Antifungal Activity, Biocompatibility, and Anti-Inflammatory Effects of Zerumbone-Enhanced Antimicrobial Photodynamic Inactivation in a Three-Dimensional Oral Tissue Model

High Resolution Image Download MS PowerPoint Slide This study investigated the antifungal activity, biocompatibility, and anti-inflammatory effects of zerumbone (ZER; monocyclic sesquiterpene from the essential oil of ginger with antibiofilm activity) combined with antimicrobial photodynamic inactivation (aPDI) using a three-dimensional (3D) coculture model infected with Candida albicans biofilms fluconazole-sensitive (CaS; ATCC 90028) and fluconazole-resistant (CaR; ATCC 96901). For this purpose, a 3D coculture model involving two cell lines (fibroblasts and keratinocytes) was developed and infected with CaS and CaR biofilms. After biofilm establishment, treatments were performed using ZER (1172 or 2344 μM) and aPDI [ (PDZ;154 μM) and LED light (660 nm; 50 J/cm 2; 44.5 mW/cm 2 )], either individually or in combination. Control groups included a conventional antifungal drug, a negative control (PBS), and a cell death-induced control (Triton X-100). Regarding uninfected 3D oral tissues, ZER (1172 μM) + aPDI exhibited minimal cytotoxicity by alamarBlue, with viability reduction under 18%. In the infected 3D coculture model, ZER (1172 μM) + aPDI resulted in reductions in viable colonies counting of 2.36 log 10 for CaS and 2.15 log 10 for CaR. Cell damage measured by lactate dehydrogenase release was 7.41% for CaS and 9.43% for CaR. The confocal laser scanning microscopy imaging supported these findings, suggesting that ZER + aPDI may impair the penetration of the fungal infection. Furthermore, ZER combined with aPDI reduced proinflammatory cytokines (IL-6 and IL-8) induced by C. albicans infection. Overall, evaluation of the 3D coculture model showed that the combination of ZER and aPDI was considered noncytotoxic and effective against fluconazole-resistant C. albicans biofilms. Therefore, this combination of approaches presents a promising strategy for combating resistant fungal infections, offering a potential alternative to conventional antifungal agents.