Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh
Flow perfusion culture of marrow stromal osteoblasts in titanium fiber mesh
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DOI：
10.1002/jbm.a.10365
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发表时间：
2003-02-01
影响因子：
4.9
通讯作者：
Mikos, AG
中科院分区：
文献类型：
--
作者：
van den Dolder, J;Bancroft, GN;Mikos, AG
本研究的目的是评价静态和流动两种细胞培养技术对种植在钛纤维网中的大鼠骨髓细胞成骨表达的影响，时间长达16天。将大鼠骨髓基质成骨细胞悬液(5×10(5)个/300微升)接种到网状材料中。此后，将构建物在静态条件下或在流动灌流系统中培养4、8和16天。为了评估细胞的增殖和分化，对构建的DNA、钙含量和碱性磷酸酶活性进行了检测。标本还进行了扫描电子显微镜(SEM)和塑料包埋组织切片检查。结果显示，从第4天到第8天，流动灌流系统的DNA含量增加。在第8天，观察到血流灌注的DNA含量显著增加。培养条件与静态培养条件比较，但在16天时，两种培养系统的细胞数相似。钙测量显示，流动灌流培养16天后，细胞内钙含量明显增加。扫描电子显微镜观察显示，流动灌流培养16天后，细胞层和矿化基质完全覆盖。此外，这种基质深入到了支架中。相比之下，在静态条件下培养的网状物在网状物的上表面只有一层薄薄的基质。对光学显微镜切片的评价证实了扫描电子显微镜的观察结果。根据我们的结果，我们得出结论，流动灌流系统可以促进种植在钛纤维网中的骨髓基质成骨细胞的早期增殖、分化和矿化基质的产生。(C)2002年威利期刊公司。
The objective of this study was to evaluate the effect of two cell culture techniques, static and flow perfusion, on the osteogenic expression of rat bone marrow cells seeded into titanium fiber mesh for a period up to 16 days. A cell suspension of rat bone marrow stromal osteoblasts (5 x 10(5) cells/300 muL) was seeded into the mesh material. Thereafter, the constructs were cultured under static conditions or in a flow perfusion system for 4, 8, and 16 days. To evaluate cellular proliferation and differentiation, constructs were examined for DNA, calcium content, and alkaline phosphatase activity. Samples were also examined with scanning electron microscopy (SEM) and plastic-embedded histological sections. Results showed an increase in DNA from day 4 to day 8 for the flow perfusion system. At day 8, a significant enhancement in DNA content was observed for flow perfusion. culture compared with static culture conditions, but similar cell numbers were found for each culture system at 16 days. Calcium measurements showed a large increase in calcium content of the meshes subjected to flow perfusion at day 16. The SEM examination revealed that the 16-day samples subjected to flow perfusion culture were completely covered with layers of cells and mineralized matrix. In addition, this matrix extended deep into the scaffolds. In contrast, meshes cultured under static conditions had only a thin sheet of matrix present on the upper surface of the meshes. Evaluation of the light microscopy sections confirmed the SEM observations. On the basis of our results, we conclude that a flow perfusion system can enhance the early proliferation, differentiation, and mineralized matrix production of bone marrow stromal osteoblasts seeded in titanium fiber mesh. (C) 2002 Wiley Periodicals, Inc.