Faculty Bibliography

OBJECTIVES/HYPOTHESIS.: Prostaglandin (PG)E(2) has been implicated in a variety of disease processes. It has been described as antifibrotic in the lower airway, yet scar-inducing in the skin. We seek to describe the effects of PGE(2) on vocal fold fibroblasts and its interactions with transforming growth factor (TGF)-beta1. In addition, we describe a novel organotypic model, a critical step in the development of therapeutic trials. STUDY DESIGN.: In vitro, ex vivo. METHODS: Collagen secretion by human vocal fold fibroblasts (HVFF) was assayed in response to TGF-beta1, PGE(2) , and specific EP receptor agonists. Basal HVFF migratory rate was also quantified in response to PGE(2) . TGF-beta1 induced COX-2 mRNA expression/PGE(2) secretion was assayed. Excised vocal folds were subjected to exogenous IL-1beta; PGE(2) secretion into the supernatant was then assayed. RESULTS: TGF-beta1-induced collagen secretion was blunted in a dose-dependent manner in response to PGE(2) . This effect appears to be mediated primarily through the EP1 and EP2 receptors. TGF-beta1 induced COX-2 mRNA expression and PGE(2) secretion. In our organ culture model, IL-1beta stimulated PGE(2) secretion in a dose-dependent manner. CONCLUSIONS: PGE(2) is antifibrotic; this finding suggests that the upper airway response to this inflammatory mediator differs significantly from the lower airway. These data have important clinical implications for a variety of pathological processes. Furthermore, exogenous TGF-beta1 elicits induction of COX-2, suggesting inherent complexity regarding these processes and PGE(2) signaling, specifically. In addition, our organ culture model may prove useful as a means to quantify biological phenomena in the vocal folds

Although Reinke's space, or layer, is a critical laryngeal structure, and the eponym is in current use in both clinical and research milieus, little is known about the life of the eponymist, German anatomist Friedrich Berthold Reinke. Extensive investigation of the archives at the University of Rostock and other sources, as well as multinational collaboration, has yielded unique insight into the personal and professional life of this pioneer who, among other things, identified and characterized the subepithelial space of the vocal fold and structures in the Leydig cells of the testicles and ovaries. This breadth of investigation reflects Reinke's intellectual curiosity and broad-ranging interest as well as his scientific environment. Without question, Reinke's observations of the human vocal fold are substantive contributions, without which modern laryngology could not have evolved. In 2009, at the 90th anniversary of Reinke's death, we summarize his achievements to express our appreciation for his singular brilliance and fundamental contribution to laryngology.

Perception of the impact of voice disorders may differ across different cultural backgrounds. This study investigated the difference in the perception of the impact of voice disorders between the American (Pittsburgh) and Chinese (Hong Kong) cultures. Study Design and Setting: Sixty dysphonic subjects from Hong Kong, China, and 60 dysphonic subjects from Pittsburgh, USA, were recruited to complete the Chinese and English versions of the Voice Activity and Participation Profile (VAPP), respectively. Data analyses using independent t tests were conducted on (1) the total profile scores; (b) total activity limitation score (ALS) and total participation restriction score (PRS); (c) section scores of job, daily communication, and social communication; and (d) section ALS and section PRS. Results: Hong Kong subjects showed significantly higher scores than the Pittsburgh subjects in total profile and other subsections except in the job PRS. Conclusion: Results support the possible influence of individualist and collectivist cultures on the perceived impact of voice disorders on the activity and participation.

OBJECTIVES/HYPOTHESIS: In response to chronic cigarette smoke exposure, a subset of patients present with edematous vocal folds, characteristically referred to as Reinke's edema. This phenotype differs from the tissue changes associated with prolonged smoke exposure in the lower airway, and the mechanism underlying Reinke's edema remains poorly described. We hypothesize that the effects of smoke are diffuse and involve both the epithelium and mucosa. STUDY DESIGN: In vitro, ex vivo experiment. METHODS: Transepithelial resistance (R(T) ) was quantified in an ex vivo, viable, porcine vocal fold model. Excised tissue was exposed to cigarette smoke condensate (CSC) and R(T) was computed at baseline and 1 and 4 hours after exposure. In vitro, human vocal fold fibroblasts were exposed to CSC. Cyclooxygenase 2 (COX-2), microsomal prostaglandin E synthase-1, and 15-hydroxyprostaglandin dehydrogenase mRNA expression were assessed at 4 hours. Prostaglandin E2 (PGE2) synthesis was quantified via immunoassay following 24 hours of CSC exposure. RESULTS: CSC had no effect on R(T) . CSC did, however, induce COX-2 mRNA expression as well as its downstream lipid mediator PGE2. PGE2 metabolism appears to be regulated via both synthetic and degradative enzymes in response to cigarette smoke. CONCLUSIONS: In vitro, CSC initiates an inflammatory response in vocal fold fibroblasts. However, in isolation, the epithelial resistance is not altered by CSC, at least acutely. These data may suggest a role for the interaction between the inflammatory response in the mucosa and compromised epithelial barrier function, as has been shown in other tissues. Laryngoscope, 2011

OBJECTIVES/HYPOTHESIS: Although the potassium titanyl phosphate (KTP) laser is versatile, the variability in laser parameters for laryngeal pathologies and the lack of clinical efficacy data remain problematic. We provide preliminary data regarding these parameters for benign lesion regression. In addition, we describe a novel method for the quantification of the effects of the KTP laser on vocal fold (VF) lesions. STUDY DESIGN: Retrospective chart review. METHODS: Images were captured from examinations before and after in-office KTP treatment in patients with a range of benign lesions. Laser settings were noted for each patient. Imaging software was then used to calculate a ratio of lesion area to VF length. Ten percent of images were requantified to determine inter-rater reliability. RESULTS: Thirty-two patients underwent 47 procedures for lesions including hemorrhagic polyp, nonhemorrhagic polyp, vocal process granuloma, Reinke's edema, cyst/pseudocyst, leukoplakia, and squamous cell carcinoma in situ. No statistically significant differences were observed with regard to the laser parameters used as a function of lesion type. Regardless, by 1 month following treatment, all lesions had significantly decreased in size, except nonhemorrhagic polyps. Similar data were obtained at 2-month follow-up. We then compared the pre-KTP lesion size with the smallest lesion size quantified during the 1-year follow-up period. All lesions were significantly smaller, with the exception of Reinke's edema. Inter-rater reliability was quite good. CONCLUSIONS: KTP laser effectively reduced VF lesion size, irrespective of the laser parameters used. In addition, our quantification method for lesion size appeared to be both viable and reliable. Laryngoscope, 2011

OBJECTIVE/HYPOTHESIS: The 532-nm KTP laser is clinically useful to induce benign vocal fold lesion regression without a fibrotic response. Previously, we described an in vivo model for KTP-induced injury in the rat larynx. This study uses this model to correlate the KTP-induced histologic and biochemical changes with the absence of long-term vocal fold fibrosis seen in clinical scenarios. STUDY DESIGN: In vivo. METHODS: Unilateral vocal fold injury was induced via KTP laser at 10W (20mS pulse width) as described by our laboratory previously. Animals were subjected to serial endoscopic imaging from postoperative days 1 through 3. Animals were euthanized at 1 day, 4 weeks, and 12 weeks posttreatment and subjected to histologic analyses via hematoxylin and eosin and trichrome staining, as well as RT-PCR analyses for MMP-3, 9, transforming growth factor-beta (TGF-beta), and COX-2 mRNA expression. Uninjured vocal folds were used as controls. RESULTS: Our study revealed gross healing of the vocal fold mucosa by 3 days posttreatment, and an immediate, moderate inflammatory infiltrate with no subsequent ultrastructural changes on histology. MMP-3 and COX-2 expression increased transiently, although no changes were seen in expression of MMP-9, an MMP involved in extracellular matrix remodeling, or TGF-beta, a profibrotic cytokine. CONCLUSIONS: These data suggest that the KTP laser induces a modest inflammatory response, selective MMP expression, and no long-term fibrotic processes in a clinically relevant simulation. Laryngoscope, 2011

INTRODUCTION: The current study seeks to provide preliminary data regarding this central, adaptive response during tongue motor tasks utilizing functional magnetic resonance imaging (fMRI) before and after glossectomy. METHODS: Six patients, with confirmed histological diagnoses of oral tongue cancer, underwent fMRI before and 6 months after partial glossectomy. These data were compared to nine healthy controls. All subjects performed three tongue motor tasks during fMRI: tongue tapping (TT), dry swallow (Dry), and wet swallow (Wet). RESULTS: Following surgery, increased activation was subjectively observed in the superior parietal lobule, supplementary motor area, and anterior cingulate. Region of interest (ROI) analysis of the precentral gyrus confirmed increased cortical activity following surgery. In addition, comparisons between pre-surgical scans and controls suggested the dry swallow task was sensitive to elicit tongue-related activation in the precentral gyrus (p </= 0.05). CONCLUSIONS: The adaptive changes in the cortex following partial glossectomy reflect recruitment of the parietal, frontal, and cingulate cortex during tongue motor tasks. In addition, post-operative activation patterns more closely approximated control levels than the pre-operative scans. Furthermore, the dry swallow task appears most specific to elicit tongue-related cortical activity

OBJECTIVE/HYPOTHESIS: We hypothesize that the central response during swallowing rehabilitation is critical and may be exploited to maximize the therapeutic benefit. We seek to provide preliminary data regarding the neural networks associated with commonly employed rehabilitation strategies to increase our understanding of the neural bases underlying these maneuvers. STUDY DESIGN: Case series. METHODS: Ten healthy adults (five males, five females), ranging in age from 20 to 30 years (mean age = 25 years, SD = 2.5 years) with no previous history of neurologic illness or swallowing complaint were subjected to a single-trial functional magnetic resonance imaging (fMRI) paradigm. The experimental tasks consisted of three swallow maneuvers, dry swallow, Effortful swallow, and the Mendelsohn maneuver. RESULTS: Multiple regions including the cingulate gyrus, inferior frontal gyrus, insula, pre- and postcentral gyrus, inferior parietal lobe, superior frontal gyrus, supramarginal gyrus thalamus, were detected. The Effortful swallow, when compared to the dry swallow, elicited significant differential activation in the left superior temporal gyrus, left insula, left inferior parietal lobe, bilateral medial frontal gyrus, and right anterior cingulate. The Mendelsohn maneuver, when compared to the dry swallow, elicited significant activation in the bilateral postcentral gyrus, bilateral precentral gyrus, bilateral cingulate gyrus, bilateral medial frontal gyrus, left inferior parietal lobe, left supramarginal gyrus, and right insula. CONCLUSIONS: Our findings suggest that a single-trial design is sensitive to delineate a widespread neural network of activation in both hemispheres associated with rehabilitation tasks. Both the Effortful swallow and Mendelsohn maneuvers elicited significantly higher responses in regions related to swallowing, suggestive of enhanced cortical activation during these tasks

OBJECTIVES/HYPOTHESIS: Inflammation and its role in a coordinated fibroplastic response, which disrupts the structure of the vocal folds following injury, is critical. Cyclooxygenase-2 (COX-2) is an important enzyme involved in both inflammation and fibrosis; in addition, it is a prime target for therapeutic intervention. We sought to study this pathway in vocal fold fibroblasts to provide a foundation for future interventional studies. STUDY DESIGN: In vitro. METHODS: Human vocal fold fibroblasts were incubated with IL-1 beta to determine the effects on COX-2 signaling, along with upstream regulatory mechanisms and downstream mediators of wound healing. In vitro methods to assess mRNA expression, as well as intracellular and secreted protein (sodium dodecyl sulfate polyacrylamide gel electrophoresis and enzyme-linked immunosorbent assay) were employed. RESULTS: IL-1 beta regulation of COX-2 mRNA and protein levels was dose and time dependent and IL-1 beta altered PGE(2) metabolism, via regulation of both synthetic and degradative enzymes. IL-1 beta increased nuclear factor (NF)-kappaB activation and nuclear translocation. Inhibition of the p50 and p65 subunits of NF-kappaB decreased IL-1 beta-induced COX-2 transcription. IL-1 beta also altered mRNA expression of four cell-surface prostaglandin receptors. CONCLUSIONS: Inflammation and fibrosis are important in the vocal fold pathophysiologic response to injury. Our data suggest that COX-2 and PGE(2) are inducible in human vocal fold fibroblasts, and this response appears to be NF-kappaB-dependent. We purport this fundamental investigation will lead to increased insight regarding injury and repair in the vocal folds, with the ultimate goal of developing novel clinical care paradigms

OBJECTIVES/HYPOTHESIS: Although the diagnostic accuracy of paradoxical vocal fold movement disorder and chronic cough has improved, the underlying pathophysiology remains relatively unknown. We hypothesize that one potential etiological factor in these patients is an aberrant laryngeal sensory response and sought to determine if respiratory retraining in addition to antireflux therapy alters this aberrant response. STUDY DESIGN: Retrospective, outcomes. METHODS: Sixteen patients who had been on at least 3 months of twice-daily proton pump inhibitors with no subjective improvement in their primary complaint of cough, self-reported symptoms of gastroesophageal and laryngopharyngeal reflux, and concurrent paradoxical vocal fold movement (PVFM) were included in the current study. In addition to continuing twice daily pharmacological therapy, subjects underwent a course of respiratory retraining. Outcome measures including the Reflux Symptom Index (RSI), transnasal flexible laryngoscopy, and laryngopharyngeal sensory discrimination thresholds were obtained prior to and following a course of respiratory retraining. RESULTS: Mean bilateral laryngeal sensory response improved significantly after combined respiratory retraining and aggressive proton pump inhibitor therapy (P = .01). In addition, mean RSI score decreased significantly following treatment (P = .02). Specifically, 13 of 16 patients experienced improved sensory response, corresponding with patient reports of improved PVFM symptoms following treatment. CONCLUSIONS: Aberrant laryngeal sensation was identified in patients with PVFM and chronic cough. This response, however, normalized following a limited course of respiratory retraining, corresponding with improved patient symptoms