Department of Biological Sciences, Center for Bone and Tissue Repair and Regeneration, Missouri University of Science and Technology, Rolla, Missouri 65409, USA.
J Biomed Mater Res A. 2012 Oct;100(10):2593-601. doi: 10.1002/jbm.a.34195. Epub 2012 Apr 24.
This in vitro study was performed to evaluate the ability of two types of porous bioactive glass scaffolds to support the growth and differentiation of an established osteogenic cell line. The two scaffold types tested included 13-93 glass fiber and trabecular-like scaffolds seeded with murine MLO-A5 cells and cultured for intervals of 2 to 12 days. Culture in MTT-containing medium showed metabolically active cells both on the surface and within the interior of the scaffolds. Scanning electron microscopy revealed well-attached cells on both types of scaffolds with a continual increase in cell density over a 6-day period. Protein measurements also showed a linear increase in cell density during the incubation. Activity of alkaline phosphatase, a key indicator of osteoblast differentiation, increased about 10-fold during the 6-day incubation with both scaffold types. The addition of mineralization media to MLO-A5 seeded scaffolds triggered extensive formation of alizarin red-positive mineralized extracellular material, additional evidence of cell differentiation and completion of the final step of bone formation on the constructs. Collectively, the results indicate that the 13-93 glass fiber and trabecular scaffolds promote the attachment, growth, and differentiation of MLO-A5 osteogenic cells and could potentially be used for bone tissue engineering applications.
本体外研究旨在评估两种多孔生物活性玻璃支架的能力，以支持已建立的成骨细胞系的生长和分化。测试的两种支架类型包括 13-93 玻璃纤维和小梁样支架，接种了鼠 MLO-A5 细胞，并在 2 至 12 天的时间间隔内进行培养。在含有 MTT 的培养基中进行培养显示出代谢活跃的细胞，这些细胞位于支架的表面和内部。扫描电子显微镜显示两种支架上的细胞都附着良好，细胞密度在 6 天内持续增加。蛋白测量也显示细胞密度在孵育期间呈线性增加。碱性磷酸酶的活性，成骨细胞分化的关键指标，在两种支架类型的 6 天孵育期间增加了约 10 倍。向接种 MLO-A5 的支架中添加矿化培养基会触发大量茜素红阳性矿化细胞外基质的形成，这是细胞分化和构建物中骨形成最后一步完成的进一步证据。总的来说，这些结果表明 13-93 玻璃纤维和小梁支架促进了 MLO-A5 成骨细胞的附着、生长和分化，并且可能可用于骨组织工程应用。
