Osteocyte Characterization on Polydimethylsiloxane Substrates for Microsystems Applications


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In the body, osteocytes reside in lacunae, lenticular shaped cavities within mineralized bone. These cells are linked to each other and surface-residing osteoblasts via physical channels known as gap junctions. It has been suggested that osteocytes sense mechanical load applied to bone and relay that signal to osteoclasts and osteoblasts. Current in vitro and in vivo models of mechanotransduction face temporal and spatial barriers. Recent advances in polydimethylsiloxane (PDMS) based microfabrication techniques may be able to overcome some of these hurdles. However, before the bone research field can effectively utilize microsystems techniques, fundamental groundwork must be completed. This study characterized the behaviour of osteocytes on PDMS coated with collagen type I (CTI) and provides the framework for bone cell mechanotransduction studies using microsystems. The goal was to determine whether osteocytes were adversely affected by the substrate material by comparing their behaviour to a standard glass substrate. In addition, optimal culture conditions and time points for growing osteocytes on PDMS substrates were determined. Results of this study suggested that use of PDMS does not adversely affect osteocyte behaviour. Furthermore, the results demonstrated that osteocytes should be cultured for no less than 72 hours prior to experimentation to allow the establishment and maintenance of phenotypic characteristics. These results completed essential groundwork necessary for further studies regarding osteocytes in microsystems modelling utilizing PDMS.







S. L. York et al., "Osteocyte Characterization on Polydimethylsiloxane Substrates for Microsystems Applications", Journal of Biomimetics, Biomaterials and Tissue Engineering, Vol. 16, pp. 27-42, 2012

Online since:

December 2012




[1] M. Bauer, G. Su, D. Beebe, A. Friedl. 3D microchannel co-culture: method and biological validation, Integr. Biol (Camb. ), (2010), 2 (7-8), 371-8.

DOI: 10.1039/c0ib00001a

[2] A. Paguirigian, D. Beebe. Microfluidics meet cell biology: bridging the gap by validation and application of microscale techniques for cell biological assays, Bioessays, (2008), 30 (9), 811-21.

DOI: 10.1002/bies.20804

[3] Y. Kato, J. J Windle, B. A Koop, G. R Mundy, L. F Bonewald. Establishment of an osteocyte-like cell line, MLO-Y4, J Bone Miner. Res., (1997), 12 (12), 2014-23.

DOI: 10.1359/jbmr.1997.12.12.2014

[4] L. You, S. Temiyasathit, S. R Coyer, E. Tao, F. Prinz, C. R Jacobs. 3D microfluidic approach to mechanical stimulation of osteocyte processes, Cell. Mol. Bioeng., (2008), 1 (1), 103-7.

DOI: 10.1007/s12195-008-0010-1

[5] L. You, S. Temiyasathit, S. R Coyer, A. J Garcia, C. R Jacobs. Bone cells grown on micropatterned surfaces are more sensitive to fluid shear stress, Cell. Mol. Bioeng., (2008), 1 (2-3), 182-8.

DOI: 10.1007/s12195-008-0017-7

[6] H. H Donahue. Gap junctions and biophysical regulation of bone cell differentiation, Bone, (2000), 26 (5), 417-22.

DOI: 10.1016/s8756-3282(00)00245-3

[7] J. Rosser, L. F Bonewald. Studying osteocyte function using the cell lines MLO-Y4 and MLO-A5, Methods Mol. Biol., (2012) 816, 67-81.

DOI: 10.1007/978-1-61779-415-5_6

[8] S. E Papanicolaou, R. J Phipps, D. P Fyhrie, D. C Genetos. Modulation of sclerostin expression by mechanical loading and bone morphogenetic proteins in osteogenic cells, Biorheology, (2009), 46 (5), 389-99.

[9] A. Santos, A. D Bakker, B. Zandieh-Doulabi, C. M Semeins, J. Klein-Nulend. Pulsating fluid flow modulates gene expression of proteins involved in Wnt signalling pathways in osteocytes, J Orthop. Res., (2009), 27 (10), 1280-87.

DOI: 10.1002/jor.20888

[10] C. Galli, G. Passeri, G. M Macaluso. Osteocyte and WNT: the mechanical control of bone formation, J Dent. Res., (2010), 89 (4), 331-43.

[11] X. Li, Y. Zhang, H. Kang, W. Liu, J. Zhang, S. E Harris, D. Wu. Sclerostin binds to LRP5/6 and antagonizes canonical Wnt signalling, J Biol Chem., (2005), 280 (20), 19883-87.

DOI: 10.1074/jbc.m413274200

[12] R. L van Bezooijen, P. ten Dijke, S. E Papapoulos, C. W Lowik. SOST/sclerostin, an osteocyte-derived negative regulator of bone formation, Cytokine Growth Factor Rev., (2005), 16 (3), 319-27.

DOI: 10.1016/j.cytogfr.2005.02.005

[13] C. Lin, X. Jiang, Z. Dai, X. Guo, T. Weng, J. Wang, Y. Li, G. Feng, X. Gao, L. He. Sclerostin mediates bone response to mechanical unloading through antagonizing Wnt/beta-catenin signalling, J Bone Miner. Res., (2009), 24 (10), 1651-61.

DOI: 10.1359/jbmr.090411

[14] P. ten Dijke, C. Krause, D. J de Gorter, C. W Lowik, R. L van Beezooijen. Osteocyte-derived sclerostin inhibits bone formation: its role in bone morphogenetic protein and Wnt signalling, J Bone Joint Surg. Am., (2008), 90 (sup 1), 31-35.

DOI: 10.2106/jbjs.g.01183

[15] A. G Robling, P. J Niziolek, L. A Baldridge, K. W Condon, M. R Allen, I. Alam, S. M Mantila, J. Gluhak-Heinrich, T. M Bellido, S. E Harris, C. H Turner. Mechanical stimulation of bone in vivo reduces Osteocyte expression of Sost/sclerostin, J Biol. Chem., (2008).

DOI: 10.1074/jbc.m705092200

[16] M. M Saunders, J. You, J. E Trosko, H. Yamasaki, H. J Donahue, C. R Jacobs. Gap junctions and fluid flow response in MC3T3-E1 cells, Am. J Physiol. Cell Physiol., (2001), 281 (6) C1917-25.

[17] M. M Saunders, J. You, Z. Zhou, Z. Li, C. E Yellowley, E. L Kunze, C. R Jacobs, H. J Donahue. Fluid-flow induced prostaglandin E2 response of osteoblastic ROS 17/2. 8 cells is gap junction-mediated and independent of cytosolic calcium, Bone, (2003).

DOI: 10.1016/s8756-3282(03)00025-5

[18] M. M Saunders, M. J Seraj, Z. Li, Z. Zhou, C. R Winter, D. R Welch, H. J Donahue. Breast cancer metastatic potential correlates with a breakdown in homospecific and heterospecific gap junctional intercellular communication, Cancer Res., (2001).

[19] J. S Davidson, I. M Baumgarten, E. H Harley. Reversible inhibition of intercellular junctional communication by glycyrrhetinic acid, Biochem. Biophys. Res. Commun., (1986), 134 (1), 29-36.

DOI: 10.1016/0006-291x(86)90522-x

[20] J. C McDonald, S. J Metallo, G. M Whitesides. Fabrication of a configurable, single-use microfluidics device, Anal. Chem., (2001), 73 (23), 5645-50.

DOI: 10.1021/ac010631r

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