Objectives: As a primary objective, we examined the gingival crevicular fluid (GCF) levels of the endogenously released platelet-derived growth factor (PDGF-BB) and vascular endothelial growth factor (VEGF) following the use of minimally invasive surgical flap reflection (MIS) and compared them with those in traditional open flap debridement (OFD). The secondary objective was to determine if any correlation exists between the expression of growth factors (GF), indicated by their levels, and associated clinical outcomes. Methods: Twenty-eight non-smoking individuals with severe chronic periodontitis were recruited in this prospective, randomized and single-blinded trial. Each person presented one interproximal defect that was randomly assigned to either the experimental MIS group (14 sites) or the open flap debridement (OFD) control group (14 sites). Plaque index, gingival index, probing depth (PD), clinical attachment level (CAL) and the intrabony depth of the defect (IBD) were measured at baseline for the patient’s enrollment. Gingival crevicular fluid (GCF) samples were collected on days 1, 3, 7, 14, 21 and 30 after therapy. Clinical follow-ups were scheduled at 6 and 9 months following the therapy. Results: In both groups, the highest levels of VEGF and PDGF-BB concentrations were found in the GCF during the period from 1 to 14 days. During the early stages of heal- ing (1, 3, 7 and 14 days), the mean VEGF and PDGF-BB levels at sites treated with MIS were significantly higher than those at the OFD-treated sites. Growth factor levels decreased sharply in the samples obtained at days 21 and 30 in both groups, with non-significant differences between the patient and control groups. MIS-treated sites reported signifi cantly more PD reduction and attachment gain 9 months following therapy. No significant difference in intrabony defect reduction was reported between the patient and control groups. Conclusions: Within the limits of the present study, we can surmise that MIS treatment of periodontal defects is associated with initially higher GCF levels of the studied growth factors. These increased GF levels are well correlated with the improved soft-tissue parameters of the periodontal defects.