Hyaluronic Acid Concentration in Female Vocal Folds With Reinke's Edema.

OBJECTIVE: The aim of the present study was to investigate hyaluronic acid (HA) concentrations in vocal folds among patients with Reinke's edema. STUDY DESIGN: Prospective and experimental study. SETTING: Single tertiary center. METHODS: An HA binding protein isolated from bovine nasal cartilage was used to identify and isolate the HA from samples. Plates coated with biotin-conjugated binding protein and streptavidin-europium conjugate were sequentially incubated with 18 Reinke's edema samples and 11 female vocal fold cover samples from cadavers (the superficial layer of the lamina propria; control group). After the release of europium from streptavidin in enhancement solution, final fluorescence was measured in a fluorometer. RESULTS: The mean HA concentration in Reinke's edema vocal folds was significantly higher than that in the control vocal folds (9.2 × 103 vs 0.9 × 103µg/g). CONCLUSION: Vocal fold covers affected by Reinke's edema present a higher concentration of HA than do vocal fold covers with no edema.

Sulfated glycosaminoglycans in human vocal fold lamina propria / Glicosaminoglicanos sulfatados na lâmina própria de prega vocal humana

Abstract Introduction: The distribution, concentration and function of glycosaminoglycans in the various vocal fold tissues are still unclear. Objective: To evaluate the distribution and concentration of sulfated glycosaminoglycans in different layers of the human vocal fold according to gender and age. Methods: We used 11 vocal folds obtained from cadavers (7 men and 4 women) with no laryngeal lesion, less than 12 h after death, and aged between 35 and 98 years. The folds underwent glycosaminoglycans extraction from the cover and ligament, and post-electrophoresis analysis. Data were compared according to the layer, age and gender. Results: The concentration of dermatan sulfate was significantly higher in all layers. No differences were observed in the total concentrations of glycosaminoglycans in layers studied according to gender. It is significantly lower in the cover of individuals aged below 60 years. Conclusion: Dermatan sulfate, chondroitin sulfate, and heparan sulfate were observed in the human vocal folds cover and ligament of both genders, with the concentration of dermatan sulfate being significantly higher in all layers. Glycosaminoglycans concentration on the cover is significantly lower in individuals below 60 years compared with elderly.

Resumo Introdução: A distribuição, concentração e função dos glicosaminoglicanos nos diversos tecidos da prega vocal ainda não está esclarecida. Objetivo: Avaliar a distribuição e concentração dos glicosaminoglicanos sulfatados nas diferentes camadas da prega vocal humana de acordo com o sexo e a idade. Método: Foram usadas 11 pregas vocais obtidas de cadáveres (sete homens e quatro mulheres) sem lesão de laringe, com menos de 12 horas de óbito e entre 35 e 98 anos. As pregas foram submetidas à extração de glicosaminoglicanos da cobertura e ligamento e leitura pós-eletroforese. Os dados foram comparados segundo camada, idade e sexo. Resultados: A concentração de dermatan sulfato foi significativamente maior em todas as camadas. Não foram observadas diferenças nas concentrações totais de glicosaminoglicanos nas camadas estudadas quanto ao gênero. É significantemente menor em indivíduos abaixo de 60 anos na cobertura. Conclusão: Dermatam sulfato, condroitim sulfato e heparam sulfato foram observados na cobertura e no ligamento de pregas vocais humanas, de ambos os sexos, sendo a concentração de dermatam sulfato foi significativamente maior em todas as camadas. A concentração de glicosaminoglicanos na cobertura é significativamente menor em indivíduos abaixo de 60 anos em comparação com idosos.

Sulfated glycosaminoglycans in human vocal fold lamina propria.

INTRODUCTION: The distribution, concentration and function of glycosaminoglycans in the various vocal fold tissues are still unclear. OBJECTIVE: To evaluate the distribution and concentration of sulfated glycosaminoglycans in different layers of the human vocal fold according to gender and age. METHODS: We used 11 vocal folds obtained from cadavers (7 men and 4 women) with no laryngeal lesion, less than 12h after death, and aged between 35 and 98 years. The folds underwent glycosaminoglycans extraction from the cover and ligament, and post-electrophoresis analysis. Data were compared according to the layer, age and gender. RESULTS: The concentration of dermatan sulfate was significantly higher in all layers. No differences were observed in the total concentrations of glycosaminoglycans in layers studied according to gender. It is significantly lower in the cover of individuals aged below 60 years. CONCLUSION: Dermatan sulfate, chondroitin sulfate, and heparan sulfate were observed in the human vocal folds cover and ligament of both genders, with the concentration of dermatan sulfate being significantly higher in all layers. Glycosaminoglycans concentration on the cover is significantly lower in individuals below 60 years compared with elderly.

GLYCOSAMINOGLYCANS AND PROTEOGLYCANS IN PALMAR FASCIA OF PATIENTS WITH DUPUYTREN.

OBJECTIVE: : To evaluate and compare the behavior of glycosaminoglycans (GAGs) in Dupuytren disease (DD). METHODS: : This is an experimental study with 23 patients diagnosed with DD. Tissue collected through fasciectomy with incision type Brunner or McCash were evaluated by electrophoresis for identification of GAGs. The quantification was carried out by immunofluorescence and dosage of proteins for different types of glycosaminoglycans. The results were expressed in percentage and statistically evaluated. RESULTS: : A significant increase was observed through eletrophoresis in GAGs, as compared to the control (p<0.05). Immunofluorescence of hyaluronic acid was reduced (23 times) when compared to the control (p<0.0001). CONCLUSION: : An increase of sulfated GAGs in Dupuytren's disease, mainly dermatan sulfate, was evident from our results, as well as a pronounced decrease of hyaluronic acid in the palmar aponeurosis from the same patients. Level of Evidence III, Case-Control Study.

Concentration of hyaluronic acid in human vocal folds in young and old subjects.

OBJECTIVE: To compare and evaluate concentrations of hyaluronic acid (HA) in normal vocal folds of both young and elderly people of both sexes. STUDY DESIGN: Experimental study. SETTING: The research was carried out at the ENT, Head and Neck Surgery Department and the Pharmacology Institute of São Paulo Federal University. SUBJECTS AND METHODS: Enzyme-linked immunosorbent assay plates coated with biotin-conjugated binding protein and streptavidin-europium conjugate were sequentially incubated with samples from the middle portion of the cover, vocal ligament, and vocal muscle of 19 young and 20 elderly subjects of both sexes. After the release of europium in an enhancement solution, final fluorescence was measured using a fluorometer. RESULTS: Statistically significant comparisons: (1) all subject groups combined exhibited a higher HA concentration in the cover and ligament compared with the muscle samples; (2) women subjects had a significantly lower HA concentration in muscle compared with the ligament, and male subjects showed significantly lower HA in muscle compared with the cover; and (3) both genders of young subjects showed a higher HA concentration in ligament samples compared with the muscle. CONCLUSION: HA concentrations are higher in the cover and ligament compared with the muscle in both genders and age groups, and there is a higher HA concentration in ligament compared with the muscle of young subjects of both genders. HA levels in the cover samples of younger women showed great variability that may relate to ovarian hormone levels, reflecting different phases of the menstrual cycle.

Analysis of glycosaminoglycans in the parametrium and vaginal apex of women with and without uterine prolapse.

BACKGROUND: Pelvic organ prolapse (POP) is a downward descent of pelvic organs that results in protrusions of the vagina, the uterus, or both. The cause of this disorder is likely to be multifactorial, attributable to a combination of risk factors, especially connective tissue disorders. Our objective was to characterize and quantify a component of the extracellular matrix (ECM)-sulfated glycosaminoglycan (GAG)-in the parametrium and vaginal apex of women with and without uterine prolapse. METHODS: Parametrium and vaginal apex tissue was obtained from 42 women who underwent surgery. Patients underwent a physical examination and were divided into groups according to the type of genital prolapse. Standard biopsies were taken during surgery and were assessed by biochemical methods. GAGs were obtained by proteolysis. The relative concentration of GAGs was determined by densitometry. Data were compared using an independent sample t-test or chi(2) test. RESULTS: In both groups (with and without prolapse) and in both types of tissue, dermatan sulfate (DS) was the most predominant glycosaminoglycan, followed by chondroitin sulfate (CS) and heparan sulfate (HS). We did not observe significant differences in the total amounts of GAGs, DS, CS, or HS. CONCLUSIONS: This study did not show altered biochemical characteristics in the ECM of parametrium and vaginal apex tissue of women either with or without uterine prolapse.

[Evaluation of glycosaminoglycans of periurethral tissue in patients with and without pelvic organ prolapse]. / Avaliação dos glicosaminoglicanos do tecido periuretral de pacientes com e sem prolapso genital.

OBJECTIVE: To characterize and quantify periurethral tissue sulphated glycosaminoglycans (GAGs) in women with and without pelvic organ prolapse. STUDY DESIGN: Periurethral tissue was obtained from 35 women who underwent surgery for pelvic organ prolapse, for stress urinary incontinence, or for other gynecological benign conditions. Patients were submitted to a clinical history, physical and urodynamic examination and were divided in two groups according to genital prolapse. The standard biopsy with 1.0 x 1.0 cm was taken from periurethral tissue during surgery and assessed by biochemical methods. The GAGs were obtained by proteolysis and precipitated by trichloroacetic acid. The relative concentration of sulfated GAGs was determined by densitometry of toluidine blue stained gel using a spectrophotometer with a 525 nm wavelength. Data were compared using analysis of variance (ANOVA). RESULTS: In the two groups dermatan sulphate (DS) was the predominant glycosaminoglycan (85%), followed by chondroitin sulphate (CS) and heparan sulphate (HS). Women with pelvic organ prolapse had significantly more total GAGs, DS and HS. Differences in CS were not observed. CONCLUSIONS: This study showed altered biochemical characteristics in the extracellular matrix of periurethral tissue and also accumulation of GAGs, DS and CS, in women with pelvic organ prolapse.

Avaliação dos glicosaminoglicanos do tecido periuretral de pacientes com e sem prolapso genital / Evaluation of glycosaminoglycans of periurethral tissue in patients with and without pelvic organ prolapse

OBJETIVOS: Caracterizar e quantificar os subtipos de glicosaminoglicanos sulfatados (GAGs) existentes no tecido peri-uretral de pacientes com e sem prolapso genital. METODOS: Foram incluídas 35 pacientes que se submeteram a cirurgia vaginal para correção de distopias genitais e/ou incontinência urinária de esforço ou por outra condição benigna. As pacientes foram avaliadas por anamnese padronizada, exame físico e urodinâmico e agrupadas segundo a existência do prolapso genital. Durante o procedimento cirúrgico, amostras de aproximadamente 1,0 x 1,0 cm do tecido periuretral foram retiradas para avaliação. Os GAGs foram extraídos do tecido por proteólise e precipitação por ácido tricloroacético e caracterizados por eletroforese em gel de agarose. A quantificação foi feita por meio de densitometria a 525 nm do gel corado com azul de toluidina. Compararam-se os dados pela análise de variância (ANOVA). RESULTADOS: Nos grupos estudados, houve maior predomínio de dermatam sulfato (DS), em torno de 85 por cento do total de GAGs, seguido do condroitim sulfato (CS) e do heparam sulfato (HS). Observou-se aumento significativo dos GAGs totais, do DS e do HS em mulheres com prolapso genital. Não se observou diferença significante com relação ao CS. CONCLUSÃO: Este estudo demonstrou diferenças na matriz extracelular do tecido periuretral com aumento de GAGs totais, DS e HS nas mulheres com prolapso genital.

OBJECTIVE: To characterize and quantify periurethral tissue sulphated glycosaminoglycans (GAGs) in women with and without pelvic organ prolapse. STUDY DESIGN: Periurethral tissue was obtained from 35 women who underwent surgery for pelvic organ prolapse, for stress urinary incontinence, or for other gynecological benign conditions. Patients were submitted to a clinical history, physical and urodynamic examination and were divided in two groups according to genital prolapse. The standard biopsy with 1.0 x 1.0 cm was taken from periurethral tissue during surgery and assessed by biochemical methods. The GAGs were obtained by proteolysis and precipitated by trichloroacetic acid. The relative concentration of sulfated GAGs was determined by densitometry of toluidine blue stained gel using a spectrophotometer with a 525 nm wavelength. Data were compared using analysis of variance (ANOVA). RESULTS: In the two groups dermatan sulphate (DS) was the predominant glycosaminoglycan (85 percent), followed by chondroitin sulphate (CS) and heparan sulphate (HS). Women with pelvic organ prolapse had significantly more total GAGs, DS and HS. Differences in CS were not observed. CONCLUSIONS: This study showed altered biochemical characteristics in the extracellular matrix of periurethral tissue and also accumulation of GAGs, DS and CS, in women with pelvic organ prolapse.