Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (51): 8292-8296.doi: 10.3969/j.issn.2095-4344.2014.51.017

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Novel cationic phosphorylcholine polymer acts as an anti-sense oligonucleotide gene delivery carrier

Lu Yong-zhi1, Sun Tian-tian2, Cao Xi-chuan2, Zhang Zhuo-qi1, Wang Zhi-rong1, Xu Wu1, Hao Zhan-jun1, Wang Xiang-dong1, Zhang Chao-qun1, Xia Yong1, Li Dong-ye1   

  1. 1Department of Cardiology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu Province, China; 2College of Materials Science and Engineering, Chinese University of Mining and Technology, Xuzhou 221116, Jiangsu Province, China
  • Online:2014-12-10 Published:2014-12-10
  • Contact: Zhang Zhuo-qi, Department of Cardiology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu Province, China
  • About author:Lu Yong-zhi, Master, Attending physician, Department of Cardiology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 30800219, 30670557; Natural Science Foundation of Jiangsu Universities, No. 08KJD320013; Special President Talent Fund of Xuzhou Medical College, No. 07KJZ26

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Abstract:

BACKGROUND: Although viral gene vector has high transfection efficiency, its safety has been questioned.

OBJECTIVE: To study the effect of gene-based drug delivery with the carrier of cationic phosphorylcholine polymers on the load and transport capacity of human c-myc anti-senseoligodeoxynucleotide, and to evaluate the safety and efficacy.
METHODS: Highly biocompatible diblock phosphorylcholine copolymer MPC30-DEA70 was synthesized using atom transfer radical polymerization and the copolymer solutions were characterized. Under different conditions, DNA complexes were formed between MPC30-DEA70 and c-myc anti-senseoligodeoxynucleotide at different N/P ratios, which were characterized by DNA gel electrophoresis. The MPC30-DEA70/AS-OD gene complexes  
were transfected into the in vitro cultured HEK293 cells to detect the cell compatibility, transfection efficiency and mechanism of intracellular transport.
RESULTS AND CONCLUSION: The results showed that the material MPC30-DEA70 at pH=4.0-11.0, showed a high degree of water-soluble property in 0.01 mol/L PBS solution, and the positive charge was increased with the decreasing of solution pH value. MPC30-DEA70/AS-ODN complexes showed different degrees of gel retardation in DNA gel electrophoresis, and was increased significantly with the increasing of N/P ratios. Methylthiazolyldiphenyl-tetrazolium bromide assay showed that the MPC30-DEA70 inhibited the viability of the HEK293 in a dose-dependent manner, and cell toxicity was significantly increased in high dose MPC30-DEA70. Flow cytometry study showed the transfection of the complexes was significantly enhanced with the increasing of N/P ratios. Confocal microscopy study found the transferred anti-senseoligodeoxynucleotide molecules in the cell nucleus, suggesting its effective nuclear localization. The results indicate that novel cationic phosphorylcholine copolymer can load and transport anti-senseoligodeoxynucleotide, which may act as an efficient and safe non-viral gene transfer vector.


中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


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Key words: gene therapy, genetic vectors, oligonucleotides, antisense, cations, polymers

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