Pepsin Inhibitors Prevent Inflammation and Loss of Laryngeal Barrier Function in Mice with Gastroesophageal Reflux.

OBJECTIVE: This study aimed to evaluate the role of pepsin inhibitors in the inflammatory response and their effects on laryngeal mucosal integrity during gastroesophageal reflux (GERD) under in vivo conditions. METHODS: A surgical model of GERD was used, in which mice were treated with pepstatin (0.3 mg/kg) or darunavir (8.6 mg/kg) for 3 days. On the third day after the experimental protocol, the laryngeal samples were collected to assess the severity of inflammation (wet weight and myeloperoxidase activity) and mucosal integrity (transepithelial electrical resistance and paracellular epithelial permeability to fluorescein). RESULTS: The surgical GERD model was reproduced. It showed features of inflammation and loss of barrier function in the laryngeal mucosa. Pepstatin and darunavir administration suppressed laryngeal inflammation and preserved laryngeal mucosal integrity. CONCLUSION: Pepsin inhibition by the administration of pepstatin and darunavir improved inflammation and protected the laryngeal mucosa in a mouse experimental model of GERD. LEVEL OF EVIDENCE: NA Laryngoscope, 134:3080-3085, 2024.

Laryngeal Mucosa Alterations in Mice Model of Gastroesophageal Reflux: Effects of Topical Protection.

OBJECTIVES/HYPOTHESIS: The objectives of this study were to evaluate laryngeal inflammation and mucosal integrity in a murine model of reflux disease and to assess the protective effects of topical agents including alginate, hyaluronic acid, and cashew gum. STUDY DESIGN: Animal study. METHODS: A surgical murine model of reflux disease was evaluated at 3 or 7 days postsurgery, and laryngeal samples were collected to measure inflammation (wet weight and myeloperoxidase [MPO]) and mucosal integrity (transepithelial resistance [TER] and mucosal permeability to fluorescein). Additional groups of animals were administered one of several topical agents (alginate, hyaluronic acid, or cashew gum) daily, and laryngeal inflammation and mucosal integrity were evaluated at 3 days postsurgery. RESULTS: At 3 days, and not 7 days postsurgery, we observed increased laryngeal wet weight and MPO, decreased laryngeal TER, and increased laryngeal mucosa permeability. Alginate partially decreased laryngeal inflammation (wet weight and not MPO) and dramatically improved laryngeal mucosal integrity. Conversely, hyaluronic acid eliminated the inflammation; however, it had no effect on laryngeal mucosal integrity impairment. Cashew gum eliminated laryngeal inflammation as well as the impairment in laryngeal mucosal integrity. CONCLUSIONS: This study shows that a surgical model of reflux disease induced laryngeal inflammation and impairment in laryngeal barrier function. These observed alterations were partially attenuated by alginate and hyaluronic acid and completely reversed by cashew gum. LEVEL OF EVIDENCE: NA Laryngoscope, 2020.

Comparative study of the behavior of p53 immunoexpression in smoking associated lesions: Reinke's edema and laryngeal carcinoma.

OBJECTIVE: To assess the behavior of the immunoexpression of protein p53 in Reinke's edema and laryngeal squamous cell carcinoma. STUDY DESIGN: retrospective. METHODS: we recovered the histological paraffin blocks of patients who were subjected to Reinke's edema and laryngeal squamous cell carcinoma surgery in 2000-2011. The paraffin blocks were cut into 3-µm sections; the specimens were prepared in silanized slides (one slide for each paraffin block) and subjected to immunohistochemical reaction according to the Avidin Biotin Peroxidase method. Monoclonal primary anti-p53 antibodies were used at 1:50 dilution. Slides were examined under a light microscope at different magnitudes and results were interpreted based on the degree of brown staining in the nuclei of epithelial cells and in the extent of the fragment by using a semi-quantitative score from 0 to 3. RESULTS: 67 slides of Reinke's edema and 60 slides of laryngeal squamous cell carcinoma were included. Scores 2 and 3 for staining of the nuclei of epithelial cells were recorded for 46 slides of Reinke's edema (68.65%) and for 57 slides of laryngeal squamous cell carcinoma (95%). As to the extent of the fragment, scores 2 and 3 were recorded for 74% slides of Reinke's edema and for 95% slides of carcinomas. CONCLUSION: the positive immunoexpression for protein p53, positive in 95% carcinomas and 74% Reinke's edemas, makes us aware of the possible preneoplastic condition of the latter lesion. Further studies are needed to identify and reveal the genetic changes that lead to these results.

Elastic fibers in Reinke's edema.

OBJECTIVES: We describe the distribution of elastic system fibers in the superficial layer of the lamina propria of Reinke's edema, as compared with normal vocal folds. METHODS: Weigert's resorcin-fuchsin stain after oxidation with 10% oxone was used to study the arrangement of elastic fibers. The findings were categorized and afterward compared with the severity of Reinke's edema. RESULTS: Analysis of 20 specimens of vocal folds with Reinke's edema showed that the network of thin elastic fibers in Reinke's space lost their undulated appearance and had a tangled aspect. In addition, these fibers were no longer parallel to the epithelial basement membrane, but had a random distribution scattered throughout Reinke's space. The elastic fiber network immediately below the epithelial basement membrane also appeared more fragmented in Reinke's edema because of some alteration in organization combined with the 5-microm-thick histologic sectioning plane. No significant difference in the degree of elastic system fiber disarrangement was observed between severity grades II and III (p = 0.382). Large areas of disarrangement were predominant (80% of cases). CONCLUSIONS: The disarrangement of elastic fibers in Reinke's edema may cause insufficient tissue resistance and resilience, contributing to the hypermobility observed in Reinke's edema.

Sulcus mucosal slicing technique.

PURPOSE OF REVIEW: To present the accurate surgical indication for the slicing mucosal technique, the case selection, surgical aspects, rehabilitation concerns, and the characteristics of immediate and long-term outcomes. RECENT FINDINGS: The literature is still scarce; few cases are submitted to the slicing mucosa technique due to its specific indication; an alternative procedure was designed for cases where mucosal movement is strongly reduced, the inner section of the vocal ligament or submucosal scar tissue, which can eventually be associated with fat inclusion. Some selected cases may require thyroplasty type III to optimize functional results. SUMMARY: Slicing technique is an aggressive powerful resource for the surgical treatment of severe cases of sulcus striae major, in which mucosal wave is absent and glottic chink is moderate to severe; voice is intensely deviated immediately postoperation; vocal rehabilitation is mandatory and an intensive regimen is usually required for the first 2 months; final results can mostly be achieved up to 6 months.

Angiogenesis, fibrinogenesis and presence of synechiae after exeresis of a swine vocal fold mucosal microflap and use of topical mitomycin-C.

PURPOSE: To evaluate swine vocal fold healing in a period of 30 days after topical mitomycin-C application. METHODS: Twelve swine underwent exeresis of mucosal flaps from the free edge of the anterior third of both vocal folds with a cold instrument (laryngeal scissors). The animals were divided into two groups: EG (Experimental Group), consisting of 6 animals undergoing topical MMC application (4 mg/dL) on the operated area for 4 minutes; CG (Control Group), 6 animals undergoing topical saline solution application on the operated area for 4 minutes. After 30 days, the animals were sacrificed and the larynges were collected and examined for the presence of synechiae as well as a histological immunohistochemical assessment of immature and mature collagen deposition, number of blood vessels and myofibroblasts. RESULTS: Mature collagen deposition in the EG was 452.12 microm(2) and 1332.31 microm(2) in the CG; immature collagen deposition was 1511.73 microm(2) in the EG and 1020.61 microm(2) in the CG. The number of myofibroblasts was 1.556 in the EG and 3.583 in the CG. The number of blood vessels was 2.565 in EG and 6.917 in the CG. There were no synechiae in the two studied groups. CONCLUSIONS: There was an increase in immature collagen deposition in the experimental group when compared with the control group. There was a decrease in mature collagen deposition in the experimental group when compared with the control group. There was a decrease in the number of myofibroblasts in the experimental group when compared with the control group. A decrease in blood vessels was observed in the experimental group when compared with the control group. There was no synechia formation in either studied group.

Angiogenesis, fibrinogenesis and presence of synechiae after exeresis of a swine vocal fold mucosal microflap and use of topical mitomycin-C / Estudo da angiogênese, fibrogênese e presença de sinéquias após exérese de fragmento de mucosa de pregas vocais de suínos utilizando instrumental frio e mitomicina-C tópica

PURPOSE: To evaluate swine vocal fold healing in a period of 30 days after topical mitomycin-C application. METHODS: Twelve swine underwent exeresis of mucosal flaps from the free edge of the anterior third of both vocal folds with a cold instrument (laryngeal scissors). The animals were divided into two groups: EG (Experimental Group), consisting of 6 animals undergoing topical MMC application (4 mg/dL) on the operated area for 4 minutes; CG (Control Group), 6 animals undergoing topical saline solution application on the operated area for 4 minutes. After 30 days, the animals were sacrificed and the larynges were collected and examined for the presence of synechiae as well as a histological immunohistochemical assessment of immature and mature collagen deposition, number of blood vessels and myofibroblasts. RESULTS: Mature collagen deposition in the EG was 452.12 μm² and 1332.31μm² in the CG; immature collagen deposition was 1511.73μm² in the EG and 1020.61μm² in the CG. The number of myofibroblasts was 1.556 in the EG and 3.583 in the CG. The number of blood vessels was 2.565 in EG and 6.917 in the CG. There were no synechiae in the two studied groups. CONCLUSIONS: There was an increase in immature collagen deposition in the experimental group when compared with the control group. There was a decrease in mature collagen deposition in the experimental group when compared with the control group. There was a decrease in the number of myofibroblasts in the experimental group when compared with the control group. A decrease in blood vessels was observed in the experimental group when compared with the control group. There was no synechia formation in either studied group.

OBJETIVO: Avaliar a cicatrização de pregas vocais de suínos utilizando mitomicina-C tópica, em 30 dias. MÉTODOS: Doze suínos foram submetidos à exérese de mucosa do bordo livre do terço anterior de ambas as pregas vocais com instrumental frio (tesoura curva) e divididos em dois grupos: Grupo E (GE), composto por seis animais com utilização tópica de mitomicina-C(4 mg/dL) na área cruenta por quatro minutos e Grupo Controle (GC), composto por seis animais com utilização tópica de soro fisiológico na área cruenta por quatro minutos. Após 30 dias os animais foram sacrificados e coletadas amostras das pregas vocais para avaliação de sinéquias e análise histológica com quantificação da deposição de colágeno maduro e imaturo, quantificação do número de vasos sanguíneos e miofibroblastos, por método imunoistoquímico. RESULTADOS: A deposição do colágeno maduro no GE foi de 452,12 μm² e 1332,31μm² no GC. A do colágeno imaturo foi de 1511,73μm² no GE e de 1020,61μm² no GC. O número de miofibroblastos foi de 1,556 no GE e de 3,583 no GC. O número de vasos sanguíneos foi de 2,565 no GE e de 6,917 no GC. Não houve formação de sinéquias nos grupo s estudados. CONCLUSÕES: No grupo experimento, quando comparado ao controle, houve significativamente aumento da deposição do colágeno imaturo e diminuição da deposição do colágeno maduro, do número de miofibroblastos e de vasos sanguíneos. Não houve formação de sinéquias em ambos os grupos.

Is Reinke's edema a precancerous lesion? Histological and electron microscopic aspects.

The objectives of this study were to evaluate morphologic alterations and precancerous lesions in Reinke's edema. Patients included were 54 smokers with bilateral Reinke's edema submitted to surgery in the Otolaryngology Department, Botucatu Medical School, São Paulo State University, Brazil, between 2002 and 2006. All specimens were evaluated by light microscopy and five contralateral lesions were also evaluated by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The main histological alterations were edema (100%), inflammation (81.48%), basal membrane (bm) thickening (77.77%), and vessel proliferation (75.92%). Epithelium alterations were classified as grade 0 (11.11%), grade 1 (70.37%), grade 2 (14.81%), and grade 3 (3.70%). In the case included in grade 3 classification, microinvasive carcinoma was detected. SEM showed epithelial surface with some cellular desquamation, few microridges, and a polished and impermeable surface aspect. TEM showed epithelial hyperplasia, basal cells with different sizes, widening of the intercellular spaces, abnormal desmosome architecture, thickening of the bm, some electron-dense vesicles, and points of interruption. The morphological alterations presented in this study are not specific to Reinke's edema but this lesion can be the site of different grades of dysplasia and carcinoma, justifying the importance of periodic laryngeal endoscopic exams and meticulous histological analysis.

Morphological and functional aspects of the senile larynx.

OBJECTIVE: To analyze the senile larynx, considering morphological aspects during respiration and functional aspects during phonation, comparing it to the young adult larynx. PATIENTS AND METHODS: A retrospective study was conducted of 100 laryngeal images of adults without vocal complaints, 50 laryngeal images of young adults aged 20-45 years and 50 laryngeal images of elderly individuals aged 65-85 years, of both genders. Images were produced by means of a rigid endoscope and stroboscopy. The morphological aspects analyzed were: laryngeal symmetry, vocal fold bowing, prominence of the vocal process and glottic proportion. The functional aspects analyzed were glottic closure, laryngeal vestibule behavior, phase (timing of closure) and amplitude symmetry of mucosal wave and tremor of laryngeal structures. RESULTS: Significant differences between the young adult larynx and the geriatric larynx were found for the following parameters: vocal fold bowing, prominence of the vocal process, glottic proportion, phase and amplitude symmetry of mucosal wave and tremor of laryngeal structures. CONCLUSION: The data suggests that the geriatric larynx differs from the young adult larynx in many characteristic aspects.

Lesöes precoces da intubaçäo endotraqueal / Early injuries of the endotracheal intubation

Os autores submeteram 30 cäes à intubaçäp endotraqueal com cânulo de alta complacência, durante período de quatro horas, com o objetivo de detetar lesöes precoces laringotraqueais secundárias à intubaçäo. Após este período, realizaram-se biópsias em cordas vocais, aritenóides, cricóide e anéis traqueais. As alteraçöes histológicas mais frequentemente observadas foram: infiltrado de células polimorfonucleares, tanto no epitéleio como no córion, congestäo vascular e necrose de células epiteliais. Os anéis traqueais, que se encontravam em íntimo contacto com o balonete da cânula, foram os locais mais comprometidos. Os autores descrevem os principais fatores lesivos ao epitélio respiratório e preconizam a utilizaçäo de cânulas com balonetes de alta complacência, além do seguimento endoscópico dos pacientes após extubaçäo

Tabagismo e mucosa da laringe / Smoke and laringeal mucous-membrane

Tabagismo e mucosa laringeana, säo discutidas, sob os aspectos patogênicos. Os pólipos da laringe säo descritos como uma reaçäo mucosa ao fumo