By definition, osseointegration means close contact between bone and implant. Bone response is related to implant surface properties. Various surfaces have been studied and applied to improve the biological properties of the implant and thereby favor the mechanism of osseointegration. This strategy aims to promote osseointegration by means of a faster and stronger bone formation, improving stability during the healing process, and thus allowing for earlier loading of the implant. Dental implant osseointegration has so far been studied in various animal models. The development of a method based on tissue engineering for assessing the osseointegration process in vitro could prove a valid biomimetic alternative to sacrificing animals. In this study, flat cylindrical dental implants with moderately rough surfaces and machined implants were set in bovine bone blocks. Then, adipose-derived stem cells (ADSCs) were three dimensionally cultured onto these blocks in osteo-endothelial medium for up to 30 days to mimic the osseointegration process in vitro. Scanning electron microscopy (SEM) and gene expression were used to examine stem cell commitment. Mechanical pull-out tests were also performed. SEM analysis identified cells with an osteoblast morphology adhering to the surface of the implants after their removal. Gene expression analysis showed that ADSCs seeded onto the bone blocks were able to express osteoblast and endothelial markers. The implants with the moderately rough surface generated higher pull-out strengths when compared with the machined implants. Nevertheless, the pull-out test values were higher for implants placed in bone blocks with ADSCs than for those set in scaffolds without stem cells. Our results demonstrate the validity of the method adopted and its potential for use in the in vitro assessment of the biological behavior of dental implant surfaces.