Recent research has determined bioactive compounds as responsible for pharmacological effects of plants. Among all, the Eugenia dysenterica has shown antioxidant, anti-fungal and anti-diarrheal effects, inhibitory activity on the enzyme tyrosinase and potential effect on gastric treatments. This study aimed to evaluate the effects of Eugenia dysenterica in primary culture of gingival mucosa and periodontal ligament cells. First, the primary culture of cells were established and MTT assay was performed both cell lines. Nine concentrations of Eugenia dysenterica extract were used. The concentrations of 41.5 and 83μg/mL showed less cytotoxicity in gingival cells (p <0.05), but were cytotoxic to the periodontal ligament samples. The concentration of 83μg/mL Eugenia dysenterica extract had no antifungal effects on Candida albicans. As a positive control, chlorhexidine digluconate was used at concentrations of 0.12% and 0.2%. Chlorhexidine was severe cytotoxic to gingival cells when compared to the Eugenia dysenterica extract (p <0.05). The healing assay with gingival and periodontal ligament cells showed no induction of wound closure after treatment with Eugenia dysenterica, as well as decreased the motility of the cells after treatment. Alizarin Red S and Von Kossa staining were used to evidence the mineralization of periodontal ligament cells treated with the extract Eugenia dysenterica but there was no or mineral deposits on these cells after the treatment. Based on the results, it was concluded that the Eugenia dysenterica extract were not cytotoxic to gingival cells. However, the concentrations used inhibited proliferation and motility of cells, modified cellular morphology and did not have osteogenic effect for periodontal ligament cells.