Boehrs Jessica, Zaharias Rebecca S, Laffoon John, Ko Y Joon, Schneider Galen B
University of Iowa College of Dentistry, Iowa City, IA 52242-1001, USA.
J Prosthodont. 2008 Oct;17(7):517-21. doi: 10.1111/j.1532-849X.2008.00330.x. Epub 2008 Jun 20.
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.
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).
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.
These results may lead to novel bone engineering techniques associated with dental treatment.
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培养的三维（3D）细胞能够增强类骨聚集体的形成并加速前成骨细胞的分化。因此，如下所述，我们假设聚集体的形成和矿化所需的细胞数量会比之前描述的少于10×10⁶个细胞。
将人前成骨细胞以不同浓度接种于旋转壁式生物反应器中，模拟微重力环境培养7天。评估聚集体大小，并用冯库萨染色和马松三色染色检测矿化和胶原蛋白表达情况。采用扫描电子显微镜进行结构和元素分析。用免疫组织化学法检测成骨标志物骨涎蛋白II（BSPII）和骨桥蛋白（OP）的表达。
聚集体大小和钙表达取决于培养的细胞数量（p < 0.01）。在整个聚集体中均检测到钙和胶原蛋白表达，但组织结构与细胞数量无关。聚集体具有相似的微观结构模式，表明其发育具有组织性。BSPII和OP的存在表明聚集体与体内骨形成共享共同的分化蛋白。
这些结果可能会带来与牙科治疗相关的新型骨工程技术。
