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J Appl Physiol 90: 1385-1391, 2001;
8750-7587/01 $5.00
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Vol. 90, Issue 4, 1385-1391, April 2001

Mechanical compression influences intracellular Ca2+ signaling in chondrocytes seeded in agarose constructs

Susan R. Roberts1, Martin M. Knight2,3, David A. Lee2,3, and Dan L. Bader2,3

1 IRC in Biomedical Materials, Institute of Orthopaedics, University College London Medical School, Brockley Hill, Stanmore, Middlesex HA7 4LP; 2 IRC in Biomedical Materials, Queen Mary and Westfield College, University of London, Mile End; and 3 Medical Engineering Division, Department of Engineering, Queen Mary and Westfield College, University of London, Mile End, London E1 4NS, United Kingdom

Ca2+ signaling forms part of a possible mechanotransduction pathway by which chondrocytes may alter their metabolism in response to mechanical loading. In this study, a well-characterized model system utilizing bovine articular chondrocytes embedded in 4% agarose constructs was used to investigate the effect of physiological mechanical compressive strain applied after 1 and 3 days in culture. The intracellular Ca2+ concentration was measured by use of the ratiometric Ca2+ indicator indo 1-AM and confocal microscopy. A positive Ca2+ response was defined as a percent increase in Ca2+ ratio above a preset threshold. A significantly greater percentage of cells exhibited a positive Ca2+ response in strained constructs compared with unstrained controls at both time points. In strained constructs, treatment with either Ga3+ or EGTA significantly reduced the number of positive Ca2+ responders compared with untreated controls. These results represent an important step in understanding the physiological role of intracellular Ca2+ in chondrocytes under mechanical compression.

mechanotransduction; microscopy; gadolinium; chondrocyte deformation; strain


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