The aim of this study was to evaluate whether the application of tension or compression forces exerted on the periodontium during the early phase of orthodontic tooth movement is reflected by differences in the composition of the gingival crevicular fluid (GCF), at the level of interleukin-1beta (IL-1beta), substance P (SP), and prostaglandin E(2) (PGE(2)). Eighteen children (mean age 10.8 yr) starting orthodontic treatment were included in the study. Molar elastic separators were inserted mesially to two first upper or lower molars. One of the antagonist molars served as the control. GCF was collected from the mesial and distal sites of each molar, before (-7 d, 0 d) and immediately after (1 min, 1 h, 1 d, and 7 d) the placement of separators. The levels of IL-1beta, SP, and PGE(2) were determined by enzme-linked immunosorbent assay. At the orthodontically moved teeth, the GCF levels of IL-1beta, SP, and PGE(2) were significantly higher than at the control teeth in both tension and compression sides, and at almost all occasions after insertion of separators. The increase, relative to baseline values, was generally higher in tension sides. For the control teeth, the three mediators remained at baseline levels throughout the experiment. The results suggest that IL-1beta, SP, and PGE(2) levels in the GCF reflect the biologic activity in the periodontium during orthodontic tooth movement.