Background There is a continuing search for compuounds to improve the chemical plaque inhibitory action of oral hygiene products. Although the antibacterial effects of chlorhexidine (CHX) and essential oils components, citral/myrcene, have been described, there is contradictory information regarding their cytotoxic effects in host tissues. This study aimed to evaluate the cytotoxic activity of the major components of the oil C. citratus , citral and myrcene on human periodontal ligament fibroblast (HPLF) cultures and their antimicrobial effect on different bacterial species present in supragingival biofilm. Material and Methods Cytotoxicity of the compounds to HPLF was determined by MTT assay. Antimicrobial activity was tested against reference strains of Enterococcus faecalis , Streptococcus mutans and Lactobacillus rhamnosus and for S. mutans clinical strains by broth microdilution assay. One-way analysis of variance (ANOVA) with Games-Howell post-hoc multiple comparison or unpaired t tests were used for inter- and intragroup comparisons. Results Overall, all of the compounds under study showed a cytotoxic effect to HPLF which varied in a dose-dependant manner. Whilst myrcene did not show bacteriostatic activity at tested concentrations, both citral and CHX exhibited bacteriostatic/bactericidal effects to all strains at specific concentrations, being CHX most effective to inhibit bacterial growth at lower concentrations than what observed for citral. Conclusions Based on these findings, it would possible to conclude that whereas myrcene might be ineffective to control bacterial growth, citral could have a promising antimicrobial activity against dental colonizers with low cytotoxicity, and may be useful for preventing the onset and progression of oral diseases. Key words: Antimicrobial activity, citral, cytotoxicity, chlorhexidine, myrcene.