Project description:Vocal cord healing is a dynamic process, and many genes and proteins are involved, which play varying roles at different regeneration stages after injury. Previous studies have shown that inflammatory responses occur at the early stage of vocal cord injury, where the fibroblasts proliferate exuberantly with intensive secretion and deposition of ECM. These activities reach the peak at 3-7 days and their intensity begins to decline 15 days later. A study based on the dermal system has shown that ECM remodeling during the repair of injury can last for several months. However, few studies have been conducted as to the dynamic changes of gene expressions and signaling pathway during the healing process of vocal cord injury. Plotting these changes will facilitate the understanding about the physiological changes during healing and the identification of key time points and target genes in fibrosis formation.
Project description:Vocal cord healing is a dynamic process, and many genes and proteins are involved, which play varying roles at different regeneration stages after injury. Previous studies have shown that inflammatory responses occur at the early stage of vocal cord injury, where the fibroblasts proliferate exuberantly with intensive secretion and deposition of ECM. These activities reach the peak at 3-7 days and their intensity begins to decline 15 days later. A study based on the dermal system has shown that ECM remodeling during the repair of injury can last for several months. However, few studies have been conducted as to the dynamic changes of gene and microRNA expressions during the healing process of vocal cord injury. Plotting these changes will facilitate the understanding about the physiological changes during healing and the identification of key time points and target genes and microRNAs in fibrosis formation.
Project description:We used microarrays to characterize transcriptome profiles of rat vocal fold tissue following surgical injury (vs. naive tissue); rat vocal fold fibroblasts harvested from scar tissue at the 60 d time point (vs. naive fibroblasts); rat vocal fold scar fibroblasts treated with siRNA against the collagen chaperone protein rat gp46 (vs. scramble siRNA). Adult Fischer 344 rat vocal fold tissue was harvested at 3, 14, and 60 days following surgical injury (control = age-matched naive tissue); rat vocal fold scar fibroblasts were obtained via explant culture of tissue obtained 60 days following surgical injury and harvested at 80% confluence during passage 1 (control = age-matched naive rat vocal fold fibroblasts); rat vocal fold scar fibroblasts were treated for 1 h with 50 nM liposome-delivered siRNA against rat gp46 when 80% confluent at passage 1, cultured for an additional 24 h in fresh media, then harvested (control = rat vocal fold scar fibroblasts treated with 50 nM liposome-delivered scramble siRNA).
Project description:We used microarrays to characterize transcriptome profiles of rat vocal fold tissue following surgical injury (vs. naive tissue); rat vocal fold fibroblasts harvested from scar tissue at the 60 d time point (vs. naive fibroblasts); rat vocal fold scar fibroblasts treated with siRNA against the collagen chaperone protein rat gp46 (vs. scramble siRNA).
Project description:The purpose of this study is to investigate if acute systemic dehydration impacts the vocal folds at the mRNA level. Rabbits were used as animal model to enable a systematic and rigorous assessment of the pathobiology of vocal fold hydration since they can be manipulated in a physiologically realistic manner. We anticipate that the results of this study will help to elucidate the effects of altered hydration state in response to furosemide on vocal fold epithelium, lamina propria and muscle tissue at the transcriptome level. Together with cellular, structural, and functional techniques that are part of this project, we expect to provide validated scientific recommendations on the adverse effects of dehydration and the therapeutic benefits of hydration.
Project description:We present here the first study evaluating the dynamic proteome changes in the remodeling process of decellularized tissue by reseeded cells using vocal fold mucosa as the model system.
Project description:Reinke’s edema is a cigarette associated, benign, mostly bilateral lesion of the vocal folds leading to dysphonia and dyspnea. Until today information about pathophysiology is only scarce, treatment is only surgical. To explore the pathophysiology of Reinke’s edema, we exposed near primary human vocal fold fibroblasts to cigarette smoke extract enriched medium or air bubbled control for 24 hrs. followed by a proteomic analysis. Proteomic analyses revealed an increase of proteins involved in oxidative stress response. We furthermore found that a significant number of essential extracellular matrix proteins was altered by culturing human VFF with CSE enriched medium: UDP-glucose 6-dehydrogenase, a protein involved in hyaluronan synthesis was upregulated, while the expression of several fibrillar collagen types was significantly reduced. The current findings corroborate previous studies but revealed new insights in possible disease mechanisms of Reinke’s edema too. We postulate that changes in the composition -reduction of collagen fibrils, increase of hyaluronan- of the vocal folds’ ECM may lead to the clinical findings.
Project description:This dataset brief is about the descriptive proteomic comparison of human oral mucosa and vocal fold tissues by high-resolution mass spectrometry (CID-MS/MS). The vast majority of voice disorders is associated with changes of the unique but delicate tissue of the human vocal folds, wheras the ability to develop new effective treatment methods is significantly limited by the physical inaccessibility and the extremely rare occasion to gather healthy tissue biopsies. Therefore, oral mucosa reached specific interest for laryngological research, as the tissue harvesting process is less invasive and accompanied with faster healing and less scarring. Proteomic analysis of both tissues will provide a fundamental laryngological resource for the research community. Our study identifies a total of 1575 proteins detected within both tissues that are highly consistent in several crucial biological processes, cellular components, and molecular functions.
Project description:Development of treatments for vocal dysphonia has been inhibited by lack of human vocal fold (VF) mucosa models because of difficulty in procuring VF epithelial cells, epithelial cells’ limited proliferative capacity and absence of cell lines. We report development of engineered VF mucosae from hiPSC, transfected via TALEN constructs for green fluorescent protein, that mimic development of VF epithelial cells in utero. Modulation of FGF signaling achieves stratified squamous epithelium from definitive and anterior foregut derived cultures. Robust culturing of these cells on collagen-fibroblast constructs produces three-dimensional models comparable to in vivo VF mucosa. Second, we demonstrate mucosal inflammation upon exposure of these constructs to 5% cigarette smoke extract. Upregulation of pro-inflammatory genes in epithelium and fibroblasts leads to aberrant VF mucosa remodeling. Collectively, our results demonstrate that hiPSC-derived VF mucosa is a versatile tool for future investigation of genetic and molecular mechanisms underlying epithelium-fibroblasts interactions in health and disease.