转化生长因子β3/聚乳酸-羟基乙酸微球对干细胞的调控  

Regulation of stem cells by transforming growth factor β3/polylactic acid-glycolic acid microspheres

作　　者：杨振 李浩 高仓健[1,2] 付力伟 田广招 查康康 孙志强 李旭 郭维民[2] 眭翔[2] 黄靖香[2] 刘舒云[2] 卢世璧[2] 郭全义[2] Yang Zhen;Li Hao;Gao Cangjian;Fu Liwei;Tian Guangzhao;Zha Kangkang;Sun Zhiqiang;Li Xu;Guo Weimin;Sui Xiang;Huang Jingxiang;Liu Shuyun;Lu Shibi;Guo Quanyi(Medical College of Nankai University,Tianjin 300071,China;Institute of Orthopedics,Chinese PLA General Hospital,Beijing Key Laboratory of Regenerative Medicine in Orthopedics,Key Laboratory of Musculoskeletal Trauma&War Injuries,PLA,Beijing 100853,China)

机构地区：[1]南开大学医学院,天津市300071 [2]解放军总医院骨科研究所,骨科再生医学北京市重点实验室,全军骨科战创伤重点实验室,北京市100853

出　　处：《中国组织工程研究》2020年第28期4540-4546,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家重点研发计划课题(2019YFA0110600),项目负责人:郭全义;国家自然科学基金(81772319),项目负责人:郭全义.

摘　　要：背景:转化生长因子β3/聚乳酸-羟基乙酸缓释微球系统可使药物在作用部位维持有效药物浓度,提高生长因子的利用率。目的:优化转化生长因子β3/聚乳酸-羟基乙酸缓释微球制备工艺,探究其对脂肪间充质干细胞增殖和迁移的影响。方法:采用乳化-溶剂挥发法制备转化生长因子β3/聚乳酸-羟基乙酸缓释微球,并对微球的形态、粒径大小、药物空间分布、包封率、载药量和缓释性能进行表征。将转化生长因子β3/聚乳酸-羟基乙酸缓释微球溶解于PBS中,于相应的时间点检测上清液中转化生长因子β3浓度,对应时间点扫描电镜观察微球形态。将脂肪间充质干细胞分6组培养,分别加入培养基(阴性对照)、含转化生长因子β3的培养基、含空白聚乳酸-羟基乙酸微球的培养基、含10,100,1000 g/L转化生长因子β3/聚乳酸-羟基乙酸微球的培养基,于对应的时间点CCK-8法检测增殖。将脂肪间充质干细胞分别与培养基(阴性对照)、含转化生长因子β3的培养基、含聚乳酸-羟基乙酸微球的培养基、含10,100,1000 g/L转化生长因子β3/聚乳酸-羟基乙酸微球的培养基以非接触方式共培养24 h,检测细胞迁移数量。结果与结论:①转化生长因子β3/聚乳酸-羟基乙酸微球呈球形,表面光滑,无粘连,粒径均匀分布,微球直径2-50μm,微球内的蛋白药物分布均匀,具有较高的包封率与载药量;②缓释微球具有良好的降解性能,体外可于6个月后完全降解;同时具有良好的缓释性能,体外可缓慢释放转化生长因子β3长达45d;③空白微球及含转化生长因子β3的缓释微球对脂肪间充质干细胞增殖无影响;④空白微球对脂肪间充质干细胞的迁移无影响,转化生长因子β3及含转化生长因子β3的缓释微球可促进脂肪间充质干细胞的迁移,不同质量浓度缓释微球间的促进效果无差异;⑤结果表明,转化生长因子β3/聚乳酸-羟基乙酸缓�BACKGROUND: Transforming growth factor β3/polylactic acid-glycolic acid(TGF-β3/PLGA) sustained-release microspheres can maintain the effective drug concentration at the site of action and provide the feasibility for efficient utilization of growth factors. OBJECTIVE: To optimize the manufacturing process of TGF-β3/PLGA sustained-release microspheres, and investigate their effects on the proliferation and migration of rabbit adipose-derived mesenchymal stem cells(ADSCs). METHODS: TGF-β3/PLGA sustained-release microspheres were prepared by emulsification-solvent evaporation method. The morphology, particle size, drug spatial distribution, encapsulation efficiency, drug loading, and sustained release properties of the microspheres were characterized. The TGF-β3/PLGA sustained-release microspheres were dissolved in phosphate buffered saline. The concentration of TGF-β3 in the supernatant was detected at the corresponding time points. The microsphere morphology was observed by scanning electron microscopy at the corresponding time point. Adipose-derived mesenchymal stem cells were divided into six groups and then cultured with single culture medium(negative control) or culture medium containing TGF-β3 or blank PLGA, or culture medium containing 10,100,1 000 g/L TGF-β3/PLGA microspheres. Cell proliferation was detected by CCK-8 assay at the corresponding time point. Cells in each group were cultured for 24 hours with corresponding medium in a non-contact manner. The number of migratory cells was counted. RESULTS AND CONCLUSION:(1) TGF-β3/PLGA sustained-release microspheres were spherical with smooth surface, no adhesion, and evenly distributed particle size. The microspheres had a diameter of 2-50 μm, and the protein drugs in the microspheres were evenly distributed, with high encapsulation efficacy and encapsulation dose.(2) The TGF-β3/PLGA sustained-release microspheres had good degradation properties and were completely degraded after 6 months in vitro. At the same time, these microspheres had good sustai

关 键 词：转化生长因子Β3 聚乳酸-羟基乙酸 缓释微球 兔脂肪间充质干细胞 增殖 迁移 软骨损伤 组织工程 

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