Temporal effects of cell adhesion on mechanical characteristics of the single chondrocyte

Cell adhesion to material surfaces is a fundamental phenomenon in tissue response to implanted devices, and an important consideration in tissue engineering. For example, elucidation of phenomena associated with adhesion of chondrocytes to biomaterials is critical in addressing the difficult problem of articular cartilage regeneration. The first objective of this study was to measure the mechanical adhesiveness characteristics of individual rabbit articular chondrocytes as a function of seeding time to provide further understanding of the cell adhesion process. The second objective was to quantify the force required to separate the plasma membrane from the underlying cytoskeleton as a function of seeding time. After culturing chondrocytes on glass coverslips for 1, 2, 4, 6 h, two biomechanical tests were performed on single chondrocytes: (i) mechanical adhesiveness measurement by the cytodetacher; and (ii) plasma membrane tether formation force measurement by optical tweezers. Cell mechanical adhesiveness increased from 231+/-149 Pa at 1 h to 1085+/-211 Pa at 6 h. The cell contact area with the substrata increased from 161+/-52 microm(2) at 1 h to 369+/-105 microm(2) at 6 h. The tether formation force increased from 232+/-23 pN at 1 h to 591+/-17 pN at 6 h. Moreover, fluorescence staining by rhodamine-phalloidin demonstrated the process of actin spreading within the cytoskeleton from 0.5 to 6 h and allowed for measurement of cell height which was found to decrease from 12.3+/-2.9 microm at 0.5 h to 6.2+/-0.9 microm at 6 h.