Background/Aim. Titanium-prepared platelet-rich fibrin (T-PRF) has superior properties with respect to resorption time, thickness, growth factor holding capacity, and gradual release at the surgical site. The aim of this study was to evaluate the cell viability and osteogenic differentiation of an osteoblastic cell line MG-63 following exposure to eluates obtained from T-PRF clots injected with antibiotic gels [amoxicillin/clavulanic acid – amoxiclav gel (AMX), metronidazole – metronidazole gel (MET)], or herbal extracts (neem gel). Methods. In this in vitro study, blood was collected, and T-PRF clots were prepared. Antibiotic/herbal gels along with T-PRF clots were added to Dulbecco’s Modified Eagle Medium – DMEM to obtain eluate. This eluate was later added to the cultured osteoblastic cells to check for cell viability using the [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide] (MTT) assay on days 1, 3, and 7. Gene expression [alkaline phosphatase (ALP), osteopontin (OSP), bone morphogenetic protein-2 (BMP-2)] was checked on day 7 using quantitative polymerase chain reaction (qPCR), and alizarin red S (ARS) staining was used for late-stage osteogenic differentiation and matrix mineralization on day 21. Statistical analysis was performed using GraphPad Prism software. One-way analysis of variance (ANOVA) was used for the MTT assay and for ARS staining, with Tukey’s post-hoc test applied only for significant ANOVA results (on day 7 MTT and day 21 ARS data). The Kruskal-Wallis test was used for qPCR analysis. Results. Cell viability was significantly higher in the T-PRF + neem group on day 3 (p = 0.0229) and day 7 (p = 0.0034), while the T-PRF + AMX group showed significantly higher cell viability on day 7 (p = 0.0142) compared with T-PRF alone. The qPCR analysis showed significantly increased expression of OSP (p = 0.0422), ALP (p = 0.0203), and BMP-2 (p = 0.0290) in all tested groups (T-PRF + AMX/MET/neem) compared with the control group (T-PRF alone). Intergroup comparisons showed no significant differences, except between the T-PRF + neem group and the control group. ARS analysis at 21 days revealed the highest mineralization in T-PRF + neem, followed by T-PRF + MET and T-PRF + AMX. Conclusion. Both antibiotic- and herbal-extract-incorporated T-PRF clots demonstrated superior osteogenic potential compared to plain T-PRF alone. Therefore, T-PRF can serve as a sustained drug delivery system.