Although concentrated growth factor (CGF) is known to promote gingival regeneration and improve the outcomes of clinical treatment, the mechanisms underlying its effects remain unknown. Therefore, this study aimed to elucidate the effects of CGF on gingival thickening. To this end, gingival mesenchymal stem cells (GMSCs) were treated with different concentrations of CGF, and the effects of CGF on cell proliferation and migration; collagen-1 (Col-1), fibronectin (FN), vascular endothelial growth factor (VEGF), and angiopoietin-1 (Ang-1) expression; and the AKT, Wnt/b-catenin, and Yes-associated protein (YAP) signalling pathways were investigated. The effects of CGF in vivo were also investigated in a rat buccal gingival injection model. GMSCs cultured with CGF showed improved cell proliferation and migration. Moreover, CGF treatment improved the levels of FN, Col-1, VEGF, and ANG-1. These effects of CGF were mediated by the AKT/Wnt and YAP pathways, with the AKT pathway possibly functioning upstream of the Wnt/b-catenin and YAP pathways. YAP was also shown to be overexpressed in the in vivo model. Thus, CGF can promote gingival regeneration, and YAP transport into the nucleus may be a key factor underlying this activity, which provides a novel perspective for gingival regeneration and further promotion of the clinical application of CGF.