Peroxiredoxin I expression in epithelial cells of buccal mucosa from patients exposed to panoramic X-rays: influence of the age.

OBJECTIVES: The aim of this study was to evaluate peroxiredoxin I (Prx I) participation in the cellular antioxidant response to low-dose X-rays through the analysis of its expression in buccal mucosa cells from patients of different ages following panoramic dental radiography. MATERIALS AND METHODS: Of the 50 patients included in this study, oral mucosa cells from six adults were collected for the immunofluorescence cytological analysis. The other 44 patients, 11 patients aged below 20 years; 22 patients aged between 20 and 50 years; and 11 patients aged above 50 years, were submitted to panoramic dental radiography, and oral mucosa cells were collected for the gene expression analysis before and 1 hour after exposure. RESULTS: The results demonstrated Prx I expression in the cytoplasm of oral mucosa cells either before or after radiation exposure. The quantitative analysis showed that in oral mucosa cells from patients aged below 50 years the mRNA levels of PRDX1 were significantly increased after radiation exposure. On the other hand, the cells from patients aged above 50 years presented significantly lower PRDX1 transcript levels after radiation exposition. CONCLUSIONS: Panoramic radiography leads to increased Prx I expression in buccal mucosa cells, probably as an adaptive response to eliminate X-ray-induced ROS, except in cells from elderly people. CLINICAL RELEVANCE: Even low doses of radiation employed for dental purposes are capable to provoke stress to cells, which was demonstrated via the induction of the antioxidant gene PRDX1. In elderly patients, such mechanism was demonstrated to be impaired.

FGF-2 is overexpressed in myoepithelial cells of carcinoma ex-pleomorphic adenoma in situ structures.

Increasing emphasis has been placed on the role of myoepithelial cells, the contractile components of secretory glands, in the in situ to invasive carcinoma transition. These cells are placed at the interface between luminal epithelial cells and the stromal compartment, which favors their cross-talk with all other cell types comprising the tumor micro-environment. To obtain some clues about this cross-talk and also to better understand our previous immunoprofile study of myoepithelial cells in salivary gland carcinoma ex-pleomorphic adenoma (CXPA), we investigated FGF-2 expression in CXPA in situ structures as well as in cells cultured under conditions attempting to simulate the cellular interactions of this tumor stage. We have observed by immunohistochemistry that myoepithelial cells of CXPA in situ structures overexpress FGF-2. In addition, our results supported by qPCR and Western blotting, demonstrated that the expression of FGF-2 in the benign myoepithelial cells was in fact increased by stimulation with the conditioned medium from malignant cells. Low molecular weight FGF-2, known to be primarily released from the cells to exert its biological activity through receptors, was the predominant FGF-2 form detected in the benign myoepithelial cells. Specific FGF-2 receptors were found in the malignant epithelial but not in the benign myo-epithelial cells of CXPA, indicating a paracrine role for benign myoepithelial cell-derived FGF-2. Abnormal paracrine myo-epithelial/epithelial cell interactions and also myoepithelial/ stromal cell interactions could favor tumor growth, invasion and metastasis.

Glucose transporter protein 1 expression in mucoepidermoid carcinoma of salivary gland: correlation with grade of malignancy.

Mucoepidermoid carcinoma (MEC), the most common primary salivary malignancy, shows great variability in clinical behaviour, thus demanding investigation to identify of prognostic markers. Since Warburg's studies, unrestricted cell growth during tumorigenesis has been linked to altered metabolism, implying hypoxic stimulation of glycolysis and diminished contribution of mitochondrial oxidative phosphorylation to cellular ATP supply. Hypothesizing that the study of MEC metabolic status could lead to the discovery of prognostic markers, we investigated by immunohistochemistry the expression of glucose transporter 1 (Glut-1), mitochondrial antigen and peroxiredoxin I (Prx I) in samples of MEC from different histological grades. Our results showed that mitochondrial antigen and Prx I were expressed in the majority of the MEC cases independent of the histological grade. In contrast Glut-1 expression increased significantly as the tumours became more aggressive. These results suggested that oxidative phosphorylation may contribute to ATP supply in all stages of MEC progression, and that the relative contribution of glycolysis over mitochondria for cellular ATP supply increases during MEC progression, favouring growth under low oxygen concentration. In addition, the observed high Prx I protein levels could provide protection to tumour cells against reactive oxygen species generated as a consequence of mitochondrial function and hypoxia-reoxygenation cycling. Altogether our findings suggest that upregulation of Glut-1 and Prx I constitute successful adaptive strategies of MEC cells conferring a growth advantage over normal salivary gland cells in the unstable oxygenation tumour environment.

Collagen type I may influence the expression of E-cadherin and beta-catenin in carcinoma ex-pleomorphic adenoma.

Carcinoma ex-pleomorphic adenoma (CXPA) is an aggressive salivary gland malignancy, usually derived from a long-standing or a recurrent benign tumor, the pleomorphic adenoma (PA). In the context of dynamic reciprocity, changes in the composition and structure of extracellular matrix proteins and cell surface receptors have been frequently associated with dysfunctional adhesion and invasive behavior of tumor cells. It is not fully understood if these changes are involved in the conversion of PA to CXPA. In this study, different progression stages of CXPA were investigated regarding the expression of the major extracellular matrix proteins, collagen type I, and of E-cadherin and beta-catenin, the components of adherens junctions. By immunohistochemical analysis, we have demonstrated that direct contact of tumor cells with fibrillar type I collagen, particularly near the invasive front and in invasive areas prevailing small nests of CXPA cells, could be associated with reduced expression of the E-cadherin and beta-catenin adhesion molecules and with invasive behavior of epithelial, but not of CXPA with myoepithelial component. Our results also suggested that this association could depend on the organization of collagen molecules, being prevented by high-order polymeric structures. These findings could implicate the local microenvironment in the transition from the premalignant PA to invasive CXPA.

Antigen-presenting cells in human immunosuppressive drug-induced gingival enlargement.

An immunoperoxidase technique was used to compare the number of CD1a+ and factor XIIIa+ dendritic cells (DCs), and CD68+ Macrophages (M) in 30 gingival samples from subjects with clinically healthy periodontitium (HP) and 10 samples from subjects with drug-induced gingival enlargement (DIGE). Fewer CD1a+ and factor XIIIa+ DCs were found in areas with inflammatory infiltration (II) of the lamina propria (LP) in the group with immunosuppressed DIGE (IDIGE) compared to the group with HP. In the sulcular and junctional/pocket epithelia, the number of CD1a+ DCs was decreased in the group with IDIGE (p<0.05). There was a tendency toward a reduced number of CD1a+ DCs and CD68+ M in areas without inflammatory infiltrate of the LP in the group with IDIGE. The alterations in the number of antigen-presenting cells (APCs) may be the reason for the decreased periodontal inflammation and breakdown clinically observed in subjects who are immunosuppressed.

Use of anorganic bovine-derived hydroxyapatite matrix/cell-binding peptide (P-15) in the treatment of class II furcation defects: a clinical and radiographic study in humans.

BACKGROUND: This study compared clinical and radiographic findings for the treatment of Class II furcation defects in human mandibular molars using anorganic bovine-derived hydroxyapatite matrix (ABM)/cell-binding peptide (P-15) or open flap debridement (OFD). METHODS: Twelve subjects showing two comparable Class II furcation defects in their mandibular molars were enrolled. The defects in each subject were assigned randomly to the test (ABM/P-15) or the control (OFD) group. Clinical measurements and standardized radiographs were taken at baseline and 6 to 7 months after surgery. RESULTS: There were no statistically significant differences between the test and control groups for any clinical or radiographic parameter (P >0.05). On comparing the baseline and final measurements, the gain in horizontal clinical attachment level and reduction in gingival recession were significant only in the test group (P < or =0.02), whereas the gain in the vertical clinical attachment level was significant in both groups (P < or =0.04). In the test group, four of 12 sites showed complete closure, and five showed partial closure; in the control group, three defects showed complete closure, and four showed partial closure (P = 0.42). Subtraction radiography revealed similar gains in bone height and increases in mean bone density with both treatments (P >0.05). CONCLUSIONS: ABM/P-15 yielded favorable results in the treatment of Class II furcation defects over a 6-month evaluation period; however, there was no difference compared to OFD. Further studies using a larger sample size are needed to confirm the present findings.

Hydrocortisone affects the expression of matrix metalloproteinases (MMP-1, -2, -3, -7, and -11) and tissue inhibitor of matrix metalloproteinases (TIMP-1) in human gingival fibroblasts.

BACKGROUND: There is a positive correlation between the course of periodontal disease and psychosocial stress status. Stress leads to activation of the hypothalamic-pituitary-adrenal axis, resulting in increased cortisol release. The present study evaluates the effect of two different hydrocortisone concentrations on mRNA expression of matrix metalloproteinases (MMPs) and tissue inhibitor of matrix metalloproteinases (TIMPs) in cultured, human gingival fibroblasts. METHODS: Gingival fibroblasts were stimulated with 10(-7) or 10(-9) M hydrocortisone for 24 hours; untreated cells served as controls. Alterations in the expression of MMP-1, -2, -3, -7, -11 and TIMP-1 and -2 were evaluated using real-time polymerase chain reaction and Western blotting. beta-actin mRNA expression was used as a reference to normalize gene expression. RESULTS: Although the higher hydrocortisone concentration upregulated MMP-1, -2, -7, -11, and TIMP-1 (P <0.05) expression, the lower concentration induced downregulation or diminished upregulation. The lower hydrocortisone concentration induced a 23-fold increase in MMP-3 gene expression, whereas the higher concentration induced less upregulation; however, protein expression was regulated similarly by both hydrocortisone concentrations. The effect of hydrocortisone on TIMP-2 expression was not significant (P >0.05). CONCLUSIONS: Hydrocortisone produced a dose-dependent regulation of MMP and TIMP expression. The higher hydrocortisone concentration significantly upregulated expression of MMP-1, -2, -7, and -11 and TIMP-1 in human gingival fibroblasts, which may constitute a mechanism underlying the increased periodontal breakdown associated with psychosocial stress status.

Effect of tumor necrosis factor-alpha gene polymorphism on peri-implant bone loss following prosthetic reconstruction.

PURPOSE: The present study investigates the association between a specific polymorphism in the tumor necrosis factor (TNF)-alpha gene, consisting of allele 2 of TNF-alpha-308, and peri-implant bone loss following prosthetic reconstruction. MATERIALS AND METHODS: This case-control study included 36 patients (20 women, 16 men; mean age 46 years) who had used implant-supported prostheses for a minimum of 6 months and a maximum of 31 months. The patients were nonsmoking, white Caucasian Brazilians, in good general health, and were not receiving medication. In the case group, patients exhibited 1 or more implants with a diagnosis of peri-implant bone loss following prosthetic reconstruction; control patients had 1 or more healthy implants. RESULTS: Polymorphism in the TNF-alpha gene, allele 2 of TNF-alpha, was not associated with an increased risk for peri-implant bone loss following prosthetic reconstruction (P=0.19; chi2=1.71; df=1), although 21.1% of the subjects carried allele in the control group 2, and 41.2% carried allele 2 in the case group. CONCLUSIONS: Polymorphism in allele 2 of the TNF-alpha-308 gene is not associated with an increased risk for peri-implant bone loss following prosthetic reconstruction. However, further studies based on a greater number of patients are necessary.

The effect of epidermal growth factor on matrix metalloproteinases and tissue inhibitors of metalloproteinase gene expression in cultured human gingival fibroblasts.

OBJECTIVE: Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play a role in the breakdown of the extracellular matrix during normal physiological processes, and in pathological processes, including periodontitis. The aim of this study was to evaluate the effect of epidermal growth factor (EGF) on the expression of MMPs and TIMPs in cultured human gingival fibroblasts. METHODS: Fibroblasts were stimulated with 10(-3), 10(-6) or 10(-12)M EGF for 24h; untreated fibroblasts served as controls. Alterations in the expression of MMP-1, 2, 3, 7, 11, TIMP-1 and 2 were evaluated using real-time PCR and Western blotting. beta-Actin expression was used as a reference to normalize gene expression. RESULTS: Increased MMP-1, 3, 7 and 11 expressions were observed at all EGF concentrations (p<0.05). At the lowest EGF concentration, MMP-1, 3 and 7 presented the lowest expression and MMP-11 presented the greatest expression; at higher EGF concentrations, MMP-1, 3 and 7 presented greater up-regulation, and MMP-11 lower up-regulation (p<0.05). Protein expression was similarly regulated by EGF: increased up-regulation of MMP-1, 3 and 7 was observed with increasing EGF concentrations, except for MMP-11 that exhibited greater up-regulation at the lower EGF concentration. The gene expression of MMP-2, TIMP-1 and 2 was not affected by EGF (p<0.05). CONCLUSIONS: We conclude that EGF regulates expression for MMP-1, 3, 7 and 11 in a dose-dependent manner, suggesting that EGF may play a role in periodontal destruction and wound repair.

Comparative study between the effects of photodynamic therapy and conventional therapy on microbial reduction in ligature-induced peri-implantitis in dogs.

BACKGROUND: Progressive peri-implant bone losses, which are accompanied by inflammatory lesions in the soft tissues, are referred to as peri-implantitis. The aim of this study was to compare the effects of photodynamic therapy (PDT) and conventional technique on microbial reduction in ligature-induced peri-implantitis in dogs. METHODS: Eighteen third premolars from nine Labrador retriever dogs were extracted and the implants were submerged. After osseointegration, peri-implantitis was induced. After 4 months, ligature was removed and natural bacterial plaque was allowed to form for another 4 months. The animals were then randomly divided into two groups. In the conventional group, they were treated using mucoperiosteal flaps for scaling the implant surface and chlorexidine (conventional) irrigation. In the PDT group, only mucoperiosteal scaling was carried out before photodynamic therapy. Inside the peri-implant pocket, a paste-based azulene photosensitizer was placed and then a GaAlAs low-power laser (lambda=660 nm, P=40 mW, E=7.2 J for 3 minutes) was used. Microbiological samples were obtained before and immediately after treatment. Before treatment, one implant was removed and analyzed by scanning electron microscopy to validate the contamination. RESULTS: The results of this study showed that Prevotella sp., Fusobacterium sp., and S. Beta-haemolyticus were significantly reduced for both groups. After treatment, no significant differences were observed between the groups. CONCLUSION: These findings suggest that photodynamic therapy is a non-invasive method that could be used to reduce microorganisms in peri-implantitis.

Expressão imunoistoquímica da ciclina D1 e do p16 em carcinoma epidermóide de boca: correlação com sistema TNM e localização / Cyclin D1 and p16 immunohistochemical expression in oral squamous cell carcinoma: correlation with TNM classification and location

Objetivo: Avaliar a expressão imunoistoquímica da ciclina D1 e do p16 (proteínas envolvidas nas vias de proliferação celular e utilizadas para determinar o prognóstico de neoplasias malignas) em carcinoma epidermóide de boca.Correlacionar a imunomarcação com o sistema TNM (Tamanho do tumor, presença de linfonodo metastático e metástase à distância) e com sua localização anatômica. Métodos: Trinta e quatro (34) blocos de parafina contendo fragmentos de biópsia incisional de carcinomas epidermóides bucais primários foram obtidos no Hospital Araújo Jorge da Associação de Combate ao Câncer em Goiás. Os dados dos pacientes quanto à localização anatômica eo Sistema TNM foram coletados dos prontuários. A expressão das proteínas ciclina D1 e p16 foi verificada através da técnica imunoistoquímica utilizando a Streptoavidina-Biotina no Laboratório de Patologia Bucal da Faculdade de Odontologia da Universidade de São Paulo (FOUSP). Resultados e Conclusões: Os resultados não revelaram diferença estatisticamente significativa entre o número médio de núcleos positivos para a ciclina D1 e os dados clínicos dos pacientes. Porém, uma menor porcentagem de marcação nos carcinomas de lábio inferior e menor expressão nos tumores classificados clinicamente como T1 foi encontrada. Não houve diferença estatisticamente significativa entre o número médio de núcleos p16 positivos e os dados clínicos dos pacientes com carcinoma epidermóide de boca. Pode-se sugerir que houve um acúmulo nuclear de p16, mas este resultado não tem significância no prognóstico. Entretanto, os resultados da ciclina D1 não mostraram que ela é um marcador absoluto de prognóstico, mas sugerem que o aumento do nível de ciclina D1 contribui junto com outros oncogenes no processo de progressão tumoral.

Comparison between subtraction radiography and conventional radiographic interpretation during long-term evaluation of periodontal therapy in Class II furcation defects.

BACKGROUND: Limited information comparing digital subtraction radiographic assessment with conventional radiographic interpretation is available from longitudinal clinical trials. The aim of this study was to evaluate the ability to detect periodontal bone changes during the long-term maintenance of Class II furcation defects by conventional radiographic interpretation compared to interpretation of digital subtraction images. METHODS: Standardized radiographs of 18 Class II furcation defects in mandibular molars were taken at baseline and at 6, 12, 18, and 24 months after non-resective periodontal surgery. Conventional radiographic and digital subtraction interpretations were performed masked, respectively, by two and three experienced examiners, according to the following categories: bone gain; bone loss; unchanged appearance; and impossible to visualize. Percent concordance and the kappa statistic value (kappa) were computed. RESULTS: Conventional radiographic and digital subtraction interpretation images resulted in 72 decisions for each examiner. The visual interpretation of digital subtraction images by two examiners revealed the same results. The interpretation of conventional radiographic images showed a low concordance between examiners (kappa < 0.40) at all examinations. The concordance between subtraction radiography and conventional radiographic interpretation was also low for all examiners (kappa < 0.36) at all examinations. Using subtraction radiography as a reference, bone changed and bone unchanged were diagnosed correctly in 47.2% of cases by examiner A, in 43.1% by examiner B, and in 38.9% by examiner C. CONCLUSION: It can be concluded that conventional radiographic interpretation is a more subjective and inaccurate method of detecting periodontal bone changes in Class II furcation defects in mandibular molars when compared with subtraction radiography.

Solitary fibrous tumor of the oral mucosa--morphological and immunohistochemical profile in the differential diagnosis with hemangiopericytoma.

The objective was to investigate two cases of solitary fibrous tumor (SFT) of oral mucosa, emphasizing the differential diagnosis with one case of oral hemangiopericytoma (HPC), in terms of their morphological and immunohistochemical features. Solitary fibrous tumors showed cellularity and collagenization varying from area to area, focal perivascular hyalinization, scattered giant nuclei cells and abundant mast cells throughout the tumor. The hemangiopericytoma case exhibited thin-walled and dilated vessels lined with flat endothelial cells, identified by "staghorn appearance". Tumoral cells of solitary fibrous tumor exhibited immunohistochemical positivity for CD34, as well as endothelial cells. The hemangiopericytoma was positive only in endothelial cells. In solitary fibrous tumor, alpha-smooth muscle actin, h-caldesmon and laminin stained the wall vessels. In hemangiopericytoma, on the other hand, the wall vessels were positive only for laminin, which staining was also observed in perivascular tumoral cells. The morphological and immunohistochemical differences observed allowed us to infer these lesions constitute distinct entities.