不同工艺制备的PCL-R_(15)/siRNA纳米复合物体外生物学效应比较评价

Comparative Evaluations of in Vitro Biological Effects of PCL-R_(15)/siRNA Nanoplexes Prepared by Different Processes

目的以两亲性聚己内酯-聚精氨酸聚合物(PCL-R_(15))为载体,采用不同工艺流程制备得到粒径电位均不同的2种纳米颗粒,并以静电结合包载siRNA得到2种纳米复合物(NR_(60)/siRNA和NR_(160)/siRNA),比较两者在体外细胞水平的效应差异。方法通过对比考察2种纳米复合物的粒径、电位、siRNA包载保护能力、细胞毒性、细胞摄取机制和基因沉默效率等方面的差异。结果 2种胶束复合物虽然具有类似的siRNA保护能力和细胞毒性,但NR_(160)/siRNA复合物表现出较高的粒径电位值,两者粒径相差约100 nm和电位相差约20 mV。并且NR_(160)/siRNA复合物具有更高的细胞摄取效率和更为复杂的摄取途径,表现出更强的基因沉默效率。结论不同工艺制得的2种PCL-R_(15)/siRNA纳米复合物可在细胞水平上引起siRNA转染效率的差异。

OBJECTIVE To prepare amphiphilic polycaprolactone-poly （arginine polymer） （PCL-R15）/siRNA Nanopexes, and two kinds of nanoparticles with different particle size were prepared by different process. After encapsulated siRNA with electrostatic interaction, both of two nanoplexes （NR60/siRNA and NR160/siRNA） were used to compare the effects in vitro cell levels. METHODS The particle size and Zeta potential, siRNA loading and protection ability, cytotoxicity, cellular uptake mechanism and gene silencing efficiency of the two nanoplexes were investigated. RESULTS The RESULTS show that the two nanoplexes have similar siRNA protection ability and cytotoxicity, but the difference between the two sizes is about 100 nm and the potential difference is about 20 mV. Moreover, NR160/siRNA complexes have higher cell uptake efficiency, more complex uptake pathways, and show greater gene silencing efficiency. CONCLUSION These nanoplexes with different particle sizes can cause different transfection efficiency for siRNA delivery in cells.