Research data and clinical observations have proven that mechanical loading is essential for the development and maintenance of normal cartilage tissue architecture. During the past decade numerous pivotal factors controlling cartilage development and growth have been identified, such as Sox9, fibroblast growth factor receptor 3 (FGFR3) and vascular endothelial growth factor (VEGF). The aim of this study was to examine the biochemical response of mandibular condylar cartilage chondrocytes in different mechanical loading conditions and whether Sox9, FGFR3 and VEGF are involved in the signaling pathway. One hundred 14-day-old female Wistar rats were divided to two groups: one group was provided with whole pellet food (hard food) and the other group was provided with soft powdery food. On day 21 (weaning day and initiation day of the experiment), ten animals from each group were sacrificed after 2, 6, 12, 24 and 48 hours. Their condyles were dissected and the cartilage tissue proceeded for immunostaining using commercially available antibodies against Sox9, FGFR3 and VEGF. In the proliferating zone statistical differences were observed in stain intensity for both Sox9 and FGFR3 in relation to time while in the early hypertrophic zone statistical differences in stain intensity were evident in relation to food type and time for Sox9, FGFR3 and VEGF. The results of the present study clearly demonstrate that different mechanical loading conditions in condylar chondrocytes trigger differentiation/ maturation related processes by affecting the expression levels of crucial factors, ultimately regulating condylar cartilage growth.