The ultimate goal of periodontal therapy is to achieve successful periodontal regeneration. The effects of different biomaterials, allogenic and alloplastic, used in periodontal surgeries to achieve regeneration have been studied in vitro on periodontal ligament (PDL) cells and MC3T3-E1 cells. The materials tested included cryopreserved bone allograft (CBA), coralline hydroxyapatite (CH), demineralized freeze-dried dentin (DFDD), and cementum. CBA and CH revealed an increase in initial PDL cell attachment, whereas CH resulted in an increase in long-term PDL cell attachment. Mineral-like nodule formation was observed significantly higher in DFDD compared to other materials tested for osteoblasts. Based on the results of this in vitro study, we conclude that the materials used are all biocompatible with human PDL cells and osteoblasts, which have pivotal importance in periodontal regeneration.