title: Biologic assessment of antiseptic mouthwashes using a three-dimensional human oral mucosal model.
abstract: BACKGROUND
  The biologic safety profile of oral health care products is often assumed on the basis of simplistic test models such as monolayer cell culture systems. We developed and characterized a tissue-engineered human oral mucosal model, which was proven to represent a potentially more informative and more clinically relevant alternative for the biologic assessment of mouthwashes. The aim of this study was to evaluate the biologic effects of alcohol-containing mouthwashes on an engineered human oral mucosal model.
  
  
  METHODS
  Three-dimensional (3D) models were engineered by the air/liquid interface culture technique using human oral fibroblasts and keratinocytes. The models were exposed to phosphate buffered saline (negative control), triethylene glycol dimethacrylate (positive control), cola, and three types of alcohol-containing mouthwashes. The biologic response was recorded using basic histology; a cell proliferation assay; 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tissue-viability assay; transmission electron microscopy (TEM) analysis; and the measurement of release of interleukin (IL)-1beta by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  Statistical analysis showed that there was no significant difference in tissue viability among the mouthwashes, cola, and negative control groups. However, exposure to the positive control significantly reduced the tissue viability and caused severe cytotoxic epithelial damage as confirmed by histology and TEM analysis. A significant increase of IL-1beta release was observed with the positive control and, to a lesser extent, with two of the tested mouthrinses.
  
  
  CONCLUSIONS
  The 3D human oral mucosal model can be a suitable model for the biologic testing of mouthwashes. The alcohol-containing mouthwashes tested in this study do not cause significant cytotoxic damage and may slightly stimulate IL-1beta release.
authors.affiliation: Department of Adult Dental Care, School of Clinical Dentistry, University of Sheffield, Sheffield, UK. k.moharamzadeh@sheffield.ac.uk
authors.forename: Keyvan
authors.initials: K
authors.lastname: Moharamzadeh
authors.forename: Kirsty L
authors.initials: KL
authors.lastname: Franklin
authors.forename: Ian M
authors.initials: IM
authors.lastname: Brook
authors.forename: Richard
authors.initials: R
authors.lastname: van Noort
mesh_terms: D049109:Cell Proliferation; D004396:Coloring Agents; D004338:Drug Combinations; D000431:Ethanol / Q000633:toxicity; D006801:Humans; D053583:Interleukin-1beta / Q000096:biosynthesis; D046529:Microscopy, Electron, Transmission; D008954:Models, Biological; D009061:Mouth Mucosa / Q000187:drug effects* / Q000378:metabolism; D009067:Mouthwashes / Q000633:toxicity*; D012459:Salicylates / Q000633:toxicity; D013729:Terpenes / Q000633:toxicity; D013778:Tetrazolium Salts; D013844:Thiazoles; D023822:Tissue Engineering / Q000379:methods*; D014022:Tissue Survival / Q000187:drug effects*
chemical_list: D004396:Coloring Agents; D004338:Drug Combinations; D053583:Interleukin-1beta; D009067:Mouthwashes; D012459:Salicylates; D013729:Terpenes; D013778:Tetrazolium Salts; D013844:Thiazoles; D000431:Ethanol; C057473:Listerine; C022616:thiazolyl blue