Chondrocyte mechanotransduction in response to mechanical loading is essential for the health and homeostasis of articular cartilage. The actin cytoskeleton has been implicated in cell mechanics and mechanotransduction. This study tests the hypothesis that loading modulates actin dynamics and organisation with subsequent changes in gene expression for actin associated proteins. Chondrocytes were transfected with eGFP-actin, seeded in agarose and subjected to cyclic compression (10 cycles, 1 Hz, 0-15% strain) on the stage of a confocal microscope. Compression resulted in a subsequent reduction in cortical eGFP-actin intensity and a reduction in fluorescence recovery after photobleaching (FRAP), suggesting net cortical actin de-polymerisation, compared to unloaded controls. Cyclic compression for 10 min up-regulated gene expression for the actin depolymerising proteins, cofilin and destrin. Thus mechanical loading alters cortical actin dynamics, providing a potential mechanism through which chondrocytes can adapt their mechanical properties and mechanosensitivity to the local mechanical environment.