Three-dimensional culture environments enhance osteoblast differentiation.
Three-dimensional culture environments enhance osteoblast differentiation.
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DOI：
10.1111/j.1532-849x.2008.00330.x
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发表时间：
2008-10-01
期刊：
影响因子：
--
通讯作者：
Schneider, Galen B
中科院分区：
文献类型：
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作者：
Boehrs, Jessica;Zaharias, Rebecca S;Schneider, Galen B
目的：在我们实验室之前的工作中，我们证明了在模拟微重力环境中开发的三维（3D）细胞培养物增强了骨样聚集体的形成并加速了前成骨细胞的分化。因此，如此处所述，我们假设如前所述，用少于 10 x 10(6) 个细胞即可发生聚集体形成和矿化。 材料和方法：在旋转壁容器中以不同浓度培养人类前成骨细胞，以模拟微重力 7 天。评估聚集体大小，并使用 Von Kossa 和 Masson Trichrome 染色检测矿化和胶原蛋白表达。扫描电子显微镜用于结构和元素分析。使用免疫组织化学检测成骨标记物 BSPII 和骨桥蛋白 (OP) 的表达。结果：大小和钙表达取决于培养的细胞数量 (p < 0.01)。在整个聚集体中检测到钙和胶原蛋白的表达，但组织与细胞数量无关。聚集体具有相似的微观结构模式，表明有组织的发展。 BSPII 和 OP 的存在表明，这些聚集体在体内骨形成方面具有共同的分化蛋白。结论：这些结果可能会带来与牙科治疗相关的新型骨工程技术。
PURPOSE: In previous work from our laboratory, we demonstrated that the three-dimensional (3D) cell cultures developed in simulated microgravity environments enhanced osseous-like aggregate formation and accelerated preosteoblast cell differentiation. Thus, as described here, we hypothesize that aggregate formation and mineralization would occur with fewer than 10 x 10(6) cells as previously described.MATERIALS AND METHODS: Human preosteoblastic cells were cultured at different concentrations in a rotary wall vessel to simulate microgravity for 7 days. Aggregate size was assessed, and mineralization and collagen expression detected using Von Kossa and Masson Trichrome staining. Scanning electron microscopy was used for structural and elemental analysis. Immunohistochemistry was used to detect expression of the osteogenic markers BSPII and osteopontin (OP).RESULTS: Size and calcium expression were dependent upon cultured cell number (p < 0.01). Calcium and collagen expression were detected throughout the aggregate, but organization was independent of cell number. Aggregates had similar microscopic structural patterns demonstrating organized development. Presence of BSPII and OP showed that the aggregates share common differentiation proteins with in vivo bone formation.CONCLUSIONS: These results may lead to novel bone engineering techniques associated with dental treatment.