羟基磷灰石纳米颗粒:一种新型基因转染载体材料

采用化学共沉淀-水热合成法制备羟基磷灰石纳米颗粒。将羟基磷灰石纳米颗粒与SGC-7901细胞混合培养,用MTT法测定细胞增殖率,显示在10~100μg/ml浓度范围内细胞贴壁生长良好,方差分析各组之间的OD值无明显差异(t>0.05),证明羟基磷灰石纳米颗粒与细胞有良好的生物相容性。琼脂糖凝胶电泳分析发现羟基磷灰石纳米颗粒在酸性与中性条件下能与质粒PEGFP-N1结合,用结合了PEGFP-N1的羟基磷灰石纳米颗粒转染胃癌SGC-7901细胞,荧光显微镜下观察发现其转染效率约为对照组脂质体的80%。用纳米颗粒及纳米颗粒-DNA复合体悬液分别尾静脉注射小白鼠,两周内动物无死亡或其它急性毒性反应,取动物的肝、肾和脑组织进行透射电镜和冰冻切片检查,发现上述组织器官有纳米颗粒分布和绿色荧光蛋白表达。实验证明,羟基磷灰石纳米颗粒是一种新型基因转染载体材料。

Hydroxyapatite Nanoparticles:A Novel Material of Gene Carrier

Hydroxyapatite nanoparticles were prepared in low Ca/P ratio by a kind of electrodeposition-hydrothermal process. The suspension of nanoparticles was cultured with SGC-7901 cells; metabolically active cells were evaluated by MTT analysis. Cells grew well and the nanoparticles in the concentration range of 10-100 microg/ml had no adverse effect on the cell viability. The results show that the nanoparticles have excellent biocompatibility with cells. Agrose gel electrophoresis analysis demonstrated that the nanoparticles had the potential to adsorb EGFP-N1 at the pH ranging between 2 to 7. Nanoparticle-DNA complex could transfer EGFP-N1 into the SGC-7901 cells, and the confocal microscopy analysis revealed that the cells with green fluorescence showed the efficiency of nanoparticle uptake to be about 80% of the efficiency of the Lipofectmine TM 2 000 uptake. In vivo, nanoparticles and DNA-nanoparticle complex were injected into mice respectively via tail-vein, and the mice grew well in two weeks. The liver, kidney, and brain of the mice were sampled and detected with electron microscopy, and all of these exhibited biodistribution of nanoparticles. This study demonstrates that Hydroxyapatite nanoparticles could be used as gene carriers.