The aim of the present study was to determine whether occlusal hypofunction and its recovery affected the structure of the Periodontal Ligament (PDL) and expression of Vascular Endothelial Growth Factor (VEGF) in mice. Five-week-old C57BL/6J mice (Jackson Laboratory, Bar Harbor ME, USA) were used and randomly divided into three groups: a Hypofunctional Group (HG), Recovery Group (RG), and Control Group (CG). In HG and RG, appliances were attached to the maxillary and mandibular incisors. These appliances were set for 11 weeks in HG and 7 weeks in RG. Appliances were then removed offer 0, 7, 14, and 28 days. Untreated mice served as CG. Histological sections were prepared and immunohistochemically stained for VEGF. The PDL area and the number of VEGF immunopositive cells in the PDL were evaluated in the 3 groups. The number of VEGF immunopositive cells and PDL area were significantly larger in CG white the RG than in HG, while the PDL area in RG was similar to that in CG. In the recovery process, the PDL area and number of VEGF positive cells in the PDL increased from days 0 to 7 and decreased from days 7 to 28. The results of the present study suggest that occlusal stimuli regulate the PDL area through the expression of VEGF in the mouse PDL. The clinical relevance of occlusal stimuli is their ability to regulate the expression of VEGF in PDL cells and these growth factors may lead to alveolar bone remodeling in the PDL.