Background: Relapse of previously moved teeth, is major clinical problem in orthodontics with respect to the goals of successful treatment. This study investigated the effect of orthodontic relapse on the proliferation of fibroblast and epithelial rests of Malassez cells in periodontal ligament of rat molars. Materials and Methods: Sixteen ten-week- old male Wister rats were randomly divided into four groups composed of four animals each: Group I received no orthodontic force (control). In both Group II and Group III, uniform standardized expansive springs were used for moving the maxillary first molars buccally for periods of one and three weeks respectively. The spring initially generated an average expansive force of 20 g on each side. In Group IV the springs were left for three weeks, until the maxillary first molars moved buccally, after that the springs were removed and the animals were scarified after three weeks of relapse tooth movement. After the humanly scarification of animals, each maxilla in all groups was dissected into two halves each half including the three maxillary molars and processed for histological examination. The number of both fibroblast and ERM cells in each cluster was counted in the PDL of the pressure side of the mesio-buccal roots of the maxillary right and left first molars in all groups and the surface areas of the ERM clusters were also measured in all groups. Results: The number of fibroblast was significantly increased at the end of active movement (Group III) and significantly very highly increased during the relapse period (Group IV). Regarding the ERM cells there were statistically significant increase in both the number of cells in each ERM cluster and the surface areas of the ERM clusters in Group III and highly significant increase in Group IV, while Group II showed no significant differences regarding all measurements. Conclusions: It was concluded that fibroblast and ERM cells may play an important role during orthodontic relapse