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Objectives To investigate the efficacy of both platelet-rich fibrin (PRF) membrane isolated and injectable PRF (i-PRF) mixed with deproteinized bovine bone mineral (DBBM) in the treatment of rat tibiae non-critical defects. Materials and methods Non-critical bone defects in the tibiae (3.5mm x 1.5mm) of sixty-four rats were made and filled with different materials according to each experimental group: CO: bone defects filled with blood clot; PRF: PRF membranes obtained by fibrin protocol; BO: DBBM isolated and SB: i-PRF mixed with DBBM. Micro–computed tomography (µCT), histological/ histomorphometric analysis, and molecular markers expression were conducted after 15 and 45 days. Results Cortical area analyses by µCT showed superior results for bone formation and cortical thickness in PRF group in comparison to the control group. The PRF exhibited comparable results with BO and SB groups in early periods. However, the DBBM isolated seems to promote more new bone formation in later periods. Histomorphometry analysis demonstrated similar percentage of newly formed bone among the groups at 15 and 45 days. The i-PRF mixed to DBBM bone not enhance the bone formation in comparison to DBBM alone in both µCT and histomorphometry analyses. Bone molecular markers expression demonstrated higher expression of Alpl, Bglap, and Runx2 genes for BO compared to the other groups. Conclusions In conclusion, the PRF, DBBM, and SB groups promoted higher bone formation in rat tibiae non-critical defects in comparison to the control group. However, the addition of i-PRF to DBBM did not improve the DBBM potential in bone healing. Clinical relevance: This pre-clinical study produced insights about the use of modified protocols for obtaining PRF by a low-speed concept which has been associated with more concentration of growth factors and better results than standard PRF protocol.