Exploring the role of the WNT5A rs566926 polymorphism and its interactions in non-syndromic orofacial cleft: a multicenter study in Brazil

Abstract Associations between the WNT5A rs566926 variant and non-syndromic orofacial cleft (NSOC) have been reported in different populations. Objective This study aimed to investigate the role of the rs566926 single nucleotide polymorphism (SNP) in WNT5A and its interactions with SNPs in BMP4, FGFR1, GREM1, MMP2, and WNT3 in the occurrence of NSOC in a Brazilian population. Methodology A case-control genetic association study was carried out involving participants from four regions of Brazil, totaling 801 patients with non-syndromic cleft lip with or without cleft palate (NSCL±P), 273 patients with cleft palate only (NSCPO), and 881 health volunteers without any congenital condition (control). Applying TaqMan allelic discrimination assays, we evaluated WNT5A rs566926 in an ancestry-structured multiple logistic regression analysis, considering sex and genomic ancestry as covariates. Interactions between rs566926 and variants in genes involved in the WNT5A signaling pathway (BMP4, FGFR1, GREM1, MMP2, and WNT3) were also explored. Results WNT5A rs566926 was significantly associated with an increased risk of NSCL±P, particularly due to a strong association with non-syndromic cleft lip only (NSCLO), in which the C allele increased the risk by 32% (OR: 1.32, 95% CI: 1.04-1.67, p=0.01). According to the proportions of European and African genomic ancestry, the association of rs566926 reached significant levels only in patients with European ancestry. Multiple interactions were detected between WNT5A rs566926 and BMP4 rs2071047, GREM1 rs16969681 and rs16969862, and FGFR1 rs7829058. Conclusion The WNT5A rs566926 polymorphism was associated with NSCL±P, particularly in individuals with NSCLO and high European ancestry. Epistatic interactions involving WNT5A rs566926 and variants in BMP4, GREM1, and FGFR1 may contribute to the risk of NSCL±P in the Brazilian population.

Comparative study of two histopathological classifications for oral squamous cell carcinoma / Estudo comparativo de duas classificações histopatológicas para carcinoma de células escamosas bucal

ABSTRACT Introduction: Squamous cell carcinoma (SCC) is the most common tumor among all cancers in the oral cavity. Despite advances, the prognosis of this neoplasm remains a challenge for professionals. Faced with this situation, several studies try to associate the histopathological analysis with prognosis, so that therapeutic planning becomes more accurate. Objectives: This research aimed to conduct an epidemiological study of oral SCC and classify them histopathological assessment according to the World Health Organization (WHO) and the Budding and Depth of Invasion (BD) model. A retrospective research was conducted. Methodology: Data from medical records filed at UOPECCAN Hospital between 2009 and 2015 were analyzed. The sample consisted of 57 patients. Epidemiological data were collected and the blocks were rescued and cut for histopathological analysis. Associations were performed using the chi-square test with a significance level of 5% (p = 0.05) by the GraphPad Prism program. The two histopathological analyzes were correlated using Spearman's statistical test. Results: After analyzing the samples, we found a higher prevalence of oral SCC in male smokers aged above 40 years. There was no correlation between the BD and WHO methods. The WHO classification was significantly associated with age (p = 0.03), and follow-up care (p = 0.05). However, the BD model associated lymph node involvement (p = 0.005) and clinical staging (p = 0.005). Conclusion: The BD classification was more objective for histopathological analysis and may be an important tool for analyzing patient prognosis, assisting in the treatment decision.

RESUMEN Introducción: El carcinoma de células escamosas (CCE), denominado además carcinoma epidermoide, es el tumor más común entre todos los cánceres de la cavidad oral. A pesar de los avances, el pronóstico de esta neoplasia sigue siendo un desafío para los cirujanos/profesionales/clínicos. Ante esta situación, varios estudios intentan asociar el análisis histopatológico con el pronóstico, para que la planificación terapéutica sea más precisa. Objectivos: Esta investigación tuvo como objetivo realizar un estudio epidemiológico del CCE oral y clasificarlo histopatológicamente de acuerdo con la Organización Mundial de la Salud (OMS) y el modelo Budding and Depth of Invasion (BD). Se realizó una investigación retrospectiva. Metodología: Se analizaron los datos de las historias clínicas archivadas en el Hospital UOPECCAN entre 2009 y 2015. La muestra estuvo formada por 57 pacientes. Se recolectaron datos epidemiológicos y los bloques fueron rescatados y cortados para análisis histopatológico. Las asociaciones se realizaron mediante la prueba de chi-cuadrado con un nivel de significancia del 5% (p = 0.05) por el programa GraphPad Prism. Los dos análisis histopatológicos se correlacionaron mediante la prueba estadística de Spearman. Resultados: Tras analizar las muestras, encontramos una mayor prevalencia de CCE oral en varones fumadores mayores de 40 años. No hubo correlación entre los métodos BD y OMS. La clasificación de la OMS se asoció significativamente con la edad (p = 0,03) y seguimiento del del tratamiento (p = 0,05). Sin embargo, el modelo de BD asoció la afectación de los ganglios linfáticos (p = 0,005) y la estadificación clínica (p = 0,005). Conclusión: La clasificación BD fue más objetiva para el análisis histopatológico y puede ser una herramienta importante para analizar el pronóstico del paciente, asistiendo en la decisión del tratamiento.

RESUMO Introdução: O carcinoma de células escamosas (CCE) é o tumor mais frequente entre todos os cânceres localizados na cavidade bucal. Apesar dos avanços, o prognóstico dessa neoplasia ainda é um desafio para os cirurgiões. Diante dessa situação, vários estudos tentam associar a análise histopatológica ao prognóstico, a fim de que os planejamentos terapêuticos se tornem mais precisos. Objetivos: Esta pesquisa teve como objetivo realizar o estudo epidemiológico dos CCEs e classificá-los histopatologicamente conforme a Organização Mundial da Saúde (OMS) e o modelo "Budding and Depth of Invasion" (BD). Um estudo retrospectivo foi realizado. Metodologia: Foram analisados dados dos prontuários arquivados no Hospital UOPECCAN entre 2009 e 2015. A amostra foi composta por 57 pacientes. Os dados epidemiológicos foram coletados e os blocos resgatados e cortados para análise histopatológica. As associações foram realizadas por meio do teste qui-quadrado, com nível de significância de 5% (p = 0,05) pelo programa GraphPad Prism. As duas análises histopatológicas foram correlacionadas por meio do teste estatístico de Spearman. Resultados: Após análise das amostras, verificamos mais prevalência de CCE nos pacientes fumantes do sexo masculino com idade superior a 40 anos. Não houve correlação entre os métodos BD e OMS. A classificação da OMS apresentou associação significante com a idade (p = 0,03) e a sequência de tratamento (p = 0,05). Já o modelo BD associou comprometimento linfonodal (p = 0,005) e estadiamento clínico (p = 0,005). Conclusão: A classificação BD foi mais objetiva para a análise histopatológica e pode ser uma importante ferramenta para análise do prognóstico do paciente, auxiliando na decisão do tratamento.

Association between maternal smoking, gender, and cleft lip and palate / Associação entre tabagismo materno, gênero e fendas labiopalatinas

ABSTRACT INTRODUCTION: Cleft lip and/or palate (CL/P) represent the most common congenital anomalies of the face. OBJECTIVE: To assess the relationship between maternal smoking, gender and CL/P. METHODS: This is an epidemiological cross-sectional study. We interviewed 1519 mothers divided into two groups: Cases: mothers of children with CL/P (n = 843) and Controls: mothers of children without CL/P (n = 676). All mothers were classified as smoker or non-smoker subjects during the first trimester of pregnancy. To determine an association among maternal smoking, gender, and CL/P, odds ratios were calculated and the adjustment was made by a logistic regression model. RESULTS: An association between maternal smoking and the presence of cleft was observed. There was also a strong association between male gender and the presence of cleft (OR = 3.51; 95% CI 2.83-4.37). By binary logistic regression analysis, it was demonstrated that both variables were independently associated with clefts. In a multivariate analysis, male gender and maternal smoking had a 2.5- and a 1.5-time greater chance of having a cleft, respectively. CONCLUSION: Our findings are consistent with a positive association between maternal smoking during pregnancy and CL/P in male gender. The results support the importance of smoking prevention and introduction of cessation programs among women with childbearing potential.

RESUMO Introdução: Fendas labiais e/ou palatinas (FL/P) representam as anomalias congênitas mais comuns da face. Objetivo: Avaliar a relação entre tabagismo materno, gênero e FL/P. Método: Realizou-se um estudo epidemiológico, de corte transversal. Foram entrevistadas 1.519 mães, divididas em dois grupos: Casos: mães de crianças com FL/P (n = 843); e Controles: mães de crianças sem FL/P (n = 676). Todas as mães foram classificadas como fumantes ou não fumantes durante o primeiro trimestre de gravidez. Para determinar a associação entre tabagismo materno, gênero e FL/P, odds ratios foram calculadas e o ajuste realizado pelo modelo de regressão logística. Resultados: Observou-se associação entre tabagismo materno, e fendas. Houve também forte associação entre sexo masculino e presença de fendas (OR = 3,51; 95% IC 2,83-4,37). Regressão logística binária demonstrou que ambas as variáveis foram independentemente associadas coma ocorrência de fendas. Na análise multivariada, o sexo masculino teve 2,5 vezes mais chance de apresentar fendas e tabagismo materno teve 1,5 vez mais chance dessa ocorrência. Conclusão: Os resultados são consistentes com a associação positiva entre tabagismo materno durante a gravidez e a ocorrência de FL/P no gênero masculino. Os resultados suportam a importância da prevenção do tabagismo e a aplicação de programas entre mulheres com potencial de gravidez.

Uncommon Oral Cleft in Wolf-Hirschhorn Syndrome

Wolf-Hirschhorn syndrome (WHS) is a syndrome with craniofacial and systemic abnormalities, which is related to 4p deletion. A 3-month old girl with an undiagnosed syndrome was referred for evaluation of the cleft lip and palate. Hypotonia, short stature, cardiac malformation, hypertrophied clitoris, and atypical thumb of both hands was observed. Microcephaly, low-set ear, prominent glabella, downslanting palpebral fissures, a characteristic "Greek warrior helmet" appearance, micrognathia, ears with pits/tags and bilateral incomplete cleft lip apart from incomplete cleft palate were observed as craniofacial findings. With clinical diagnosis of WHS, blood was subjected to karyotyping, which showed a 4p15.2 deletion, consistent with the condition. Here is reported the case of this WHS patient with an uncommon oral cleft extending the phenotypic spectrum of the disorder. The child was referred to a multidisciplinary team to reparative surgery of the cleft lip and palate. The patient is on regular medical follow-up and will be further assisted by dentists, physical therapists, occupational therapists and psychologists. The genotype-phenotype correlation of the affected patient with previous WSH syndrome reports is described.

A síndrome de Wolf-Hirschhorn (WHS) é uma condição genética caracterizada por anomalias craniofaciais e sistêmicas, causada por deleção cromossômica na região 4p. Paciente de 3 meses de idade, gênero feminino, foi encaminhada para avaliação de fissura de lábio e fissura palatina, associada a uma síndrome não diagnosticada. A paciente apresentava-se com hipotonia, baixa estatura, malformação cardíaca, clitóris hipertrofiado e implantação atípica do polegar nas duas mãos. Microcefalia, baixa implantação da orelha, glabela proeminente, inclinação baixa das fissuras palpebrais, aparência característica de capacete de guerreiro grego, micrognatia, fossetas em orelhas, fissura labial bilateral incompleta e fissura palatina incompleta foram observadas como características craniofaciais. Com um diagnóstico clínico de WHS, foi realizado o cariótipo, que mostrou a deleção 4p15.2, consistente com a condição. Esse relato de caso apresenta um caso de WHS, com uma fissura oral incomum, ampliando o espectro fenotípico da doença. A paciente foi encaminhada a tratamento com equipe multidisciplinar para correção cirúrgica da fissura labial e palatina. Encontra-se em acompanhamento médico bem como odontológico, fisioterapêutico e em terapia ocupacional e psicológica. Uma correlação entre genótipo e fenótipo pode ser observada nesse relato da síndrome de WHS.

Minichromosome maintenance 2 and 5 expressions are increased in the epithelium of hereditary gingival fibromatosis associated with dental abnormalities

INTRODUCTION: Gingiva fibromatosis is a relatively rare condition characterized by diffuse enlargement of the gingiva, which is caused by expansion and accumulation of the connective tissue. OBJECTIVE: The aim of the present study was to investigate proliferative and apoptotic biomarker expression in normal gingiva and two forms of gingival fibromatosis. METHODS: Archived tissue specimens of hereditary gingival fibromatosis, gingival fibromatosis and dental abnormality syndrome and normal gingiva were subject to morphological analysis and immunohistochemical staining. The results were analyzed statistically. RESULTS: Proteins associated with proliferation were found in the nuclei of epithelial cells from the basal and suprabasal layers, whereas apoptotic proteins were detected in the cytoplasm of the upper layers of the epithelium. Increased expressions of minichromosome maintenance proteins 2 and 5 were observed in the gingival fibromatosis and dental abnormality syndrome samples. In contrast, geminin expression was higher in normal gingiva samples. No difference in the expression of apoptotic proteins was observed among the groups. CONCLUSION: Our findings support a role for augmented proliferation of epithelial cells within the overgrown tissues associated with gingival fibromatosis or dental abnormality syndrome. However, our data suggest that different biological mechanisms may account for the pathogenesis of different types of gingival fibromatosis.

Goldenhar syndrome: clinical features with orofacial emphasis

OBJECTIVES: Goldenhar syndrome (GS) is a relatively common developmental disorder characterized by craniofacial anomalies in association with vertebral, cardiac, renal, and central nervous system defects. This paper describes GS features with special emphasis on oral characteristics. MATERIAL AND METHODS: The clinical features of 6 patients with GS aged 3 months to 12 years are described, and a brief review of the literature about this genetic disorder is presented. RESULTS: All patients demonstrated the classical triad of GS, including mandibular hypoplasia resulting in facial asymmetry, ear and/or eye malformation, and vertebral anomalies. In addition, renal and gastrointestinal abnormalities were observed in 2 patients. Regarding the oral involvement, 2 patients presented cleft lip and palate, and 1 patient had temporomandibular joint malformation. Malocclusion was found in all patients. CONCLUSION: Our orofacial findings correlate with the reported cases in the literature, and point out that after diagnosis GS patients need to be examined for systemic abnormalities.

Oral manifestations leading to the diagnosis of familial tuberous sclerosis.

Clinical manifestations of tuberous sclerosis (TS) are variable, and oral involvement occurs in less than 10% of the affected patients. We reported herein a nine-year-old boy with dental enamel pits and gingival nodular lesions, histologically diagnosed as angiofibromas that leaded to the diagnosis of a family with TS. In this report, we demonstrated the multiprofessional importance in the diagnosis of TS. Early diagnosis of TS is essential for an appropriate treatment of the affected patients and genetic counseling.

Aumento gengival induzido por ciclosporina A: revisão dos aspectos clínicos, histopatológicos, biológicos e modalidades de tratamento / Gingival overgrowth induced by cyclosporin A: review of clinical, histopathological and biological aspects and treatment modalities

Ciclosporina A é um medicamento imunossupressor utilizado na prevenção da rejeição de órgãos transplantados e no tratamento de doenças auto-imunes. O uso da ciclosporina A está relacionado com o desenvolvimento de inúmeros efeitos colaterais, dentre os quais o de maior interesse para a Odontologia é o aumento gengival. Neste artigo os autores apresentam uma revisão da literatura sobre as principais características clínicas, histopatológicas e modalidades de tratamento dos aumentos gengivais induzidos por ciclosporina A. Os mecanismos biológicos associados a esta alteração também são enfatizados.

Valor prognóstico de p53 e PCNA em carcinomas espinocelulares da cavidade oral / Prognostic value of p53 and PCNA in squamous cell carcinoma of oral cavity

Embora o carcinoma espinocelular (CEC) seja a neoplasia mais comum da cavidade oral, os fatores prognósticos deste tumor ainda não são completamente conhecidos. O objetivo do presente estudo foi analisar o padrão de expressão de PNCA e p53 e suas influências na sobrevida de pacientes em CECs da cavidade oral. Cinqüenta e uma amostras de CECs foram analisadas por imuno-histoquímica utilizando-se anticorpos monoclonais contra as proteínas PCNA e p53, e a porcentagem de expressão nuclear foi determinada e correlacionada com as informações clínicas dos pacientes. Todas as amostras foram positivas para PCNA, com uma média de células positivas de 66+/-16,3%, enquanto que 36 das 51 amostras (70,6%) foram positivas para a proteína p53, com uma média de 31.1+/-27,7%. Pacientes com neoplasias com metástases regionais ou com tumores primários grandes (T3 e T4) apresentaram índices de expressão para PCNA significantemente maiores que neoplasias menores ou sem metástases regionais (p<0,05). A expressão de p53 não foi correlacionada com nenhum dos parâmetros analisados. Os resultados do presente estudo demonstraram que a detecção imuno-histoquímica de p53 não apresenta um valor prognóstico para pacientes com CECs orais, enquanto que a expressão de PCNA correlaciona com o tamanho do tumor primário e a presença de metástases regionais

Expression and activity of matrix metalloproteinase-2 (MMP-2) in the development of rat first molar tooth germ

Tooth germ development is associated with morphological and biochemical changes of the dental papilla and enamel organ. Enzymes with gelatinolytic activities were studied by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and enzymography in tooth germ of newborn to 15-day-old rats. Three major bands with gelatinolytic activity were detected at all periods and characterized as the latent and active forms of MMP-2 using their molecular weight and activity dependent on Zn++ and Ca++ ions as criteria. Expression and activity of MMP-2 increased progressively from 0 to 15 days after birth. Mechanical separation of the tooth germ from 10-day-old rats showed that the gelatinolytic activity was localized mainly in the dental papilla and not the dental organ. These data indicate that the expression and activity of MMP-2 varies during the development and maturation of rat first molar tooth germ.