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质粒包裹的聚乙二醇接枝聚乙烯亚胺纳米颗粒用于向大鼠间充质干细胞递送基因。

Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells.

机构信息

Department of Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.

出版信息

Int J Nanomedicine. 2011;6:843-53. doi: 10.2147/IJN.S17155. Epub 2011 Apr 21.

DOI:10.2147/IJN.S17155
PMID:21589652
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3090281/
Abstract

BACKGROUND

Mesenchymal stem cell transplantation is a promising method in regenerative medicine. Gene-modified mesenchymal stem cells possess superior characteristics of specific tissue differentiation, resistance to apoptosis, and directional migration. Viral vectors have the disadvantages of potential immunogenicity, carcinogenicity, and complicated synthetic procedures. Polyethylene glycol-grafted polyethylenimine (PEG-PEI) holds promise in gene delivery because of easy preparation and potentially targeting modification.

METHODS

A PEG8k-PEI25k graft copolymer was synthesized. Agarose gel retardation assay and dynamic light scattering were used to determine the properties of the nanoparticles. MTT reduction, wound and healing, and differentiation assays were used to test the cytobiological characteristics of rat mesenchymal stem cells, fluorescence microscopy and flow cytometry were used to determine transfection efficiency, and atomic force microscopy was used to evaluate the interaction between PEG-PEI/plasmid nanoparticles and mesenchymal stem cells.

RESULTS

After incubation with the copolymer, the bionomics of mesenchymal stem cells showed no significant change. The mesenchymal stem cells still maintained high viability, resettled the wound area, and differentiated into adipocytes and osteoblasts. The PEG-PEI completely packed plasmid and condensed plasmid into stable nanoparticles of 100-150 nm diameter. After optimizing the N/P ratio, the PEG-PEI/plasmid microcapsules delivered plasmid into mesenchymal stem cells and obtained an optimum transfection efficiency of 15%-21%, which was higher than for cationic liposomes.

CONCLUSION

These data indicate that PEG-PEI is a valid gene delivery agent and has better transfection efficiency than cationic liposomes in mesenchymal stem cells.

摘要

背景

间充质干细胞移植是再生医学中很有前途的方法。基因修饰的间充质干细胞具有特定组织分化、抗凋亡和定向迁移的优异特性。病毒载体具有潜在的免疫原性、致癌性和复杂的合成程序等缺点。聚乙二醇接枝的聚乙烯亚胺（PEG-PEI）在基因传递方面有很大的应用前景，因为其易于制备并且具有潜在的靶向修饰能力。

方法

合成了 PEG8k-PEI25k 接枝共聚物。琼脂糖凝胶阻滞试验和动态光散射用于确定纳米粒子的性质。MTT 还原、划痕和愈合以及分化试验用于测试大鼠间充质干细胞的细胞生物学特性，荧光显微镜和流式细胞术用于确定转染效率，原子力显微镜用于评估 PEG-PEI/质粒纳米颗粒与间充质干细胞的相互作用。

结果

与共聚物孵育后，间充质干细胞的生物特性没有明显变化。间充质干细胞仍然保持高活力，重新定居伤口区域，并分化为脂肪细胞和成骨细胞。PEG-PEI 完全包裹质粒并将质粒浓缩成 100-150nm 直径的稳定纳米颗粒。优化 N/P 比后，PEG-PEI/质粒微胶囊将质粒递送入间充质干细胞，并获得 15%-21%的最佳转染效率，高于阳离子脂质体。

结论

这些数据表明，PEG-PEI 是一种有效的基因传递剂，在间充质干细胞中的转染效率优于阳离子脂质体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/f8002a14c41c/ijn-6-843f8.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/dcb320931069/ijn-6-843f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/f8002a14c41c/ijn-6-843f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/1032051d97fc/ijn-6-843f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/c34d8bfdd561/ijn-6-843f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/b8ede9fa4a28/ijn-6-843f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/4c57b0f59849/ijn-6-843f4.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/45ead7aebe46/ijn-6-843f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/dcb320931069/ijn-6-843f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/efb7/3090281/f8002a14c41c/ijn-6-843f8.jpg

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