PEN载ODN MT01复合物性能检测及其对成骨细胞增殖的影响

Performance of Complex PEN Loading ODN MT01 and Its Effect on the Proliferation of Osteoblast

目的：检测PEI衍生物PEN装载ODN MT01复合物性能及PEN/ODN MT01复合物转染MG63细胞对细胞增殖影响。方法：以PEN为载体静电装载ODN MT01,Nanodrop测定PEN与ODN MT01不同质量比时装载效率;琼脂糖凝胶电泳检测PEN装载ODN MT01能力;马尔文粒度电位仪检测PEN/ODN MT01复合物粒度及电位;PEN装载ODN MT01-Cy5转染MG63细胞,观察荧光强度及进行FCM检测;MTT检测转染PEN/ODN MT01复合物细胞增殖率。结果：PEN/ODN MT01复合物随质量比增加表面由负电转为正电,粒径大小90-110nm;ODN MT01与PEN质量比为1∶6时,装载率高达77.67%,且转染MG63细胞荧光强度最大;PEN/ODN MT01复合物与对照组相比明显促进MG63细胞增殖。结论：PEN与ODN MT01质量比为1∶6时,可以高效装载ODN MT01,且可以明显促进MG63细胞增殖。

Objective： To determinate the performance of PEI derivatives of PEN loading ODN MT01 and discuss the proliferation of MG63 cells transfected with PEN/ODN MT01 complex. Methods： The following methods use Nanodrop to determinate the loading efficiency and Agarose gel electrophoresis to determinate the capability of PEN with ODN MT01 by different mass ration. We use the Malvin particle potential to test the particle size and potential of PEN/ODN MT01 complex. By FCM to ensure which is the best ration at the same time by the fluorescence observation of PEN loading ODN MT01--Cy5 which transfects MG63. We detemine the cells of proliferation rate of PEN/MT01 complex by MTT detection. Results： The surface of PEN/MT01 complex is positively charged. Particle size is 90-110 nm,low cytotoxicity,and the quality ration of ODN MT01 and PEN is 1 ： 6, the loading ration is 77.67%. What is more,the transfection of MG63 cell florescence intensity is the largest. Conclusion： The mass ration of PEN and ODN MT01 is 1 ： 6; the ODN MT01 can be efficiently loaded,and can obviously promote the pro-liferation of MG 6a cells.