Characterization of Terpinen-4-ol on fungi and bacteria: evaluation of synergic effect/additive, cytotoxicity, adhesion in co-culture and change of virulence factors

Abstract

Fungal and bacterial infections mouth has been increasingly reported in the literature. Alternative therapies for microbial control are being developed particularly for resistant microorganisms. Plant extracts and phytochemicals compounds open new perspectives for prevention and efficient control of infectious diseases in the search for favorable therapeutic effects. Terpinen-4-ol is a monoterpene that works in induction of membrane loss, affecting the integrity and the physiology of the cell and organism presents broad spectrum of antimicrobial activity and anti-inflammatory. The objectives of this study are: i) to evaluate the synergistic activity of terpinen-4-ol and nystatin, the cytotoxic potential, antifungal property of terpinen-4-ol in nanoparticles form applied in normal oral immortalized cells (NOK), and cell adhesion of Candida albicans after treatment with terpinen-4-ol; ii) to analyze the antibacterial action and the expression of virulence factors of Terpinen-4-ol against cariogenic bacteria; iii) to analyze the decontamination of titanium surfaces with Terpinen-4-ol on the properties of wettability, surface free energy, chemical structure and roughness. Biofilms are prepared using static model microtitre plate and evaluated for cell viability (XTT assay), counting of colony forming units (CFU / mL) and confocal laser microscopy. The antimicrobial activity of Terpinen-4-ol will be determined by microdilution broth method for planktonic cells, biofilms are evaluated by MTT assay for cytotoxicity and NOK cell lines will be assessed by assay of 0.1% sulforodamide B (SRB). In addition, scanning electron microscopy will be used for observation of biofilms and laser confocal microscopy architecture. The synergism of Terpinen-4-ol will be evaluated by the checkerboard methodology. The decontamination protocols of titanium surfaces will be evaluated by atomic force microscopy and photoelectron spectroscopy X-ray. (AU)