Avaliação clínica, morfológica e molecular do carcinoma epidermoide oral com envolvimento mandibular / Clinical, morphological and molecular evaluation of oral squamous cell carcinoma with mandibular invasion

O carcinoma epidermóide (CEC) é a neoplasia maligna mais frequente em cavidade oral, representando aproximadamente 90% de todos os cânceres da boca. Esta neoplasia tem origem no epitélio de revestimento bucal, e a depender do subtipo e da sua localização, pode apresentar tendência marcante de invasão do tecido ósseo adjacente. Existe uma interação entre osso e neoplasia, e atenção principal deve ser dada aos reguladores relacionados à osteoclastogênese. As moléculas RANK/RANKL/OPG são fundamentais neste processo, e juntamente com a IL-6, promovem formação e ativação de osteoclastos, com consequente promoção da invasão mandibular. Neste sentido, o objetivo do estudo foi avaliar as características clínicas, radiográficas e histopatológicas dos pacientes submetidos a mandibulectomia para tratamento do carcinoma epidermóide oral, e correlacionar/associar com a expressão gênica na neoplasia de marcadores reguladores da invasão óssea. Foi realizado um estudo retrospectivo de pacientes tratados com algum tipo de mandibulectomia no A.C. Camargo Cancer Center durante o período de1990 a 2015. Um total de 220 casos foi selecionado, e destes, foram extraídos 40 casos, posteriormente submetidos as etapas de análise molecular. A análise molecular foi realizada por PCR quantitativa em tempo real (qPCR), por meio da mensuração de mRNA para os genes estudados, calibrados pelo gene endógeno GAPDH e por um pool de controle composto por RNA não neoplásico. Dos 220 pacientes incluídos na pesquisa, 161 eram do sexo masculino, a idade média foi de 60 anos, e 53,6% eram etilistas e 66,8% tabagistas. A invasão óssea da mandíbula foi observada através do laudo anatomopatológico em 77 pacientes, e foi associada à presença de margens comprometidas/exíguas e a localização da lesão (gengiva inferior, rebordo e região retro-molar). Ao final do estudo, 127 pacientes foram a óbito, e a média de sobrevida foi de 96,92 meses. Parâmetros clínicos, dos exames de imagem e histopatológicos foram significantemente associados com a sobrevida global, principalmente - tamanho do tumor, infiltração perineural, invasão óssea (imagem e anatomopatológico). Na análise multivariada, apenas invasão óssea e infiltração perineural aumentaram risco de óbito nos pacientes. Na análise molecular de expressão gênica foi possível avaliar que das 36 amostras do gene RANK, 16 eram downreguladas. Para RANKL, das 37 amostras, 28 estavam super expressas. Das 35 do gene da IL-6, 19 amostras eram downreguladas. E por fim, das 37 amostras do gene da OPG, 16 tinham expressão normal. A avaliação de associação entre a expressão dos genes analisados demostrou que o gene da IL-6 está associado a presença de invasão óssea, e que existe uma associação significativa entre as expressões do RANK, RANKL, OPG e IL-6. Maior sobrevida dos pacientes foi associada significativamente a uma expressão mais baixa de IL-6 (108,5 meses). Conclui-se que a invasão mandibular interfere na sobrevida global dos pacientes, e que este mecanismo de invasão óssea está associado com a expressão gênica dos genes RANK, RANKL, OPG e IL-6. A IL-6 downregulada nas células malignas está associada com maior sobrevida

Oral Squamous Cell Carcinoma (OSCC) is the most frequent malignant neoplasm in the oral cavity, representing 90% of all oral cancers. The tumor usually arises within the epithelium lining of the oral cavity, and in cases of certain subtype or location may reveal a tendency for invasion of the surrounding bone tissue. There is a relation between tumor and bone, and more attention should be given to the events related to osteoclastogenesis. The molecules RANK/RANKL/OPG are crucial in this process, and together with IL-6, promotes the formation and activation of osteoclasts, with influence on the mandibular invasion process. In this way, the aim of this study was to evaluate the clinical, demographic, imaging, and histopathology data of patients diagnosed with OSCC treated by mandibulectomy. Also, analyze the gene expression condition of the genes RANK, RANKL, OPG, and IL-6 on the tumor. A retrospective study was conducted on patients treated with mandibulectomy at the A.C. Camargo Cancer Center, between 1990 and 2015. Two hundred and twenty cases were retrieved, and 40 of them were selected for gene expression analysis. The mRNA expressions of the selected genes were determined by real-time polymerase chain reaction, extracted from the tumor, and calibrated by endogenous GAPDH and a control pool of non-neoplastic RNA. Among 220 patients, 161 were male, with a mean age of 60 years. 53.6% were alcohol users and 66.8% tobacco users. Mandible bone invasion was reported on 77 histopathology reports and associated with compromised margins and the site of the lesion (gingiva, alveolar ridge, and retromolar region). One hundred and twenty-seven patients died of the disease, and the mean overall survival rate was 96.92 months. Overall survival was associated with clinical, imaging, and pathological parameters, especially, tumor size, perineural growth, and bone invasion (both imaging and pathologic). Multivariate analysis revealed higher risk of death only to bone invasion and perineural growth. Gene expression analysis revealed - 16 downregulation of 36 RANK samples evaluated; 28 overexpression of 37 RANKL samples evaluated; 19 downregulation of 35 IL-6 analyzed, and 16 normal expression of 37 OPG samples evaluated. Association analysis revealed that IL-6 expression is associated with bone invasion, and there is also a significant association among the expression of RANK, RANKL, OPG, and IL-6. Higher overall survival rate was associated with an IL-6 down expression (108;5 months). In conclusion, mandibular invasion interferes with the patient's overall survivall rate, and the bone invasion mechanism is associated with gene expression of RANK, RANKL, OPG, and IL-6 on the tumor. IL-6 in downregulation expressed by malignant cells is associated with higher overall survivor rates

Low Intensity laser therapy in patients with burning mouth syndrome: a randomized, placebo-controlled study

Abstract The aim of this study was to assess the effectiveness of low intensity laser therapy in patients with Burning Mouth Syndrome (BMS). Thirty BMS subjects were randomized into two groups - Laser (LG) and Placebo (CG). Seven patients dropped out, leaving 13 patients in LG and 10 patients in CG. Each patient received 4 irradiations (laser or placebo) twice a week, for two consecutive weeks (blinded to the type of irradiation received). Infrared laser (AsGaAI) irradiations were applied to the affected mucosa in scanning mode, wavelength of 790 nm, output power of 20 mW and fluence of 6 J/cm2. A visual analogue scale (VAS) was used to assess the therapeutic effect before and after each irradiation, and at all the control time periods: 7, 14, 30, 60 and 90 days after the last irradiation. One researcher delivered irradiation and another recorded the results. Both researchers were blinded, the first to the results, and the second to the type of radiation applied. The results were categorized according to the percentage of symptom level variation, and showed a statistically better response in LG in only two categories of the control checkpoints (p=0.02; Fisher's Exact Test). According to the protocol used in this study, low intensity laser therapy is as beneficial to patients with BMS as placebo treatment, indicating a great emotional component of involvement in BMS symptomatology. Nevertheless, there were positive results in some statistical analyses, thus encouraging further research in BMS laser therapy with other irradiation parameters.

Análise da expressão gênica do FOXP3, MIP-3­­­­? e Interleucinas 2, 10 e 35 em pacientes com ulceração aftosa recorrente / Analysis of gene expression of FOXP3, MIP-3? and interleukins 2, 10 and 35 in patients with recurrent aphthous ulcers

A ulceração aftosa recorrente (UAR) é considerada a doença ulcerativa mais frequente da cavidade bucal. Sua etiopatogenia ainda não está plenamente esclarecida, embora inúmeros fatores locais e sistêmicos já tenham sido a ela associados. Recentemente, a resposta imune anormal do tipo celular tem sido considerada a responsável pela lesão bucal na UAR, favorecendo uma resposta imunológica pró-inflamatória do tipo Th1, em conjunto com alterações em linfócitos T regulatórios. Sendo assim, o objetivo do presente estudo foi realizar análise da expressão gênica da FOXP3, MIP-3? e Interleucinas 2, 10 e 35 em pacientes com ulceração aftosa recorrente, por meio de estudo caso-controle. Os pacientes do grupo caso apresentavam quadros frequentes de UAR com pelo menos um ano de manifestação de surtos ulcerativos e história negativa de condições sistêmicas ou locais interferentes com a expressão das UAR. Estes foram submetidos a biópsia de lesão ulcerativa recente para a análise molecular. Os pacientes do grupo controle apresentavam história negativa de UAR, mucosa clinicamente saudável, e doaram voluntariamente fragmento de mucosa saudável para análise molecular, quando submetidos a procedimentos cirúrgicos como exodontia de terceiros molares ou biópsias ósseas. Todos os pacientes foram incluídos no grupo de pesquisa apenas após anuência

com termo de consentimento livre e esclarecido. Submeteram-se a exame clínico, realizaram exames complementares para controle da saúde geral e suporte diagnóstico. Onze pacientes UAR e três controles voluntários compuseram a casuística estudada, sendo submetidos a biópsia de lesões de UAR ou de mucosa de revestimento sadia. As amostras de tecido bucal foram submetidas aos procedimentos laboratoriais de extração do RNA e análise da expressão gênica da FOXP3, MIP-3? e Interleucinas 2, 10 e 35 por meio da técnica de RT-PCR em tempo real. Não houve diferença significativa na expressão dos genes estudados entre as amostras de portadores de UAR e controles sadios. Concluímos que os genes aqui avaliados não parecem desempenhar papel distintivo na fase ulcerativa inicial das UAR, entretanto estudos adicionais são recomendados a fim de se verificar a real participação desses agentes da inflamação na expressão da doença.

Recurrent aphthous ulcers (RAU) is the most common ulcerative disease of the oral cavity. Its pathogenesis is poorly understood yet, although numerous local and systemic factors have been associated with it. Recently, abnormal immune response of cellular type has been considered responsible for the RAU oral lesions, promoting a pro-inflammatory immune response Th1-type, in conjunction with changes in regulatory T cells. Thus, the aim of this study was to analyze the gene expression of FOXP3, MIP-3? and interleukins 2, 10 and 35 in patients with recurrent aphthous ulceration through a case-control study. The case group of patients presented frequent RAU bouts with at least one year of manifestation of ulcerative outbreaks and negative history of local or systemic conditions interfering with the RAU expression. These patients were submitted to a biopsy procedure of a recent ulcerative lesion for molecular analysis. Patients in the control group presented no history of RAU, and agreed with a donation of a healthy mucosa fragment for molecular analysis when undergoing surgical procedures such as extraction of third molars or bone biopsies.

All patients were included in the research group only after agreement with an informed consent. All subjects underwent clinical examination and were submitted to additional lab tests to check overall health and support diagnosis. Eleven RAU patients and three control volunteers composed the sample size and undergone biopsy of RAU lesions or healthy mucosal lining. The oral tissue samples were submitted to the laboratory procedures of RNA extraction and analysis of gene expression of FOXP3, MIP-3? and interleukins 2, 10, 35 by real time RT-PCR. There was no significant difference in gene expression between the studied samples of patients with RAU and healthy controls. It was concluded that the genes evaluated do not seem to play distinctive role in the initial ulcerative phase of RAU, however further studies are recommended in order to verify the actual participation of these inflammation agents in RAU expression.