IGF-I明胶微球对骨髓基质干细胞增值分化作用的实验性研究

Fabrication of IGF-I Gelatin Microspheres and the Effect on the Proliferation Differentiation of Bone Marrow Stromal Cells

目的制备胰岛素样生长因子-I明胶微球(IGF-I-GMs),观察其一般性质、体外释药性及对兔骨髓基质干细胞(BMSCs)增值和分化效应。方法采用乳化交联法制备IGF-I-GMs,观察其性质;ELISA试剂盒测定IGF-I的含量,计算微球包封率、载药率以及体外释药性。体外分离培养兔BMSCs,四唑盐比色法(MTT)检测IGF-I和IGF-I-GMs对BMSCs增值的影响。用PNPP偶氮法检测碱性磷酸酶活性,反应IGF-I和IGF-I-GMs对BMSCs向成骨细胞定向分化影响。结果①所制备的微球表面光滑圆整、均匀分布,微球载药量和包封率高,体外药物缓释作用好;②IGF-I和IGF-I-GMs均促进BMSCs增值,且IGF-I-GMs效果更加显著;③IGF-I和IGF-I-GMs均促进BMSCs向成骨细胞分化,且IGF-I-GMs效果更加显著。结论 IGF-I-GMs在7天内对IGF-I有良好的缓慢释放作用;BMSCs形态和活性良好;IGF-I-GMs与BMSCs有良好的生物相容性,且促进BMSCs增值及成骨作用均明显优于单独使用IGF-I。

Objective The preparation of the experimental insulin-like growth factor-1 gelatin microspheres, the obselvation of a general nature and in vitro release studies of bone marrow stem cell proliferation and differentiation effects. Method Emulsification cross-linking method prepare IGF-I-GMs and observe its nature; ELISA kit was used to determine the content of IGF-I and calculate the microsphere encapsulation efficiency, drug loading and release pharmaceutical properties in vitro. Bone marrow stem cells were isolated and cultured in vitro application of the MTT assay IGF-1 and IGF-I-GMs value- added role of bone marrow stromal cells. Alkaline phosphatase activity the PNPP detection reaction IGF-I and IGF-I-GMs promote the differentiation of bone marrow stromal cells into osteoblasts directed. Result （1）The smooth surface of the microspheres prepared, evenly distributed and microsphere drug loading and high encapsulation efficiency of microspheres in vitro drug release good results. （2） Both IGF-I and IGF-I-GMs group can promote BMSCs value-added, and IGF-I-GMs group was more significant; （3）Both IGF-I and IGF-I-GMs can promote BMSCs forming osteoblasts, compared to IGF-I, IGF- i-GMs group of the more significant results. Conclusion IGF-I-GMs in 7 days can slow the release of IGF-I meet experimental requirements; IGF-I-GMs good biocompatibility with BMSCs and promote BMSCs appreciation and osteogenic better than the use of IGF-I alone.