Evaluation of the Effects of Ethanol on Monolayer and 3D Models of Human Oral Mucosa

Abstract

The aim of this study was to examine and compare the effects of different ethanol-containing solutions on a 3D oral mucosal model (OMM) with those of fresh oral mucosal biopsies and monolayer keratinocyte cultures. 3D OMMs were engineered by the air/liquid interface culture technique using human oral fibroblasts and keratinocytes. Fresh tissue biopsies were also obtained from patients and divided into small pieces. OMMs, fresh biopsies, and monolayer cultures were exposed to serial dilutions of ethanol in PBS (5% to 50%) for 30 secondss and 60 secondss. Biological response was evaluated using basic histology and the Alamar Blue tissue viability assay. Histological examination showed that the effects of ethanol on the OMMs resembled the fresh tissue biopsies. The 3D model was also able to detect the concentration-dependent and time-dependent effects of ethanol. The TC50 values for the monolayer cultures were significantly lower than those for the 3D models. It can be concluded that ethanol in high concentrations significantly affects the oral mucosal model’s viability and histological appearance. The response of the OMM to ethanol exposure was similar to the response of the fresh clinical biopsies. Monolayer keratinocytes are more sensitive to ethanol than 3D models or biopsies.