Aim The current randomized controlled trial assessed for the first time the effect of a low-speed platelet-rich fibrin (PRF) with open flap debridement (OFD) versus OFD alone in the treatment of periodontal intra-osseous defects of stage-III periodontitis patients. Methods Twenty-two periodontitis patients with ≥ 6 mm probing depth (PD) and ≥ 3 mm intra-osseous defects were randomized into test (PRF + OFD; n = 11) and control (OFD; n = 11) groups. Clinical attachment level (CAL)–gain (primary outcome), PD-reduction, gingival recession depth (GRD), full-mouth bleeding scores (FMBS), full-mouth plaque scores (FMPS), radiographic linear defect depth (RLDD), and radiographic bone fill (secondary-outcomes) were examined over 9 months post-surgically. Results Low-speed PRF + OFD and OFD demonstrated significant intra-group CAL-gain and PD- and RLDD-reduction at 3, 6, and 9 months (p < 0.01). Low-speed PRF + OFD exhibited a significant CAL-gain of 3.36 ± 1.12 mm at 6 months (2.36 ± 0.81 mm for the control group; p < 0.05), and a significantly greater PD-reduction of 3.36 ± 1.12 mm at 3 months, of 3.64 ± 1.12 mm at 6 months and of 3.73 ± 1.19 mm at 9 months (2.00 ± 0.89 mm, 2.09 ± 1.04 mm, and 2.18 ± 1.17 mm in the control group respectively; p < 0.05). No significant differences were notable regarding GRD, FMPS, FMBS, RLDD, or bone fill between both groups (p > 0.05). Conclusions Within the current clinical trial’s limitations, the use of low-speed PRF in conjunction with OFD improved CAL and PD post-surgically, and could provide a cost-effective modality to augment surgical periodontal therapy of intra-osseous defects of stage-III periodontitis patients. Clinical relevance Low-speed PRF could provide a cost-effective modality to improve clinical attachment gain and periodontal probing depth reduction with open flap debridement approaches.