北京生物医学工程

巯基烷基化壳聚糖载绿色荧光蛋白质粒基因纳米粒子的制备与研究

Preparation and Studies of Thiolated N Alkylated Chitosan Green Fluorescent Protein pDNA Nanoparticles

作者：慕卿赵伟亮白欣李树宝卢迪方月娥赵宇

单位：中国科学技术大学（合肥230026）

分类号：

出版年·卷·期（页码）：2010·29·1（6-10）

摘要：

合成了新型的基因载体巯基烷基化壳聚糖（TACS），采用复凝聚法制备了巯基烷基化壳聚糖基因纳米粒子，并表征其形态和粒径。利用体外转染实验定性评估了纳米粒子的转染效率。结果表明，粒子形态不很均一，粒径在390nm左右；凝胶电泳实验证明了载体能有效地包裹和保护基因不受DNase Ⅰ酶的消化；溶解实验证实了巯基化增强了TACS的水溶性；体外基因转染实验证实了TACS能够在人胚胎肾细胞（HEK293）转染基因，并且其转染效率远高于壳聚糖纳米粒子。这些结果表明，巯基烷基化壳聚糖有望成为有价值的基因载体。

Thiolated Nalkylated chitosan （TACS） was developed as a novel tool for gene delivery and the modified chitosanpDNA nanoparticles were prepared by complex coacervation. The morphology of nanoparticles was irregular， observed by transmission electronic microscopy （TEM）.The average particle size was 390nm determined by nanoparticle size analyzer. The TACSpDNA nanoparticles could partially protect the encapsulated plasmid pDNA from nuclease degradation as shown by electrophoretic mobility analysis. The solubility test showed sulfhedrylated reaction enhanced the water solubility of TACS. The gene transfection experiment in vitro suggested that the TACSpDNA nanoparticles could transfect HEK293 effectively and the transfection efficiency of TACSpDNA nanoparticles was much higher than that of chitosanpDNA nanoparticles. All these identified TACS as a promising vector for gene delivery.

参考文献：

［1］Huang L， Viroonchatapan E. Introduction// Huang M. Wagner M， eds. Nonviral vector for gene therapy ［C］.New York: Academic Press， 1999.
［2］Marxhall J， Yew NS， Eastman SJ，et al. Cationic lipidmediated gene delivery to the airway ［A］// Huang M， Wagner M， eds. Nonviral vectors for gene therapy ［C］. New York: Academic Press， 1999.
［3］Miller AD. Human genetherapy comes of age ［J］. Nature， 1992， 357: 455-450.
［4］Kim JS， Maruyama A， Akaike T et al. In vitro gene expression on smooth muscle cells using a terplex delivery system ［J］. J Control Rel， 1997， 47: 51-59.
［5］Borchard G. Chitosans for gene delivery ［J］.Adv Drug Deliver Rev， 2001， 52（2）: 145-150.
［6］Mumper RJ， Wang J， Claspell JM，et al. Proc Intl Symp Controlled Rel Bioact Mater， 1995， 22: 178.
［7］Mao HQ， Roy K， Vu L，et al. ChitosanDNA nanoparticles as gene carriers synthesis， characterization and transfection efficiency ［J］. J Control Rel， 2001， 70: 399-421.
［8］Sato T， Ishii T， Okahata Y. In vitro gene delivery mediated by chitosan. Effect of pH， serum， and molecular mass of chitosan on the transfection efficiency ［J］. Biomaterials， 2001， 22: 2075-2080.
［9］Strand SP， Danielsen S， Christensen BE，et al. Influence of chitosan structure on the formation and stability of DNAchitosan polyelectrolyte complexes ［J］. Biomacromolecules， 2005， 6:3357-3366.
［10］Kawamura K， Oishi J， Kang JH，et al. Intracellular signalresponsive gene carrier for cellspecific gene expression ［J］. Biomacromolecules， 2005， 6（2）: 908-913.
［11］Liu WG， Zhang Xin ， Sun SJ，et al. NAlkylated chitosan as a potential nonviral vector for gene transfection ［J］. Bioconjugate Chem， 2003， 14: 782-789.
［12］Martinovich GG， Cherenkevich SN， Sauer H. Intracellular redox state: towards quantitative description ［J］. Eur Biophys J， 2005， 34:937-942.
［13］Schmitz T， BravoOsuna I， Vauthier C，et al. Development and in vitro evaluation of a thiomerbased nanoparticulate gene delivery system ［J］. Biomaterials， 2007， 28: 524-531.
［14］Loretz B， Thaler M， BernkopSchnurch A. Role of Sulfhydryl Groups in Transfection A Case Study with ChitosanNAC Nanoparticles ［J］. Bioconjugate Chem， 2007， 18:1028-1035.
［15］Liu WG， Yao KD， Liu QG. Formation of a DNA/NDodecylated Chitosan Complex and SaltInduced Gene Delivery ［J］. J Appl Polym Sci， 2001， 82: 3391-3395.
［16］Dai Zhao， Sun Duo Xian， Guo Yao. Preparation of cetylchitosan nanoparticles as carricers for paracetamol ［J］.Transaction of Tianjin University， 2002， 8（4）:235-238.
［17］Li Fang， Liu WG， Yao KD. Preparation of oxidized glucosecrosslinked Nalkylated chitosan membrane and in vitro studies of pHsensitive drug delivery behaviour ［J］. Biomaterials， 2002， 23: 343-347.
［18］Kast CE， Frick W， Losert U，et al. Chitosanthioglycolic acid conjugate: a new scaffold material for tissue engineering ［J］.Int J Pharm， 2003， 256: 183-189.
［19］Kast CE， BernkopSchnurch A. Thiolated polymersthiomers: development and in vitro evaluation of chitosanthioglycolic acid conjugates ［J］. Biomaterials， 2001， 22: 2345-2352.
［20］王荣祺，张军，王常勇，等.壳聚糖-hVEGF165基因微粒的制备及体外转染的实验研究［J］. 中国生物医学工程学报， 2006， 25（1）: 14-18.

服务与反馈：

【文章下载】【加入收藏】

提示：您还未登录，请登录！点此登录