Peptide dendrimers as efficient and biocompatible gene delivery vectors: Synthesis and in vitro characterization.
Peptide dendrimers as efficient and biocompatible gene delivery vectors: Synthesis and in vitro characterization.
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DOI：
10.1016/j.jconrel.2010.10.006
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发表时间：
2011-10
期刊：
影响因子：
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通讯作者：
Kui Luo;Caixia Li;G. Wang;Yu Nie;B. He;Yao Wu;Z. Gu
中科院分区：
文献类型：
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作者：
Kui Luo;Caixia Li;G. Wang;Yu Nie;B. He;Yao Wu;Z. Gu
我们报道了不同世代的树状多(L-赖氨酸)载体的合成和表征，以及它们在体外基因转染中的应用。凝胶滞留分析表明，树枝状大分子能够与质粒DNA形成复合体，G3、G4和G5代树枝状细胞分别在N/P比为0.5、1和2时抑制pDNA的迁移。DNase I分析表明，通过与树枝状大分子络合获得的PDNA对核酸酶催化的降解具有保护作用，G5的保护作用甚至强于聚乙二亚胺(PEI)。原子力显微镜(AFM)显示，所研究的4代树枝状大分子/DNA复合体的颗粒大小在100-200 nm之间。Zeta电位测量表明，随着N/P比从1增加到25，所有树状大分子/PDNA络合物逐渐从负电荷转变为正电荷。世代越高，产生的正电势越大，表明该复合体与带负电荷的细胞膜相互作用的能力越强。体外和体内细胞毒性评价表明，在所研究的不同N/P比下，树枝状大分子及其配合物具有良好的生物相容性。体外基因转染显示G5的效率高于其他树状大分子，且对血清的变化不敏感。考虑到其与PEI相似的转染率，但对培养细胞的毒性较低，树枝状大分子G5可能是更好的基因输送候选。
We report the synthesis and characterization of different generations of dendritic poly(l-lysine) vectors, and their use for in vitro gene transfection. Gel retardation assay revealed that the dendrimers could form complexes with plasmid DNAs (pDNAs), evident from the inhibition of the migration of pDNA at the N/P ratios of 0.5, 1 and 2 by G3, G4 and G5 dendritic generations, respectively. DNase I assay revealed the protection of pDNA acquired from the complexation with dendrimers from nuclease-catalyzed degradation, with the protection capacity of G5 being even stronger than poly(ethyleneimine) (PEI). Atomic force microscopy (AFM) revealed that all 4 generations of dendrimer/DNA complexes studied were of similar particle sizes within 100–200nm. Zeta potential measurements showed that as the N/P ratio increased from 1 to 25, all dendrimer/pDNA complexes gradually changed from negative to positive charges. The higher generations tended to produce the greater positive potentials, indicating a stronger potency of the complexes to interact with negatively charged cell membranes. In vitro and in vivo cytotoxicity evaluations showed good biocompatibility of the dendrimers and their complexes over the different N/P ratios studied. In vitro gene transfection revealed higher efficiency of G5 than other dendrimers and insensitive variation to the presence of serum. Given its similar transfection efficiency to PEI but lower toxicity to cultured cells, dendrimer G5 could be a better candidate for gene delivery.