Because osteoblasts play a key role in bone remodeling and the influence of low-level laser therapy on this process is not clear, Saos-2 human osteoblast-like cells were irradiated by a gallium–aluminum–arsenide diode laser (915 nm) for 10, 48, 96, 193, and 482 s using doses 1, 5, 10, 20, and , respectively. A control group was not irradiated. Morphology, viability, and cytotoxicity analyses were carried out after 1 hr, 1 day, and 3 days. Deoxyribose nucleic acid (DNA) content and release of vascular endothelial growth factor (VEGF), receptor activator of nuclear factor kappa B ligand (RANKL), and osteoprotegerin (OPG) were evaluated. Viability was modulated by laser irradiation in a dose-dependent manner, with inducing a biostimulatory response and 20 to determining a bioinhibitory and cytotoxic effect. Accordingly, DNA content was generally increased for the dose and decreased for the dose. A rapid and transitory trend toward increased RANKL/OPG ratio and a tendency toward a delayed increase in VEGF release for doses of 1 to was found. Further investigations using the biostimulatory dose of emerged from this study are needed to establish the ideal treatment regimens in the laboratory as well as in clinical practice.