Dataset Information

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 ?g/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-?B 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?

SUBMITTER: King SN

PROVIDER: S-EPMC4300178 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Lipopolysaccharide responsiveness in vocal fold fibroblasts.

King Suzanne N SN Berchtold Craig M CM Thibeault Susan L SL

Journal of inflammation (London, England) 20141219 1

<h4>Background</h4>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 t ...[more]

PMID: 25606025

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Project description:Injuries of the vocal folds frequently heal with scar formation, which can have lifelong detrimental impact on voice quality. Current treatments to prevent or resolve scars of the vocal fold mucosa are highly unsatisfactory. In contrast, the adjacent oral mucosa is mostly resistant to scarring. These differences in healing tendency might relate to distinct properties of the fibroblasts populating oral and vocal fold mucosae. We thus established the in vitro cultivation of paired, near-primary vocal fold fibroblasts (VFF) and oral mucosa fibroblasts (OMF) to perform a basic cellular characterization and comparative cellular proteomics. VFF were significantly larger than OMF, proliferated more slowly, and exhibited a sustained TGF-β1-induced elevation of pro-fibrotic interleukin 6. Cluster analysis of the proteomic data revealed distinct protein repertoires specific for VFF and OMF. Further, VFF displayed a broader protein spectrum, particularly a more sophisticated array of factors constituting and modifying the extracellular matrix. Conversely, subsets of OMF-enriched proteins were linked to cellular proliferation, nuclear events, and protection against oxidative stress. Altogether, this study supports the notion that fibroblasts sensitively adapt to the functional peculiarities of their respective anatomical location and presents several molecular targets for further investigation in the context of vocal fold wound healing.Biological significanceMammalian vocal folds are a unique but delicate tissue. A considerable fraction of people is affected by voice problems, yet many of the underlying vocal fold pathologies are sparsely understood at the molecular level. One such pathology is vocal fold scarring - the tendency of vocal fold injuries to heal with scar formation -, which represents a clinical problem with highly suboptimal treatment modalities. This study employed proteomics to obtain comprehensive insight into the protein repertoire of vocal fold fibroblasts, which are the cells that predominantly synthesize the extracellular matrix in both physiological and pathophysiological conditions. Protein profiles were compared to paired fibroblasts from the oral mucosa, a neighboring tissue that is remarkably resistant to scarring. Bioinformatic analyses of the data revealed a number of pathways as well as single proteins (e.g. ECM-remodeling factors, transcription factors, enzymes) that were significantly different between the two fibroblast types. Thereby, this study has revealed novel interesting molecular targets which can be analyzed in the future for their impact on vocal fold wound healing.

Dataset Information

Lipopolysaccharide responsiveness in vocal fold fibroblasts.

Publications

Lipopolysaccharide responsiveness in vocal fold fibroblasts.

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