人骨髓基质干细胞与表面置换珊瑚羟基磷灰石培养的定向诱导分化

Directional differentiation culture of human bone marrow mesenchymal stem cells on surface exchanging coralline hydroxyapatite

目的观察人骨髓基质干细胞(hMSC)与表面置换的珊瑚羟基磷灰石(SCHA)体外培养的细胞粘附、增殖和分化,寻找理想的骨修复材料。方法海南天然滨珊瑚在特定温度和压力下部分水热反应,制成表面置换的珊瑚羟基磷灰石,将SCHA薄片与人骨髓基质干细胞体外培养,经诱导后于4、8、12、16d分别用荧光显微镜和扫描电镜(SEM)观察细胞活性、粘附和分化过程。结果SCHA保留原有的珊瑚贯穿多孔的三维结构。荧光显微镜可见hMSC在SCHA的表面和孔道内生长良好,第16d达最高水平;电镜显示细胞粘附良好,分化为成骨细胞,分泌大量胶原纤维,可见合成的钙结节。结论hMSC在表面置换的珊瑚羟基磷灰石内粘附、增殖、分化良好,两者具有较好的生物相容性,SCHA是一种良好的骨组织工程支架材料。

Objective To observe the cell attachment,proliferation,and differentiation of human bone marrow mesenchymal stem cells （hMSC） on surface exchanging coralline hydroxyapatite in in vitro culture. Methods The Hainan coralline was treated into surface exchanging coralline hydroxyapatite（SCHA） by ＂Hydrothermal exchange＂at specific condition. The cell viability and characterization was observed by fluorescence microscope and scanning electronic microscope （SEM） at day 4,8,12,16 after directional culture of hMSC on SCHA slices in vitro. Results SCHA has the three-dimensional and interporons structure same as the nature coral, hMSCs grew well and got highest level at day 16 within the surface and channels of CHA from fluorescent microscope. The SEM pictures also revealed that the cells preferentially attached to the rough surfaces hydroxyapatite crystals,induced to osteoblast and secrete a lots of collagen fiber. Calcium nodes were observed. Conclusion hMSC has a better adhesion,proliferation, differentiation to the SCHA. There are good biocompatibility between hMSC and SCHA. It is a good osseous tissue engineering scaffold.