载基因纳米球对原代肌腱细胞转染效率优化和生物相容性评价  

Optimization of transfection efficiency and biocompatibility evaluation of gene loaded nanospheres on primary tendon cells

作　　者：杨纤纤 金静 周友浪 Yang Qianqian;Jin Jing;Zhou Youlang(Research Center of Clinical Medicine,Research Center of Hand Surgery,Afiliated Hospital of Nantong University,Medical School of Nantong University,Nantong 226001,China)

机构地区：[1]南通大学附属医院临床医学研究中心,南通大学附属医院手外科研究中心,南通大学医学院,南通226001

出　　处：《中华手外科杂志》2023年第5期474-478,共5页Chinese Journal of Hand Surgery

基　　金：国家自然科学基金(81871763)。

摘　　要：目的筛选非病毒载体纳米球对原代肌腱细胞的最佳转染效率以及生物相容性评价。方法制备不同剂量PEI修饰的纳米球, 原代培养鸡、大鼠、小鼠肌腱细胞, 转染不同剂量的siRNA, 转染12 h后对细胞进行荧光拍照与流式细胞仪分析, CCK-8检测转染24、48和72 h后细胞的活力情况, WB检测GAPDH蛋白表达水平, 未转染细胞作为阴性对照, 脂质体作为阳性对照。结果对于鸡、大鼠、小鼠三种原代肌腱细胞, 随着转染试剂剂量的增大, 脂质体的转染效率明显提高, 纳米球的转染效率未见显著改变, 但所有纳米球的转染效率均高于脂质体。相较于脂质体, 纳米球对细胞的初期细胞毒性反应较大, 随着时间的推移, 毒性影响逐渐消失。结论与脂质体相比, 纳米球对于鸡、大鼠、小鼠三种肌腱细胞的转染效率都较高, 纳米球是动物肌腱细胞转染siRNA的理想转染试剂。Objective To screen the optimal transfection efficiency and biocompatibility evaluation of nonviral vector nanospheres on primary tendon cells.Methods The nanospheres with different NP ratios were prepared.The tendon cells of chicken,rat,and mouse were performed primary culture and transfected with different doses of siRNA.And 12 hours after transfection,fluorescence photography and flow cytometry analysis were performed on the cells.CCK-8 was used to detect the viability of the cells after transfection for 24,48,and 72 hours,and WB was used to detect the expression level of GAPDH protein.Non transfected cells were used as negative controls,while liposomes were used as positive controls.Results For the three primary tendon cells of chicken,rat,and mouse,the transfection efficiency of liposomes significantly increased with the increase of transfection reagent dosage.The transfection efficiency of nanospheres did not significantly change,but the transfection efficiency of all nanospheres was higher than that of liposomes.Compared to liposomes,nanospheres have a greater cytotoxic response to cells,and the toxic effects gradually disappear over time.Conclusion Compared with liposomes,nanospheres have higher transfection efficiency for chicken,rat,and mouse tendon cells,and are ideal transfection reagents for siRNA in animal tendon cells.

关 键 词：脂质体 生物相容性 非病毒载体 纳米球 原代肌腱细胞 

分 类 号：R686[医药卫生—骨科学]