Spatiotemporal Analysis of Osteoblast Morphology and Wnt Signal-Induced Osteoblast Reactivation during Bone Modeling in Vitro.
Spatiotemporal Analysis of Osteoblast Morphology and Wnt Signal-Induced Osteoblast Reactivation during Bone Modeling in Vitro.
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DOI：
10.1002/jbm4.10689
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发表时间：
2022-11
期刊：
影响因子：
3.8
通讯作者：
Hikita, Atsuhiko
中科院分区：
文献类型：
--
作者：
Tsuji, Naoki;Sakamoto, Tomoaki;Hoshi, Kazuto;Hikita, Atsuhiko
通过分化成骨细胞形成骨结节被认为是一种模拟骨模型的体外模型。然而，骨结节形成过程中成骨细胞行为和基质生成的细节尚不清楚。在这里，我们提出了一个时空分析系统，用于通过双光子显微镜评估骨体外建模过程中成骨细胞的形态和基质的产生。利用该系统，在矿化结节形成过程中观察到成骨细胞形态从立方体到扁平的变化，并对这种变化进行了量化。骨成形率高的区域密集分布着立方体成骨细胞，其特征是细胞膜上有气泡和突出的结构。带泡的长方体成骨细胞流动性强，成骨细胞泡呈极性分布。此外，模拟romosozumab治疗，当分化的扁平成骨细胞被BIO（一种GSK3β抑制剂）刺激时，它们被重新激活，获得具有膜上气泡的立方体形态，并且比未刺激的细胞产生更多的基质。我们的分析系统是骨建模过程中评估成骨细胞形态和功能的有力工具。©2022作者。JBMR Plus由Wiley期刊有限责任公司代表美国骨骼和矿物研究协会出版。
Bone nodule formation by differentiating osteoblasts is considered an in vitro model that mimics bone modeling. However, the details of osteoblast behavior and matrix production during bone nodule formation are poorly understood. Here, we present a spatiotemporal analysis system for evaluating osteoblast morphology and matrix production during bone modeling in vitro via two‐photon microscopy. Using this system, a change in osteoblast morphology from cuboidal to flat was observed during the formation of mineralized nodules, and this change was quantified. Areas with high bone formation were densely populated with cuboidal osteoblasts, which were characterized by blebs, protruding structures on their cell membranes. Cuboidal osteoblasts with blebs were highly mobile, and osteoblast blebs exhibited a polar distribution. Furthermore, mimicking romosozumab treatment, when differentiated flattened osteoblasts were stimulated with BIO, a GSK3β inhibitor, they were reactivated to acquire a cuboidal morphology with blebs on their membranes and produced more matrix than nonstimulated cells. Our analysis system is a powerful tool for evaluating the cell morphology and function of osteoblasts during bone modeling. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.