Xu Zhi Ping, Walker Tara L, Liu Kerh-lin, Cooper Helen M, Lu G Q Max, Bartlett Perry F
ARC Centre for Functional Nanomaterials, Australian Institute of Bioengineering and Nanotechnology, The University of Queensland, Brisbane.
Int J Nanomedicine. 2007;2(2):163-74.
We prepared stable homogeneous suspensions with layered double hydroxide (LDH) nanoparticles for in vitro gene delivery tests. The viability of HEK 293T cells in the presence of LDH nanoparticles at different concentrations was investigated. This revealed 50% cell viability at 500 microg/mL of LDH nanoparticles that is much higher than 50-100 microg/mL used for the delivery tests. The supercoiled pEF-eGFP plasmid (ca. 6100 base pairs) was mixed with LDH nanoparticle suspensions for anion exchange at a weight ratio of DNA/LDH between 1:25 and 1:100. In vitro experiments show that GFP expression in HEK 293T cells starts in the first day, reaches the maximum levels by the second day and continues in the third day. The GFP expression generally increases with the increase in DNA loading in DNA-LDH nanohybrids. However, the delivery efficiency with LDH nanoparticles as the agent is low. For example, the relative efficiency is 7%-15% of that of the commercial agent FuGENE 6. Three to 6% of total cells expressed GFP in an amount detectable by the FACS cytometry 2 days after transfection at 1 microg/mL of plasmid DNA with 25 microg/mL of LDH nanomaterial. The lower delivery efficiency could be attributed to the aggregation of LDH nanoparticles caused by the long-chain plasmid DNA.
我们制备了含层状双氢氧化物（LDH）纳米颗粒的稳定均匀悬浮液，用于体外基因递送测试。研究了不同浓度的LDH纳米颗粒存在下HEK 293T细胞的活力。结果显示，在500μg/mL的LDH纳米颗粒时细胞活力为50%，这远高于用于递送测试的50 - 100μg/mL。将超螺旋pEF - eGFP质粒（约6100个碱基对）与LDH纳米颗粒悬浮液混合，以进行阴离子交换，DNA/LDH的重量比为1:25至1:100。体外实验表明，HEK 293T细胞中的GFP表达在第一天开始，第二天达到最高水平，并在第三天持续。GFP表达通常随着DNA - LDH纳米杂化物中DNA负载量的增加而增加。然而，以LDH纳米颗粒作为载体的递送效率较低。例如，相对效率为商业载体FuGENE 6的7% - 15%。在用25μg/mL的LDH纳米材料和1μg/mL的质粒DNA转染2天后，通过FACS细胞仪检测，有3%至6%的总细胞表达了可检测量的GFP。较低的递送效率可能归因于长链质粒DNA导致的LDH纳米颗粒聚集。
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