Immunoexpression of HSP27 does not seem to influence the prognosis of oral tongue squamous cell carcinoma

Abstract New methods of early detection and risk assessment have been studied aiming to predict the prognosis of patients and directing a specialized treatment of the oral tongue squamous cell carcinoma (OTSCC). In this context, several molecular biomarkers have been investigated for this purpose, and, among them, the heat shock protein 27 (HSP27) can be named. The study aimed to analyze whether heat shock protein 27 (HSP27) exerts any influence on OTSCC, correlating its immunoexpression with clinicopathological parameters, and patient survival. The sample comprised 55 OTSCC cases and 20 normal oral mucosa specimens. The malignancy grading systems proposed by the WHO in 2005, Brandwein-Gensler et al., and Almangush et al. were applied in a histomorphological study. HSP27 expressions were evaluated through the Immunoreactivity Score System (IRS). Significant values were considered at p <0.05 for all statistical tests. Higher IRS results were observed for normal oral mucosa specimens when compared to OTSCC cases (p <0.001). No significant associations between HSP27 immunostaining, the analyzed clinicopathological parameters and patient survival were observed. The results of the present study indicate lower HSP27 expression in OTSCC cases compared to normal oral mucosa specimens. Thus, HSP27 expression does not seem to influence patient prognosis.

Resumo Novos métodos de detecção precoce e avaliação de risco estão sendo estudados com o intuito de predizer o prognóstico dos pacientes e direcionar um tratamento diferenciado. Neste contexto, vários biomarcadores moleculares têm sido investigados com esta finalidade, dentre eles a heat shock protein 27 (HSP27). Esta pesquisa objetivou analisar se a HSP27 exerce alguma influência nos carcinomas de células escamosas de língua oral (CCELO), correlacionando a sua imunoexpressão com parâmetros clinicopatológicos e com a sobrevida dos pacientes. A amostra foi constituída por 55 casos de CCELO e 20 espécimes de mucosa oral normal. Os sistemas de gradação de malignidade propostos pela OMS em 2005, Brandwein-Gensler et al. e Almangush et al. foram aplicados em um estudo histomorfológico. A expressão da HSP27 foi avaliada através do Sistema de Escore de Imunorreatividade (IRS). Para todos os testes estatísticos foram considerados valores significativos com p<0,05. Foi observado um maior IRS para a mucosa oral normal quando comparado aos casos de CCELO (p<0,001). Não foram encontradas associações significativas entre a imunomarcação da HSP27 com os parâmetros clinicopatológicos analisados e com a sobrevida dos pacientes. Os resultados do presente estudo indicam uma menor expressão da HSP27 nos casos de CCELO quando comparados aos espécimes de mucosa oral normal. Assim, a expressão da HSP27 parece não influenciar o prognóstico dos pacientes.

EGFR, VEGF, and angiogenesis promote the development of lipoma in the oral cavity

Abstract This study aimed to detect, quantify and compare the immunohistochemical expression of EGFR and VEGF and microvessel count (MVC) in oral lipomas, and to correlate the findings with clinical and morphological characteristics of the cases studied. The sample consisted of 54 oral lipomas (33 classic and 21 non-classic) and 23 normal adipose tissue specimens. Cytoplasmic and/or nuclear immunohistochemical staining of EGFR and VEGF was analyzed. The angiogenic index was determined by MVC. Cells were counted using the Image J® software. The Statistical Package for the Social Sciences was used for data analysis, adopting a level of significance of 5% for all statistical tests. A statistically significant difference in EGFR immunoexpression (p=0.047), especially, between classic lipomas and normal adipose tissue. There was a significant difference in MVC between non-classic lipomas and normal adipose tissue (p=0.022). In non-classic lipomas, only VEGF immunoexpression showed a significant moderate positive correlation (r=0.607, p=0.01) with MVC. In classic lipomas, the number of EGFR-immunostained adipocytes was directly proportional to the number of VEGF-positive cells, demonstrating a significant moderate positive correlation (r=0.566, p=0.005). The results suggest that EGFR, VEGF, and angiogenesis participate in the development of oral lipomas but are not primarily involved in the growth of these tumors.

Resumo Lipomas são as neoplasias mesenquimais benignas mais comuns, no entanto sua etiopatogenia ainda permanece desconhecida. Dessa forma, essa pesquisa teve como objetivo detectar, quantificar e comparar a expressão imunoistoquímica do EGFR, VEGF e contagem microvascular (MVC) dos lipomas orais, relacionando-os com as características clínicas e morfológicas dos casos estudados. A amostra foi composta por 54 lipomas orais (33 clássicos e 21 não clássicos) e 23 casos de tecido adiposo normal. A análise da expressão imunoistoquímica de EGFR e VEGF foi fundamentada na marcação citoplasmática e/ou nuclear. O índice angiogênico foi avaliado por meio da MVC. A contagem de células foi realizada utilizando software IMAGE J®. Os dados obtidos foram analisados no software Statistical Package for Social Science. O nível se significância de 5% foi adotado para os testes estatístico. A análise da imunoexpressão das proteínas revelou para o EGFR diferença estatisticamente significativa (p=0,041) entre o lipoma clássico e o tecido adiposo normal. Houve diferença significativa na MVC entre lipomas não clássicos e tecido adiposo normal (p=0,022). Nos lipomas não clássicos, apenas a imunoexpressão de VEGF apresentou correlação do tipo moderada, positiva e significativa (r=0,607; p=0,010) em relação a MVC. Ademais, nos lipomas clássicos foi percebido que os adipócitos imunomarcados para EGFR estiveram diretamente proporcionais a imunoexpressão de VEGF, apresentando correlação do tipo moderada, positiva e estatisticamente significativa (r=0,566; p = 0,005). Com base nos resultados, pode-se sugerir que o EGFR, VEGFR e MCV participam do desenvolvimento nos lipomas orais, contudo, não estão primariamente envolvidos no crescimento tumoral dessas neoplasias.

Immunohistochemical analysis of SHH, SMO and GLI-1 proteins in epithelial odontogenic lesions

Abstract The present study analyzed the expression of proteins involved in the sonic hedgehog signaling pathway (SHH, SMO, and GLI-1) in benign epithelial odontogenic lesions (odontogenic keratocyst - OKC, ameloblastoma - AB, and adenomatoid odontogenic tumor - AOT) in order to identify the role of these proteins in the pathogenesis of these lesions. The sample consisted of 20 OKCs, 20 ABs, and 10 AOTs. The Kruskal-Wallis, Mann-Whitney U, and Spearman's (r) tests were used for statistical analysis, with the level of significance set at 5% (p < 0.05). The membrane/cytoplasmic expression of SHH was significantly higher in AB compared to AOT (p = 0.022) and OKC (p = 0.02). No differences were found in the membrane/cytoplasmic expression of SMO between the lesions studied. Regarding GLI-1, significant differences were observed at the nuclear level for AB and OKC compared to AOT (p < 0.0001). In addition, significant positive correlations were found between cytoplasmic and nuclear GLI-1 in AB (r = 0.482; p = 0.031) and OKC (r = 0.865; p < 0.0001), and between membrane/cytoplasmic SMO and cytoplasmic GLI-1 in AOT (r = 0.667; p = 0.035) and OKC (r = 0.535; p = 0.015). The results of this study confirm the participation of the sonic hedgehog signaling pathway in the pathogenesis of the lesions studied. Overexpression of SHH in ABs and nuclear expression of GLI-1 in ABs and OKCs indicate that these proteins contribute to the more aggressive behavior of these two lesions when compared to AOT.

Resumo O presente estudo analisou a expressão de proteínas envolvidas na via de sinalização Sonic Hedgehog (SHH, SMO e GLI-1) em lesões benignas do epitélio odontogênico de comportamento biológico distintos, tais como ceratocistos odontogênicos (CO), ameloblastomas (AMB) e tumores odontogênicos adenomatoides (TOA), com o intuito de identificar o papel destas proteínas na patogênese destas lesões. A amostra foi constituída de 20 CO, 20 AMB e 10 TOA, analisada pela técnica da imuno-histoquímica de forma semiquantitativa por compartimento celular, onde foi feita uma análise da membrana e citoplasma das células nas proteínas SHH e SMO, enquanto que para a proteína GLI-1, foi feita uma análise nuclear e/ou citoplasmática. Para análise estatística, foram utilizados os testes de Kruskal-Wallis (KW), Mann-Whitney (U) e Spearman (r), com o nível de significância estabelecido em 5% (p < 0,05). Ao analisar a proteína SHH, observou-se que o AMB demonstrou expressão membranar/citoplasmática significativamente maior em comparação ao TOA (p = 0,022) e CO (p = 0,020). Com relação à análise membranar/citoplasmática da SMO, não foram identificadas diferenças entre as lesões estudadas. Para a proteína GLI-1, foram constatadas diferenças estatisticamente significativas, em nível nuclear, para o AMB e CO em comparação ao TOA (p< 0,0001). Além disso, foram observadas correlações positivas com significância estatística entre GLI-1 citoplasmático e GLI-1 nuclear para o AMB (r = 0,482; p = 0,031) e CO (r = 0,865; p< 0,0001), e entre o SMO membranar/citoplasmático e o GLI-1 citoplasmático para o TOA (r = 0,667; p = 0,035) e o CO (r = 0,535; p = 0,015). Os resultados deste estudo confirmam a participação da via de sinalização Sonic Hedgehog na patogênese das lesões estudadas e, a superexpressão de SHH em AMBs e GLI-1 nuclear em AMBs e COs, indica que estas proteínas contribuem com o comportamento biológico mais agressivo destas duas lesões quando comparado ao TOA.

Differential protein expression of osteoclastogenic factors in odontogenic cysts and tumors

Abstract: The osteolytic activity of odontogenic cysts and tumors is directly associated with their growth and aggressiveness. The influence of proteins expressed by epithelial and mesenchymal cells on this biological event differs between indolent cystic lesions, aggressive cystic lesions, and odontogenic tumors. The objective of this study was to compare the immunohistochemical expression of factors that stimulate (receptor activator of nuclear factor kappa-Β ligand - RANKL, cathepsin K - CatK and matrix metallopeptidase 8 - MMP-8) and inhibit (osteoprotegerin - OPG) osteoclastogenesis between dentigerous cyst (DC), glandular odontogenic cyst (GOC), odontogenic keratocyst (OKC), and ameloblastoma (AB). Paraffin-embedded sections of nine DCs, nine GOCs, 20 OKCs, 21 ABs, and four dental follicles (DFs) were subjected to immunohistochemistry. Immunoreactivity was analyzed semiquantitatively and quantitatively in epithelium and connective tissue, respectively. The proteins were immunoexpressed in epithelial and mesenchymal cells of all lesions studied. The expression of RANKL and CatK was higher in OKC, AB, and GOC (p<0.005). Higher expression of OPG was found in DF and DC compared to the other markers (p<0.005). MMP-8 expression was high in GOC and OKC. This study demonstrated the differential expression of factors that inhibit and stimulate bone resorption during the development of DC, GOC, OKC, and AB. Higher expression of RANKL and CatK was observed in more aggressive lesions. OPG appears to be one of the molecules responsible for the slower growth of DC.

Involvement of tryptase-positive mast cells and angiogenesis in the growth of inflammatory odontogenic cysts

Abstract: Inflammatory periapical lesions are characterized by infiltration of different immune cell types, the functions of which depend on an effective vascular network. This study aimed to evaluate the mast cells density (MCD) in inflamatory odontogenic cysts capsules concerning microvascular density (MVD), microvascular area (MVA), and microvascular perimeter (MVP), and correlate such findings with the type of lesion, intensity of the inflammatory infiltrate, and thickness of the epithelial lining. Twenty inflamatory dentigerous cysts (IDCs), twenty radicular cysts (RCs), and twenty residual radicular cysts (RRCs) were submitted to immunohistochemical analysis using anti-tryptase and anti-CD34 antibodies. RCs exhibited the highest MCD, MVD, MVA, and MVP indexes (p = < 0.001, p = 0.008, p = 0.003 and p = < 0.001, respectively), and lesions with inflammatory infiltrate grade III showed the highest MVD (p = 0.044). Considering epithelial thickness, a higher MVP index was identified in lesions with hyperplastic epithelium (p = 0.018). In IDCs, RCs, and RRCs, a strong positive correlation was observed between MVA and MVP (r = 0.950 and p = < 0.001; r = 0.914 and p = < 0.001; r = 0.713 and p = < 0.001, respectively). In IDCs, a moderate correlation was observed between MCD and both MVA and MVP (r = 0.660 and p = 0.002; r = 0.634 and p = 0.003, respectively). These results suggest that tryptase-positive mast cells might play an important role in the angiogenic activity of IDCs, while RCs had the highest indexes. Our findings also confirmed that the intensity of the inflammatory infiltrate and epithelial thickness influence angiogenesis.

Polymorphisms of matrix metalloproteinase-7 and -9 are associated with oral tongue squamous cell carcinoma

Abstract Matrix degradation is an important event in the progression, invasion and metastasis of malignant head and neck lesions. Imbalances, mutations and polymorphisms of MMPs and their inhibitors are observed in several cancer subtypes. The aim of this study was to evaluate the association of the MMP-7 gene promoter (181 A/G) and MMP-9 (-1562 C/T) polymorphisms in oral tongue squamous cell carcinoma (OTSCC). MMP-7 (rs11568818) and MMP-9 (rs3918242) single-nucleotide polymorphisms (SNPs) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in 71 cases of OTSCC. Normal tissue specimens were obtained from 60 healthy volunteers to serve as the control. The MMP-7 G allele and MMP-9 T allele were more frequent in the OTSCC group than the control group, but only when these two SNPs were taken together was a significant association found with the nodal metastasis of OTSCC (p < 0.001). Based on our results, SNPs in the promoter region of MMP-7 and MMP-9 appear to be associated with greater risk of developing OTSCC, and with a higher propensity to form metastatic tumors. In this respect, molecular studies investigating polymorphisms may be useful in predicting tumor behavior.

Immunohistochemical expression of DNA repair proteins in oral tongue and lower lip squamous cell carcinoma

Abstract The DNA repair system involves genes and proteins that are essential for the maintenance of genome integrity and the consequent control of various cellular processes. Alterations in these genes and proteins play a role in tumor development and progression and might be associated with prognosis. The aims of this study were to analyze the immunoexpression of two DNA repair proteins, XPF and XRCC1, in lower lip squamous cell carcinoma (LLSCC) and oral tongue squamous cell carcinoma (OTSCC), and to investigate possible associations with clinical and histopathological parameters. The immunohistochemical expression of XPF and XRCC1 was analyzed semi-quantitatively in 40 cases each of LLSCC and OTSCC. The chi-squared test or Fisher's exact test, when appropriate, was used to investigate the association between expression of the proteins and clinicopathological characteristics. The cytoplasmic immunoexpression of XPF was high in OTSCC (95% of the cases analyzed) but low in LLSCC (52.5%). Among the clinicopathological parameters evaluated, a statistically significant association was observed between high nuclear expression of XRCC1 and the absence of regional lymph node metastasis in patients diagnosed with OTSCC (p=0.006). The high protein expression of XPF and XRCC1 in OTSCC and LLSCC suggests an important role in the development and progression of these tumors. Our study found an association between high nuclear expression of XRCC1 and the absence of loco-regional metastasis in cases diagnosed as OTSCC, suggesting a role of this protein in tumor progression.

Immunohistochemical study of fibronectin, tenascin and type I collagen in tongue and lip carcinomas

AIM: The aim was to compare the immunoexpression of extracellular matrix proteins in squamous cell carcinomas of tongue (SCCTo) and lower lip (SCCLi). METHODS: Eleven SCCTo and 11 SCCLi were selected and examined according to Bryne's method (1998). For immunohistochemical study utilized antibodies to fibronectin, tenascin and type I collagen. Histopathologic and immunohistochemical analysis were performed on the tumor invasive front. RESULTS: All SCCTo were classified in high score malignant grade and all SCCLi in lower score. Fibronectin showed strong immunorreactivity in the peritumoral basement membrane (BM) in 91% of SCCTo and all cases of SCCLi, while in the tumor stroma (TS) all cases of SCCTo and SCCLi had strong intensity. Tenascin had strong expression in BM of 91% cases of SCCTo and 63.4% of SCCLi and in TS had strong expression in 91% cases of SCCTo and 54.6% of SCCLi. Type I collagen demonstrated weak immunoreactivity in the TS of 72.7% cases of SCCTo and 63.4% of SCCLi. CONCLUSION: These results may suggest that the strong expression of fibronectin and tenascin proteins and the weak expression of type I collagen could play a role in the invasive process of oral SCC (AU)

Immunohistochemical Analysis of Cell Proliferation and Bcl-2 Expression in Drug-Induced Gingival Overgrowth

Objective: To compare the rate of cell proliferation and expression of antiapoptotic protein Bcl-2 between drug-induced gingival overgrowth (DIGO) and clinical healthy gingiva (CHG) and to establish associations with histopathological features. Material and Methods: Twenty specimens of DIGO and 20 CHG specimens were submitted to morphological and immunohistochemical analysis by light microscopy. Cell proliferation (Ki-67) and the expression of Bcl-2 were evaluated in epithelial cells and spindle-shaped mononuclear cells of the connective tissue by establishing the labeling index (LI). Results: In epithelial tissue, the mean LI for Ki-67 was 17.2% in DIGO and 21.71% in CHG (p = 0.137). The mean LIs for Bcl-2 in epithelial tissue were 14.67% and 10.24% in DIGO and CHG, respectively (p = 0.026). In connective tissue, DIGO and CHG specimens exhibited low LIs for Ki-67 and Bcl-2, with mean values of less than 0.5% in both groups. No significant differences in the LIs for Ki-67 or Bcl-2 in epithelial tissue were observed according to the degree of collagenization, degree of vascularization and intensity of inflammatory infiltration (p > 0.05). No significant correlations were observed between the LIs for Ki-67 and Bcl-2 (p > 0.05). Conclusion: The present results suggest that the pathogenesis of DIGO does not involve increased proliferation or decreased apoptosis of fibroblasts. On the other hand, the morphological pattern of elongated epithelial cristae observed in DIGO could mainly be due to the inhibition of keratinocyte apoptosis and not to increased proliferation of these cells.

Histochemical and immunohistochemical differences between solitary oral fibroma and fibrous papule of the face

Abstract: Background: The morphological similarities between fibrous papules of the face and multiple sporadic oral fibromas were mentioned long ago and a relationship between them has been reported in the literature. Objective: The aim of this study was to evaluate the participation of mast cells, elastin and collagen in a series of oral fibromas and fibrous papules of the face in order to better understand the possible role of these factors in fibrosis and the formation of these lesions. Methods: Thirty cases of oral fibroma involving the buccal mucosa and 30 cases of fibrous papules of the face were selected. Tissue samples were submitted to picrosirius red staining and immunohistochemistry using anti-elastin and anti-tryptase antibodies. Results: The percentage of tryptase-positive mast cells and expression of elastin were higher in cases of fibrous papules of the face (p < 0.05). In contrast, a higher intensity of collagen deposition was observed in oral fibromas. The results showed mast cell accumulation and higher elastin synthesis in fibrous papules of the face, and mast cell accumulation with higher collagen fiber synthesis in oral fibromas. Conclusion: These findings support the hypothesis that mast cells influence the development and growth of these lesions through different mechanisms.

Immunoexpression of HIF-1α and VEGF in Periodontal Disease and Healthy Gingival Tissues

Abstract Hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) are proteins that stimulate the proliferation and migration of endothelial cells. These proteins have been described in many pathologic and inflammatory conditions, but their involvement in the development of periodontitis has not been thoroughly investigated. This study compared the immunohistochemical expression of these proteins, involved in angiogenesis and hypoxia, by imunnostained inflammatory and endothelial cells in periodontal disease and healthy gingival tissues. Gingival tissue samples were divided as follows: 30 samples with chronic periodontitis, 30 with chronic gingivitis, and 30 of healthy gingiva. Results were analyzed statistically by the Kruskal-Wallis, Mann-Whitney and Spearman correlation tests (p=0.01). Inflammatory and endothelial cells were found to express these proteins. Periodontitis showed median percentage of HIF-1α-positive cells of 39.6%, 22.0% in cases of gingivitis and 0.9% in the healthy gingiva group (p=0.001). For VEGF, median percentage of immunopositive cells was 68.7% for periodontitis, 66.1% in cases for gingivitis, and 19.2% for healthy gingival specimens (p<0.001). Significant correlation between VEGF and HIF-1α was also observed in healthy gingiva (p<0.001).The increased expression of HIF-1αα and VEGF in periodontitis, compared to gingivitis and healthy gingiva, suggests possible activation of the HIF-1α pathway in advanced periodontal disease. The correlation between HIF-1α and VEGF expression in healthy gingiva suggests a physiological function for these proteins in conditions of homeostasis. In periodontal disease, HIF-1 and VEGF expression may be regulated by other factors, in addition to hypoxia, such as bacterial endotoxins and inflammatory cytokines.

Resumo O fator induzível por hipóxia 1 alfa (HIF-1α) e o fator de crescimento endotelial vascular (VEGF) são proteínas que estimulam a proliferação e a migração de células endoteliais. Estas proteínas têm sido descritas em muitas condições patológicas e inflamatórias, mas o envolvimento dessas no desenvolvimento de periodontite não foi completamente investigado. Este estudo comparou a expressão imunohistoquímica destas proteínas, envolvidas na angiogênese e hipóxia, na doença periodontal e em tecidos gengivais saudáveis por meio de contagem de células inflamatórias e endoteliais imunomarcadas. As amostras de tecido gengival foram divididas da seguinte forma: 30 amostras com periodontite crônica, 30 com gengivite crônica e 30 de gengiva saudável. Os resultados foram analisados estatisticamente pelos testes de Kruskal-Wallis e Mann-Whitney (p=0.01). As células inflamatórias e endoteliais foram positivas para estas proteínas. A porcentagem média de células positivas para HIF-1α foi de 39,6% nos casos de periodontite, 22,0% nos casos de gengivite e de 0,9% no grupo de gengiva saudável (p = 0,001). A porcentagem média de células imunopositivas para o VEGF foi de 68,7% nos casos de periodontite, 66,1% nos casos de gengivite, e 19,2% em espécimes gengivais saudáveis (p<0,001). Correlação significativa entre o VEGF e HIF-1α foi observada em gengival. A expressão elevada do HIF-1α e VEGF em periodontite, comparada a gengivite e gengiva saudável, sugere ativação da via do HIF-1α, na doença periodontal avançada. A correlação entre o HIF-1α e expressão de VEGF na gengiva saudável, sugere uma função fisiológica para estas proteínas em condições de homeostase. Na doença periodontal, a expressão de HIF-1α e VEGF pode ser regulada por outros fatores, além da hipóxia, tais como endotoxinas bacterianas e citocinas inflamatórias.

Collagen IV and laminin expression in squamous cellcarcinomas of lower lip and tongue / Expressão de colágeno IV e laminina em carcinomas de células escamosas de lábio inferior e língua

In this study, the expression of extra cellular matrix proteins was immunohisto chemically studied and compared with the histological grading of squamous cell carcinomas of the lower lip and tongue. Material and Methods: The lower lip carcinomas (n =12) and the tongue carcinomas (n = 12) were histopathologically graded according to Bryne’smethod. The immunohisto chemical technique used specific antibodies for collagen IV and laminin. Histopathological and immunohisto chemical analysis were carried-out at the invasive tumor front. Results: Most of the lower lip carcinomas(91.7%) were classified with lower scores and all tongue carcinomas (100%) with high-grade malignant scores (p < 0.01). Collagen type IV expression was absent in the peri tumoral basement membrane in 50% of lower lip carcinomas and in66. 7% of tongue carcinomas (p = 0.09). Laminin expression was absent in the peritumoral basement membrane in 66.7% of lower lip carcinomas and in 58.3% of tongue carcinomas (p = 0.48). When these two glycoproteins were expressed, they showed alinear, thin and discontinuous pattern and a weak intensity of expression. Conclusion: The high gradem alignancy score of the tongue carcinomas was associated with the pattern of expression of the matrix proteins studied. This suggested that tonguesquamous cell carcinomas have more invasive potential and more aggressive biological behaviorthan the lower lip carcinomas...

Neste estudo, a expressão das proteínas da matriz extracelular foi estuda da imunoisto quimicamente e comparada com a classificação histológica dos carcinomas de células escamosas do lábio inferior e língua. Material e Métodos: Os carcinomas de lábio inferior (n = 12)e os carcinomas de língua (n = 12) foram graduados histopatologicamente de acordo com o método de Bryne. A técnica de imunoistoquímica utilizou anticorpos específicos para colágeno IV e laminina. Análises histopatológica e imunoistoquímica foram conduzidas na frente invasiva tumoral. Resultados: A maioria dos carcinomas de lábio inferior (91,7%) foi classificada em baixo grau e todos os carcinomas de língua (100%) em alto grau de malignidade (p < 0,01). A expressão de colágeno tipo IV estava ausente na membrana basal peri tumoral em 50% dos carcinomas de lábio inferior e em 66,7% dos carcinomas de língua (p = 0,09). A expressão de laminina estava ausente na membrana basal peri tumoral em 66,7%dos carcinomas do lábio inferior e em 58,3% dos carcinomas de língua (p = 0,48). Quando estas duas glicoproteínas foram expressas, mostraram-se comum padrão linear, fino e descontínuo e uma fraca intensidade de expressão. Conclusão: O alto grau de malignidade dos carcinomas de língua associou-se com o padrão de expressão das proteínas de matriz estudadas. Isso sugere que carcinomas de células escamosas de língua têm comportamento biológico mais agressivo e potencial mais invasivo do que os carcinomas de lábio inferior...

Influência dos herpes-vírus na patogênese da doença periodontal / Influence of herpesvirus in pathogenesis of disease periodontal – a review of literature

A etiologia da doença periodontal está estreitamente relacionada à presença de microrganismos patogênicos, principalmente bactérias anaeróbias gram-negativas. Estudos recentes sugerem que os herpes-vírus (HHV) podem estar associados à ocorrência e progressão de diferentes formas da doença periodontal. Diante disso, este trabalho fez uma investigação na literatura em relação à presença dos vírus Epstein-Barr (EBV), Citomegalovírus (HCMV) e Herpes simples (HSV) em pacientes com doença periodontal. A análise dos estudos mostra que estes herpes-vírus podem contribuir para a destruição periodontal através da infecção e replicação viral, liberação de mediadores pró-inflamatórios e citotoxinas, ou como resultado de uma alteração na defesa do hospedeiro. Um número crescente de pesquisas sugere que, dentre os HHVs, o EBV e o HCMV são os que estão mais comumente envolvidos no desenvolvimento da doença periodontal. De acordo com a literatura revisada, embora ainda não tenha sido estabelecida uma relação causal entre a infecção por herpes-vírus e o desenvolvimento da doença periodontal, tem sido destacada uma associação positiva entre ambas.

Manifestações periodontais associadas às doenças auto-imunes: uma revisão de literatura / Periodontal manifestations associated with autoimmune diseases: a literature review

As doenças auto-imunes envolvem a formação de auto-anticorpos direcionados contra alguns elementos teciduais, particularmente os da pele ou superfícies das mucosas, como forma sistêmica. Dentre estas doenças podemos destacar o pênfigo vulgar, o lúpus eritematoso sistêmico e o penfigóide das membranas mucosas. Estas estão, muitas vezes, associadas com sintomas bucais, podendo apresentar em diversos sítios orais a presença de lesões vesículo-bolhosas e gengivite descamativa. Baseado nisto, este artigo realizou uma revisão de literatura que teve como objetivo associar as manifestações periodontais ao pênfigo vulgar, lúpus eritematoso sistêmico e penfigóide das membranas mucosas com o intuito fornecer aos profissionais um melhor entendimento sobre essas doenças, de modo que eles sejam capazes de oferecer um atendimento clínico mais adequado para estes pacientes. Os resultados puderam mostrar que há uma associação dessas doenças auto-imunes a manifestações periodontais, especialmente a gengivite descamativa. Além disso, a literatura as associa à perda óssea alveolar, ao aumento do sangramento gengival, a perda de inserção e a outras desordens periodontais.

Th2 cells and the ifn-γ R1 subunit in early and advanced experimental periodontitis in rats; an immunohistochemical study

Aim: To evaluate the involvement of Th2 cells in different periods of the active phase of experimental periodontal disease and expression of the R1 subunit of the receptor for IFN-γ during the early and advanced progression of the disease. Methods: Experimental periodontitis was induced in 30 male Wistar rats by placing cotton ligatures around the mandibular first molars. The rats were then randomly assigned into two groups: G1=15 and G2=15, in group G1, ligatures were maintained for 2 days, a period that corresponds to the initial stage of periodontal disease in rats, in G2 ligatures were left for 15 days, a period that corresponds to the advanced stage of periodontal disease. The contra-lateral teeth served as controls (unligated). An immunohistochemical investigation of the gingival tissue was performed to detect the presence of the Th2 specific transcription factor (GATA3). Results Light microscopy analysis revealed a decreased expression of GATA-3-positive cells when bone loss progressed. IFN-γ R1 was detected at an early stage during the active phase of disease, but the expression of positive cells remained unaltered during the remaining period of the study.

Low-level laser irradiation induces in vitro proliferation of mesenchymal stem cells / Laser de baixa intensidade induz à proliferação in vitro de células-tronco mesenquimais

Objective : To evaluate the effect of low-level laser irradiation on the proliferation and possible nuclear morphological changes of mouse mesenchymal stem cells. Methods : Mesenchymal stem cells derived from bone marrow and adipose tissue were submitted to two applications (T0 and T48 hours) of low-level laser irradiation (660nm; doses of 0.5 and 1.0J/cm2). The trypan blue assay was used to evaluate cell viability, and growth curves were used to analyze proliferation at zero, 24, 48, and 72 hours. Nuclear alterations were evaluated by staining with DAPI (4’-6-diamidino-2-phenylindole) at 72 hours. Results : Bone marrow-derived mesenchymal stem cells responded to laser therapy in a dose-dependent manner. Higher cell growth was observed when the cells were irradiated with a dose of 1.0J/cm2, especially after 24 hours (p<0.01). Adipose-derived mesenchymal stem cells responded better to a dose of 1.0J/cm2, but higher cell proliferation was observed after 48 hours (p<0.05) and 72 hours (p<0.01). Neither nuclear alterations nor a significant change in cell viability was detected in the studied groups. Conclusion : Low-level laser irradiation stimulated the proliferation of mouse mesenchymal stem cells without causing nuclear alterations. The biostimulation of mesenchymal stem cells using laser therapy might be an important tool for regenerative therapy and tissue engineering. .

Objetivo : Avaliar o efeito da terapia com laser de baixa intensidade sobre a proliferação e as possíveis alterações morfológicas nucleares em células-tronco mesenquimais de camundongos. Métodos : Células-tronco mesenquimais derivadas da medula óssea e do tecido adiposo foram submetidas a duas aplicações (T0 e T48 horas) de laser de baixa intensidade (660nm; doses de 0,5 e 1,0J/cm2). O ensaio de azul de tripan foi utilizado para a avaliação da viabilidade celular, e curvas de crescimento foram usadas para avaliar a proliferação das células em zero, 24, 48, e 72 horas. Alterações nucleares foram avaliadas por coloração com DAPI (4-6-diamidino-2-fenilindolo) em 72 horas. Resultados : As células-tronco mesenquimais derivadas da medula óssea responderam a terapia com laser de forma dose-dependente. Um maior crescimento celular foi observado quando as células foram irradiadas com dose de 1,0J/cm2, especialmente depois de 24 horas (p<0,01). As células-tronco mesenquimais derivadas do tecido adiposo responderam melhor à dose de 1,0J/cm2, com maior proliferação após 48 (p<0,05) e 72 horas (p<0,01). Nem alterações nucleares nem a mudança significativa na viabilidade celular foi detectada nos grupos estudados. Conclusão : Laser de baixa intensidade estimulou a proliferação de células-tronco mesenquimais sem causar alterações nucleares. A bioestimulação de células-tronco mesenquimais por laserterapia pode ser uma ferramenta importante para a terapia regenerativa e a engenharia tecidual. .

Maxillary brown tumor associated with chronic kidney failure: a case report / Tumor marrom na maxila associado à insuficiência renal crônica: relato de caso

The brown tumor is a bone lesion that may affect the entire skeleton, including the maxillary bones. These tumors are characterized as focal giant cell lesions that may be associated with primary or secondary hyperparathyroidism (HPT). Brown tumors are invasive in some cases and an association with chronic renal failure (CRF) has been reported. With the aim to facilitate the differential diagnosis of bone lesions that may affect dialysis patients, this paper describes a case of brown tumor in a 36- year old patient with CRF, secondary HPT carrier, who had a lesion on the right maxilla for approximately five months.

O tumor marrom é uma lesão óssea que pode acometer todo o esqueleto, inclusive os ossos maxilares. Apresenta-se como uma lesão focal de células gigantes associada ao hiperparatireoidismo (HPT) primário ou secundário; é invasiva em alguns casos e pode associar-se à insuficiência renal crônica (IRC). O presente trabalho descreve um caso de tumor marrom em uma paciente de 36 anos com insuficiência renal crônica (IRC), portadora de HPT secundário, que apresentava lesão, com duração de aproximadamente cinco meses, localizada na maxila direita, com o objetivo de auxiliar o diagnóstico diferencial das lesões ósseas que podem afetar pacientes dialíticos.

Influence of dialysis duration and parathyroid hormone on the clinical and radiographic oral conditions of pre-transplant patients with chronic kidney disease

Aim: To evaluate the oral conditions of patients with chronic kidney disease undergoing dialysisand to determine the influence of dialysis duration and bone metabolism on the prevalence andseverity of the alterations found. Methods: The simplified oral hygiene index (OHI-S), prevalenceof dental caries (decayed, missing filled teeth index, DMFT), and the periodontal screening andrecording (PSR) index were evaluated in 154 patients. Parathyroid hormone (PTH), calcium,phosphorus and urea measurements, as well as panoramic radiographs, were obtained from allpatients. To evaluate the effect of duration of dialysis treatment on oral health, the patients weredivided into two groups: (1) <5 years and (2) > 5 years. Regarding blood levels of PTH, patientswere divided into three groups: (1) 0-149 pg/mL, (2) 150-584 pg/mL, and (3) >585 pg/mL. TheOHI-S identified the accumulation of biofilm and calculus around the teeth. Results: Gingivalinflammation was found in 100% of dentate patients, with 2 being the predominant PSR score(72.3%). The DMFT index was high (17.52). Dental calculus was the most common radiographicfinding (70.8%). No significant correlation was observed between dialysis duration, biochemicalalterations, and oral health. Conclusions: Most patients undergoing dialysis presented precariousoral hygiene, periodontal inflammation, and bone alterations. However, these manifestationswere not influenced by the duration of dialysis or bone metabolism.

Immunohistochemical expression of protein 53, murine double minute 2, B-cell lymphoma 2, and proliferating cell nuclear antigen in odontogenic cysts and keratocystic odontogenic tumor.

Introduction: Even though odontogenic cysts share a similar histogenesis, they show different growth and differentiation profile due to differences in the proliferative cellular activity. Aims: We perform an immunohistochemical assessment of protein 53 (p53), proliferating cell nuclear antigen (PCNA), B-cell lymphoma 2 (bcl-2), and murine double minute 2 (MDM2) expression in odontogenic cysts and keratocystic odontogenic tumor analyzing their correlation with the biological behavior of these lesions. Materials and Methods: By the streptavidin-biotin-peroxidase method with antibodies against p53, PCNA, bcl-2, and MDM2 proteins, 11 radicular cysts, 11 dentigerous cysts, and 11 keratocystic odontogenic tumor were analyzed. The non-parametric Mann-Whitney U-test and Kruskall-Wallis test (P ≤ 0.05) were used to analyze the data. Results: Immunopositivity for PCNA was observed in all cases appraised, predominantly in the suprabasal layer of keratocystic odontogenic tumor epithelial lining (SD ± 19.44), but no significant differences were found among the groups of lesions. Bcl-2 immunoexpression was observed especially in the basal layer of keratocystic odontogenic tumor. PCNA LI was significantly higher than bcl-2 LI in keratocystic odontogenic tumor. MDM2 and p53 immunoexpression were not detected in the lesions studied. Among the evaluated lesions, the keratocystic odontogenic tumor showed different immunoexpression of the proliferation and apoptosis markers. Conclusion: The results of this study suggest that the keratocystic odontogenic tumor presents distinct biological behavior of the odontogenic cysts, as for the processes of proliferation, apoptosis, and differentiation, reinforcing the information in favor of the neoplastic nature of this lesion.

Expressão de MMPs, marcadores angiogênicos e proliferação celular em tumores odontogênicos / Expression of MMPs, angiogenic and proliferation cell markers in odontogenic tumors

INTRODUÇÃO E OBJETIVO: O conhecimento do comportamento biológico de lesões de natureza odontogênica é essencial para tornar a abordagem terapêutica adequada e estabelecer um prognóstico. A produção de metaloproteinases da matriz extracelular (MMPs), a angiogênese e a proliferação celular fornecem subsídios para o crescimento tumoral. O presente artigo tem como objetivo fazer uma revisão de literatura de pesquisas em tumores odontogênicos (TOs) selecionados a partir da nova classificação da Organização Mundial da Saúde (OMS) de 2005 sobre a expressão de MMPs, marcadores angiogênicos e proliferação celular e verificar, nestes estudos, a relação desses marcadores quanto ao comportamento biológico dessas lesões. RESULTADOS: Nota-se que as MMPs -1, -2, -7, -9 e -26 encontram-se mais expressas no componente epitelial e estroma e, particularmente, a -13 em estroma. Uma maior angiogênese é observada em TOs mais agressivos. CD 105 foi mais expresso no TO ceratocístico (TOC) e CD34 em ameloblastomas sólidos (ASs). Relata-se elevada expressão do Ki-67 e p53 no TOC e no AS e baixo índice de proliferação celular no TO adenomatoide (TOA). CONCLUSÃO: Esses resultados mostram que as MMPs participam no processo de invasão e recorrência de algumas lesões odontogênicas, estando associadas ao comportamento biológico desses tumores. A angiogênese é fundamental para fornecer suporte à proliferação celular e esses dois eventos em conjunto estão correlacionados com diferentes níveis de comportamento biológico nos TOs, quando comparados com cistos de natureza odontogênica, o que pode sugerir o uso de inibidores angiogênicos como provável abordagem terapêutica nessas lesões.

INTRODUCTION AND OBJECTIVE: The study of biological behavior of odontogenic lesions is essential to the establishment of appropriate therapeutic approach and prognosis. The production of extracellular matrix metalloproteinases (MMPs), angiogenesis and cell proliferation contribute to tumor growth. This paper aims to review the literature on odontogenic tumors (OT) selected according to the new World Health Organization classification (WHO- 2005) by evaluating the expression of MMPs, angiogenic and cell proliferation. Furthermore, it aims to verify the relation between these markers and the biological behavior of these lesions. RESULTS: it was found that MMPs -1, -2, -7, -9 and -26 had a higher expression in both epithelial component and stroma, and 13 particularly in the stroma. Increased angiogenesis was observed in more aggressive OT. CD105 expression was higher in keratocystic odontogenic tumour (KOT) and CD34 in solid ameloblastomas (SA). It was observed a higher expression of Ki-67 and p53 in SA and KOT and a low cell proliferation rate in the adenomatoid odontogenic tumour (AOT). CONCLUSION: These results show that MMPs are involved in invasion and recurrence of some odontogenic lesions and are associated with the biological behavior of these tumors. Angiogenesis is critical to provide support to cell proliferation and these concomitant events are correlated with different levels of biological behavior in OT when compared to odontogenic cysts, hence the use of angiogenic inhibitors may be a potential therapeutic approach in these lesions.