Purpose: The objective of this study is to evaluate the effects of gradually and non-gradually surface-roughened dental implants with identical macro-design and anodized surface topography on osteogenic differentiation in vitro in three-dimensional cell culture. Materials and Methods: The study compared two groups of implants, gradually (TiUltra) and non-gradually (TiUnite) anodized surfaces. Mouse-derived bone marrow stem cells (MSCs) were cultivated up to passage three. Subsequently, the GelXA BONE bioink, which had been homogeneously combined with the cells, was transferred into poly-lactic acid cages that had been previously rinsed in phosphate-buffered saline. To assess cell viability and proliferation, metabolic activity was measured. The adhesion of cells was evaluated through the use of a DNA-binding probe. The expression of osteogenic differantiation genes was analysed through quantitative polymerase chain reaction (qPCR) analysis on days 7, 14, and 21. The osteogenic gene expression markers were Type-I collagen, fibronectin, alkaline phosphatase (ALP), osteopontin (OSP), osteocalcin (OC), with ß-Actin serving as the housekeeping gene. Results: No statistically significant differences were observed in cell viability and proliferation values. Nevertheless, the qPCR values for osteogenic differentiation markers were notably elevated in the TiUltra group comparison to both theTiUnite and the control groups (p< 0.05). ALP values were significantly lower in the TiUltra group (p< 0.05). Conclusions: It can be concluded that the TiUltra surface demonstrated higher osteogenic gene expression than the TiUnite surface. However, further experimental and clinical studies are required.