title: Lipopolysaccharide responsiveness in vocal fold fibroblasts.
abstract: BACKGROUND
  Vocal fold fibroblast's (VFF) strategic location in the lamina propria and their ability to respond to external stimuli by producing inflammatory molecules suggest their possible direct involvement in innate immunity. Toll-like receptors (TLRs) are an essential signaling component to this response, as they allow for recognition of various microorganisms, leading to subsequent induction of pro-inflammatory genes. The objective of this study was to elucidate the role of VFF in the host immune response and subsequent influence on inflammatory cytokine secretion.
  
  
  METHODS
  VFF derived from polyp, scar, and normal tissue were treated with 5 μg/ml lipopolysaccharide (LPS). TLR1 through 9, CD14, and MD-2 were measured during stable conditions by polymerase chain reaction (PCR). Expression of TLR4 and IL-1R type-1 genes were quantified after 24 hrs LPS stimulation by reverse transcription-PCR. LPS responsiveness was determined by NF-κB nuclear translocation as measured by subunit p65 expression in nucleus with immunocytochemistry. Downstream effects were confirmed with immunoassay measuring IL-8 concentrations in supernatant after 8 hrs.
  
  
  RESULTS
  All VFFs constitutively expressed TLR1 to 6, TLR9, CD14, and MD-2 mRNA. Polyp VFF exhibited significantly higher TLR4 transcript levels (p < 0.001) in comparison to scar and normal VFF. LPS stimulated scar and polyp VFF exhibited increased levels of p65 in the nucleus (p < 0.01) and secreted greater IL-8 protein (p < 0.0001) compared to normal VFF.
  
  
  CONCLUSIONS
  VFF constitutively express genes for the receptors essential to the host immune response. Scar and polyp VFF produced greater LPS responsiveness resulting in over-activated inflammatory patterns. These findings support VFF role in the pathogenesis of inflammatory vocal fold disorders and suggests their presence in the wound bed could lead to chronic inflammation.
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Wisconsin-Madison, WIMR 5107 1111 Highland Avenue, Madison, WI 53705-2725 USA.
authors.forename: Suzanne N
authors.initials: SN
authors.lastname: King
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Wisconsin-Madison, WIMR 5107 1111 Highland Avenue, Madison, WI 53705-2725 USA.
authors.forename: Craig M
authors.initials: CM
authors.lastname: Berchtold
authors.affiliation: Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Wisconsin-Madison, WIMR 5107 1111 Highland Avenue, Madison, WI 53705-2725 USA.
authors.forename: Susan L
authors.initials: SL
authors.lastname: Thibeault
keywords: Fibroblasts; Lipopolysaccharide; Polyp; Scar; Toll-like receptors; Vocal fold