Terpinen-4-ol: study of synergistic / additive effect, adhesion in co-culture and the modification of virulence factors of Candida spp

Author(s)

Caroline Coradi Tonon

• Document type: Master's Dissertation
• Press: Araraquara. 2016-05-24.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Odontologia. Araraquara
• Defense date: 2016-03-22
• Advisor: Denise Madalena Palomari Spolidorio

Abstract

The genus Candida can be found in up to 50% of healthy individuals without causing apparent damage, however, under predisposing conditions such as systemic diseases or physiological conditions, can become pathogenic causing inflammation and tissue destruction. Candida spp. in the form of biofilms are important in the development of infections because they are associated with high levels of resistance to antimicrobial agents. Alternative therapies with natural extracts open new perspectives for prevention and control of oral diseases in search of favorable therapeutic effects. The Terpinen-4-ol is a monoterpene engaged in membrane loss of induction and presents broad spectrum of antimicrobial and anti-inflammatory activity. The aim of this study was to evaluate the antifungal effect, synergistic / additive inhibition of adhesion on oral cells and modification of virulence factors of Terpinen-4-ol of Candida albicans and Candida tropicalis. Thus, the identification of Minimum Inhibitory Concentration was performed (MIC) and Minimum Fungicidal Concentration (MFC) of Terpinen-4-ol on standard strains C. albicans (ATCC 90028) and C. tropicalis (ATCC4563), using the method microdilution broth. Biofilms mono and dual-species were prepared using the microtiter plate static model and quantitated by colony forming units (CFU / mL). The effect of Terpinen-4-ol in the adhesion on C. tropicalis and C. albicans was carried out in co-culture with oral cells NOK Si as well as virulence factors (proteases, phospholipases and hemolysin) were evaluated in vitro. The MIC for Terpinen-4-ol was 2.31 mg / ml for both species and CFM 0.002 mg / ml and 0.004 mg / ml respectively for Candida albicans and Candida tropicalis. When tested in combination there was synergism and additive effect (T-4-ol 1.06 mg / ml and 0.00012 mg Nis / ml) for Candida albicans. In the biofilm, the results presented aditismo for both species. The Terpinen-4-ol was a reduction in adhesion of Candida tropicalis on cells NOK Si, Nystatin already had most effective for Candida albicans and for Candida tropicalis. For enzymatic activity, Terpinen-4-ol and Nystatin showed no action on the production of proteinase, phospholipase and hemolysin when compared to the control group (no treatment). The combination of Terpinen-4-ol and Nystatin presented antifungal effect potentiated on the two Candida species evaluated, showing synergistic / modulatory effect on planktonic and biofilm cultures. Terpinen-4-ol reduces the adhesion of C. tropicalis and Nystatin reduces the adhesion of both species in oral immortalized cells (NOK Si). The sub-inhibitory concentrations (MIC50) of Terpinen-4-ol and Nystatin used did not show activity in the production of proteinase, phospholipase and hemolysin. (AU)

• FAPESP's process: 15/00155-0 - Terpinen-4-ol: study of synergistic/additive effect, adhesion in co-culture and change of virulence factors of Candida spp
• Grantee: Caroline Coradi Tonon
• Support Opportunities: Scholarships in Brazil - Master