Análise do comportamento das linhagens celulares SCC-25, SAS e HSC-3 frente à presença dos exossomos derivados dos macrófagos (TAMs) dos subtipos M1 e M2 / Analysis of the behavior of cell lines SCC-25, SAS and HSC-3 against the presence of exosomes derived from macrophages (TAMs) of subtypes M1 and M2

O carcinoma epidermóide (CE) de língua representa uma das lesões malignas mais comuns em cavidade oral e caracteriza-se por apresentar um comportamento localmente invasivo e agressivo. Os exossomos são responsáveis pela comunicação célula-célula e podem influenciar na progressão tumoral, metástase e eficácia terapêutica. Dentre as células capazes de secretar exossomos estão as células tumorais e as células imunes. Sabe-se que a presença das células imunes é importante para erradicar os tumores. No entanto, achados recentes demonstram que a inflamação pode promover o crescimento tumoral. Os macrófagos associados a tumores (TAMs) são conhecidos por apresentarem diferentes subtipos, M1 e M2, capazes de secretarem exossomos. O presente estudo se propôs a observar o comportamento dos exossomos derivados dos TAMs, dos subtipos 1 e 2, frente a cultura de células humanas SCC-25, HSC-3 e SAS derivadas de CE de língua, por meio da análise da capacidade de invasão, proliferação e viabilidade das células tumorais na presença dos exossomos. Observou-se que as microvesículas derivadas dos TAMs apresentam positividade para CD63, caracterizando-as como exossomos. Os exossomos dos TAMs do subtipo M2 foram os únicos a apresentarem marcação para TGF-ß, quando em comparação com os exossomos M1, THP1 e das linhagens celulares de CE, sugerindo que os exossomos M2 podem ser responsáveis pela expressão de TGF-ß nas células tumorais, uma vez que são internalizados. Nos ensaios de migração, observou-se que as células SCC-25 em presença de meio de cultura DMEM F/12, apresentaram maior capacidade de invasão frente aos exossomos M2 (p≤0,001), para concentração de 0,1 µg/ml. Para as células HSC-3 e SAS, não foi observada relação estatisticamente significante entre a presença de exossomos cultivados juntamente com as células tumorais e a capacidade de invasão celular (p>0,05). Quando os exossomos foram colocados no compartimento inferior do transwell, as células HSC-3 em presença dos exossomos M2 (1,0 µg/ml) apresentaram maior capacidade de invasão (p≤0,001). O teste de viabilidade demonstrou que as células HSC-3 tornam-se mais viáveis frente à presença dos exossomos M2 (p≤0,001) na concentração de 50 µg/ml. Para as células SCC-25, o resultado foi o mesmo (p≤0,05). A imunofluorescência demonstrou a internalização dos exossomos nas linhagens celulares estudadas. Os achados sugerem que a presença de exossomos M2, frente às culturas de células de CE de língua, pode ser um campo de pesquisa importante para futuros estudos com terapias-alvo (AU).

Squamous cell carcinoma (SCC) of the oral tongue is one of the most common malignant lesions in the oral cavity and is characterized by presenting a locally invasive and aggressive behavior. The exosomes are responsible for cell-cell communication and may influence tumor progression, metastasis and therapeutic efficacy. Among the cells that can secrete exosomes are tumor cells and immune cells. It is known that the presence of immune cells is important to eradicate tumors. However, recent findings suggest that inflammation may promote tumor growth. The tumor associated macrophages (TAMs) are known to have subtypes, M1 and M2, that secrete exosomes. This study's goal was to observe the behavior of derivatives TAMs exosomes, subtypes 1 and 2, against human cell culture SCC-25, HSC-3 and SAS derived from SCC of oral tongue, through the analysis of invasiveness, proliferation and viability of tumor cells in the presence of exosomes. It was observed that the microvesicles derived from TAMs are positive for CD63, characterizing them as exosomes. The exosomes of the M2 subtype TAMs were the only ones to present TGF-ß marking, as compared with M1 exosomes, THP1 and SCC cell lines, suggesting that M2 exosomes may be responsible for TGF-ß expression in tumor cells. In the migration tests, it was found that the SCC-25 cells in the presence of culture medium DMEM F / 12 showed higher invasion capacity in the presence of M2 exosomes (p≤0,001) to a concentration of 0.1 µg/ml. For HSC-3 and SAS cells, there was no significant statistical relationship between the presence of exosomes and invasiveness (p> 0.05) for the exosomes derived from TAMs. When the exosomes were placed in the lower compartment of the transwell, the HSC-3 cells in the presence of M2 exosomes (1.0 µg/ml) had higher invasiveness (p≤0,001). The viability test showed that HSC-3 cells became more viable in the presence of M2 exosomes (p≤0.001) at a concentration of 50 µg/ml. For SCC-25 cells, the result was the same (p≤0.05). Immunofluorescence showed the internalization of exosomes in the cell lines studied. These findings suggest that the presence of M2 exosomes in the SCC of the oral tongue cell cultures may be an important research field for future studies of targeted therapies (AU).

Expression of glucose transporters 1 and 3 in metastatic and non-metastatic lower lip squamous cell carcinoma.

This study aimed to evaluate the immunoexpression of glucose transporters 1 (GLUT-1) and 3 (GLUT-3) in metastatic and non-metastatic lower lip squamous cell carcinoma (LLSCC). Twenty LLSCCs with regional nodal metastasis and 20 LLSCCs without metastasis were selected. The distribution of staining and the percentage of GLUT-1 and GLUT-3 staining in each tumor core and at the deep invasive front were assessed. Most tumors (70%) exhibited peripheral staining for GLUT-1 in nests, sheets and islands of neoplastic cells, whereas predominantly central staining was observed for GLUT-3 (72.5%). A high percentage of GLUT-1-positive cells was observed at the deep invasive front and in the tumor core of metastatic and non-metastatic tumors (p>0.05). The percentage of GLUT-1-positive cells was much higher than that of GLUT-3-positive cells both in the deep invasive front (p<0.001) and in the tumor core (p<0.001) of LLSCCs. No significant differences in the percentage of GLUT-1- and GLUT-3-positive cells were observed according to nodal metastasis, clinical stage or histological grade of malignancy (p>0.05). In conclusion, the results of the present study suggest an important role of GLUT-1 in glucose uptake in LLSCCs, although this protein does not seem to be involved in the progression of these tumors. On the other hand, GLUT-3 expression may represent a secondary glucose uptake mechanism in LLSCCs.

Expression of Glucose Transporters 1 and 3 in Metastatic and Non-Metastatic Lower Lip Squamous Cell Carcinoma

This study aimed to evaluate the immunoexpression of glucose transporters 1 (GLUT-1) and 3 (GLUT-3) in metastatic and non-metastatic lower lip squamous cell carcinoma (LLSCC). Twenty LLSCCs with regional nodal metastasis and 20 LLSCCs without metastasis were selected. The distribution of staining and the percentage of GLUT-1 and GLUT-3 staining in each tumor core and at the deep invasive front were assessed. Most tumors (70%) exhibited peripheral staining for GLUT-1 in nests, sheets and islands of neoplastic cells, whereas predominantly central staining was observed for GLUT-3 (72.5%). A high percentage of GLUT-1-positive cells was observed at the deep invasive front and in the tumor core of metastatic and non-metastatic tumors (p>0.05). The percentage of GLUT-1-positive cells was much higher than that of GLUT-3-positive cells both in the deep invasive front (p<0.001) and in the tumor core (p<0.001) of LLSCCs. No significant differences in the percentage of GLUT-1- and GLUT-3-positive cells were observed according to nodal metastasis, clinical stage or histological grade of malignancy (p>0.05). In conclusion, the results of the present study suggest an important role of GLUT-1 in glucose uptake in LLSCCs, although this protein does not seem to be involved in the progression of these tumors. On the other hand, GLUT-3 expression may represent a secondary glucose uptake mechanism in LLSCCs.

Este estudo objetivou avaliar a imunoexpressão dos transportadores de glicose 1 (GLUT-1) e 3 (GLUT-3) em carcinomas de células escamosas de lábio inferior (CCELI) metastáticos e não-metastáticos. Vinte CCELIs com metástase nodal regional e 20 CCELI sem metástase foram selecionados. Foram analisados a distribuição da imunomarcação e o percentual de imunorreatividade para GLUT-1 e GLUT-3 no centro tumoral e no front de invasão tumoral. A maioria dos tumores (70%) revelou marcação para GLUT-1 em áreas periféricas dos ninhos, lençóis e ilhas de células neoplásicas, ao passo que GLUT-3 revelou predomínio de marcação em áreas centrais (72.5%). Um alto percentual de células positivas para GLUT-1 foi observado no front de invasão e no centro tumoral das lesões metastáticas e não-metastáticas (p>0,05). O percentual de células positivas para GLUT-1 foi superior ao percentual de células positivas para GLUT-3, tanto no front de invasão (p<0,001) quanto no centro tumoral (p<0,001) dos CCELIs. Não foram observadas diferenças significativas no percentual de células positivas para GLUT-1 e GLUT-3 em relação à mestástase nodal, ao estádio clínico ou ao grau histológico de malignidade (p>0,05). Em conclusão, os resultados do presente estudo sugerem um importante papel para GLUT-1 na absorção de glicose nos CCELIs, embora esta proteína não pareça estar envolvida na progressão destes tumores. Por outro lado, a expressão de GLUT-3 pode representar um mecanismo secundário para a absorção de glicose nos CCELIs.

Prevalence of subgingival Staphylococcus at periodontally healthy and diseased sites.

Staphylococci are considered members of the transient oral microbiota and are seldom isolated from the oral cavity. The aim of this study was to establish the prevalence of subgingival staphylococci in healthy and periodontal disease sites. Sterile endodontic paper points were used to isolate subgingival staphylococci in periodontally healthy and periodontally diseased sites in 30 adult subjects (n=540 sites). Staphylococcus spp were identified by an automated method and confirmed by conventional biochemical tests. All the samples were identified as coagulase-negative staphylococci. The results were analyzed using Mann-Whitney U, chi-square and Fisher's exact test at 5% significance level. A total of 86.7% of the subjects harbored these microorganisms in 11.7% of their periodontal sites. The most frequently isolated species was S. auricularis, which was isolated from 31.4% of the periodontal sites, followed by S. epidermidis, isolated from 21.4% of them. There was no statistically significant difference between the frequencies of these species isolated either from the healthy and the diseased sites (p>0.153). Although staphylococci are present in the subgingival environment and contribute to the pathogenic synergism involved in periodontal diseases, the results suggest that they do not participate directly in the pathogenesis of these diseases.

Prevalence of Subgingival Staphylococcus at Periodontally Healthy and Diseased Sites

Staphylococci are considered members of the transient oral microbiota and are seldom isolated from the oral cavity. The aim of this study was to establish the prevalence of subgingival staphylococci in healthy and periodontal disease sites. Sterile endodontic paper points were used to isolate subgingival staphylococci in periodontally healthy and periodontally diseased sites in 30 adult subjects (n=540 sites). Staphylococcus spp were identified by an automated method and confirmed by conventional biochemical tests. All the samples were identified as coagulase-negative staphylococci. The results were analyzed using Mann-Whitney U, chi-square and Fisher's exact test at 5% significance level. A total of 86.7% of the subjects harbored these microorganisms in 11.7% of their periodontal sites. The most frequently isolated species was S. auricularis, which was isolated from 31.4% of the periodontal sites, followed by S. epidermidis, isolated from 21.4% of them. There was no statistically significant difference between the frequencies of these species isolated either from the healthy and the diseased sites (p>0.153). Although staphylococci are present in the subgingival environment and contribute to the pathogenic synergism involved in periodontal diseases, the results suggest that they do not participate directly in the pathogenesis of these diseases.

Os estafilococos são considerados membros da microbiota oral transiente e são pouco isolados da cavidade oral. O objetivo deste estudo foi estabelecer a prevalência de estafilococos subgengivais em sítios saudáveis e com doença periodontal. Pontas endodônticas de papel absorvente estéreis foram usadas para isolar estafilococos subgengivais de sítios periodontais doentes e saudáveis, em 30 indivíduos adultos (n=540 sítios). Os Staphylococcus spp foram identificados por método automatizado e confirmados por provas bioquímicas convencionais. Todas as amostras foram identificadas como estafilococos coagulase-negativa. Os resultados foram analisados através dos testes de Mann-Whitney U, Qui-quadrado ou Exato de Fisher para um nível de significância de 5%. Um total de 86,7% dos indivíduos albergavam estes microrganismos em 11,7% de seus sítios periodontais. A espécie mais frequentemente isolada foi S. auricularis, a qual foi isolada em 31.4% dos sítios periodontais, seguida pela espécie S. epidermidis, isolada de 21.4% dos sítios. Não houve associação significativa na freqüência de isolamento das espécies, seja nos sitios periodontais saudáveis ou doentes (p>0.153). Embora os estafilococos estejam presentes no ambiente subgengival e contribuam para o sinergismo patogênico envolvido em doenças periodontais, sugere-se que eles não participem diretamente na patogênese destas doenças.

Fibromas Ossificantes Centrais e Displasias Fibrosas dos Maxilares: Estudo Clínico, Radiográfico e Histopatológico de 28 Casos / Central Ossifying Fibroma and Fibrous Dysplasia of the Jaws: A Clinical, Radiographic and Histopathologic Study of 28 Cases

Histopatológicos em uma série de casos de fibromas ossificantes centrais (FOCs) e displasias fibrosas (DFs) dos ossos maxilares. Método: Análise retrospectiva de FOCs e DFs dos ossos maxilares diagnosticados no Serviço de Patologia Oral do Departamento de Odontologia da UFRN, no período compreendido entre janeiro de 1970 e janeiro de 2007. Foram avaliados dados clínicos referentes ao sexo e idade dos pacientes e localização anatômica das lesões. Através de exames radiográficos panorâmicos, FOCs e DFs foram avaliados quanto à densidade radiográfica e ao limite da lesão em relação ao tecido ósseo circunvizinho. Para o estudo histopatológico, FOCs e DFs foram avaliados quanto à morfologia, grau de maturação e atividade osteoblástica do tecido mineralizado, bem como quanto à celularidade, vascularização e colagenização do tecido conjuntivo. Resultados: Foram constatados 15 FOCs e 13 DFs, os quais revelaram marcante predileção pelo sexo feminino. Em relação à localização anatômica, 66,7% dos FOCs acometeram a mandíbula e 69,2% das DFs se apresentaram em maxila. Em particular, 90% dos FOCs de mandíbula e 66,7% das DFs localizadas em maxila acometeram as regiões posteriores destes ossos. Radiograficamente, FOCs e DFs demonstraram densidade predominantemente mista ou radiopaca. Todos os FOCs se apresentaram como lesões circunscritas e 100% das DFs revelaram limites radiográficos difusos.