Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles.
Delivery of a transforming growth factor β-1 plasmid to mesenchymal stem cells via cationized Pleurotus eryngii polysaccharide nanoparticles.
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DOI：
10.2147/ijn.s28010
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发表时间：
2012
影响因子：
8
通讯作者：
Yu JN
中科院分区：
文献类型：
--
作者：
Deng WW;Cao X;Wang M;Qu R;Su WY;Yang Y;Wei YW;Xu XM;Yu JN
本研究旨在探讨阳离子化侧耳多糖（CPEPS）作为非病毒基因载体，将转化生长因子β 1（pTGF-β1）质粒DNA转染骨髓间充质干细胞（MSCs）。对杏鲍菇粗多糖进行纯化，然后将精胺接枝到多糖骨架上进行阳离子化。使用琼脂糖凝胶电泳、透射电子显微镜和Nano Sense Zetasizer（马尔文仪器，马尔文，英国）表征CPEPS-pTGF-β1纳米颗粒。细胞毒性分析结果表明，当CPEPS/pTGF-β1重量比≥ 10：1时，琼脂糖凝胶电泳中观察到更大的凝胶阻滞效应。重量比分别为20：1的CPEPS-pTGF-β1纳米颗粒具有直径为80.8 nm的平均粒径和+17.4 ± 0.1 mV的zeta电位。值得注意的是，这些CPEPS-pTGF-β1纳米颗粒显示出比聚乙烯亚胺（25 kDa）（P = 0.006，学生t检验）和LipofectamineTM 2000（P = 0.002，学生t检验）更低的细胞毒性和更高的转染效率。转染CPEPS-pTGF-β1纳米粒的MSCs中TGF-β1 mRNA的表达水平明显高于转染游离质粒DNA的MSCs，但略高于转染Lipofectamine 2000的MSCs。流式细胞仪分析显示，转染CPEPS-pTGF-β1纳米粒后，92.38%的MSCs阻滞于G1期，显示有分化趋势。综上所述，本研究的结果表明，在本工作中制备的CPEPS-pTGF-β1纳米粒具有优异的转染效率和低毒性。因此，它们有可能发展成为一种很有前途的非病毒载体，用于体外基因传递。
The objective of this study was to investigate the use of cationized Pleurotus eryngii polysaccharide (CPEPS) as a nonviral gene delivery vehicle to transfer plasmid DNA encoding transforming growth factor beta-1 (pTGF-β1) into mesenchymal stem cells (MSCs) in vitro. Crude P. eryngii polysaccharide was purified, and then cationized by grafting spermine onto the backbone of the polysaccharide. Agarose gel electrophoresis, transmission electron microscopy, and a Nano Sense Zetasizer (Malvern Instruments, Malvern, UK) were used to characterize the CPEPS-pTGF-β1 nanoparticles. The findings of cytotoxicity analysis showed that when the nanoparticles were formulated with a CPEPS/pTGF-β1 weight ratio ≥ 10:1, a greater gel retardation effect was observed during agarose gel electrophoresis. The CPEPS-pTGF-β1 nanoparticles with a weight ratio of 20:1, respectively, possessed an average particle size of 80.8 nm in diameter and a zeta potential of +17.4 ± 0.1 mV. Significantly, these CPEPS-pTGF-β1 nanoparticles showed lower cytotoxicity and higher transfection efficiency than both polyethylenimine (25 kDa) (P = 0.006, Student’s t-test) and LipofectamineTM 2000 (P = 0.002, Student’s t-test). Additionally, the messenger RNA expression level of TGF-β1 in MSCs transfected with CPEPS-pTGF-β1 nanoparticles was significantly higher than that of free plasmid DNA-transfected MSCs and slightly elevated compared with that of Lipofectamine 2000-transfected MSCs. Flow cytometry analysis demonstrated that 92.38% of MSCs were arrested in the G1 phase after being transfected with CPEPS-pTGF-β1 nanoparticles, indicating a tendency toward differentiation. In summary, the findings of this study suggest that the CPEPS-pTGF-β1 nanoparticles prepared in this work exhibited excellent transfection efficiency and low toxicity. Therefore, they could be developed into a promising nonviral vector for gene delivery in vitro.