Cartilage tissue engineering typically involves the combination of a biodegradable polymeric support material with chondrocytes. The culture environment in which cell-material constructs are created and stored is an important factor. The aim of the present study was to investigate the effects of combined stimuli on cartilage zonal organization which is important to maintain cartilage functions such as lubrication and cushion. For that purpose, we developed a joint mimicking loading system which was composed of compression and shear stress. To mimic the joint loading condition, we manufactured a stimuli system that has a device similar to the shape of a femoral condyle in human knee. The fibrin/hyaluronic acid mixture with chondrocytes were dropped into support made of silicon, and placed under the device. The cartilage explants were stimulated with the joint mimicking loading system for 1 hour per day over the course of 4 weeks. The amounts of GAG and collagen in the stimulated tissue were more than that of the static cultured tissue. Cells and collagen were arranged horizontally paralleled to the surface by stimuli, while it did not happen in the control group. The results of this study suggests that mechanical load exerting in the joint play a crucial role in stimulation of extracellular matrix (ECM) production as well as its functional rearrangement.