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Fluid Flow Induced Calcium Response in Bone Cell Network

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Abstract

In our previous work, bone cell networks with controlled spacing and functional intercellular gap junctions had been successfully established by using microcontact printing and self assembled monolayers technologies [Guo, X. E., E. Takai, X. Jiang, Q. Xu, G. M. Whitesides, J. T. Yardley, C. T. Hung, E. M. Chow, T. Hantschel, and K. D. Costa. Mol. Cell. Biomech. 3:95–107, 2006]. The present study investigated the calcium response and the underlying signaling pathways in patterned bone cell networks exposed to a steady fluid flow. The glass slides with cell networks were separated into eight groups for treatment with specific pharmacological agents that inhibit pathways significant in bone cell calcium signaling. The calcium transients of the network were recorded and quantitatively evaluated with a set of network parameters. The results showed that 18α-GA (gap junction blocker), suramin (ATP inhibitor), and thapsigargin (depleting intracellular calcium stores) significantly reduced the occurrence of multiple calcium peaks, which were visually obvious in the untreated group. The number of responsive peaks also decreased slightly yet significantly when either the COX-2/PGE2 or the NOS/nitric oxide pathway was disrupted. Different from all other groups, cells treated with 18α-GA maintained a high concentration of intracellular calcium following the first peak. In the absence of calcium in the culture medium, the intracellular calcium concentration decreased slowly with fluid flow without any calcium transients observed. These findings have identified important factors in the flow mediated calcium signaling of bone cells within a patterned network.

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Acknowledgments

This work was supported by NIH grant R21 AR052417 (X. Edward Guo). The authors wish to thank Mr. Andrew D. Baik for editing of this paper.

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Authors and Affiliations

  1. Bone Bioengineering Laboratory, Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 1210 Amsterdam Avenue, Mail Code 8904, New York, NY, 10027, USA

    Bo Huo, Xin L. Lu & X. Edward Guo

  2. Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100080, P.R. China

    Bo Huo

  3. Cellular Engineering Laboratory, Department of Biomedical Engineering, Columbia University, New York, NY, 10027, USA

    Clark T. Hung

  4. Cardiac Cell Mechanics Laboratory, Department of Biomedical Engineering, Columbia University, New York, NY, 10027, USA

    Kevin D. Costa

  5. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, 02138, USA

    Qiaobing Xu & George M. Whitesides

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  1. Bo Huo
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Correspondence to X. Edward Guo.

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Huo, B., Lu, X.L., Hung, C.T. et al. Fluid Flow Induced Calcium Response in Bone Cell Network. Cel. Mol. Bioeng. 1, 58–66 (2008). https://doi.org/10.1007/s12195-008-0011-0

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  • DOI: https://doi.org/10.1007/s12195-008-0011-0

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