The aim of this study was to investigate the difference in the biological response of osteoblasts when stretched and compressed. A cellular cyclic tension and compression apparatus (CCTCA) was designed to stretch and compress cells under the same conditions. After stretching or compressing MC3T3-E1 with continuously increased strain for 5 hours, cellular cytoskeletal modulation was detected by immunohistochemical assay with actin antibody. Real-time polymerase chain reaction was performed at 1, 3, and 5 hours to detect local factors related to bone remodelling. Statistical analysis was undertaken with analysis of variance and the Kruskal-Wallis. Following stretching or compression for 5 hours, MC3T3-E1 attached to the culture dishes grew well. Compared with the control, the microfilaments orientated parallel with each other and were clearly observed by laser scanning confocal microscope after 5 hours of stretching. The morphology of MC3T3-E1 cells was thinner and longer than the control. However, microfilaments presented a disordered arrangement after 5 hours of compression, and the MC3T3-E1 cells decreased in size. Gene expression of Wnt10b and Lrp5 increased during tension but more in the compression groups at 1, 3, and 5 hours. The ratio of osteoprotegerin to receptor activator for nuclear factor kappa B ligand increased in the tension group compared with the control but decreased in the compression group at 5 hours.