In this in vitro study ovine osteoblast-like cells were cultured on seven different alloplastic biomaterials used for augmentation and for reconstruction of bone defects in dental and craniomaxillofacial surgery. The aim of this study was to examine the growth behaviour (viability, cell density and morphology) of ovine osteoblast-like cells on the investigated biomaterials to get knowledge which biomaterial is qualified to act as a cell carrier system in further in vivo experiments. The biomaterials were either synthetically manufactured or of natural origin. As synthetically manufactured biomaterials Ethisorb®, MakroSorb®, Palacos®R, and PDS® film were used. As biomaterials of natural origin Beriplast®P, Bio-Oss® and Titanmesh were investigated. The cell proliferation and cell colonization were analyzed by a proliferation assay and scanning electron microscopy. Osteoblast-like cells proliferated and attached on all biomaterials, except on Beriplast®. On Ethisorb® the highest cell proliferation rate was measured followed by Palacos®R. Both biomaterials offer suitable growth and proliferation conditions for ovine osteoblast-like cells. The proliferation rates of Bio-Oss®, MakroSorb®, PDS®-film and Titanmesh were low and SEM examinations of these materials showed less spread osteoblast-like cells. The results showed that ovine osteoblast-like cells appear to be sensitive to substrate composition and topography. This in vitro study provides the basis for further in vivo studies using the sheep model to examine the biocompatibility and the long-term interaction between the test material and tissue (bone regeneration).