚、同时支化和AAM取代反应制备了Sb-TCO。该产物在生理pH条件下完全溶解，形成胶体稳定性和物理稳定性均较高的复合体。SB-TCO在HEK293细胞中显示出较高的转染率，最高可达70%，并产生了大量的转基因。在HepG2细胞中证实了基因转移的有效性，SB-TCO介导的基因表达水平分别比未取代的和取代的线性寡聚体高出10倍和4倍。转基因在两种细胞系中的快速表达表明从SB-TCO复合体中有效地释放和转录了DNA。与以22 kDa线性PEI为基础的转染剂相比，SB-TCO具有更高的基因转移效率、更低的细胞毒性和更好的血清相容性。
Chitosan is a promising biomaterial with an attractive safety profile; however, its application potential for gene delivery is hampered by poor compatibility at physiological pH values. Here we have tailored the molecular architecture of chitosan to improve the functional properties and gene transfer efficacy of chitosan oligomers and have developed self-branched glycosylated chitosan oligomer (SB-TCO) substituted with a trisaccharide containing N-acetylglucosamine, AAM. SB-TCO was prepared by controlled depolymerization of chitosan, followed by simultaneous branching and AAM substitution. The product was fully soluble at physiological pH and complexed plasmid DNA into polyplexes of high colloidal and physical stability. SB-TCO displayed high transfection efficacy in HEK293 cells, reaching transfection efficiencies of up to 70%, and large amounts of transgene were produced. Gene transfer efficacy was confirmed in HepG2 cells, where gene expression levels mediated by SB-TCO were up to 10 and 4 times higher than those obtained with unsubstituted and substituted linear oligomers, respectively. The rapid onset of transgene expression in both cell lines indicates efficient DNA release and transcription from SB-TCO polyplexes. In comparison with 22 kDa linear PEI-based transfection reagent used as the control, SB-TCO possessed higher gene transfer efficacy, significantly lower cytotoxicity, and improved serum compatibility.