Low-magnitude, high-frequency vibration (LMHFV) with rest days (particularly seven rest days) was considerably effective in improving the morphological and mechanical properties of rat proximal femur. However, current knowledge is limited regarding the possible benefit of this mechanical regimen to other bone sites and whether the optimal rest days are the same. This study followed our previous experiment on LMHFV loading with rest days for three-month-old male Wistar rats. The experiment involved seven groups, namely, vibrational loading for X day followed with X day rest (X=1, 3, 5, 7), daily vibrational loading, tail suspension and baseline control. Micro-computed tomography (micro-CT) scanning was used to evaluate the microarchitecture of the distal femoral trabecular bone. Micro-CT image-based microfinite element analysis was performed for each distal femoral metaphysis. LMHFV with rest days substantially changed the trabecular arrangement from remarkably plate-like to rod-like. Vibrational loading with 1 day rest was substantially effective in improving the architecture and apparent- and tissuelevel mechanical properties of the rat distal femoral metaphysis. This study may provide an improved understanding of the sitespecific responses of bone tissue to LMHFV with rest days for a substantially effective therapy of a targeted bone site.