This study investigated the relationship between acoustic spectral/cepstral measures and listener severity ratings in normal and disordered voice samples. CAPE-V sentence samples and the vowel /a/were elicited from eight normal speakers and 24 patients with varying degrees of dysphonia severity. Samples were analysed for measures of the cepstral peak prominence (CPP), the ratio of low-to-high spectral energy, and their respective standard deviations. Perceptual ratings of overall severity were also obtained for all samples. Results showed that all acoustic variables combined in a four-factor model which correlated with perceived severity with R = 0.81 (R(2) = 0.65). For the vowel /a/, a five-factor model incorporating all acoustic variables and gender correlated with perceived severity with R = 0.96 (R(2) = 0.91). Results indicate that a strong relationship between perceptual and acoustic estimates of dysphonia severity can be achieved in both continuous speech and vowel contexts using a model incorporating spectral/cepstral measures.

Recent evidence suggests that fibroblasts play a critical role in regulating inflammation during wound healing because they express several inflammatory mediators in response to bacteria. The objective of this study was to analyze the effects of lipopolysaccaride (LPS) on the immunomodulatory properties of vocal fold fibroblasts (VFF) derived from polyps, scar and normal tissue co-cultured with macrophages, to provide insight into their interactions during the inflammatory process. Fibroblasts were co-cultured with CD14+ monocytes and after 7 days, wells were treated with LPS for 24 and 72 hours. Culture supernatants were collected and concentrations of TNF-α, IL-6, IL-8, IL-10, IL-12, IL-1β, and MCP-1 were quantified by ELISA. Normal VFF and CD14+ monocultures were used as controls. Twenty-four hours after LPS activation, macrophages co-cultured with polyp VFF had significantly increased expression of TNF-α, IL-1β, IL-12, and IL-10 compared to controls (p<0.0001). In contrast, macrophages co-cultured with scar VFF had significantly lower expression of TNF-α, IL-1β and IL-12 with significantly higher IL-10 compared to control (p<0.0001). After 72 hours, macrophages co-cultured with polyp VFF increased expression of TNF-α, IL-1β, IL-10, IL-6, IL-8, MCP-1 and TGF-β (p<0.01) and macrophages co-cultured with scar VFF significantly decreased their expression of IL-1β and IL-12 compared to control (p<0.0001). Scar VFF at both time points produced significantly lower levels of IL-8, MCP-1, IL-6 and TGF-β compared to controls (p<0.05). Based on our findings, VFF and macrophages secrete several inflammatory mediators that modify their diverse functions. Polyp and scar VFF may play a role in regulating abnormal inflammatory responses, which could result in excessive ECM deposition that disrupts the function of the vocal folds.

Objectives/Hypothesis In vitro modeling of cell-matrix interactions that occur during human vocal fold scarring is uncommon, as primary human vocal fold scar fibroblast cell lines are difficult to acquire. The purpose of this study was to characterize morphologic features, growth kinetics, contractile properties, α-smooth muscle actin (α-SMA) protein expression and gene expression profile of human vocal fold fibroblasts derived from scar (sVFF) relative to normal vocal fold fibroblasts (nVFF). Study Design In vitro. Methods We successfully cultured human vocal fold fibroblasts from tissue explants of scarred vocal folds from a 56-year-old female and compared these to normal fibroblast cells from a 59-year-old female. Growth and proliferation were assessed by daily cell counts and morphology was compared at 60% confluence for 5 days. Gel contraction assays were evaluated after seeding cells within a collagen matrix. Alpha-SMA was measured using western blotting and immunocytochemistry (ICC). Quantitative RT-PCR was used to assess differential extracellular matrix gene expression between the two cell types. Results sVFF were morphologically indistinguishable from nVFF. sVFF maintained significantly lower proliferation rates relative to nVFF on days 3-6 (day 3: p = 0.0138, days 4, 5, and 6: p < 0.0001). There were no significant differences in contractile properties between the two cell types at any time point (0h: p = 0.70, 24h: p = 0.79, 48h: p = 0.58). ICC and western blot analyses revealed increased expression of α-SMA in sVFF as compared with nVFF at passages 4 and 5, but not at passage 6 (passage 4: p = 0.006, passage 5: p = 0.0015, passage 6 = 0.8860). Analysis of 84 extracellular matrix genes using qRT-PCR revealed differential expression of 15 genes (p < 0.01). Conclusions nVFF and sVFF displayed differences in proliferation rates, α-SMA expression, and gene expression, whereas no differences were observed in contractile properties or morphology. Further investigation with a larger sample size is necessary to confirm these findings.

Prolonged diffuse laryngeal inflammation from smoking and/or reflux is commonly diagnosed as chronic laryngitis and treated empirically with expensive drugs that have not proven effective. Shifts in microbiota have been associated with many inflammatory diseases, though little is known about how resident microbes may contribute to chronic laryngitis. We sought to characterize the core microbiota of disease-free human laryngeal tissue and to investigate shifts in microbial community membership associated with exposure to cigarette smoke and reflux. Using 454 pyrosequencing of the 16S rRNA gene, we compared bacterial communities of laryngeal tissue biopsies collected from 97 non-treatment-seeking volunteers based on reflux and smoking status. The core community was characterized by a highly abundant OTU within the family Comamonadaceae found in all laryngeal tissues. Smokers demonstrated less microbial diversity than nonsmokers, with differences in relative abundances of OTUs classified as Streptococcus, unclassified Comamonadaceae, Cloacibacterium, and Helicobacter. Reflux status did not affect microbial diversity nor community structure nor composition. Comparison of healthy laryngeal microbial communities to benign vocal fold disease samples revealed greater abundance of Streptococcus in benign vocal fold disease suggesting that mucosal dominance by Streptococcus may be a factor in disease etiology.