The presence of an appropriate number of reparative cells in an articular cartilage defect is probably necessary for consistent and successful repair. Following the transplantation of chondrocytes into a defect, cell proliferation may modulate local defect cellularity. Transplanted cells can be compressed during cartilage repair as a result of joint-loading or press-fitting a graft into a cartilage defect. The objective of this study was to characterize the proliferative response of chondrocytes after attachment to cartilage and application of static compressive stress between cartilaginous surfaces in an ex vivo model. The chondrocytes were isolated from adult bovine cartilage, cultured in high-density monolayer, resuspended, and then transplanted onto the surface of devitalized cartilage at a density of 250,000 cells/cm2. The total DNA content of transplanted cell layers increased steadily to a plateau by 5 days and represented a 4-fold increase in cell number during incubation in medium including serum and ascorbate. Over the culture period, the level of DNA synthesis ([3H]thymidine incorporation), on a per cell basis, decreased steadily (88% between days 0 and 6). The application of 24 hours of static compressive stress (0.06-0.4 MPa) to the adherent cells at 1 and 4 days after transplantation inhibited overall DNA synthesis by 70-approximately 87% compared with unloaded controls. After release from load, cell proliferation generally remained at low levels. The marked proliferation of chondrocytes when attached to cartilage without applied load and the inhibition of this proliferation by relatively low-amplitude static compressive stress may be relevant to the occasional overgrowth of tissue in some chondrocyte transplantation procedures. The dosimetry of these effects suggests that the in vivo mechanical environment may have a marked effect on proliferation of transplanted chondrocytes.