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Mechanical Properties and Gene Expression of Chondrocytes on Micropatterned Substrates Following Dedifferentiation in Monolayer

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Abstract

Chondrocytes in articular cartilage normally exhibit high expression of collagen II and aggrecan but rapidly dedifferentiate to a fibroblastic phenotype if passaged in culture. Previous studies have suggested that the loss of chondrocyte phenotype is associated with changes in the structure of the F-actin cytoskeleton, which also controls cell mechanical properties. In this study, we examined how dedifferentiation in monolayer influences the mechanical properties of chondrocytes isolated from different zones of articular cartilage. Atomic force microscopy was used to measure the mechanical properties of superficial and middle/deep zone chondrocytes as they underwent serial passaging and subsequent growth on fibronectin-coated, micropatterned self-assembled monolayers that restored a rounded cell shape in 2D culture. Chondrocytes exhibited significant increases in elastic and viscoelastic moduli with dedifferentiation in culture. These changes were only partially ameliorated by the restoration of a rounded shape on micropatterned surfaces. Furthermore, intrinsic zonal differences in cell mechanical properties were rapidly lost with passage. These findings indicate that cell mechanical properties may provide additional measures of phenotypic expression of chondrocytes as they undergo dedifferentiation and possibly redifferentiation in culture.

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Acknowledgments

This work was supported in part by NIH grants AR53448 (EMD), EB01630, AG15768, AR48182, AR50245, and EB002025 (RS) and NSF grant CMS-0507151 (RS). The authors would like to thank Drs. Ashutosh Chilkoti, Dominic Chow, David Dumbauld, and Andrés Garcia for discussions and advice on the development of micropatterned substrates. The authors would also like to acknowledge the Shared Materials Instrumentation Facility at Duke University for providing resources for SEM imaging.

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  1. Eric M. Darling

    Present address: Department of Molecular Pharmacology, Physiology, & Biotechnology, Brown University, Providence, RI, 02912, USA

Authors and Affiliations

  1. Department of Surgery, Orthopaedic Research Laboratories, Duke University Medical Center, 375 MSRB, Box 3093, Durham, NC, 27710, USA

    Eric M. Darling, Poston E. Pritchett & Farshid Guilak

  2. Department of Biomedical Engineering, Duke University Medical Center, Durham, NC, 27710, USA

    Eric M. Darling & Farshid Guilak

  3. Department of Mechanical Engineering & Materials Science, Duke University Medical Center, Durham, NC, 27710, USA

    Stefan Zauscher & Farshid Guilak

  4. Department of Physics, Elon University, Elon, NC, 27244, USA

    Benjamin A. Evans

  5. Department of Physics and Astronomy, University of North Carolina, Chapel Hill, Chapel Hill, NC, 27599, USA

    Richard Superfine

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Correspondence to Farshid Guilak.

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Darling, E.M., Pritchett, P.E., Evans, B.A. et al. Mechanical Properties and Gene Expression of Chondrocytes on Micropatterned Substrates Following Dedifferentiation in Monolayer. Cel. Mol. Bioeng. 2, 395–404 (2009). https://doi.org/10.1007/s12195-009-0077-3

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  • DOI: https://doi.org/10.1007/s12195-009-0077-3

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