Zhong Zhi-rong, Liu Ji, Deng Yong, Zhang Zhi-rong, Song Qing-guo, He Qin
West China School of Pharmacy, Sichuan University, Chengdu 610041, China.
Yao Xue Xue Bao. 2007 Feb;42(2):216-20.
A novel transferrin modified non-viral gene delivery system Tf-PLPD was developed and the related characteristics was investigated. Blank procationic liposomes were prepared by film dispersion-filteration method. PLPD was prepared as follows by first mixing the plasmid DNA and protamine together, then the resulted polyplexes were incubated for 10 min at room temperature, followed by addition of preformed blank procationic liposomes. Transferrin was adsorbed at the surface of PLPD via electrostatic interactions to form Tf-PLPD. Central composite design (CCD) was employed to optimize the formulation. The HepG2 cells were transfected using lacZ as reporter gene and characteristics such as the morphology, the mean particle size, the zeta potential and the transfection efficiency in HepG2 cells were further investigated by different methods. The resulting PLPD had a regular spherical surface with an average size of (228. 9 +/- 8. 0) nm (polydispersity index, PDI = 0. 122 +/- 0. 02, n = 3) , a zeta potential of ( - 25. 08 +/-2. 50) mV (n = 3) and a transfection efficiency of (12. 18 +/- 3. 80) mU x mg(-1) (protein). The Tf-PLPD had an average size of (240 +/- 12) nm (polydispersity index, PDI = 0. 150 +/- 0. 03, n = 3), a zeta potential of ( - 24. 10 +/- 2. 50) mV ( n = 3) and a transfection efficiency of (24. 26 +/- 2. 60) mU x mg(-1) (protein) , 20 times greater than that of the naked plasmid DNA. The presence of serum didn' t affect the tansfection activity of PLPD or Tf-PLPD. Compared to one kind of cationic liposomes (liposome-protamine-DNA, LPD), the PLPD and Tf-PLPD had much less cytotoxicity to three hepatic cell lines (including HepG2, SMMC7721 and Chang' s normal hepatocyte). The results indicated that the Tf-PLPD is a perspective non-viral vector for gene delivery systems.
开发了一种新型转铁蛋白修饰的非病毒基因递送系统Tf-PLPD，并对其相关特性进行了研究。采用薄膜分散-过滤法制备空白阳离子脂质体。PLPD的制备方法如下：首先将质粒DNA和鱼精蛋白混合在一起，然后将所得的多聚体在室温下孵育10分钟，接着加入预先形成的空白阳离子脂质体。通过静电相互作用将转铁蛋白吸附在PLPD表面以形成Tf-PLPD。采用中心复合设计（CCD）优化配方。以lacZ作为报告基因转染HepG2细胞，并通过不同方法进一步研究其在HepG2细胞中的形态、平均粒径、zeta电位和转染效率等特性。所得的PLPD表面呈规则球形，平均大小为(228.9±8.0)nm（多分散指数，PDI = 0.122±0.02，n = 3），zeta电位为(-25.08±2.50)mV（n = 3），转染效率为(12.18±3.80)mU·mg⁻¹（蛋白质）。Tf-PLPD的平均大小为(240±12)nm（多分散指数，PDI = 0.150±0.03，n = 3），zeta电位为(-24.10±2.50)mV（n = 3），转染效率为(24.26±2.60)mU·mg⁻¹（蛋白质），是裸质粒DNA的20倍。血清的存在不影响PLPD或Tf-PLPD的转染活性。与一种阳离子脂质体（脂质体-鱼精蛋白-DNA，LPD）相比，PLPD和Tf-PLPD对三种肝细胞系（包括HepG2、SMMC7721和张氏正常肝细胞）具有更低的细胞毒性。结果表明，Tf-PLPD是一种有前景的基因递送系统非病毒载体。
