Increased bone formation resulting from mechanical loading is well documented;​ however, the interactions of the mechanotransduction pathways are less well understood. Endothelin-1, a ubiquitous autocrine/paracrine signaling molecule promotes osteogenesis in metastatic disease. In the present study, it was hypothesized that exposure to big endothelin-1 (big ET1) and/or mechanical loading would promote osteogenesis in ex vivo trabecular bone cores. In a 2 × 2 factorial trial of daily mechanical loading (− 2000 με, 120 cycles daily, “jump” waveform) and big ET1 (25 ng/mL), 48 bovine sternal trabecular bone cores were maintained in bioreactor chambers for 23 days. The bone cores' response to the treatment stimuli was assessed with percent change in core apparent elastic modulus (ΔE_app), static and dynamic histomorphometry, and prostaglandin E2 (PGE2) secretion. Two-way ANOVA with a post hoc Fisher's LSD test found no significant treatment effects on ΔE_app (p = 0.25 and 0.51 for load and big ET1, respectively). The ΔE_app in the “no load + big ET1” (CE, 13 ± 12.2%, p = 0.56), “load + no big ET1” (LC, 17 ± 3.9%, p = 0.14) and “load + big ET1” (LE, 19 ± 4.2%, p = 0.13) treatment groups were not statistically different than the control group (CC, 3.3% ± 8.6%). Mineralizing surface (MS/BS), mineral apposition (MAR) and bone formation rates (BFR/BS) were significantly greater in LE than CC (p = 0.037, 0.0040 and 0.019, respectively). While the histological bone formation markers in LC trended to be greater than CC (p = 0.055, 0.11 and 0.074, respectively) there was no difference between CE and CC (p = 0.61, 0.50 and 0.72, respectively). Cores in LE and LC had more than 50% greater MS/BS (p = 0.037, p = 0.055 respectively) and MAR (p = 0.0040, p = 0.11 respectively) than CC. The BFR/BS was more than two times greater in LE (p = 0.019) and LC (p = 0.074) than CC. The PGE2 levels were elevated at 8 days post-osteotomy in all groups and the treatment groups remained elevated compared to the CC group on days 15, 19 and 23. The data suggest that combined exposure to big ET1 and mechanical loading results in increased osteogenesis as measured in biomechanical, histomorphometric and biochemical responses.