负载siRNA胶原/生物活性玻璃复合材料的成骨作用  

Osteogenic effect of collagen/bioglass composites carrying noggin siRNA

作　　者：陈燕玲 陈良娇[2] 李正茂[2] 兰泽栋 CHEN Yanling1, CHEN Liangjiao2, LI Zhengmao2, LAN Zedong3(1Department of Orthodontics, Stomatological Hospital Affiliated to Fujian Medical University, Fuzhou 350004, China; 2.Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatology Hospital of Guangzhou Medical University, Guangzhou 510140, China; 3.Shenzhen Stomatological Hospital Affiliated to Southern Medical University, Shenzhen 518000, Chin)

机构地区：[1]福建医科大学附属口腔医院正畸科,福建福州350004 [2]广州医科大学附属口腔医院//广州口腔病研究所//口腔医学重点实验室,广东广州510140 [3]南方医科大学深圳口腔医院,广东深圳518000

出　　处：《南方医科大学学报》2018年第1期106-111,共6页Journal of Southern Medical University

基　　金：广州市医药卫生科技项目(20161A011090)

摘　　要：目的探讨负载si RNA胶原/生物活性玻璃复合材料的成骨作用。方法冷冻干燥法分别制备胶原/生物玻璃、负载阴性对照si RNA胶原/生物玻璃和负载noggin胶原/生物玻璃3组复合材料。用CCK8试验检测空白组上述3组不同支架材料浸提液对细胞增殖的影响,ALP活性检测、q-PCR检测、茜素红染色评估3组不同支架对MC3T3细胞矿化的影响。结果材料浸提液共培养MC3T3细胞3、5 d后,胶原/生物玻璃复合材料、负载阴性对照si RNA胶原/生物玻璃复合材料和负载noggin胶原/生物玻璃支架组细胞增殖数目均明显高于空白组,差异具有统计学意义(P<0.05)。培养14 d后,负载si RNA胶原/生物玻璃支架组ALP活性高于单纯支架组,差异具有统计学意义(P<0.05)。培养14 d后,负载si RNA胶原/生物玻璃支架组ALP、Runx2、BSP表达量高于单纯支架组,差异具有统计学意义(P<0.05)。茜素红染色结果表明负载si RNA胶原/生物玻璃支架组较其余组有更多矿化结节形成,差异具有统计学意义(P<0.05)。结论负载si RNA胶原/生物玻璃复合材料支架具有良好的生物相容性,胶原/生物玻璃复合材料和si RNA noggin可协同增效促进成骨。Objective To investigate osteogenic effect of collagen/bioglass composites loaded with a small interfering RNA（si RNA） targeting noggin. Methods The collagen/bioglass composites loaded with the negative control si RNA or noggin si RNA were prepared by freeze-drying method. CCK8 test was used to evaluate the proliferation of MC3 T3 cells exposed to the aqueous extracts of collagen/bioglass composites and the si RNA-loaded collagen/bioglass composites. ALP activity assay,quantitative real-time PCR and Alizarin Red staining were used to assess the effect of the 3 composites on mineralization in MC3 T3 cells. Results MC3 T3 cells cultured for 3 and 5 days in the presence of the extracts of the 3 composites all showed significantly more active proliferation than the blank control cells（P〈0.05）. Compared with the cells seeded on the scaffold without si RNA, MC3 T3 cells seeded on collagen/bioglass scaffold loaded with noggin si RNA showed a significantly higher ALP activity at 14 days after seeding（P〈0.05） with significantly increased expression of ALP, Runx2 and BSP m RNAs（P〈0.05）.Alizarin Red staining showed that the cells seeded on the noggin si RNA-loading collagen/bioglass scaffold contained significantly more mineralized nodules than the other cells（P〈0.05）. Conclusion The collagen/bioglass composites loaded with noggin si RNA have a good biocompatibility, and the collagen/bioglass composites and noggin si RNA show a synergistic effect in promoting osteogenesis.

关 键 词：SIRNA 胶原 生物活性玻璃 成骨 

分 类 号：R318.08[医药卫生—生物医学工程] R68[医药卫生—基础医学]