摘要
目的:利用静电纺丝技术构建骨形态发生蛋白2(bone morphogenetic protein,BMP-2)缓释纳米微球(nanospheres,NPs)的聚己内酯(polycaprolactone,PCL)复合纳米纤维支架材料,评价其对MC3T3-E1细胞增殖、分化的影响。方法:采用去溶剂法和静电自组装技术制备载BMP-2的壳聚糖纳米微球,检测其形貌、粒径及成分,BMP-2蛋白包封率及体外缓释情况。利用静电纺丝技术制备含BMP-2纳米微球的PCL复合支架材料,检测其形貌、亲水性能及BMP-2蛋白的缓释情况。进行体外细胞学实验,观察细胞在材料中的生长情况,检测材料促细胞成骨分化过程中碱性磷酸酶(alkaline phosphatase,ALP)、相关基因和矿化结节的形成情况。采用SPSS 21.0软件包进行统计学分析。结果:成功制备出结构稳定的纳米微球结构,微球载药率高并可实现BMP-2的持续释放。PCL/BNPs支架材料组有着良好的亲水性能并有利于细胞增殖、分化。体外细胞实验结果显示,细胞在支架上铺展良好,黏附数量增加。ALP和相关成骨基因COL1、OPN和RUNX2均表达增高,钙结节大小和数量明显增加。结论:BMP-2缓释纳米微球的聚己内酯复合纤维支架材料可为骨组织工程的发展提供新的材料选择。
PURPOSE:Electrospinning technology was used to construct PCL composite nanofiber scaffold material of BMP-2 sustained-release nanospheres,and the effect of this nanospheres on proliferation and differentiation of MC3T3-E1 cells was evaluated.METHODS:Solvent removal method and electrostatic self-assembly technology were used to prepare BMP-2 loaded chitosan nanospheres,the morphology,particle size and composition,BMP-2 protein encapsulation efficiency,and in vitro sustained release were tested.Electrospinning technology was ued to prepare PCL composite scaffold material containing BMP-2 nanospheres,and its morphology,hydrophilicity,and sustained release of BMP-2 protein were examined.In vitro cytology experiment was conducted to observe the growth of cells in the material,and the formation of ALP,related genes,and mineralized nodules during the process of osteogenic differentiation of the material were detected.SPSS 21.0 software package was used for statistical analysis.RESULTS:The nanospheres structure with stable structure was successfully prepared,with a high drug loading rate and sustained release of BMP-2.The PCL/BNPs scaffold material group had good hydrophilic properties and was conducive to cell proliferation and differentiation.The results of in vitro cell experiments showed that the cells spread well on the scaffold and the number of adhesions increased.ALP and related osteogenic genes COL1,OPN,RUNX2 increased,and the size and number of calcium nodules increased signifi cantly.CONCLUSIONS:The polycaprolactone composite fiber scaffold material of BMP-2 sustained-release nanospheres can provide a new choice for the development of bone tissue engineering.
作者
段昕
刘菲
刘珂珂
王松松
张云涛
DUAN Xin;LIU Fei;LIU Ke-ke;WANG Song-song;ZHANG Yun-tao(Department of Prosthodontics,Affiliated Hospital of Binzhou Medical University,Binzhou 256600,Shandong Province,China)
出处
《上海口腔医学》
CAS
北大核心
2022年第5期476-482,共7页
Shanghai Journal of Stomatology
基金
山东省医药卫生科技发展计划面上项目(2017WS231)。
关键词
静电纺丝
骨形态发生蛋白2
骨组织工程
成骨分化
双重缓释
Electrospinning
Bone morphogenetic protein 2
Bone tissue engineering
Osteogenic differentiation
Dual release
