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

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.

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.

Macrophages subpopulations in chronic periapical lesions according to clinical and morphological aspects.

The aim of this study was to evaluate macrophage M1 and M2 subpopulations in radicular cysts (RCs) and periapical granulomas (PGs) and relate them to clinical and morphological aspects. M1 macrophages were evaluated by the percentage of CD68 immunostaining associated with the inflammatory cytokine TNF-α, and M2 macrophages, by its specific CD163 antibody. The CD68+/CD163+ ratio was adopted to distinguish between the two macrophage subpopulations. Clinical, radiographic, symptomatology, treatment, and morphological parameters of lesions were collected and a significance level of p = 0.05 was adopted for statistical analysis. The results showed that the CD68+/CD163+ ratio was higher in the RCs (median = 1.22, p = 0.002), and the highest TNF-α immunostaining scores were found in RCs (p = 0.018); in PGs, the CD68+/CD163+ ratio was lower and associated with a greater CD163+ immunostaining (median = 1.02, p <0.001). The TNF-α in cyst epithelium had a score of 3 in 10 cases and predominance of M1 macrophages by CD68+/CD163+ (median = 2.23). In addition, CD68+ cells had higher percentage of immunostaining in smaller RCs (p = 0.034). Our findings suggest that increased CD68 immunostaining associated with TNF-α cytokine in RCs results in a greater differentiation of the M1 phenotype. The higher CD163 immunostaining in PGs results in greater differentiation of the M2 phenotype. Therefore, the inflammatory state promoted by M1 macrophages is related to growth and progression of RCs; on the other hand, the immunomodulatory state of M2 macrophages is related to maintenance of PGs.

Macrophages subpopulations in chronic periapical lesions according to clinical and morphological aspects

Abstract: The aim of this study was to evaluate macrophage M1 and M2 subpopulations in radicular cysts (RCs) and periapical granulomas (PGs) and relate them to clinical and morphological aspects. M1 macrophages were evaluated by the percentage of CD68 immunostaining associated with the inflammatory cytokine TNF-α, and M2 macrophages, by its specific CD163 antibody. The CD68+/CD163+ ratio was adopted to distinguish between the two macrophage subpopulations. Clinical, radiographic, symptomatology, treatment, and morphological parameters of lesions were collected and a significance level of p = 0.05 was adopted for statistical analysis. The results showed that the CD68+/CD163+ ratio was higher in the RCs (median = 1.22, p = 0.002), and the highest TNF-α immunostaining scores were found in RCs (p = 0.018); in PGs, the CD68+/CD163+ ratio was lower and associated with a greater CD163+ immunostaining (median = 1.02, p <0.001). The TNF-α in cyst epithelium had a score of 3 in 10 cases and predominance of M1 macrophages by CD68+/CD163+ (median = 2.23). In addition, CD68+ cells had higher percentage of immunostaining in smaller RCs (p = 0.034). Our findings suggest that increased CD68 immunostaining associated with TNF-α cytokine in RCs results in a greater differentiation of the M1 phenotype. The higher CD163 immunostaining in PGs results in greater differentiation of the M2 phenotype. Therefore, the inflammatory state promoted by M1 macrophages is related to growth and progression of RCs; on the other hand, the immunomodulatory state of M2 macrophages is related to maintenance of PGs.

Estaniocalcina 2 modula eventos importantes para a tumorigênese oral e é um marcador prognóstico para pacientes com carcinoma de células escamosas oral / Staniocalcin 2 modulates important events for oral tumorigenesis and is a prognostic marker for patients with oral squamous cell carcinoma

O hormônio glicoproteico estaniocalcina 2 (STC2) está envolvido na carcinogênese e progressão de muitos tipos de câncer. No entanto, seu significado clínico e mecanismos moleculares no carcinoma de células escamosas oral (CCEO) foram pouco estudados e permanecem incertos. O presente estudo investigou associações da expressão da STC2 com parâmetros clinicopatológicos e de sobrevida em pacientes com CCEO. Além disso, foram avaliados os efeitos biológicos causados pela redução dos níveis de STC2 em linhagens celulares de CCEO e fibroblastos associados ao câncer (do inglês CAF ­ carcinoma associated fibroblasts). A análise imunoistoquímica em 100 casos de CCEOs primários indicou que a superexpressão da STC2 foi associada com o parâmetro N do sistema TNM e foi um fator de risco independente para sobrevida específica da doença e sobrevida livre de doença em pacientes com CCEO. Usando ensaios in vitro, foi demonstrado que o silenciamento da STC2 em linhagens de CCEO promoveu a apoptose e reduziu a proliferação celular, migração, invasão e transição epitélio-mesenquimal. Análises adicionais revelaram que o CAF expressa maiores níveis de STC2 do que as células de CCEO. O silenciamento da STC2 no CAF reduziu a invasão celular do CCEO, sugerindo que a STC2 liberada por CAFs contribui para um fenótipo mais invasivo no CCEO. Esses resultados sugerem que a STC2 modula eventos importantes para a tumorigênese oral e pode ser um biomarcador prognóstico para pacientes com CCEO (AU).

The glycoprotein hormone stanniocalcin 2 (STC2) is involved in carcinogenesis and progression of several cancer types. However, its clinical significance and molecular mechanisms in oral squamous cell carcinoma (OSCC) have been partially studied and remain uncertain. In the present study, we investigated associations of STC2 expression with clinicopathological and survival parameters of OSCCs patients. We also determined the biological effects caused by STC2 downregulation in OSCC and cancer associated fibroblasts (CAF) cell lines. Immunohistochemical analysis in 100 cases of primary OSCC indicated that STC2 overexpression was associated with N stage (TNM staging) and was an independent risk factor for disease-specific survival and disease-free survival in patients with OSCC. Using in vitro assays, we demonstrated that STC2 knockdown in OSCC cell lines promoted apoptosis, and reduced cell proliferation, migration, invasion and epithelial-mesenchymal transition. Further analysis revealed that CAF expresses higher levels of STC2 than OSCC cells. Knockdown of STC2 in CAF reduced OSCC cell invasion, suggesting that STC2 released by CAF contributes to a more invasive phenotype in OSCC. These results suggest that STC2 modulates important events for oral tumorigenesis and can be a prognostic biomarker for OSCC (AU).

Expressão imunoistoquímica de EGFR e PTEN em displasias epiteliais orais / Immunohistochemical expression of EGFR and PTEN in oral epithelial dysplasia

A displasia epitelial (DE) oral é uma desordem potencialmente maligna (DPM), cujo diagnóstico e gradação histológica se baseiam nas suas alterações arquiteturais e citológicas. Para avaliar o risco de transformação maligna dessas lesões de forma mais precisa é fundamental entender e localizar alterações genéticas e epigenéticas nas células displásicas, as quais podem ajudar a compreender melhor a progressão para a malignidade. Dessa forma, o presente estudo objetivou avaliar a imunoexpressão de EGFR e PTEN nas DEs orais e relacionar esse aspecto com as características clínicas e gradação histológica pelo sistema binário (baixo e alto risco de transformação maligna). Para tanto, foram selecionados 20 casos de DE de alto risco e 20 de baixo risco para serem submetidos à análise imunoistoquímica para os biomarcadores supracitados. A imunomarcação de cada caso foi avaliada semiquantitativamente através de escores e quanto à localização nos estratos epiteliais. A análise estatística foi realizada através dos testes de Mann-Whitney, Qui-quadrado de Pearson, Exato de Fisher e de correlação de Spearman com nível de significância estabelecido em 5%. Os resultados mostraram que 57,5% dos pacientes eram do gênero feminino, a média de idade foi de 57,5 anos, 42,5% foram diagnosticados clinicamente como leucoplasia e a maioria dos casos foi proveniente de lesões localizadas na língua (32,5%). De forma geral, gênero e idade não exerceram influência na imunoexpressão do EGFR e PTEN. A expressão do EGFR foi observada em 100% dos casos, nos quais houve predomínio do escore 3 (75%) e imunoreatividade em todas as camadas epiteliais (55%), independente da gradação histológica (p = 0,453 e p = 0,204, respectivamente). O PTEN revelou positividade de marcação em 87,5% dos casos, nos quais observou-se predomínio do escore 0 (55%) e imunoreatividade limitada à camada basal (40%), porém sem diferenças significativas entre os grupos histológicos (p = 0,904 e p = 0,915, respectivamente). Por fim, quando analisados, em conjunto, os 40 casos de DEs, foi observada uma fraca correlação positiva, estatisticamente significativa, entre os padrões de imunoexpressão do EGFR e do PTEN (r = 0,317; p = 0,046). Com base nesses resultados, alterações no padrão de expressão do EGFR e PTEN sugerem que essas proteínas participam de processos moleculares relacionados com a carcinogênese em mucosa oral. (AU)

The oral epithelial dysplasia (ED) is a potentially malign disorder (PMD) which the diagnosis and histological gradation are based on their architectural and cytological alterations. To evaluate the risk of malign transformation of these lesions in a more precise manner it is fundamental to understand and localize genetic and epigenetic alterations on the dysplastic cells, which can help to comprehend better the progression to malignancy. This way, this study has as an objective to evaluate the immunostaining of EGFR and PTEN in the oral EDs and relate it with the clinical characteristics and histological gradation using the binary system (low and high risk of malign transformation). To this end, 20 cases of high risk ED and 20 of low risk ED were selected to be subject to immunochemical analysis to the aforementioned biomarkers. The immunostaining of each case was evaluated semiquantitatively by scores and it was also analyzed regarding the epithelial strata. Statistical analysis was performed using the Mann-Whitney, Pearson's Chi Square, Fisher's exact and Spearman's correlation tests with significance level set at 5%. The results showed that 57,5% of the patients were female, the mean age was 57,5 years, 42,5% were clinically diagnosed as leucoplasia and most of the cases were from tongue lesions (32.5%). Gender and age did not influence on the immunoexpression of EGFR and PTEN. The expression of EGFR was observed in 100% of the cases, in which were predominance of score 3 (75%) and immunoreactivity limited to the epithelial layers (55%), regardless of the histological gradation (p=0,453 and p= 0,204, respectively). The PTEN revealed marking positivity in 87,5% of the cases, in which it was observed the predominance of score 0 (55%) and immunoreactivity limited to the basal layer (40%), but without significant differences between the histological groups (p= 0,904 and p= 0,915, respectively). Lastly, when the 40 cases of EDs were analyzed collectively, it was observed a weak positive correlation, statistically significant, among the patterns of immunoexpression of EGFR and PTEN (r= 0,317; p=0,046). Based on those results, the alterations in the pattern of immunoexpression of EGFR and PTEN suggest that these proteins participate on the molecular processes related to oral carcinogenesis. (AU)