Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus, OH 43210, USA.
Int J Pharm. 2010 Jun 15;392(1-2):201-8. doi: 10.1016/j.ijpharm.2010.03.012. Epub 2010 Mar 7.
While calcium-phosphate has been used to deliver plasmid DNA (pDNA) for decades, the method is typically characterized by low and irreproducible transfection efficiency relative to the other non-viral approaches, such as liposomes and polymers. Here we report a novel gene transfer vector comprising lipid-coated nano-calcium-phosphate (LNCP) that provides consistently efficient and satisfactory pDNA delivery. It is based on core-shell nanoparticles comprising a calcium-phosphate core and a cationic lipid shell. This method, in contrast to the solution precipitation methods used in the past, yields colloidally stable calcium-phosphate nanoparticles inside the cationic liposomes. Our results indicate that the particle size and the size distribution of the LNCP remain virtually unchanged even after 21 days of storage. Atomic force microscopy measurements reveal that the LNCP have a 5-fold higher rigidity than common cationic liposomes. The LNCP transfected pDNA 24 times greater than the naked pDNA and 10-fold greater relative to the standard calcium-phosphate precipitation preparations, suggesting that the LNCP may have potential as a novel transfection agent for gene therapy.
几十年来，钙磷一直被用于递送质粒 DNA（pDNA），但与其他非病毒方法（如脂质体和聚合物）相比，该方法的转染效率通常较低且不可重复。在这里，我们报告了一种由脂质包覆的纳米钙磷（LNCP）组成的新型基因传递载体，该载体可提供始终如一的高效和令人满意的 pDNA 递送。它基于包含钙磷核和阳离子脂质壳的核壳纳米粒子。与过去使用的溶液沉淀方法相比，该方法在阳离子脂质体内部产生胶体稳定的钙磷纳米粒子。我们的结果表明，LNCP 的粒径和粒径分布在储存 21 天后几乎没有变化。原子力显微镜测量表明，LNCP 的刚性比普通阳离子脂质体高 5 倍。LNCP 转染的 pDNA 是裸露 pDNA 的 24 倍，相对于标准的钙磷沉淀制剂增加了 10 倍，这表明 LNCP 可能有作为基因治疗的新型转染剂的潜力。
