Objective: The objective of the study was to compare the effect of mineral trioxide aggregate and other endodontic materials in contact with osteoblast-like cell over a time period of 20 days. Materials and methods: Human PDL cells and osteoblasts were gathered, cultured and permitted to standardized protocols. The cell populations were characterized with the conforming surface markers following standardized processes. The specimens were produced with special concern to constant dimensions and volume in the different groups. Cell attachment and proliferation were assessed morphologically after Richardson staining and cell count was performed after 1d, 7d, 14d and 21d. All trials were done in triplets. The outcomes were statistically analyzed using the ANOVA- and Tukey-test (p < 0.05). Results: Morphological examination showed good proliferation and cell attachment in both cements. A remarkable outcome was the prepared increasing and parallel alignment of the PDL cells in connection with MTA and particularly Biodentine. From 7d forward Biodentine indicated the highest quantity of PDL cells (p < 0.05). Biodentine and MTA caused in a significantly greater cell density in osteoblast and PDL cell culture. The other groups presented a lower PDL cell density from 7d and a lower osteoblast cell density from 14d when matched to control and cement samples (p < 0.05). Conclusions: MTA and Biodentine showed a good biocompatibility in contact with the human osteoblasts and cells of the periodontal ligament. Regarding cell survival and proliferation particularly of PDL cells Biodentine showed good results and can be considered as a well-tolerated bioactive endodontic material.