title: Development of an in vitro periodontal biofilm model for assessing antimicrobial and host modulatory effects of bioactive molecules.
abstract: BACKGROUND
  Inflammation within the oral cavity occurs due to dysregulation between microbial biofilms and the host response. Understanding how different oral hygiene products influence inflammatory properties is important for the development of new products. Therefore, creation of a robust host-pathogen biofilm platform capable of evaluating novel oral healthcare compounds is an attractive option. We therefore devised a multi-species biofilm co-culture model to evaluate the naturally derived polyphenol resveratrol (RSV) and gold standard chlorhexidine (CHX) with respect to anti-biofilm and anti-inflammatory properties.
  
  
  METHODS
  An in vitro multi-species biofilm containing S. mitis, F. nucleatum, P. gingivalis and A. actinomycetemcomitans was created to represent a disease-associated biofilm and the oral epithelial cell in OKF6-TERT2. Cytotoxicity studies were performed using RSV and CHX. Multi-species biofilms were either treated with either molecule, or alternatively epithelial cells were treated with these prior to biofilm co-culture. Biofilm composition was evaluated and inflammatory responses quantified at a transcriptional and protein level.
  
  
  RESULTS
  CHX was toxic to epithelial cells and multi-species biofilms at concentrations ranging from 0.01-0.2%. RSV did not effect multi-species biofilm composition, but was toxic to epithelial cells at concentrations greater than 0.01%. In co-culture, CHX-treated biofilms resulted in down regulation of the inflammatory chemokine IL-8 at both mRNA and protein level. RSV-treated epithelial cells in co-culture were down-regulated in the release of IL-8 protein, but not mRNA.
  
  
  CONCLUSIONS
  CHX possesses potent bactericidal properties, which may impact downstream inflammatory mediators. RSV does not appear to have bactericidal properties against multi-species biofilms, however it did appear to supress epithelial cells from releasing inflammatory mediators. This study demonstrates the potential to understand the mechanisms by which different oral hygiene products may influence gingival inflammation, thereby validating the use of a biofilm co-culture model.
authors: Emma
authors: E
authors: Millhouse
authors: Anto
authors: A
authors: Jose
authors: Leighann
authors: L
authors: Sherry
authors: David F
authors: DF
authors: Lappin
authors: Nisha
authors: N
authors: Patel
authors: Andrew M
authors: AM
authors: Middleton
authors: Jonathan
authors: J
authors: Pratten
authors: Shauna
authors: S
authors: Culshaw
authors: Infection and Immunity Research Group, Glasgow Dental School, School of Medicine, College of Medical, Veterinary and Life Sciences, University of Glasgow, 378 Sauchiehall Street, Glasgow G2 3JZ, UK. gordon.ramage@glasgow.ac.uk.
authors: Gordon
authors: G
authors: Ramage
mesh_terms: D016976:Aggregatibacter actinomycetemcomitans / Q000187:drug effects; D000900:Anti-Bacterial Agents / Q000494:pharmacology* / Q000633:toxicity; D000891:Anti-Infective Agents, Local / Q000494:pharmacology / Q000633:toxicity; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000633:toxicity; D018441:Biofilms / Q000187:drug effects; D002460:Cell Line; D002710:Chlorhexidine / Q000494:pharmacology / Q000633:toxicity; D018920:Coculture Techniques; D015536:Down-Regulation; D004847:Epithelial Cells / Q000187:drug effects / Q000382:microbiology; D016967:Fusobacterium nucleatum / Q000187:drug effects; D054884:Host-Pathogen Interactions / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D018836:Inflammation Mediators / Q000276:immunology; D016209:Interleukin-8 / Q000187:drug effects / Q000276:immunology; D015603:Keratinocytes / Q000187:drug effects / Q000382:microbiology; D059013:Microbial Consortia / Q000187:drug effects / Q000502:physiology*; D010510:Periodontal Diseases / Q000382:microbiology*; D016966:Porphyromonas gingivalis / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects; D000077185:Resveratrol; D012464:Saliva, Artificial; D013267:Stilbenes / Q000494:pharmacology / Q000633:toxicity; D034361:Streptococcus mitis / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D000891:Anti-Infective Agents, Local; D000893:Anti-Inflammatory Agents; C508606:CXCL8 protein, human; D018836:Inflammation Mediators; D016209:Interleukin-8; D012333:RNA, Messenger; D012464:Saliva, Artificial; D013267:Stilbenes; D000077185:Resveratrol; D002710:Chlorhexidine