Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The voice on congenital isolated growth hormone deficiency (IGHD) is commonly refered high pitched voice . However, vocal and laryngeal data are rare within literature. In Itabaianinha County, Northeast Brazil, it had been described the most extend kindred with severe IGHD due to a GH-realising hormone receptor (GHRHR) gene mutation. The aim of this descriptive transversal study was to evaluate vocal and laryngeal affections in this group. 23 subjects with IGHD were studied, being 6 males and 17 females They were submitted to an otorhinolaryngological evaluation and strobovideolaryngoscopic exam. Besides, a speech evaluation like hearing perception analysis (GRBAS scale) and objective voice evaluation (maximum phonatory time, s/z ratio and acoustic analysis) were also used. The results were performed using qui-square, exact Fisher and unpaired t test (p<0,05). The main vocal complaints were vocal fatigue (36,36%), hoarsenesses (31,81%), sore throat (27,27%), neckache (27,27%), phonatory pain (22,72%) and aphonia (18,18%). No male presented any complaint. Based on findings, vocal abuse (72,72%), etilism (45,45%) and smoking (36,36%) were considered like nocive habits to normal voice. 72,7% of the subjects presented abnormal voice and 27,3% were the normal ones. Amongst the abnormal voice, the mild/moderate grade was prevalent, with the presence of roughness, breathiness and strain. The strobovideoalaryngoscopic exam in 21 subjects showed laryngopharyngeal reflux signs (66,66%), functional dysphonia (52,38%), vocal nodules (23,80%) and vocal cysts (9,52%). There was significant association between vocal complaints and grade of abnormal voice. There weren t association between abnormal voice and laryngeal diseases; nocive habits and abnormal voice; nocive habits and laryngeal diseases; laryngopharyngeal reflux signs and functional dysphonia; vocal complaint and laryngeal diseases. The maximum phonatory time was lower than 10s and s/z ratio was 1,19. In acoustic analysis, fundamental frequency was 221,06 Hz in females and 204,67 Hz in males. There wasn t statistical dependency between genders showing high pitch and dominance IGHD effect above gender. There wasn t statistical dependency between normal and abnormal voice in acoustic analysis neither. Conclusions: Voice presents high pitch in both genders and the most commonly abnormalities were roughness, breathiness and strain. The mild/moderate grade was prevalent. Laryngopharyngeal reflux signs and functional dysphonia were the most reached diagnostics using strobovideolaryngoscopic exam. / A voz na deficiência isolada do hormônio de crescimento (DIGH) de início na infância é comumente referida como voz de caráter agudo, porém dados laríngeos e vocais são escassos na literatura. Na cidade de Itabaianinha-SE, no nordeste brasileiro, foi descrito o maior agrupamento familiar com DIGH severa, devido a uma mutação no gene do receptor do hormônio liberador do hormônio do crescimento (GHRHR). O objetivo do presente estudo foi avaliar alterações laríngeas e vocais nessa população, realizando-se um estudo transversal descritivo. Foram, então, estudados 23 indivíduos com DIGH, sendo 6 do sexo masculino e 17 do sexo feminino. Eles foram submetidos à avaliação otorrinolaringológica geral por entrevista e exame videolaringoestroboscópico. Avaliou-se também o grau de alteração na qualidade vocal, através da análise perceptivo-auditiva (escala GRBAS) e avaliação objetiva da voz (tempo máximo de fonação, relação s/z e análise acústica). Para comparação entre os achados, utilizou-se teste do quiquadrado e exato de Fisher (p<0,05) e entre os sexos, na análise acústica, teste t para amostras independentes (p<0,05). As principais queixas vocais relatadas foram cansaço vocal (36,36%), rouquidão (31,81%), pigarro (27,27%), dor cervical (27,27%), dor à fonação (22,72%) e perda da voz (18,18%). Nenhum indivíduo do sexo masculino apresentou queixa. Os principais hábitos nocivos à saúde vocal foram abuso vocal (72,72%), etilismo (45,45%), tabagismo (36,36%). Foram detectadas 72,7% de vozes alteradas e 27,3% de vozes normais. Dentre as alteradas, o grau leve/moderado foi preponderante, com presença de rouquidão/aspereza, soprosidade e tensão. A videolaringoestroboscopia foi realizada em 21 sujeitos e evidenciou sinais sugestivos de refluxo laringo-faríngeo (66,66%), disfonia funcional (52,38%), nódulos (23,80%) e cisto (9,52%). Houve também associação significativa entre queixas vocais e grau de alteração vocal. Porém, não houve associação significativa entre tipo de voz e doenças detectadas; hábitos nocivos e tipo de voz; hábitos nocivos e doenças detectadas; sinais sugestivos de refluxo laringofaríngeo e disfonia funcional; queixa vocal e doença. Na avaliação objetiva da voz, o tempo máximo de fonação foi abaixo de 10s em todas as variáveis e relação s/z de 1,19 . Na análise acústica, a freqüência fundamental foi 221,06 Hz, em mulheres, e 204,67 Hz, nos homens, sem diferença estatística, caracterizando um patamar agudo e um efeito dominante da DIGH sobre o sexo, nesta variável. Também não houve diferença estatística entre tipo de voz alterada e normal, nas variáveis da análise acústica. Conclusões: a voz foi aguda em ambos os sexos, sendo as alterações vocais mais freqüentes rouquidão/aspereza, soprosidade e tensão, grau leve/moderado. Os sinais sugestivos de refluxo laringo-faríngeo e disfonia funcional foram os achados mais comuns na videolaringoestroboscopia.

No município de São Paulo, em 1970 foram relatados 169 casos novos de câncer de laringe, com uma taxa de incidência ajustada por idade pela população mundial de 25,5/100.000 (30-74 anos) e 299 casos em 1985, elevando a taxa para 37,6 (ambos nosexo masculino). O Tabagismo e o consumo de álcool são os fatores de risco mais bem estabelecidos para o câncer de laringe. Com relação aos fatores ocupacionais, o único carcinógeno estabelecido é a exposição à névoa de ácidos inorgânicosfortes. É proposto um estudo caso-controle de base hospitalar, para investigar e quantificar o papel da exposição ocupacional no desenvolvimento do câncer de laringe e é discutida uma nova proposta de avaliação de exposição que visa melhorar avalidade interna desse tipo de estudo / Laryngeal cancer comprises 3,1% of the new cases of cancer in men in the world, representing the tenth most common malign neoplasm for males. In São Paulo City, 169 new cases of laryngeal cancer were reported in 1970, representing 25,5/100.000 and 299 cases in 1985, increasing the rate to 37,6/100.000 (both for males, age standardized truncated rate, 30-74 years). The best established risk factors for laryngeal cancer are tobacco and alcohol. In respect to occupational factors, the only established carcinogen is exposure to strong inorganic acid mists. However, asbestos, pesticides, paints, gasoline and diesel engine emissions, dusts, among other factors are reported in the literature as occupational agents that increase the risk of laryngeal cancer. A hospital-based case-control study was conducted, in seven hospitals in São Paulo, to investigate occupational risk factors for laryngeal cancer. The study included 122 laryngeal cancers and 187 controls, selected by frequency matching on sex and age. Detailed information on smoking, alcohol consumption, and occupational history was collected. Occupational hygienists assessed the exposure to 49 agents. Odds Ratios (OR) and 95% confidence intervals (95% CI) were estimated by unconditional logistic regression, and were adjusted for sex, age, smoking and alcohol. Laryngeal cancer was associated with exposure to respirable free crystalline silica (OR 1,83, 95% CI 1,00 - 3,36), soot (from coal, coke, fuel oil, wood) (OR 1,78, 95% CI 1,03 - 3,03), fumes (OR 2,55, CI 95% 1,14 - 5,67) and to live animals (OR 1,80, 95% CI 1,02 - 3,19). This study showed that occupational exposures to: respirable free crystalline silica, soot (from coal, coke, fuel oil, wood), fumes and live animals are risk factors for laryngeal cancer.

Introdução: A ciclina D1 constitui um importante regulador do ciclo celular e pode funcionar como co-regulador de transcrição. A superexpressão da ciclina D1 tem sido associada ao desenvolvimento e progressão do câncer. A degradação irregular da ciclina D1 pode ser responsável pelos seus níveis elevados em algumas neoplasias malignas. A ciclina D1, além disso, modula indiretamente a estrutura da cromatina e transcrição de genes envolvidos na proliferação e diferenciação. Mutações, amplificação e superexpressão da ciclina D1, que alteram a progressão do ciclo celular, são observadas frequentemente em várias apresentações de neoplasias malignas, incluindo o carcinoma epidermóide de laringe, e as elucidações inferidas destas observações podem trazer melhor entendimento à oncogênese. Objetivo: O objetivo deste estudo foi comparar a expressão imunoistoquímica de ciclina D1 e a ocorrência de metástase linfática em estádios III e IV de carcinoma epidermóide de laringe. Métodos: Estudo retrospectivo, coorte longitudinal, por avaliação imunoistoquímica e quantificação digital da imunoexpressão nuclear e citoplasmática de ciclina D1 em espécimes de tumor preservados em parafina oriundos de pacientes consecutivos submetidos à cirurgia oncológica radical, entre 1999 e 2004. A sobrevida global dos pacientes foi avaliada, bem como a idade, sexo, tabagismo, estado de comprometimento linfático, grau de diferenciação e estadiamento (pTNM). A análise estatística teve como significância valores de p< 0.05. A curva de sobrevida foi elaborada utilizando o método de Kaplan-Meier. Resultados: Houve imunomarcação citoplasmática em 566 (1,2%) células, imunomarcação nuclear em 13788 (29,6%) células, a relação do IPc e IPn foi de 0,007 (0,7%), ausência de imunomarcação celular foi observada em 32210 (69,1%) células, perfazendo um total de 46554 (100%) células investigadas. Entre os 28 (59,5%) casos que não apresentaram metástase linfática, o IPn foi de 26,8 (9,7 - 46,9) e o IPc foi de 0,1 (0 0,3); naqueles 19 (40,4%) em que foi observada metástase linfática, o IPn foi de 26,7 (16,7 39,0) e o IPc foi de 0,3 (0 - 1,0). Conclusões: Não houve associação estatisticamente significante entre expressão nuclear e citoplasmática de ciclina D1, em carcinoma epidermóide primário de laringe, e ocorrência de metástase linfática cervical, graus de diferenciação histológica, bem como recidiva loco-regional e metástase hematogênica. O presente estudo não subsidia a superexpressão de ciclina D1 como fator limitante de sobrevida global / Background: Cyclin D1 is an important regulator of cell cycle progression and can function as a transcriptional co-regulator. The overexpression of cyclin D1 has been linked to the development and progression of cancer. Abnormal cyclin D1 degradation appears to be responsible for the increased levels of cyclin D1 in several cancers. Recent findings have identified novel mechanisms involved in the regulation of cyclin D1 stability. Cyclin D1 belongs to the family of cyclin proteins which function as the regulatory subunits of cyclin/cyclin dependent kinase (Cdk) holoenzymes that regulate entry into and progression through the cell cycle. Cyclin D1 expression is induced upon stimulation by growth factors (e.g. EGF, IGF-I/II), aminoacids, hormones, and oncogenes such as Ras, Src, ErbB2, and SV40 T antigen. Cdk4 and Cdk6 can partner with cyclin D1 in early to mid-G1 phase to phosphorylate and inactivate the cell cycle inhibitory function of the retinoblastoma protein (pRB) in cooperation with cyclin E/Cdk2. Cyclin D1 is also known to modulate local chromatin structure and transcription of genes involved in proliferation and differentiation through CDK-independent association with histone acetylases (e.g. CBP, P/CAF) and deacetylases. Mutations, amplification or overexpression of cyclin D1, which alters cell cycle progression, are observed frequently in a variety of tumors, including laryngeal squamous cell carcinoma, and may contribute to oncogenesis. Methods: This was a retrospective study by immunohistochemical determination of cyclin D1 in fixated and paraffin-embedded tumour especimens from 47 consecutive patients with squamous cell carcinoma in larynx treated by curative oncological surgery from 1999 to 2004. Survival of patients was related to age, gender, nodal status and stage at termination of treatment. Significant differences were considered for p<0.05. Results: Cytoplasmic immunostain was observed in 566 (1,2%) cell, nuclear immunostain in 13788 (29,6%) cell, relationship between PIc and PIn was 0,007 (0,7%), absent cell immunostain was observed in 32210 (69,1%), and total 46554 (100%). Among 28 (59,5%) cases with no lymph node metastasis, PIn was 26,8 (9,7 - 46,9) and PIc was 0,1 (0 0,3); those 19 (40,4%) with lymph node metastasis, had a PIn of 26,7 (16,7 39,0) and PIc of 0,3 (0 -1,0). Conclusions: According to these results, it has been concluded that cyclin D1 showed nuclear and cytoplasmic expression in larynx squamous cell carcinoma; however, tumor cyclin D1 expression was not significantly associated with lymph node metastasis when quantified by quantitative or semiquantitative methods. Besides, cyclin D1 expression showed no influence in overall survival

The range and use of the tenor voice in classical music has long been established since the late 19th century. It is widely accepted among pedagogues that the range is C3-C5 (with obvious exceptions depending on the fach). However, with the advent and development of the American Musical as a genre since the early 20th century, the ‘tenor’ has taken on an entirely new direction and range altogether. Several well-known sources have stated that the ‘Broadway tenor’ has a range of A2-A4. This is (as it widely accepted in the classical profession) the range of a baritone. The catalyst of these changes include vaudeville, composers, social trends, and probably most important, the invention and proliferation of the microphone. This study will analyze a cross section of repertoire in order to demonstrate this downward shift of vocal range, and demonstrate some of the main reasons why this shift occurred.

Die Tumorausdehnung und das Vorliegen regionärer Lymphknotenmetastasen in Form der pT- und pN-Kategorie der TNM-Klassifikation sind unumstrittene Prognosefaktoren bei Patienten mit Plattenepithelkarzinomen des Kopf-Hals-Bereiches. Der Einfluss der Invasion von Lymphgefäßen (L-Klassifikation), Venen (V-Klassifikation) und Perineuralscheiden (Pn-Klassifikation) ist unklar und weniger häufig untersucht. In der vorliegenden Studie wurden die Daten von 608 Patienten des Tumorzentrum Leipzig mit Plattenepithelkarzinom der Mundhöhle, des Hypopharynx und des Larynx der Jahre 2005-2012 retrospektiv ausgewertet. Es erfolgte eine statistische Erhebung der relativen Überlebenswahrscheinlichkeit und der Vergleich der Prognose der unterschiedlichen Tumorlokalisationen anhand der pT-Kategorie, pN-Kategorie, L-Klassifikation, V-Klassifikation und Pn-Klassifikation des Tumors. Die negativen Auswirkungen zunehmender pT- und pN-Kategorie, sowie L-Klassifikation auf die Prognose war bei allen Lokalisationen signifikant, während die V-Klassifikation nur bei den Patienten mit Karzinom der Mundhöhle einen signifikanten prognostischen Wert zeigten. Die Pn-Klassifikation war nur für die Patienten mit Karzinom der Mundhöhle und des Larynx prognostisch relevant.:Inhaltsverzeichnis 1 Einleitung: 1.1 Definition 1 1.2 Epidemiologie und Ätiologie 1 1.3 Prognosefaktoren 3 1.4 Aktuelle multimodale Therapiekonzepte 4 2. Ziele der Arbeit 6 3. Material und Methoden 7 3.1 Patienten 7 3.1.1 Alter und Geschlecht 9 3.1.2 Tumorlokalisationen 9 3.1.3 Tumorstadien 9 3.2 Methoden 10 3.2.1 Datenerfassung 10 3.2.2 Datenauswertung 11 3.2.3 Methode der statistischen Auswertung 11 4. Ergebnisse 12 4.1 Deskriptive Statistik: Häufigkeiten nach Lokalisationen 12 4.1.1 pT-Kategorie: 12 4.1.2 pN-Kategorie: 13 4.1.3 L-Klassifikation: 14 4.1.4 V-Klassifikation: 15 4.1.5 Pn-Klassifikation: 16 4.2 Kaplan-Meier-Überlebenszeitanalysen 17 4.2.1 Relatives 5-Jahres-Überleben gesamt nach Lokalisation 17 4.2.2 Relatives 5-Jahresüberleben nach pT-Kategorie 20 4.2.3 Relatives 5-Jahresüberleben nach pN-Kategorie 23 4.2.4 Relatives 5-Jahresüberleben nach L-Klassifikation 26 4.2.5 Relatives 5-Jahresüberleben nach V-Klassifikation 29 4.2.6 Relatives 5-Jahresüberleben nach Pn-Klassifikation 32 5. Diskussion 35 5.1 Patienten 35 5.1.1 Alter 35 5.1.2 Geschlecht 37 5.1.3 Tumorlokalisationen 38 5.1.4 TNM-Parameter 41 5.1.4.1 pT-Kategorie Lokalisation Mundhöhle 41 5.1.4.2 pT-Kategorie Lokalisation Hypopharynx 41 5.1.4.3 pT-Kategorie Lokalisation Larynx 42 5.1.4.4 pN-Kategorie Lokalisation Mundhöhle 45 5.1.4.5 pN-Kategorie Lokalisation Hypopharynx 46 5.1.4.6 pN-Kategorie Lokalisation Larynx 46 5.1.5 Fakultative Deskriptoren 48 5.1.5.1 L-Klassifikation Lokalisation Mundhöhle 48 5.1.5.2 L-Klassifikation Lokalisation Hypopharynx 48 5.1.5.3 L-Klassifikation Lokalisation Larynx 48 5.1.5.4 V-Klassifikation Mundhöhle 48 5.1.5.5 V-Klassifikation Hypopharynx 49 5.1.5.6 V-Klassifikation Larynx 49 5.1.5.7 Pn-Klassifikation Mundhöhle 49 5.1.5.8 Pn-Klassifikation Hypopharynx 49 5.1.5.9 Pn-Klassifikation Larynx 49 5.2 Prognose nach Lokalisation 50 5.2.1 Prognose Mundhöhle ICD-C02-C06 50 5.2.2 Prognose Hypopharynx ICD-C12-C13 51 5.2.3 Prognose Larynx ICD-C32.0-C32.2, C10.1 51 5.2.4 Prognostische Bedeutung Tumorlokalisation 52 5.3 Prognose nach pT-Kategorie 53 5.3.1 Prognose nach pT-Kategorie Mundhöhle 53 5.3.2 Prognose nach pT-Kategorie Hypopharynx 53 5.3.3 Prognose nach pT-Kategorie Larynx 54 5.3.4 Prognostische Bedeutung pT-Kategorie 55 5.4 Prognose nach pN-Kategorie 57 5.4.1 Prognose nach pN-Kategorie Mundhöhle 57 5.4.2 Prognose nach pN-Kategorie Hypopharynx 58 5.4.3. Prognose nach pN-Kategorie Larynx 58 5.4.4 Prognostische Bedeutung pN-Kategorie 59 5.5 Prognose nach L-Klassifikation 60 5.5.1 Prognose nach L-Klassifikation alle Lokalisationen 60 5.5.2 Prognostische Bedeutung L-Klassifikation 60 5.6 Prognose nach V-Klassifikation 61 5.6.1 Prognose nach V-Klassifikation alle Lokalisationen 61 5.6.2 Prognostische Bedeutung V-Klassifikation 62 5.7 Prognose nach Pn-Klassifikation 63 5.7.1 Prognose nach Pn-Klassifikation alle Lokalisationen 63 5.7.2 Prognostische Bedeutung Pn-Klassifikation 64 5.8 Prognosevergleich der Patienten 65 5.9 Kritik an der vorliegenden Studie 67 Zusammenfassung 69 Literaturverzeichnis Abbildungsverzeichnis Tabellenverzeichnis Literaturverzeichnis Abbildungsverzeichnis Tabellenverzeichnis Erklärung zur eigenständigen Abfassung

Several studies involving excised animal larynges have been performed to simulate the structural and physiological properties of the human larynx. The most common way to preserve the laryngeal tissue being studied is by immersing it in a 0.9% isotonic saline solution and then flash freezing it. Isotonic saline is used empirically to replenish the potential ion loss that occurs postmortem. Each larynx is flash frozen so it can be used at a more convenient time while still maintaining the integrity of the tissue. However, the preservation methods found in previous studies tend to vary and no consensus had been reached about which method of preservation is ideal. This study sought to investigate the effects of solution and storage on phonation threshold pressure (PTP). Phonation threshold pressure is commonly used to investigate mucosal wave of the vocal folds, prephonatory glottal width, and vocal fold cover. This study involved a prospective, mixed experimental design with three groups, including a control group and two experimental groups. Each group consisted of 10 bench-mounted porcine larynges. The control group was immersed in 0.9% isotonic saline, flash frozen with liquid nitrogen within 24 hours postmortem, and thawed overnight before the experiment. The second group was immersed in 0.9% isotonic saline and the third group was submersed in Ringer's solution. Each of these groups was kept in their solution in a refrigerator for approximately 15 hours and was used for the experiment within 24 hours postmortem. Each larynx was mounted on a bench on a tabletop with three micropositioners to adduct and elongate the vocal folds. A pseudolung connected to the trachea directed humidified air to the vocal folds subglottally until phonation was achieved. The larynges in all three groups underwent these phonatory trials with 5-minute desiccation trials between each until phonation could no longer be achieved. Phonation threshold pressure was then observed and compared within groups and between groups. The signals were obtained using MATLAB. The results indicated that PTP was lowest for the frozen versus fresh groups. PTP values increased slightly for the frozen group, but the frozen group demonstrated less variability across specimens as compared to the fresh groups. Collectively, these results indicate that there are substantial differences between fresh and frozen specimens. These differences should be considered when designing tissue studies for purposes of generalization to human phonation.

Animal models are used extensively in voice research to study aspects of phonation, including physiology, kinematics, structure, and histology. Animals such as dog, cow, pig, sheep, deer, monkey, ferret, and rabbit have been used in voice research, with pig being one of the most common models. It is thought that the pig larynx is highly similar to the human larynx and one of the best models used in animal translational research. As with any model, however, the pig larynx does have some limitations. Perhaps a limitation most important to the rationale of this investigation is that pigs are difficult animals to study in vivo. Maintenance for a pig is challenging due to its large size and the variability of phonation use in the animal. Therefore, viable and practical alternatives are needed for in vivo voice research. The current study collected preliminary normative data from an alternate animal model, the rabbit, which has been used more recently in studies to model human phonation. The rabbit model was chosen due to its histological similarities to humans, in vivo phonation patterns, size, and practicality. The rabbit represents a more practical model for some longitudinal designs, as well as ex vivo phonatory models with aerodynamic measures as the primary variables. The current study involved a comparison of two aerodynamic measures, specifically phonation threshold pressure (PTP) and phonation threshold flow (PTF) between two groups, pig and rabbit larynges. The purpose of this study was to determine normative aerodynamic values for rabbits and to compare these with normative values for pigs during excised larynx benchtop phonation. Each group consisted of 15 larynges that were finely dissected to reveal the true vocal folds. Each larynx was then connected to a pseudolung and humidified air was passed through it. Fifteen phonation trials were elicited and the results averaged for each larynx. The results indicated that PTP and PTF were significantly different between the two groups, with PTP and PTF being lower for the rabbit group. Additionally, PTP values for rabbits were closer than pigs to the typical human value; however, some methodological challenges to rabbit benchtop models, including size and structural integrity, also exist. But the results from this study indicate that rabbits should be considered a viable option for voice research that would be more feasible with a small animal option that translates well to humans than a large animal option.

This thesis examined the effects of combination inhaled corticosteroids (ICs) on the stability of six aerodynamic measures of phonation utilizing a traditional benchtop model with leporine larynges. The motivation for this study was based on the increase of voice disorders associated with IC use in recent years. The aerodynamic measures examined were phonation threshold pressure (PTP), phonation threshold flow (PTF), onset resistance, sustained pressure, sustained flow, and sustained resistance. Leporine larynges were selected as the model for this study due to histological similarities between leporine and human vocal folds that make them ideal for translational research. Rabbits were either exposed to saline solution or ICs for 8 weeks before being sacrificed. After being sacrificed, larynges were excised and dissected. After dissection, the larynges were mounted on a benchtop, the aerodynamic data were gathered, and stability over multiple phonation trials was calculated. The results indicate that the variation between individual rabbits across the measures did not differ significantly. However, after controlling for trial, the average variation of the groups across all trials did differ significantly. PTP and sustained pressure were more variable for the inhaler group, while PTF, sustained flow, onset resistance, and sustained resistance were more variable for the control group. These results suggest that some level of variability in aerodynamic measures both within and between subjects is to be expected when using the leporine benchtop model. Furthermore, while IC exposure does not seem to impact within-subject variability, it does influence between-subjects variability.

Combination corticosteroid inhalers are the primary treatment option for long-term pulmonary disorders including asthma, persistent bronchitis, and chronic obstructive pulmonary disease. Common side effects of these medications are xerostomia in the mouth and throat, hoarseness, and soreness in the oropharynx. Research indicates that a large percentage of the inhaler particles are deposited onto laryngeal tissue, leaving an alteration of laryngeal mucosal properties. As the first stage in a long-term project, this thesis addresses the need for baseline phonatory data that will lay groundwork for quantifying inhaler-induced phonatory changes. Excised larynx research is a powerful tool for assessing aerodynamic alterations that accompany laryngeal pathology. Porcine (pig) larynges are a traditional species employed in voice disorder research, though leporine (rabbit) larynges are an emerging species that lends itself to histologic vocal fold studies as they have the most similar vocal fold cover structure to humans compared to any other animal to date. The purpose of this study was to examine the measurement stability of six aerodynamic parameters in a traditional excised larynx benchtop model. Specifically, the current author assessed measurement stability of leporine larynges compared to porcine larynges with the following aerodynamic metrics: phonation onset pressure (PTP; cmH2O), phonation onset flow (PTF; L/m), sustained pressure (cmH2O), sustained flow (L/m), onset laryngeal resistance (cmH2O/L/m), and sustained laryngeal resistance (cmH2O/L/m). A total of 30 larynges—15 leporine and 15 porcine—were mounted on a benchtop setup; phonation was sampled over 15 trials for each larynx. Measurement stability for the above six tokens was examined using coefficient of variation (%) analyses. Leporine larynges demonstrated significantly less variation across all six aerodynamic parameters when compared to porcine larynges. The leporine PTP values were most stable as compared to leporine and porcine pressure and airflow values. Leporine airflow values were also more stable than porcine PTP and PTF values. These results indicate that leporine larynges might be a preferred excised larynx specimen for certain benchtop phonation studies. These findings are important for establishing expected measurement variability in porcine and leporine larynges, particularly when translating benchtop research to laryngeal pathology.

Laryngeal cancer affects tens of thousands worldwide every year. The standard of care of surgical resection, chemotherapy, and/or radiation therapy results in significant quality of life deficits including reliance on tracheostomy tubes, loss of voice, and inability to swallow. There are no therapeutic options that restore a functional larynx so that patients can live a more normal life. Laryngeal reconstruction using tissue engineering strategies offers the potential to solve this problem. Laryngeal anatomy is complex with multiple tissue types and therefore engineering approaches require consideration of a multi-layer, interfacial tissue design. Our strategy to overcome these challenges involves the use of advanced bio fabrication techniques where type I oligomeric collagen alone or in the presence of autologous stem cells is used to custom-make the cartilage, skeletal muscle, and mucosal layer of the larynx. This doctoral research project begins by describing the development of the tissue engineered skeletal muscle with aligned collagen matrix and autologous muscle progenitor cells induced to express motor endplates. Next, using this engineered muscle plus the cartilage layer developed by a colleague; we implanted the myochondral engineered construct in a rat hemilaryngectomy model. In this study we saw host-implant integration with no inflammatory foreign body response, neo cartilage and muscle formation, and some return of laryngeal function on the reconstructed side. Next, we worked to scale-up these technologies for use in a porcine model. The pig larynx is more similar in size and function to the human larynx and allows for a full thickness defect to be created. Using confined compression, we created 4-mm thick acellular and cellular cartilage constructs, as well as a 0.5-mm thick acellular mucosal layer. A 1-cm diameter muscle layer containing autologous muscle progenitor cells was created using flow alignment and cultured to induce expression of motor endplates before implantation. Tissue constructs were subjected to mechanical property analyses as well as PCR analysis to describe the differential gene expression by component cells within muscle and cartilage constructs. Each layer was individually sutured into a pig hemilaryngectomy model. The pigs recovered well from the surgery, were eating, had no difficulty breathing, and no aspiration events. At 2 months, respiratory epithelium had completely healed over the implant and was vascularized and had areas of submucosal gland growth. The motor endplate expressing muscle implants showed new, organized muscle ingrowth while the acellular implants showed a relative paucity of new, disorganized muscle. This work represents a significant advancement in the field of laryngeal reconstruction and is a first of its kind to use scalable tissue engineering technologies designed to specifically meet each layer’s functional criteria.