Comparative Cytotoxicity of Menthol and Eucalyptol: An In Vitro Study on Human Gingival Fibroblasts
Clara Puig-Herreros,
José Luis Sanz,
David García-Bernal,
Francisco Javier Rodríguez-Lozano,
Laura Murcia,
Leopoldo Forner,
James Ghilotti,
Ricardo E. Oñate-Sánchez,
Sergio López-García
Affiliations
Clara Puig-Herreros
Speech Therapy University Clinic, Department of Basic Psychology, Universitat de València, 46010 Valencia, Spain
José Luis Sanz
Departament d’Estomatologia, Facultat de Medicina i Odontologia, Universitat de València, 46010 Valencia, Spain
David García-Bernal
Department of Biochemistry, Molecular Biology B and Immunology, Faculty of Medicine, University of Murcia, 30120 Murcia, Spain
Francisco Javier Rodríguez-Lozano
Biomedical Research Institute (IMIB), 30120 Murcia, Spain
Laura Murcia
Department of Health Sciences, Catholic University San Antonio of Murcia, 30107 Murcia, Spain
Leopoldo Forner
Departament d’Estomatologia, Facultat de Medicina i Odontologia, Universitat de València, 46010 Valencia, Spain
James Ghilotti
Departament d’Estomatologia, Facultat de Medicina i Odontologia, Universitat de València, 46010 Valencia, Spain
Ricardo E. Oñate-Sánchez
Department of Dermatology, Stomatology, Radiology and Physical Medicine, Morales Meseguer Hospital, Faculty of Medicine, University of Murcia, 30008 Murcia, Spain
Sergio López-García
Departament d’Estomatologia, Facultat de Medicina i Odontologia, Universitat de València, 46010 Valencia, Spain
The aim of this study was to assess the influence of eucalyptol and menthol on the cell viability, migration, and reactive oxygen species production of human gingival fibroblasts (GFs) in vitro. Three different concentrations of eucalyptol and menthol were prepared following ISO 10993-5 guidelines (1, 5, and 10 mM). GFs were isolated from extracted teeth from healthy donors. The following parameters were assessed: cell viability via MTT, Annexin-V-FITC and 7-AAD staining, and IC50 assays; cell migration via horizontal scratch wound assay; and cell oxidative stress via reactive oxygen species assay. Data were analyzed using one-way ANOVA and Tukey’s post hoc test. Statistical significance was established at p < 0.05. Eucalyptol and Menthol exhibited high cytotoxicity on gingival fibroblasts, as evidenced by cytotoxicity assays. Eucalyptol showed lower levels of cytotoxicity than menthol, compared to the control group. The cytotoxicity of the tested substances increased in a concentration-dependent manner. The same occurred in a time-dependent manner, although even 10 min of exposure to the tested substances showed a high cytotoxicity to the GFs. Commercially available products for oral application with these substances in their composition should be tested for cytotoxicity before their use.
