纤维蛋白胶与异种骨复合支架中立体培养的兔骨髓基质细胞

单一的支架材料往往具有一些自身难以克服的缺点，拟构建一种具有较多优越性的复合支架，希望能够解决种子细胞与支架材料黏附的问题。 目的：应用纤维蛋白胶、异种无机骨构建骨组织工程复合支架材料，探讨兔骨髓基质细胞在这种复合支架材料中立体培养情况。 设计、时间及地点：观察实验，于2007-11/ 2008-03在吉林大学中日联谊医院骨科研究室、天津理工大学机械工程学院完成。 材料：纤维蛋白原与凝血酶按比例制备纤维蛋白胶，牛松质骨经去脂去蛋白等无机化处理后与纤维蛋白胶复合，制成复合骨支架材料。 方法：将兔骨髓基质细胞作为种子细胞进行传代培养，收集后在纤维蛋白胶、异种无机骨构成的复合支架中进行立体培养。 主要观察指标：采用相差显微镜、透射电子显微镜等手段观察骨髓基质细胞在纤维蛋白胶中的生长状况。 结果：相差显微镜下观察见骨髓基质细胞均匀分布于复合支架中，具有良好活性，细胞不断增殖、分化，培养4周骨髓基质细胞形成密集立体网状。透射电子显微镜下见复合支架中培养4周的基质细胞内细胞器结构完整，局部有细突起，胞质内可见线粒体，核糖体，粗面内质网。 结论：在纤维蛋白胶与异种无机骨构建的复合支架中骨髓基质细胞具有良好活性，可迅速扩增生长。

Marrow stromal cells cultured in a composite scaffold of fibrin glue and xenogeneic inorganic bone

BACKGROUND: A superior composite scaffold hopes be constructed to resolve adhesion between seed cell and scaffold material. OBJECTIVE: To construct composite scaffolds with fibrin glue and xenogeneic inorganic bone and to explore the three-dimensional culture of rabbit marrow stromal cells (MSCs). DESIGN, TIME AND SETTING: Observational experiment was performed at the Department of Orthopedics of China-Japan Union Hospital and the Department of Mechanical Engineering, College of Mechanical Engineering of Jilin University from November 2007 to March 2008. MATERIALS: Fibrin glue was made by a certain ratio of fibrinogen and thrombin; bovine cancellous bone following defatting and deproteinization was mixed with fibrin glue to establish composite scaffold. METHODS: Rabbit MSCs were cultured in vitro and transferred, and the MSCs were collected for three-dimensional culture with combined scaffolds made of xenogeneic inorganic bone and fibrin glue. MAIN OUTCOME MEASURES: The growth and proliferation of MSCs were examined by phase-contrast microscope and transmission electron microscopy. RESULTS: Phase contrast microscope showed that the MSCs could spread evenly in the combined scaffolds. After cultured 4 weeks, the MSCs formed into densely three-dimensional net. It could be observed under the transmission electron microscope that there were micro-protrusions in local stromal cells at 4 weeks after culture, and the mitochondrion as well as ribosomes was observed in the cytoplasm with rough endoplasmic reticulum. CONCLUSION: The MSCs cultured in the combination of fibrin glue and xenogeneic inorganic bone scaffolds show a better activity, and they can proliferate rapidly.