辛伐他汀缓释微球影响骨组织工程成骨性能的体外实验  

Simvastatin sustained-release microspheres promote osteogenesis of bone tissue engineering in vitro

作　　者：青薇 代彦君 黄丽娟 任静 郑佳俊 庹嫱 任小华 牟雁东 Qing Wei;Dai Yanjun;Huang Lijuan;Ren Jing;Zheng Jiajun;Tuo Qiang;Ren Xiaohua;Mu Yandong(School of Stomatology,Southwest Medical University,Luzhou 646000,Sichuan Province,China;School of Medicine,University of Electronic Science and Technology,Chengdu 610000,Sichuan Province,China)

机构地区：[1]西南医科大学口腔医学院,四川省泸州市646000 [2]电子科技大学医学院,四川省成都市610000

出　　处：《中国组织工程研究》2019年第2期239-244,共6页Chinese Journal of Tissue Engineering Research

基　　金：四川省科技厅项目(2016TD0008;项目负责人:牟雁东)~~

摘　　要：背景:近年来国内外研究将辛伐他汀缓释微球组装在支架材料上,发现其可完成骨缺损的修复,取得不错的效果。目的:探讨辛伐他汀缓释微球对组织工程成骨性能的影响。方法:采用薄膜分散法制作纳米脂质体辛伐他汀缓释微球,检测其粒径和多分散系数及包封率。采用透析装置检测游离辛伐他汀溶液与辛伐他汀缓释微球溶液的缓释性能。将第3代人胎盘间充质干细胞接种于多孔羟基磷灰石陶瓷支架上,待细胞贴壁生长后分2组培养,实验组加入辛伐他汀缓释微球溶液,空白组加入普通培养基,7,14,21 d后进行转录组测序,实时荧光定量PCR验证成骨基因表达水平。结果与结论:(1)辛伐他汀缓释微球平均粒径为(77.27±6.4) nm,多分散系数为0.131±0.040,包封率为85.6%;(2)游离辛伐他汀溶液在前3 d迅速释放超过了总药量的80%,辛伐他汀缓释微球溶液在前3 d的药物释放量仅约40%,此后也一直较缓慢持续释放至14d,14d累计释放量接近80%;(3)转录组测序显示,实验组骨形态发生蛋白2、骨形态发生蛋白4和血管内皮生长因子等成骨相关基因富集表达;(4)实时荧光定量PCR检测显示,实验组7,14,21 d的骨形态发生蛋白2、骨形态发生蛋白4、血管内皮生长因子基因表达均高于空白组(P <0.05);(5)结果表明,辛伐他汀缓释微球可促进组织工程成骨性能表达。BACKGROUND:In recent years,domestic and foreign studies have assembled simvastatin sustained-release microspheres onto scaffold materials.It has been found that simvastatin can be used to repair bone defects and good results have been obtained.OBJECTIVE:To investigate the effect of simvastatin sustained-release microspheres on tissue engineering osteogenesis.METHODS:Nanoliposome simvastatin sustained-release microspheres were prepared by thin-film dispersion method.The particle size,polydispersity coefficient and encapsulation efficiency were measured.The sustained release properties of free simvastatin solution and simvastatin sustained-release microsphere solution were measured by dialysis device.The third generation of human placental mesenchymal stem cells was inoculated on porous hydroxyapatite ceramic scaffolds.The cells were cultured in two groups after adherent growth.The experimental group was treated with simvastatin sustained-release microspheres,and the normal medium was added to the blank group.Transcriptome sequencing was carried out after 7,14,21 days.The expression levels of osteogenic genes were verified by real-time fluorescence quantitative PCR.RESULTS AND CONCLUSION:(1)The average particle size of simvastatin sustained-release microspheres was(77.27±6.4)nm,the polydispersity coefficient was 0.131±0.040,and the encapsulation efficiency was 85.6%.(2)The rapid release of free simvastatin solution exceeded 80%total dose in the first 3 days.While the simvastatin sustained-release microsphere solution released only about 40%of the drug in the first 3 days,and had been slowly released until the 14th day,and the cumulative release at the 14th day was close to 80%.(3)Transcriptome sequencing showed that the osteogenesis-related genes such as bone morphogenetic protein 2,bone morphogenetic protein 4 and vascular endothelial growth factor were enriched and expressed in the experimental group.(4)Real-time quantitative PCR showed that the mRNA levels of bone morphogenetic protein 2,bone morphogenetic pro

关 键 词：羟基磷灰石 辛伐他汀 缓释微球 脂质体 间充质干细胞 骨组织工程 转录组测序 成骨效能 生物材料 羟基磷灰石类 脂质体 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]