Spatiotemporal properties of intracellular calcium signaling in osteocytic and osteoblastic cell networks under fluid flow.
Spatiotemporal properties of intracellular calcium signaling in osteocytic and osteoblastic cell networks under fluid flow.
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DOI：
10.1016/j.bone.2013.01.008
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发表时间：
2013-04
期刊：
影响因子：
4.1
通讯作者：
Guo, X. Edward
中科院分区：
文献类型：
--
作者：
Jing, Da;Lu, X. Lucas;Luo, Erping;Sajda, Paul;Leong, Pui L.;Guo, X. Edward
关键词：
机械刺激可以触发骨细胞和成骨细胞的细胞内钙反应。成功构建骨细胞网络需要对网络中钙信号的时空特性进行更细致和系统的分析。本文采用一种基于独立分量分析(ICA)的无监督算法来提取网络中骨细胞的钙信号。我们证明了基于ICA的技术可以产生比人工感兴趣区域(ROI)方法更高的信号保真度。其次，对成骨细胞样MLO-Y4和成骨细胞样MC3T3-E1细胞网络在层流和稳态流刺激下钙信号的时空特性进行了系统的分析和比较。与MC3T3-E1细胞相比，MLO-Y4细胞表现出更多的钙离子瞬变，表现为更多的钙离子峰，到达第一峰的时间更短，第一和第二峰之间的时间更短。在时间特性方面，MLO-Y4细胞表现出较高的刺激率和钙振荡频率。MLO-Y4细胞网络中钙信号的空间同步性高于MC3T3-E1细胞网络，且与细胞间距离呈负相关，提示在MLO-Y4细胞网络中钙波传播速度更快。我们的研究结果表明，基于无监督ICA的信号提取技术比传统的ROI分析方法更敏感、更定量，有可能广泛应用于细胞网络中的Ca~(2+)信号提取。本研究还揭示了在处理机械刺激的过程中，成骨细胞和成骨细胞网络的钙信号在时空上的显著差异。MLO-Y4细胞较高的细胞内钙振荡行为和细胞间的协调性进一步证明，骨细胞可能是骨建模和重建过程中的主要力学传感器。
Mechanical stimuli can trigger intracellular calcium (Ca2+) responses in osteocytes and osteoblasts. Successful construction of bone cell networks necessitates more elaborate and systematic analysis for the spatiotemporal properties of Ca2+ signaling in the networks. In the present study, an unsupervised algorithm based on independent component analysis (ICA) was employed to extract the Ca2+ signals of bone cells in the network. We demonstrated that the ICA-based technology could yield higher signal fidelity than the manual region of interest (ROI) method. Second, the spatiotemporal properties of Ca2+ signaling in osteocyte-like MLO-Y4 and osteoblast-like MC3T3-E1 cell networks under laminar and steady fluid flow stimulation were systematically analyzed and compared. MLO-Y4 cells exhibited much more active Ca2+ transients than MC3T3-E1 cells, evidenced by more Ca2+ peaks, less time to the 1st peak and less time between the 1st and 2nd peaks. With respect to temporal properties, MLO-Y4 cells demonstrated higher spike rate and Ca2+ oscillating frequency. The spatial intercellular synchronous activities of Ca2+ signaling in MLO-Y4 cell networks were higher than those in MC3T3-E1 cell networks and also negatively correlated with the intercellular distance, revealing faster Ca2+ wave propagation in MLO-Y4 cell networks. Our findings show that the unsupervised ICA-based technique results in more sensitive and quantitative signal extraction than traditional ROI analysis, with the potential to be widely employed in Ca2+ signaling extraction in the cell networks. The present study also revealed a dramatic spatiotemporal difference in Ca2+ signaling for osteocytic and osteoblastic cell networks in processing the mechanical stimulus. The higher intracellular Ca2+ oscillatory behaviors and intercellular coordination of MLO-Y4 cells provided further evidences that osteocytes may behave as the major mechanical sensor in bone modeling and remodeling processes.
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