Three-dimensional cell culture for the study of nasal polyps

Abstract Objectives: Three-dimensional (3D) cell cultures have many applications such as stem cell biology research, new drug discovery, cancer, and Chronic Rhinosinusitis with Nasal Polyps (CRSwNP). This disease is characterized by a significant impact on quality of life and productivity. The diversity of factors that act in the progression of CRSwNP point to the creation of a cell culture model that allows the integration of different cell types with extracellular matrix. This work aimed to create a cell culture model in 3 dimensions (spheroids) for the study of Nasal Polyposis. Methods: Nasal polyp tissue from patients diagnosed with CRSwNP was mechanically dissociated using tweezers and a scalpel and the solution containing cells and small aggregates of nasal polyps was transferred to a Petri dish containing 5 mL of culture medium at the concentration of 106 cells/mL. Results: The spheroids were cultivated for 20 days, fixed and analyzed using confocal microscopy. In a 3D culture environment, the spheroids were formed both by clustering cells and from small tissue fragments. In the cultures analyzed, the ciliary beat was present from the dissociation of the cells up to 20 days in culture. Conclusion: Our findings also point to these characteristics showing the environment generated in our study, the cells remained differentiated for a longer time and with ciliary beating. Thus, this work shows that nasal polyp-derived cells can be maintained in a 3D environment, enabling better strategies for understanding CRSwNP in situations similar to those found in vivo. Level of evidence: Laboratory studies.

CD133, a Progenitor Cell Marker, is Reduced in Nasal Polyposis and Showed Significant Correlations with TGF-β1 and IL-8

Abstract Introduction Combination of chronic inflammation and an altered tissue remodeling process are involved in the development of Chronic Rhinosinusitis with Nasal Polyps (CRSwNP). Studies demonstrated that mesenchymal stem cells expressing the progenitor gene CD133 were involved in a significant reduction of the chronic inflammatory process in the polypoid tissue. Objective To evaluate the levels of CD133 (Prominin-1) in nasal polypoid tissue and its correlation with interleukin-8 (IL-8) and transforming growth factor β1 (TGF-β1). Methods A total of 74 subjects were divided in the following groups: control group (n = 35); chronic rhinosinusitis with nasal polyps nonpresenting comorbid asthma and aspirin intolerance (CRSwNPnonAI) group (n = 27); and chronic rhinosinusitis with nasal polyps presenting comorbid asthma and aspirin intolerance (CRSwNPAI) group (n = 12). Histologic analysis and also evaluation of the concentration of CD133, IL-8, and TGF-β1 by enzyme-linked immunosorbent assay (ELISA) kits were performed in nasal tissue obtained from nasal polypectomy or from middle turbinate tissue. Results Higher eosinophilic infiltration was found in both CRSwNP groups by histologic analysis. Lower levels of TGF-β1 and IL-8 were observed in both CRSwNP groups when compared with the control group, whereas the CD133 levels were significantly reduced only in the CRSwNPnonAI group compared with the control group. Conclusion It was demonstrated that the nasal mucosa presenting polyposis showed a significant reduction of CD133 levels, and also that this reduction was significantly correlated with the reduction of TGF-β1 levels, but not with IL-8 levels. Therefore, these findings may be involved in the altered inflammatory and remodeling processes observed in the nasal polyposis.

Evaluation of palatal dimensions in different facial patterns by using digital dental casts

ABSTRACT Objective: To analyze the variations of hard palate volume in adults with normal occlusion and different facial types and patterns, by using a three-dimensional analysis on digital casts. Methods: The dental casts of 70 Caucasian adults (28 men, 42 women), mean age of 16.4 years (SD 1.3 years), were scanned by using a tridimensional scanner (Delcam PowerSHAPE™, 2010, Birmingham, UK). Close points were selected in the gingival and cervical regions on the lingual surface of the maxillary teeth, to analyze palatal morphology. The facial patterns and types, and the measurements (width, length, height, volume) of the space on the hard palate were compared using analysis of covariance (ANCOVA), with age as the covariate, and sex as the independent variable. The significance level of 5% (p < 0.05) was adopted. Results: This study showed that the measurements of the width and length were similar among the mesofacial, dolichofacial and brachyfacial facial types, although the height and volume of the space on the hard palate were slightly smaller in dolichofacial individuals, and both Pattern I and Pattern II individuals showed no significant changes for the four measurements. The mean values among facial patterns were: Pattern I - width 38.31±2.59 mm; length 37.44±2.42 mm; height 17.03±2.42 mm and volume 10.52±1.72 mm3; Pattern II - width 37.48±2.44 mm; length 37.48±2.44 mm; height 16.79±2.42 mm and volume 10.41±1.65 mm3 (p>0.05 for all variables). Conclusion: There were no significant differences for the facial patterns and facial types of the individuals compared in the analyzed sample.

RESUMO Objetivo: Analisar as variações do volume do palato duro em adultos com oclusão normal e diferentes tipos e padrões faciais, por meio de análise tridimensional em modelos digitais. Métodos: Os modelos das arcadas dentárias de 70 adultos caucasianos (28 homens, 42 mulheres), média de idade de 16,4 anos (DP 1,3 anos), foram digitalizados usando um scanner tridimensional (Delcam PowerSHAPE™, 2010, Birmingham, Reino Unido). Pontos próximos foram selecionados nas regiões gengival e cervical na superfície lingual dos dentes superiores, para analisar a morfologia palatina. Os padrões e tipos faciais e as medidas (largura, comprimento, altura, volume) do espaço no palato duro foram comparados por meio de análise de covariância (ANCOVA), com a idade como covariável e o sexo como variável independente, a um nível de significância de 5% (p< 0,05). Resultados: O presente estudo mostrou que as medidas de largura e comprimento foram semelhantes entre os tipos faciais mesofacial, dolicofacial e braquifacial, embora a altura e o volume do espaço no palato duro tenham sido ligeiramente menores nos indivíduos dolicofaciais, e tanto os indivíduos do Padrão I quanto do Padrão II não apresentaram alterações significativas para as quatro medidas. Os valores médios entre os padrões faciais foram: Padrão I - largura 38,31±2,59 mm, comprimento 37,44±2,42 mm, altura 17,03±2,42 mm, volume 10,52±1,72 mm3; Padrão II - largura 37,48±2,44 mm, comprimento 37,48±2,44 mm, altura 16,79±2,42 mm, volume 10,41±1,65 mm3 (p>0,05 para todas as variáveis). Conclusão: Não houve diferenças significativas para os padrões faciais e tipos faciais dos indivíduos comparados na amostra analisada.

Relationship between Anxiety and Interleukin 10 in Female Soccer Players with and Without Premenstrual Syndrome (PMS) / Relação dos níveis de ansiedade e da interleucina 10 em jogadoras de futebol de campo com e sem síndrome pré-menstrual (SPM)

Abstract Objective To investigate the level of anxiety and its relationship with interleukin (IL)- 10 (anti inflammatory cytokine that modulates mood swings) in a group of female soccer players. Methods Fifty-two eumenorrheic soccer players were evaluated (age 19.8 ± 4.7 years). The presence of premenstrual syndrome (PMS) and phases of the menstrual cycle were determined by a daily symptomreport (DSR) kept for 3 consecutivemonths. The concentration of cytokine IL-10 was determined from urine samples collected at four moments: at the follicular and luteal phases of the menstrual cycle, and before (pre) and after (post) the simulated game, and it was quantified by flow cytometry (Luminex xMAP - EMDMillipore, Billerica, MA, USA). The level of anxietywas determined through the BAI anxiety questionnaire answered by all athletes at the same time of the urine collection. The Student t-test, analysis of variance (ANOVA) and Pearson correlation with significance level at 5% were used for data analysis. Results We showed that the prevalence of PMS among female soccer players is similar to that reported in the literature. In addition,we showed that the group withPMS has a higher level of anxiety compared with group without PMS (p = 0.002). Interleukin-10 analysis in players without PMS revealed that there was a significant decrease in the level of this cytokine before the game during the luteal phase when compared with the follicular phase (p < 0.05). The correlation analysis between IL-10 and anxiety showed a negative correlation post-game in the luteal phase in the group without PMS (p = 0.02; r = -0.50) and a positive correlation post-game in the luteal phase in PMS group (p = 0.04; r = 0.36). Conclusion Our results suggest that IL-10 may contribute to reduce anxiety in the group without PMS. This could be attributed to the fact that no IL-10 variation was observed in the group with PMS, which presented higher anxiety symptoms when compared with the group without PMS.

Resumo Objetivo Investigar o nível de ansiedade e a sua relação com a interleucina 10 (IL-10) em jogadoras de futebol de campo com e sem síndrome pré-menstrual (SPM). A IL-10 é uma citocina anti-inflamatória que modula o humor e a ansiedade. Métodos Foram avaliadas 52 jogadoras de futebol de campo eumenorreicas (idade 19.8 ± 4.7 anos). A síndrome pré-menstrual (SPM) e as fases do ciclo menstrual foram determinadas pelo questionário de sintomas (DSR) por 3 meses consecutivos. A concentração da interleucina (IL)-10 foi obtida das amostras de urina coletadas em 4 momentos: nas fases folicular e lútea do ciclo menstrual e antes e depois do jogo, e foi quantificada por citometria de fluxo (Luminex xMAP - EMDMillipore, Billerica,MA, USA). O nível de ansiedade foi determinado pelo questionário de ansiedade de BAI, respondido por todas as atletas nos mesmos momentos das coletas de urina. O teste t de Student, a análise de variância (ANOVA) e a correlação de Pearson com nível de significância de 5% foram utilizados para a análise dos dados. Resultados Observou-se 59,6% de SPM nas jogadoras de futebol avaliadas, similar aos dados da literatura. O grupo com SPM possui um estado de ansiedade mais elevado quando comparado ao grupo sem SPM (p = 0,002). A IL-10 apresentou diminuição significante na fase lútea antes do jogo em comparação ao mesmo momento na fase folicular nas jogadoras sem SPM (p < 0,05). A correlação entre a IL-10 e a ansiedade revelou correlação negativa na fase lútea após o jogo no grupo sem SPM (p = 0,02; r = -0,50), bem como correlação positiva na fase lútea após o jogo no grupo com SPM (p = 0,04; r = 0,36). Conclusão Os resultados no grupo sem SPM evidenciam provável controle da ansiedade com a contribuição da IL-10. O grupo com SPM; com ansiedade maior que o grupo sem SPM; não teve variação significativa na IL-10, sugerindo dificuldade maior no controle da ansiedade nessas atletas.