Immunoexpression of CXCL12 and CXCR4 in Radicular Cysts, Dentigerous Cysts, and Odontogenic Keratocysts.

The aim of this study was to evaluate the immunoexpression of chemokine CXCL12 and its receptor CXCR4 in radicular cysts (RCs), dentigerous cysts (DCs), and odontogenic keratocysts (OKCs), and to correlate the findings with morphologic parameters of RCs (inflammatory infiltrate and cystic epithelium). Twenty RCs, 20 DCs, and 20 OKCs were submitted to immunohistochemistry. The percentages of cytoplasmic (CXCL12 and CXCR4) and nuclear (CXCR4) staining in epithelial and fibrous capsule cells were determined. RCs and DCs exhibited higher epithelial expression of CXCL12 than OKCs ( P <0.05). The expression of CXCL12 in the fibrous capsule was higher in DCs than in RCs and OKCs ( P <0.05). Higher cytoplasmic expression of CXCR4 was observed in the epithelial lining and fibrous capsule of RCs and DCs compared with OKCs ( P <0.05). In the fibrous capsule, DCs exhibited higher nuclear expression of CXCR4 than OKCs ( P <0.05). No significant differences in the immunoexpression of CXCL12 or CXCR4 were observed according to the morphologic parameters of RCs ( P >0.05). Strong positive correlations were found between cytoplasmic and nuclear expression of CXCR4 in the epithelial lining of RCs and DCs and in the fibrous capsule of all groups ( P <0.05). The results suggest the participation of CXCL12 and CXCR4 in the pathogenesis of RCs, DCs, and OKCs. These proteins may be particularly relevant for the development of odontogenic cysts with less aggressive biological behavior, irrespective of their nature (inflammatory or developmental). In RCs, the expression of CXCL12 and CXCR4 may not be related to the intensity of the inflammatory infiltrate or the status of cystic epithelium.

Variability in the Epithelial Expression of Ki67 and Polarization Characteristics of Collagen in Odontogenic Cysts and Tumours

ABSTRACT Objective: To assess the proliferation of epithelium (using the Ki67 index) and the polarization pattern of collagen in selected odontogenic cysts and tumours. In addition, an exploratory analysis of the effect of inflammation on the proliferation rate was done. Material and Methods: Following immunohistochemical staining, the labelling/proliferation index of Ki67 was calculated. The thickness and corresponding polarization colour of 100 juxta-epithelial picrosirius red-stained collagen fibers were assessed using linear micrometry with an eyepiece reticule under × 1000 magnification. Inflammation was graded subjectively as mild, moderate, and severe. Results: Overall Ki-67 expression was higher in the radicular cyst, Odontogenic Keratocyst, Ameloblastoma, while suprabasal Ki-67 positivity was maximum in Odontogenic Keratocyst. The stromal collagen fibers in Ameloblastoma showed predominantly green birefringence, whereas Odontogenic Keratocyst had orange birefringence. There was no significant association of inflammation with Ki-67 expression or birefringence patterns. Conclusion: The highest Ki67 expression in the radicular cyst, followed by Odontogenic Keratocyst and Ameloblastoma. Differences in the collagen maturation pattern were noted innately in five lesions studied and were further influenced by inflammatory changes. Epithelial proliferation and concomitant expression of thickness and maturity of the stromal collagen are innate features of the lesion further influenced by inflammation in various odontogenic cysts and tumours and may, in turn, guide the clinical behavior.

Abordagem cirúrgica de cisto periapical abscedado na maxila: relato de caso / Surgical approach of absceded periapical cyst in the maxilla: case report / Abordaje quirúrgico de quiste periapical abscedido en la maxila: reporte de caso

O cisto radicular é uma lesão inflamatória associada à necrose pulpar que ocorre frequentemente em maxila. Objetivo: Descrever um caso cirúrgico detalhado de Cisto Periapical Abscedado. Relato de Caso: paciente gênero feminino, 40 anos, hipertensa, diabética Tipo II, compareceu à clínica queixando-se de dor ao ingerir alimentos frios e quentes na região da maxila, lado esquerdo. Ao exame físico, observou-se destruição coronária e presença de fístula na região do elemento dentário 23. Ao exame radiográfico, observou-se uma área radiolúcida ovalada bem circunscrita com halo radiopaco envolvendo a região apical do elemento dentário 23. Frente ao aspecto clínico e radiográfico, foram sugeridas as hipóteses diagnósticas de abscesso periapical crônico, granuloma periapical ou cisto apical abscedado. Foi realizada a exodontia do elemento 23 seguida de enucleação cística. O diagnóstico histopatológico final foi de cisto abscedado. Após 5 meses de evolução observa-se mucosa íntegra e reparo ósseo alveolar. Conclusão: É imprescindível um exame clínico cuidadoso associado ao exame radiográfico e histopatológico para analisar minuciosamente o caso a fim de oferecer ao paciente melhores condutas de tratamento. O diagnóstico de lesões intraósseas associado ao correto tratamento interrompe a evolução do processo patológico, evita danos maiores e restabelece a condição de saúde dos pacientes... (AU)

The radicular cyst is an inflammatory lesion associated with pulp necrosis that often occurs in the maxilla. Objective: To describe a detailed surgical case of Abscessed Periapical Cyst. Case Report: female patient, 40 years old, hypertensive, type II diabetic, came to the clinic complaining of pain when ingesting cold and hot foods in the left side of the maxilla. On physical examination, coronary destruction and the presence of a fistula in the region of the tooth 23 were observed. The radiographic examination showed a well-circumscribed oval radiolucent area with a radiopaque halo involving the apical region of the tooth 23. In view of the clinical and radiography, the diagnostic hypotheses of chronic periapical abscess, periapical granuloma or abscessed apical cyst were suggested. Element 23 extraction was performed followed by cystic enucleation. The final histopathological diagnosis was an abscessed cyst. After 5 months of evolution, intact mucosa and alveolar bone repair are observed. Conclusion: A careful clinical examination associated with radiographic and histopathological data is essential to systematically analyze the case in order to offer the patient better treatment. The diagnosis of intraosseous lesions associated with the correct treatment interrupts the evolution of the pathological process, avoids further damage and restores the patients' health condition... (AU)

El quiste radicular es una lesión inflamatoria asociada a necrosis pulpar que frecuentemente se presenta en el maxilar. Objetivo: Describir un caso quirúrgico detallado de Quiste Periapical Absceso. Caso Clínico: paciente femenina, de 40 años, hipertensa, diabética tipo II, acudió a la consulta quejándose de dolor al ingerir alimentos fríos y calientes en el lado izquierdo del maxilar. Al examen físico se observó destrucción coronaria y la presencia de una fístula en la región del diente 23. El examen radiográfico mostró un área radiolúcida oval bien delimitada con un halo radiopaco que involucraba la región apical del diente 23. En vista de la clínico y radiográfico, se sugirieron las hipótesis diagnósticas de absceso periapical crónico, granuloma periapical o quiste apical abscesificado. Se realizó la extracción del elemento 23 seguida de enucleación quística. El diagnóstico histopatológico final fue de quiste abscesificado. A los 5 meses de evolución se observa mucosa intacta y reparación del hueso alveolar. Conclusión: Un examen clínico cuidadoso asociado con el examen radiográfico e histopatológico es fundamental para analizar a fondo el caso con el fin de ofrecer al paciente mejores enfoques de tratamiento. El diagnóstico de lesiones intraóseas asociado al correcto tratamiento interrumpe la evolución del proceso patológico, previene mayores daños y restablece el estado de salud de los pacientes... (AU)

Imunoexpressão de ING 4, VEGF e NF-κB em lesões periapicais inflamatórias / Immunoexpression of ING 4, VEGF and NF-κB in inflammatory periapical lesions

As lesões periapicais inflamatórias (LPIs) são condições patológicas decorrentes de infecções de origem odontogênica, principalmente representadas pelos granulomas periapicais (GPs) e cistos radiculares (CRs). Sua patogênese está associada a mecanismos imunológicos e angiogênicos. O presente estudo, do tipo retrospectivo, teve como objetivo analisar, de forma semiquantitativa, a expressão imuno-histoquímica de ING-4, VEGF e NF-κB em LPIs, e correlacionar o padrão de expressão dessas proteínas. A amostra consistiu de 26 GPs, 17 CRs e 19 cistos radiculares residuais (CRRs). Foram avaliados espessura epitelial e infiltrado inflamatório, e a associação desses achados com o padrão de expressão das proteínas ING-4, VEGF e NF-κB nas LPIs selecionadas. Para a realização da análise estatística, foram utilizados os testes de qui-quadrado, Kruskal-Wallis, Mann-Whitney e correlação de Spearman (p < 0,05). O infiltrado inflamatório exibiu maior intensidade no GP, seguido pelo CR, e por último, o CRR (p < 0,05). Apesar de não haver associação estatisticamente significativa ao associar a expressão de ING-4 nas células inflamatórias do tecido conjuntivo ou cápsula fibrosa entre os grupos de LPIs, o GP e CR evidenciaram, através da média de postos, maior expressão dessa proteína. Não foi evidenciada associação estatisticamente significativa de ING-4 com a intensidade do infiltrado inflamatório. A imunoexpressão de VEGF no núcleo das células inflamatórias do tecido conjuntivo ou cápsula fibrosa exibem associação significativa com as LPIs, que ocorre maior expressão dessa proteína nos cistos (p= 0,002). A maior expressão de NF-κB foi evidenciada nos casos de GPs, tanto a nível nuclear quanto citoplasmático das células inflamatórias (p = 0,005; p = 0,002). Não houve associação estatisticamente significativa quando comparada a expressão de NF-κB entre os cistos, mas a mediana da expressão dessa proteína foi maior para os CRs. Na cápsula fibrosa, a imunoexpressão nuclear e citoplasmática de NF-κB nas células inflamatórias foi superior nas lesões periapicais com intenso infiltrado inflamatório (p < 0,001). Portanto, sugere-se que ING-4, VEGF e NF-κB participam do desenvolvimento das LPIs, e apesar de ocorrer relação diretamente proporcional entre a expressão dessas proteínas, ING-4 não exerceu atividade reguladora na inflamação associada a essas lesões (AU).

Inflammatory periapical lesions (IPLs) are pathological conditions resulting from infections of odontogenic origin, being mainly represented by periapical granulomas (PGs) and radicular cysts (RCs). Its pathogenesis is associated with immunological and angiogenic mechanisms. This retrospective study, semi-quantitative and comparative aimed to analyze the immunohistochemical expression of ING-4, VEGF and NF-κB in IPLs, and to correlate the pattern of expression of these proteins. The sample consisted of 26 were PGs, 17 RCs and 19 residual radicular cysts (RRCs). Epithelial thickness and inflammatory infiltrate were evaluated, and the correlation of these findings with the expression pattern of ING-4, VEGF and NF-κB proteins in selected IPLs. To perform the statistical analysis, the chi-square, Kruskal-Wallis, Mann-Whitney and Spearman correlation tests were used (p < 0.05). The inflammatory infiltrate exhibited greater intensity in the PG, followed by the RC, and finally, the RRC (p < 0.05). Although there was no statistically significant association when associating the expression of ING-4 in inflammatory cells of the connective tissue or fibrous capsule the groups of IPLs, the PG and RC showed higher expression of this protein. There was no statistically significant association between ING-4 and the intensity of the inflammatory infiltrate. Immunoexpression of VEGF in the nucleus of inflammatory cells in the connective tissue or fibrous capsule shows a significant association with IPLs, in which there is greater expression of this protein occurring in cysts (p= 0,002). The highest expression of NF-κB was evidenced in cases of PGs, both at the nuclear and cytoplasmic level of inflammatory cells (p=0,005; p= 0,002). There was no statistically significant association when comparing the expression of NF-κB between the cysts, but the median expression of this protein was expression was higher for the RCs. In the fibrous capsule, nuclear and cytoplasmic NF-κB immunoexpression in inflammatory cells was higher in periapical lesions with intense inflammatory infiltrate (p<0.001). Therefore, it is suggested that ING-4, VEGF and NF-κB participate in the etiopathogenesis of IPLs, and that there is a directly proportional relationship between the expression of these proteins. ING-4 did not exert regulatory activity in the inflammation associated with these lesions (AU).

Expression of epithelial growth factors and of apoptosis-regulating proteins, and presence of CD57+ cells in the development of inflammatory periapical lesions.

OBJECTIVE: The mechanisms that stimulate the proliferation of epithelial cells in inflammatory periapical lesions are not completely understood and the literature suggests that changes in the balance between apoptosis and immunity regulation appear to influence this process.To evaluate the expression of the epidermal growth factor (EGF), its receptor (EGFR) and of the keratinocyte growth factor (KGF), the presence of CD57+ cells, the epithelial cell proliferation index, and the expression of the Bcl-2 protein in inflammatory periapical lesions (IPL) at different stages of development. METHODOLOGY: Our sample was composed of 52 IPLs (22 periapical granulomas - PG - and 30 periapical cysts - PC), divided into three groups: PGs, small PCs, and large PCs. Specimens were processed for histopathologic and immunohistochemical analyses. Sections were evaluated according to the amount of positive staining for each antibody. RESULTS: We found no significant differences among the groups regarding Bcl-2 (p=0.328) and Ki-67 (p>0.05) expression or the presence of CD57+ cells (p=0.748). EGF (p=0.0001) and KGF (p=0.0001) expression was more frequent in PCs than in PGs, and CD57+ cells were more frequent in IPLs with intense inflammatory infiltrates (p=0.0001). We found no significant differences in KGF (p=0.423), Bcl-2 (p=0.943), and EGF (p=0.53) expression in relation to inflammatory infiltrates or to the type of PC epithelial lining, but observed greater KGF expression (p=0.0001) in initial PCs. EGFR expression was similar among the groups (p>0.05). CONCLUSION: More frequent EGF and KGF expression in PCs and the greater presence of CD57+ cells in lesions with intense inflammatory infiltrates suggest that these factors influence IPL development. The greater KGF expression in initial PCs suggests its importance for the initial stages of PC formation.

¿Los Quistes Periradiculares Dependen de la Infección Endodóntica? / ¿Radicular Cysts Depend on Endodontic Infection?

Los quistes periradiculares, tanto los de bolsillo como los verdaderos, son de origen inflamatorio; sin embargo, en la literatura ha existido una gran controversia asociada, en primer lugar, a la dependencia de la infección contenida dentro del sistema de conductos radiculares y, en segundo lugar, al tipo de tratamiento endodóntico llevado a cabo en este tipo de lesiones. Con el fin de dilucidar esta controversia se realizará una revisión a la literatura sobre quistes perira diculares donde se explicará cuál es la técnica más adecuada en el diagnóstico de quistes verdaderos y de bolsillo. Igualmente, se explicará la teoría de autosustentabilidad, así como el tratamiento endodóntico de grandes lesiones periradiculares asociadas a dichos quistes. Ahora bien, en la actualidad se ha demostrado que ambos quistes están asociados a infecciones contenidas en el sistema de conductos radiculares y son diferentes solamente en su morfología. Así mismo, se ha recomendado el tratamiento de conducto convencional en dientes con grandes lesiones asociadas a quistes.

Radicular cysts, either the pocket or the true cysts, are originated by an inflammation. However, in the literature, there has been great controversy surrounding, firstly, the dependence of the infection contained within the radicular conducts and, secondly, the kind of endodontic treatment performed in this sort of injury. In order to clarify this controversy, the literature concerning radicular cysts will be reviewed and it will be explained what technique is the most appropriate for diagnosing true cysts and pocket cysts. Likewise, the theory of self-sustaining, as well as the endodontic treatment for large radicular injuries associated to such cysts, will be further explained. Nonetheless, nowadays it has been demonstrated that both cysts are associated to infections contained within the system of radicular conducts and that they are only morphologically different. In addition, the root canal treatment has been recommended for teeth with great injuries associated to cysts.

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.

[Artigo Parcialmente Retratado] Prevalência da infecção por Helicobacter pylori em pacientes dispépticos e associação com fatores de riscos clínicos para o desenvolvimento de adenocarcinoma gástrico / [article partial retraction] prevalence of helicobacter pylori infection in dyspeptic patients and its association with clinical risk factors for developing gastric adenocarcinoma

ABSTRACT BACKGROUND: In Brazil, particularly in the underdeveloped localities, the prevalence of Helicobacter pylori (H. pylori) infections can range up to 90%. These rates are higher in older individuals and vary by country region. H. pylori infections are linked to the development of gastric pathologies, namely mild to moderate gastritis, gastroenteritis, peptic ulcer, intestinal metaplasia, and gastric cancer. In 1994, this organism was classified by the International Agency for Research on Cancer (IARC) as pertaining to the Group 1 carcinogen for gastric adenocarcinoma etiology. Gastric cancer represents a significant public health problem, being the fourth most common malignant tumor and the second largest cause of cancer-related deaths. OBJECTIVE: To investigate the prevalence of H. pylori infection in dyspeptic patients and determine the link between clinical risk factors and gastric adenocarcinoma diagnosis. METHODS: Polymerase chain reaction (PCR) analysis was employed for molecular diagnosis of gastric tissue biopsies collected from 113 dyspeptic patients at the University Hospital of Federal University of Goiás. Molecular analyses allowed the identification of H. pylori infections. Furthermore, histopathological examinations were performed to determine the clinical risks of developing gastric malignancies. RESULTS: The test results identified 69 individuals older than 44 years, from 75 (66.4%) positive H. pylori infection samples. The prevalence of gastric adenocarcinoma in this study was 1.3%. Among the infected patients, six (8.2%) had high risk, and 67 (91.8%) had a low risk of developing gastric cancer (P<0.05). CONCLUSION: This study shows a high prevalence of H. pylori infection and identifies its contribution to gastric inflammations, which in the long term are manifested in high-risk clinical factors for the development of gastric adenocarcinoma.

RESUMO CONTEXTO: No Brasil, particularmente nas áreas mais pobres, a prevalência da infecção por Helicobacter pylori pode variar até 90%. Esses índices aumentam com o envelhecimento da população e são distintos entre as diferentes regiões do país. Podendo manifestar diferentes sintomatologias, essa infecção está diretamente relacionada com o desenvolvimento de patologias gástricas como gastrite leve a moderada, gastroenterites, úlcera péptica, metaplasia intestinal e principalmente, o câncer gástrico. Em 1994 a bactéria foi categorizada pela International Agency for Research on Cancer (IARC) como carcinógeno do Grupo 1 para adenocarcinoma gástrico, tipo de câncer que representa um importante problema de saúde pública, sendo o quarto tumor maligno mais comum e a segunda maior causa de mortes por câncer no mundo. OBJETIVO: Analisar a prevalência da bactéria em pacientes dispépticos e avaliar a associação de fatores de risco clínicos para desenvolvimento de adenocarcinoma gástrico. MÉTODOS: Biópsias de tecido gástrico coletadas de 113 pacientes dispépticos, atendidos no Hospital das Clínicas da Universidade Federal de Goiás, foram submetidas a diagnóstico molecular por meio de Reação em Cadeia da Polimerase, para identificação da infecção por Helicobacter pylori, e exame histopatológico, para avaliar o risco clínico de desenvolvimento de adenocarcinoma gástrico. RESULTADOS: Foram diagnosticadas 75 (66,4%) amostras positivas para infecção por Helicobacter pylori, sendo 69 indivíduos maiores de 44 anos de idade. A prevalência do adenocarcinoma gástrico nesse estudo foi de 1,3% e dentre os pacientes positivos para a infecção bacteriana seis (8,2%) possuem alto risco e 67 (91,8%) baixo risco de desenvolver esse tipo de câncer (P<0,05). CONCLUSÃO: Esse estudo mostra uma alta prevalência da infecção por H. pylori na população estudada e identifica sua intrínseca contribuição para inflamações gástricas, que a longo prazo se manifestam em fatores clínicos de alto risco para o desenvolvimento de adenocarcinoma gástrico.

Estudo imunoistoquímico das proteínas CD34 e α-SMA em cistos radiculares e cistos radiculares residuais / Immunohistochemical study of CD34 and α-SMA proteins in radicular cysts and residual radicular cysts

Introdução: Os cistos radiculares são lesões inflamatórias provenientes de processos infecciosos da necrose da polpa. Os miofibroblastos (MFs) e a angiogênese são fatores importantes no desenvolvimento e progressão das lesões císticas, de modo que essas células secretam fatores de crescimento e proteases, assim como a angiogênese é importante para a produção de matriz e a migração celular, afetando o crescimento das periapicopatias. Objetivo: O presente trabalho consistiu em analisar, por meio da imunoistoquímica, as expressões do CD34 e αSMA em cistos radiculares (CRs) e cistos radiculares residuais (CRRs), com o propósito de contribuir para um maior entendimento a respeito da expansão e progressão das lesões periapicais. Materiais e Métodos: Foram realizadas análises clínicas, morfológicas e imunoistoquímicas em 30 CRs e 30 CRRs. A presente pesquisa constituiu em uma análise descritiva e quantitativa das expressões imunoistoquímicas positivas do CD34 e α-SMA nos espécimes. A expressão do αSMA foi avaliada na cápsula fibrosa das lesões em 10 campos, inicialmente a um aumento de (100x), posteriormente em (400x) e uma média final foi estabelecida, entretanto, para o índice angiogênico foi realizada a contagem de microvasos imunomarcados pelo anticorpo anti-CD34 em 10 campos de (200x). A análise estatística foi realizada através dos testes de Mann-Whitney, Qui-quadrado de Pearson e Correlação de Spearman, o nível de significância estabelecido foi p<0,05. Resultados: Os resultados demonstraram diferenças estatisticamente significativas referentes ao α-SMA e α-SMA versus CD34, todavia, com relação a imunomarcação do CD34 não revelou diferença estatística entre as lesões. Conclusão: Diante dos resultados obtidos consideramos que a correlação significativa entre a presença de miofibroblastos e o índice angiogênico entre as lesões estudadas estão relacionadas com o reparo das periapicopatias (AU).

Introduction: Radicular cysts are inflammatory lesions resulting from infectious processes of pulp necrosis. Myofibroblasts (MFs) and angiogenesis are important factors in the development and progression of cystic lesions, so that MFs secrete growth factors and proteases, as well as angiogenesis stimulates matrix deposition and cell migration, affecting the growth of periapicopathies. Objective: This study aims to evaluate the immunohistochemical expression of CD34 and α-SMA in radicular cysts (CRs) and residual radicular cysts (CRRs), in order to contribute a better understanding of expansion and progression of periapical lesions. Materials and Methods: Clinical, morphological and immunohistochemical analyzes were performed on 30 CRs and 30 CRRs. The present study constitutes a descriptive, quantitative and comparative analysis of CD34 and α-SMA positive immunohistochemical expression in the specimens. Expression of α-SMA was evaluated in the fibrous capsule, at 100x magnification 10 fields of higher immunolabeling were selected below epithelial lining, thereafter at 400X magnification, the positive cells were quantified and at the end were calculated the averages of positive cells per field. However, angiogenic index was performed by counting microvessels immunolabelled by anti-CD34 antibody in 10 (200x) fields. Statistical analysis was performed using the Mann-Whitney, Pearson's Chi-square test and Spearman's correlation, with significance level set (p <0.05). Results: The results showed that there were statistically significant differences regarding α-SMA and α-SMAxCD34, however with respect to the CD34 immunostaining, there were no statistical differences between the lesions. Conclusion: Myofibroblasts and angiogenic index among studied lesions presented a significant correlation. Thus, they are believed contribute to the repair of the periapicopathies (AU).

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.

Periodontitis apical: caracterización histológica y morfométrica de quistes radiculares y granulomas periapicales / Apical periodontitis: histological and morphometric characterization of radicular cysts and periapical granulomas

La periodontitis apical es una patología inflamatoria que afecta los tejidos periapicales de un diente desvitalizado. El objetivo de este estudio fue caracterizar histológica y morfométricamente las lesiones de quistes y granulomas utilizando microscopía óptica. Se analizaron seis biopsias obtenidas de dientes con indicación de exodoncia. El análisis histológico se realizó mediante microscopía óptica y microfotografías, con análisis de contraste de imágenes y conteo celular mediante ImageJ. Descripción de las características histológicas: en los quistes se observaron cavidades rodeadas de epitelio escamoso estratificado no queratinizado y una cápsula fibrosa compuesta de fibrocitos, fibroblastos e infiltrado inflamatorio crónico; en los granulomas se observaron capilares, una capsula fibrosa de fibrocitos/fibroblastos y un infiltrado inflamatorio de predominio linfocitario. Cuantificación del número celular de infiltrado inflamatorio: para quistes fue de 9,2 cel/10000 µm2, mientras que para granulomas fue de 20,8 cel/10000 µm2, sin diferencias estadísticas significativas entre ambos (p=0,654). Cuantificación del número celular de fibrocitos/fibroblastos: para quistes fue de 15,4 cel/10000 µm2, mientras que para granulomas fue de 18,5 cel/10000 µm2, sin diferencia estadística significativa (p=0,499). Porcentaje de colágeno tipo I: para los quistes fue de 37,8±19,2 %, mientras que para granulomas fue de 33,8±23,3 %, sin diferencias estadísticas significativas (p=0,704). Se observó una correlación negativa moderada para el infiltrado inflamatorio (R=0,637) y una correlación positiva baja para fibrocitos/fibroblastos (R=0,121), en relación a la cantidad de colágeno tipo I. Medición del área de las lesiones periapicales: el promedio total de las lesiones fue de 10,7±5,0 mm2, siendo el mayor tamaño un quiste de 18,1 mm2 y el menor un granuloma de 5,2 mm2. El análisis histológico permite realizar un diagnóstico diferencial de lesiones con características similares y así definir el tratamiento más adecuado.

Apical periodontitis is an inflammatory pathology that affects the periapical tissues of a devitalized tooth. The aim of this study was to histologically and morphometrically characterize lesions of cysts and granulomas using light microscopy. Six biopsies obtained from teeth with indication of exodontia were analyzed. The histological analysis was carried out by means of optical microscopy and microphotographs, with contrast analysis of images and cell count by ImageJ. A description of the histological characteristics was made, observing the cavities surrounded by stratified squamous non-keratinized epithelium and a fibrous capsule composed of fibrocytes, fibroblasts and chronic inflammatory infiltrate; in the granulomas, capillaries, a fibrous capsule of fibrocytes/fibroblasts and a predominantly lymphocytic inflammatory infiltrate were observed. In relation to quantification of the cellular number of inflammatory infiltrate, for cysts itwas of 9.2 cel / 10000 mm2, while for granulomas it was 20.8 cel / 10000 mm2, without significant statistical differences between both (p = 0.654). The quantification of the fibrocyte / fibroblast cell number was, for cysts, 15.4 cells / 10000 mm2, while for granulomas it was 18.5 cells / 10000 mm2, without significant statistical difference (p = 0.499). With respect to the percentage of collagen type I, for the cysts was 37.8 ± 19.2%, while for granulomas it was 33.8 ± 23.3%, without significant statistical differences (p = 0.704). A moderate negative correlation was observed for the inflammatory infiltrate (R = 0.667) and a low positive correlation for fibrocytes / fibroblasts (R = 0.121), in relation to the amount of type I collagen. Measurement of the area of the periapical lesions: the total average of lesions were 10.7 ± 5.0 mm2, the largest being a cyst of 18.1 mm2 and the smallest a granuloma of 5.2 mm2. The histological analysis allows to make a differential diagnosis of lesions with similar characteristics and thus define the most appropriate treatment.

Immunohistochemical Analysis of Cyclooxygenase-2 and Tumor Necrosis Factor Alpha in Periapical Lesions.

INTRODUCTION: The purpose of this study was to evaluate the expression of cyclooxygenase-2 (COX-2) and tumor necrosis factor alpha (TNF-α) in periapical granuloma (PG) and radicular cyst (RC) samples and to correlate it with the type of lesion, the intensity of the inflammatory infiltrate, and the thickness of the epithelial lining. METHODS: A total of 51 cases of periapical lesions (25 PGs and 26 RCs) were subjected to morphologic analysis and immunohistochemical study. The anti-COX-2 and anti-TNF-α antibodies were applied using the immunoperoxidase technique. Data were analyzed by the Mann-Whitney test, Pearson chi-square test, Fisher exact test, and Spearman correlation. RESULTS: Analysis of the inflammatory infiltrate revealed that 80% of PGs exhibited a grade III infiltrate as opposed to a 19% rate in RCs (P < .001). Morphologic evaluation of the epithelial thickness of RCs revealed the presence of atrophic epithelium in 73% of cases. The majority of PGs had a score of 1 for COX-2 immunoexpression (n = 14, 54%) and a score of 2 for TNF-α expression (n = 16, 64%), whereas in cases of RCs a score of 1 was more prevalent for COX-2 and TNF-α expression (n = 17, 65%). Significant differences in the expression scores of COX-2 and TNF-α were detected in periapical lesions (P < .001). CONCLUSIONS: Based on these findings, we emphasize that RCs and PGs have a similar expression of inflammatory mediators (COX-2 and TNF-α) although the secretion of TNF-α by macrophages and of COX-2 by several cells was higher in PGs, indicating a greater inflammatory response in these lesions.

Pro-inflammatory and anti-inflammatory cytokine expression in post-treatment apical periodontitis.

OBJECTIVE: This study evaluated the expression of pro-inflammatory (IL-1ß, IL-6, IFN-γ and TNF-α) and anti-inflammatory (IL-4 and TGF-ß) cytokines in apical periodontitis lesions. Correlations between these cytokines and clinical and cone-beam computed tomographic (CBCT) data were also assessed. MATERIAL AND METHODS: Apical periodontitis lesions' data were obtained from 27 patients subjected to periradicular surgery. Specimens were processed for histopathologic and immunohistochemical analysis. Sections were evaluated according to the amount of positive staining for each antibody. Expression levels of the target mediators were compared with clinical and CBCT data. RESULTS: Twenty lesions were diagnosed as granuloma and 7 as cyst. In granulomas, IL-4 expression was significantly higher than IL-6 (p=0.001) and TNF-α (p=0.001). There was a significant relationship between high levels of TNF-α and lesions <5 mm (p=0.017). In cysts, IL-6 expression was significant lower than IL-4 (p=0.001) and IFN-γ (p=0.004). There was a significant relationship between high levels of TGF-ß and endodontic treatment performed ≤4 years before (p=0.045). In general, IL-4 was the most expressed mediator in both cysts and granulomas. CONCLUSIONS: There was a balance between the expression of pro-inflammatory and anti-inflammatory cytokines associated with the chronic periradicular inflammatory process. TNF-α and TGF-ß were related to some clinical and CBCT data.

Comparative Expression of CD34, Intercellular Adhesion Molecule-1, and Podoplanin and the Presence of Mast Cells in Periapical Granulomas, Cysts, and Residual Cysts.

INTRODUCTION: The aim of the present study was to compare the immunoexpression of CD34, intercellular adhesion molecule-1 (ICAM-1), and podoplanin and the presence of mast cells with clinical, demographic, radiologic, and histologic features from periapical granulomas, periapical cysts, and residual cysts. METHODS: Thirty-one lesions (5 granulomas, 15 periapical cysts, and 11 residual cysts) were selected. Histologic sections in silanized slides were used for the immunohistochemical reactions. The analysis of the images was performed by using an optical microscope, and data were analyzed with 5% significance (P < .05). RESULTS: Cysts presented atrophic and hyperplastic epithelium in 11 cases (35.5%) and 15 cases (48.8%), respectively (P > .05). The intensity of the inflammatory infiltrate was similar when comparing the 3 groups (P > .05). CD34 and podoplanin expression and the presence of mast cells were similar when comparing the 3 groups; ICAM-1 expression was more intense in granulomas than cysts (P < .05). There were no statistically significant differences associated with the expression of the evaluated markers according to the intensity of the inflammatory infiltrate. CONCLUSIONS: There were no differences in the expression of CD34 and podoplanin and in the presence of mast cells when the 3 groups were compared. ICAM-1 expression was more common in periapical granulomas.

Immunohistochemical Analysis of Galectins-1, -3, and -7 in Periapical Granulomas, Radicular Cysts, and Residual Radicular Cysts.

INTRODUCTION: Galectins play important roles in immunoinflammatory responses, but their participation in the development of periapical lesions remains unclear. This study aimed to evaluate the expressions of galectins-1, -3, and -7 in periapical lesions, correlating them with the intensity of the inflammatory infiltrate and the pattern of the cystic epithelium. METHODS: Twenty periapical granulomas (PGs), 20 radicular cysts (RCs), and 20 residual radicular cysts (RRCs) were submitted to immunohistochemistry using anti-galectin-1, -3, and -7 antibodies. The percentage of immunopositive cells in epithelial and connective tissues was determined. RESULTS: In connective tissue, PGs exhibited higher cytoplasmic/membrane expression of galectins-1 and -7 than RCs and RRCs (P < .05). There was higher nuclear expression of galectin-1 in PGs compared with RCs and RRCs (P < .05). The expression of galectins-1 and -7 in connective tissue was higher in lesions with grade III inflammation (P < .05). No significant differences in galectin-3 immunoexpression were observed for any of the parameters evaluated (P > .05). In the epithelial component, a higher nuclear expression of galectin-7 was detected in RRCs (P < .05), and a higher cytoplasmic/membrane expression of this protein was found in cysts with hyperplastic epithelium (P < .05). Positive correlations were observed between the nuclear and cytoplasmic/membrane expression of galectin-1 in connective tissue (P < .05) as well as between the nuclear and cytoplasmic/membrane expression of galectin-7 in epithelial tissue of cysts (P < .05). CONCLUSIONS: Galectins-1 and -7 may play important roles in the pathogenesis of PGs, RCs, and RRCs. On the other hand, the present results suggest only a minor involvement of galectin-3 in the development of these lesions.

M2 macrophages coexist with a Th1-driven profile in periapical cysts.

AIM: To evaluate the participation of both Th1 and Th2 responses in periapical cysts by assessing the presence of M2 macrophages, as well as acute IL-1 ß, TNF-α and IL-6 cytokines. METHODOLOGY: Twenty-four cases of periapical cysts were selected. Immuno-expressions of IL-1 ß, IL-6, TNF-α and CD163 were analysed in the cystic capsules in both superficial and deeper regions. Data were analysed with paired Wilcoxon test and Spearman correlation coefficient (P ≤ 0.05). RESULTS: There was a higher expression of IL-1ß, IL-6, TNF-α and M2 macrophages in the superficial region (P < 0.001) of cystic capsules. All acute cytokines had significant positive correlations amongst them regardless of the cystic capsule region. Regarding CD163, positive correlations occurred only with TNF-α (P = 0.007; r = 0.537) and IL-6 (P = 0.018; r = 0.478) in the superficial regions of the cystic capsule. CONCLUSIONS: M2 macrophages participated actively in the inflammatory response of periapical cysts and correlated with the expression of certain acute Th1-related cytokines. This illustrates the coexistence of an acute and chronic Th2-driven immune response in these lesions. Although M2 macrophages favour the healing process, their presence is not sufficient for periapical cyst regression, once an acute active response has occurred due to an infectious stimuli.

Pro-inflammatory and anti-inflammatory cytokine expression in post-treatment apical periodontitis

Abstract Objective: This study evaluated the expression of pro-inflammatory (IL-1β, IL-6, IFN-γ and TNF-α) and anti-inflammatory (IL-4 and TGF-β) cytokines in apical periodontitis lesions. Correlations between these cytokines and clinical and cone-beam computed tomographic (CBCT) data were also assessed. Material and Methods: Apical periodontitis lesions' data were obtained from 27 patients subjected to periradicular surgery. Specimens were processed for histopathologic and immunohistochemical analysis. Sections were evaluated according to the amount of positive staining for each antibody. Expression levels of the target mediators were compared with clinical and CBCT data. Results: Twenty lesions were diagnosed as granuloma and 7 as cyst. In granulomas, IL-4 expression was significantly higher than IL-6 (p=0.001) and TNF-α (p=0.001). There was a significant relationship between high levels of TNF-α and lesions <5 mm (p=0.017). In cysts, IL-6 expression was significant lower than IL-4 (p=0.001) and IFN-γ (p=0.004). There was a significant relationship between high levels of TGF-β and endodontic treatment performed ≤4 years before (p=0.045). In general, IL-4 was the most expressed mediator in both cysts and granulomas. Conclusions: There was a balance between the expression of pro-inflammatory and anti-inflammatory cytokines associated with the chronic periradicular inflammatory process. TNF-α and TGF-β were related to some clinical and CBCT data.

Analysis of CD57+ natural killer cells and CD8+ T lymphocytes in periapical granulomas and radicular cysts.

The aim of this study was to compare the number of CD57+ natural killer (NK) cells and CD8+ T lymphocytes between periapical granulomas (PGs) and radicular cysts (RCs). Twenty-fives cases of PGs and 25 of RCs were submitted to histological analysis and immunohistochemistry using anti-CD57 and anti-CD8 biomarkers. Positive cells were counted in 10 fields (400× magnification) and the median value was calculated for each case. Statistical tests were used to evaluate differences in the number of CD57+ NK cells and CD8+ T lymphocytes according to type of lesion, intensity of the infiltrate and thickness of the lining epithelium. The number of CD57+ NK cells and CD8+ T lymphocytes was higher in PGs than in RCs (p = 0.129 and p = 0.541, respectively). Comparison of the number of CD57+ NK cells in atrophic and hyperplastic epithelium revealed a larger number of cells in the atrophic epithelium (p = 0.042). A larger number of CD57+ NK cells and CD8+ T lymphocytes were observed in grade III infiltrates compared to grade I/II (p = 0.145 and p = 0.725, respectively). CD8+ T lymphocytes were more prevalent than CD57+ NK cells in most cases when PGs and RCs were analyzed separately or in combination (p < 0.0001). CD57+ NK cells and CD8+ T lymphocytes play a key role in antiviral defense and the presence of these cells supports evidence suggesting the participation of these microorganisms in the pathogenesis of PGs and RCs. The response mediated by CD8+ T lymphocytes was more frequent, indicating greater participation of the adaptive immunity in these chronic lesions.

Participation of osteoclastogenic factors in immunopathogenesis of human chronic periapical lesions.

BACKGROUND: Chronic periapical lesions (CPLs) are common lesions of the oral cavity and are the result of caries, tooth fracture, iatrogenic causes, or factors causing contamination and pulp necrosis. Inflammatory cells participate in the expansion of CPLs by releasing factors that stimulate or inhibit osteolytic activity. The objective of this study was to investigate the participation of RANKL, TNF-α, cathepsin K, IL-33, and OPG in the development of radicular cysts (RCs) and periapical granulomas (PGs). METHODS: Paraffin-embedded sections of 30 RCs and 22 PGs were submitted to immunohistochemistry. RESULTS: Immunoexpression of the proteins studied was observed in the epithelium and capsule of RCs, as well as in connective tissue of PGs. The expression of the osteoclastogenic factors studied differed significantly in RCs and PGs (P < .001), with lower expression of OPG in RCs. In PGs, the lowest expression was observed for cathepsin K. Comparison of the 2 lesions showed a similar participation of RANKL and IL33, while a significant difference was observed for OPG (P < .001), TNF-α (P = .002), and cathepsin K (P = .016). No association of the expression of the proteins with lesions size was observed. CONCLUSIONS: This study demonstrated the participation of RANKL, TNF-α, IL-33, cathepsin K, and OPG in the development of RCs and PGs, with emphasis on the highest immunoreactivity of cathepsin in RCs and TNF-α and OPG in PGs. OPG possibly determines the slower growth of PGs compared to RCs.

Immunohistochemical expression of TGF-ß1 and MMP-9 in periapical lesions.

The objective of this study was to evaluate the expression of matrix metalloproteinase 9 (MMP-9) and transforming growth factor beta (TGF-ß1) in periapical lesion samples correlated with the intensity of the inflammatory infiltrate and thickness of the epithelial lining. Forty-five cases of periapical lesions (23 periapical granulomas and 22 radicular cysts) were subjected to morphological and immunohistochemical analyses using anti-MMP-9 and anti-TGF-ß1 antibodies. The data were analyzed using the following tests: non-parametric Mann-Whitney, chi-square, Fisher's exact test and Spearman's correlation test (P<0.05). Analysis of inflammatory infiltrate revealed that 78% of periapical granulomas presented infiltrate grade III, in contrast with 32% of radicular cysts (P<0.001). Morphological evaluation of the epithelial thickness in radicular cysts revealed the presence of atrophic epithelium in 86% of the cysts. The immunostaining of MMP-9 was score 2 in 67% of the granulomas and 77% of the cysts. Both lesions were predominantly score 1 for TGF-ß1. Significant differences were confirmed between the expression scores of TGF-ß1 and MMP-9 in periapical granulomas (p = 0.004) and in radicular cysts (p < 0.001). Expression of TGF-ß1 was different for periapical granulomas and radicular cysts. This immunoregulatory cytokine seems more representative in asymptomatic lesions. The extracellular matrix remodeling process dependent on MMP-9 seems to be similar for both periapical granulomas and radicular cysts. TGF-ß1 and MMP-9 may play an important role in the maintenance of periapical lesions.