Early initiation of endochondral ossification of mouse femur cultured in hydrogel with different mechanical stiffness.
Early initiation of endochondral ossification of mouse femur cultured in hydrogel with different mechanical stiffness.
复制标题
在具有不同机械刚度的水凝胶中培养的小鼠股骨软骨内骨化的早期启动。
DOI：
10.1089/ten.tec.2014.0475
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Matsumoto T.
中科院分区：
文献类型：
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作者：
Sathi GA;Kenmizaki K;Yamaguchi S;Nagatsuka H;Yoshida Y;Matsugaki A;Ishimoto T;Imazato S;Nakano T;Matsumoto T.
矿化是骨组织正常发育和疾病状态中最重要的过程之一。开发一种新的、标准化的体外模型系统，可以很容易地监测细胞动力学和矿化，这对于更好地了解骨组织的发育和生长至关重要。近年来的研究表明，机械环境是成矿的关键条件。我们假设不同机械刚度的水凝胶可以提供一个可以调节骨组织生长和矿化的仿生机械环境。将小鼠胚胎（胚胎第16天）股骨包埋于琼脂糖水凝胶（2-60 kPa）中，在成骨培养基中培养一周。微计算机断层扫描（μCT）结果显示，凝胶条件下培养的股骨头矿化增强，而对照（漂浮培养）条件下培养的股骨头没有矿化。矿化区与次生骨化中心区相对应。组织学和定量分析均表明，10 kPa凝胶条件下培养的股骨头矿化区最大，矿化面积明显大于2、40、60 kPa凝胶条件下培养的股骨头矿化区。免疫荧光结果显示，该区域较高的软骨分化导致矿化增强。这种增强主要与机械力有关，而与氧张力无关。由于与传统的二维或三维细胞培养系统相比，该系统可以增强和缩短矿化过程，因此该系统将成为更好地了解矿化组织发育的独特模型之一。
Mineralization is one of the most important processes in normal bone tissue development and in disease condition. Developing a novel and standardizedin vitromodel system that can readily monitor both cellular dynamics and mineralization is crucial for better understanding the bone tissue development and growth. Recent studies indicated that the mechanical environment is a critical condition in mineralization. We hypothesized that hydrogel with different mechanical stiffness can provide a biomimetic mechanical environment that can modulate bone tissue growth and mineralization. A femur of mouse embryo (embryonic day 16) was embedded in agarose hydrogel (2–60 kPa) and cultured in an osteogenic medium for a week. Microcomputed tomography (μCT) results revealed enhanced mineralization was detected in the femur head cultured in the gel condition, whereas no mineralization in the femur head cultured in the control (floating culture) condition. The mineralized region was corresponding to the region of secondary ossification center. Both histological and quantitative analyses indicated that the mineralized region of femur head cultured in 10 kPa gel condition was the highest and the mineralized area was significantly larger than that cultured in 2, 40, and 60 kPa gel condition. Immunofluorescence results indicated the enhanced mineralization caused by the higher chondrogenic differentiation at that region. This enhancement mainly relating to the mechanical forces and not to the oxygen tension was also confirmed. Since this system enhances and shortens the mineralization procedure compared with the conventional two-dimensional or three-dimensional cell culture system, this hydrogel system would be one of the unique models for better understanding the mineralized tissue development.