合成聚合物Kartogenin/Pluronic F127胶束与骨髓间充质干细胞的成骨分化  

Effect of Kartogenin/Pluronic F127 micelles on osteogenic differentiation of bone marrow mesenchymal stem cells

作　　者：陈栋[1] 蒋欣[1] Chen Dong;Jiang Xin(China-Japan Friendship Hospital,Beijing 100029,China)

机构地区：[1]中日友好医院,北京市100029

出　　处：《中国组织工程研究》2021年第34期5473-5477,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:小分子药物Kartogenin是Smad4/Smad5通路激活剂,拥有极强的促骨分化能力,可促进骨髓间充质干细胞定向分化为成骨细胞,但其药物效果较为局限。目的:设计并研制Kartogenin/Pluronic F127胶束,观察其对骨髓间充质干细胞定向成骨分化的作用。方法:将小分子药物Kartogenin溶于二甲基亚砜中,然后与Pluronic F127溶液混合均匀,通过真空旋转蒸发仪去除有机溶剂购形成一干燥的药脂膜,加入PBS制备为Kartogenin/Pluronic F127胶束。取对数生长期的大鼠骨髓间充质干细胞,分4组处理,分别加入常规细胞培养液(空白组)、含Pluronic F127胶束溶液的细胞培养液(Pluronic F127组)、含Kartogenin的细胞培养液(药物组)、含Kartogenin/Pluronic F127胶束溶液的细胞培养液(实验组),其中药物组与实验组中药物浓度均为1μmol/L,Pluronic F127组与实验组胶束浓度一致。检测细胞增殖、凋亡及成骨相关蛋白表达。结果与结论:①透射电镜显示Kartogenin/Pluronic F127胶束呈现出不规则的球状形态,粒径分布较为均匀,粒径范围为32.6-118.9 nm,多集中在77.3 nm;②MTT检测显示,随着培养时间的延长,4组细胞存活率下降,但均保持在90%以上,4组间细胞生存率比较差异无显著性意义(P>0.05);③处理24 h后的流式细胞仪检测显示,4组间的细胞凋亡率比较差异无显著性意义(P>0.05);④处理24 h后的Western blot检测显示,药物组、实验组的骨桥蛋白、碱性磷酸酶、骨形态发生蛋白2及基质金属蛋白酶2表达均高于Pluronic F127胶束组、空白组(P<0.05),实验组的碱性磷酸酶、骨形态发生蛋白2及基质金属蛋白酶2表达高于药物组(P<0.05);⑤结果表明,Kartogenin/Pluronic F127胶束可促进骨髓间充干细胞的成骨分化。BACKGROUND:The small molecule drug Kartogenin is an activator of the Smad4/Smad5 pathway.It has a strong ability to promote bone differentiation and can promote the directional differentiation of bone marrow mesenchymal stem cells into osteoblasts,but its drug effect is relatively limited.OBJECTIVE:We designed and developed Kartogenin/Pluronic F127 micelles to observe its effect on the directional osteogenic differentiation of bone marrow mesenchymal stem cells.METHODS:Kartogenin was dissolved in dimethyl sulfoxide and mixed with Pluronic F127 solution.The organic solvent was removed by vacuum rotary evaporator to form a dry drug lipid membrane.PBS was added to prepare Kartogenin/Pluronic F127 micelles.The bone marrow mesenchymal stem cells of rats in the logarithmic growth phase were divided into four groups,which were treated with conventional cell culture medium(blank group),cell culture medium containing Pluronic F127 micelle solution(Pluronic F127 group),cell culture medium containing Kartogenin(drug group),and cell culture medium containing Kartogenin/Pluronic F127 micelle solution(experimental group).Among them,the drug concentration in the drug group and the experimental group was 1μmol/L,and the micelle concentration in Pluronic F127 group was the same as that in the experimental group.Cell proliferation,apoptosis and expression of osteogenic related proteins were detected.RESULTS AND CONCLUSION:(1)Transmission electron microscope revealed that Kartogenin/Pluronic F127 micelles had irregular spherical shape and uniform particle size distribution.The particle size ranged from 32.6 nm to 118.9 nm,most of which were concentrated at 77.3 nm.(2)MTT assay showed that with the extension of culture time,the cell survival rate of the four groups decreased,but remained above 90%,and there was no significant difference in the cell survival rate among the four groups(P>0.05).(3)Flow cytometry after 24 hours of treatment showed that there was no significant difference in the cell apoptosis rate among the four groups

关 键 词：骨 材料 胶束 Pluronic F127 Kartogenin 干细胞 成骨分化 细胞增殖 蛋白 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]