Preliminary findings on the possible role of B-lymphocyte stimulator (BLyS) on diabetes-related periodontitis

Abstract The possible role of B-cell growth and differentiation-related cytokines on the pathogenesis of diabetes-related periodontitis has not been addressed so far. The aim of this study was to evaluate the effects of diabetes mellitus (DM) on the gene expression of proliferation-inducing ligand (APRIL) and B-lymphocyte stimulator (BLyS), two major cytokines associated to survival, differentiation and maturation of B cells in biopsies from gingival tissue with periodontitis. Gingival biopsies were obtained from subjects with periodontitis (n = 17), with periodontitis and DM (n = 19) as well as from periodontally and systemically healthy controls (n = 10). Gene expressions for APRIL, BLyS, RANKL, OPG, TRAP and DC-STAMP were evaluated using qPCR. The expressions APRIL, BLyS, RANKL, OPG, TRAP and DC-STAMP were all higher in both periodontitis groups when compared to the control group (p < 0.05). Furthermore, the expressions of BLyS, TRAP and RANKL were significantly higher in the subjects with periodontitis and DM when compared to those with periodontitis alone (p < 0.05). The mRNA levels of BLyS correlated positively with RANKL in the subjects with periodontitis and DM (p < 0.05). BLyS is overexpressed in periodontitis tissues of subjects with type 2 DM, suggesting a possible role of this cytokine on the pathogenesis DM-related periodontitis.

Effects of astragaloside IV on inflammation and immunity in rats with experimental periodontitis

Abstract: This study aimed to investigate the effects of astragaloside IV (AsIV) on inflammation and immunity in rats with experimental periodontitis. Periodontitis was established in 48 Wistar rats, which were then randomly divided into model and 10, 20 and 40 mg/kg AsIV groups, with 12 rats in each group. The latter 3 groups were treated with AsIV at doses of 10, 20 and 40 mg/kg, respectively. The control group (12 rats, without periodontitis) and model group were given the same amount of 5% sodium carboxymethyl cellulose. The treatment was performed once per day for 8 weeks. Before and after treatment, the tooth mobility scores of the rats were determined. After treatment, the salivary occult blood index (SOBI), plaque index (PLI), peripheral blood T lymphocyte subsets, and serum inflammatory factor and immunoglobulin levels were determined. The results showed that, after treatment, compared with that in model group, in 40 mg/kg AsIV group, the general state of rats was improved, while the tooth mobility score, SOBI and PLI were significantly decreased (p < 0.05); the peripheral blood CD4+ T cell percentage and CD4+/CD8+ ratio were significantly increased (p < 0.05), while the CD8+ T cell percentage was significantly decreased (p < 0.05); the serum tumor necrosis factor-α, interleukin-1β and interleukin-2 levels were significantly decreased (p < 0.05); the serum immunoglobulin A and immunoglobulin G levels were significantly decreased (p < 0.05). In conclusion, AsIV can alleviate inflammation and enhance immunity in rats with experimental periodontitis.

Immunolocalization of heme oxygenase-1 in periodontal diseases.

Context: Chronic periodontitis is an inflammatory condition of supporting tissues initiated by organisms in dental plaque. The reactive oxygen species and free radicals mediate connective tissue destruction in periodontitis. In order to counteract the free radical mediated tissue damage, numerous antioxidant mechanisms exist within the host. One such system is heme oxygenase enzymes. Heme oxygenase is the key enzyme involved in catabolism of heme. It cleaves the heme molecule to yield equimolar amounts of biliverdin, carbon monoxide, and iron. These end products act as important scavengers of reactive oxygen metabolites. Increased heme oxygenase expression has been identified in inflammatory condition, such as pancreatitis, diabetes, nephritis, and atherosclerosis. Since chronic periodontitis is one such inflammatory condition, we assessed the expression of heme oxygenase-1, in smokers and periodontitis group using immunohistochemistry technique. Aims: The aim of this study is to compare the expression of heme oxygenase-1 in patients with healthy periodontium, periodontitis and smokers. Materials and Methods: Gingival tissue samples were taken from 30 patients, who were divided into three groups healthy controls (n = 10), chronic periodontitis (n = 10), and smokers with chronic periodontitis (n = 10). All the samples were subjected to immunohistochemical staining using the antiheme oxygenase-1 antibody and were tested for efficiency by staining a positive control (prostate cancer tissue sections) and a negative control. The results were tabulated and analyzed. Results: Our results showed increased expression of heme oxygenase-1 in the gingival tissue samples taken from smokers compared with periodontitis and healthy tissue. Conclusion: The results of our study is an increasing evidence of involvement of antioxidant enzymes like heme oxygenase-1 in periodontal inflammation and their implication for treatment of chronic periodontitis.

Mecanismos moleculares implicados en la destrucción ósea en la periodontitis: revisión de la literatura / Molecular mechanisms involved in bone destruction in periodontitis: literature review

La periodontitis es una enfermedad inflamatoria crónica multifactorial, la cual se inicia a partir de la biopelícula que se forma alrededor de los dientes y se acumula en margen gingival, colonizando el surco gingival. La complejidad de la biopelícula madura genera estímulos para las células epiteliales e inflamatorias y sobre las demás células del tejido conectivo activando los mecanismos de la respuesta inmune innata y adaptativa. Se reconoce que la acumulación de placa dental genera de forma indefectible gingivitis, pero se desconocen las señales específicas que disparan la periodontitis. Se reconoce también que microorganismos periodontopáticos como Aggregatibacter actynomicetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Prevotella nigrescens, entre otros, poseen mediadores osteolíticos que actúan directa o indirectamente en las células del hueso y que son responsables del proceso de remodelación ósea, lo cual desequilibra el eje RANKL-RANK/OPG. Los productos microbianos y la respuesta inflamatoria inducen la secreción de citoquinas específicas como IL-1B, TNFalfa y otros mediadores pro-inflamatorios como PGE2, metalloproteinases, MMP-8, MMP-3, RANKL, además los linfocitos T y B activados inducen la pérdida de hueso alveolar al sintetizar y secretar directamente RANKL. Debido a que la pérdida de hueso alveolar es uno de los signos patognomónicos de la enfermedad periodontal, se hace importante revisar los mecanismos moleculares que explican la destrucción ósea, así como algunos avances en el tratamiento óseo.

Periodontitis is a multifactorial chronic inflammatory disease started by biofilm accumulation around the teeth and the gingival margin including the gingival sulcus. Mature biofilm is complex in microbial nature and it triggers signals to the boundary connective tissue and epithelial cells activating mechanisms of innate and acquired immune response. It is known that the dental plaque accumulation indefectibly results in gingivitis. However the specific signals that lead to periodontitis are unknown. The main periodontopathic organisms are Aggregatibacter actynomicetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, Prevotella nigrescens among others. Those microorganisms produce osteolytic mediators that act directly and indirectly on bone cells affecting the bone turnover rate, regulated by the axis RANKL-RANK/OPG. Microbial products and periodontal inflammation induce the release of specific cytokines IL-1B, TNFalpha, PGE2, metalloproteinases, MMP-8, MMP-3, RANKL, T and B lymphocytes elicit bone resorption. Indeed, alveolar bone loss is one of the most pathognomonic features of periodontal disease. Therefore it is essential to review the molecular mechanisms explaining periodontal destruction, as well as the advances in bone therapy.

Análise da expressão imuo-histoquímica da ciclofilina A, EMMPRIN e MMP-7 na doença periodontal / Expression analysis imuo staining of cyclophilin A, EMMPRIN and MMP-7 in periodontal disease

A doença periodontal é uma entidade infecciosa que resulta da resposta imuno-inflamatória do hospedeiro aos microrganismos presentes no biofilme dentário, levando à destruição tecidual. O propósito do presente estudo foi avaliar a expressão imuno-histoquímica da ciclofilina A (CYPA), do indutor de metaloproteinases da matriz extracelular (EMMPRIN) e da metaloproteinase da matriz 7 (MMP-7) em espécimes humanos de gengiva clinicamente saudável (n=32), gengivite induzida por biofilme dentário (n=28) e periodontite crônica (n=30). Foram realizadas biópsias das três condições clínicas e feita a análise imuno-histoquímica através da contagem total do número de células positivas, correlacionando-a com parâmetros clínicos. A imunopositividade da CYPA, do EMMPRIN e da MMP-7 revelou diferença estatisticamente significativa entre os três grupos, com maiores percentuais de positividade nos espécimes de periodontite crônica, seguidos pelos espécimes de gengivite crônica e de gengiva saudável (p < 0,001). Foi evidenciada maior expressão de CYPA e MMP-7 nos grupos que tinham infiltrado inflamatório mais intenso. Foram observadas correlações das imunoexpressões de EMMPRIN, MMP-7 e CYPA, tanto entre si como com parâmetros clínicos (profundidade de sondagem e perda de inserção clínica). Foram verificadas correlações positivas entre a expressão de CYPA e as expressões da MMP-7 (r = 0,831; p < 0,001) e do EMMPRIN (r = 0,289; p = 0,006). Além disso, a profundidade de sondagem revelou correlação positiva, estatisticamente significativa, com as expressões de MMP-7 (r = 0,726; p < 0,001), EMMPRIN (r = 0,345; p = 0,001) e CYPA (r = 0,803; p < 0,001). Esses resultados evidenciam que a CYPA, o EMMPRIN e a MMP-7 podem estar associadas à patogênese e progressão da doença periodontal. (AU)

Periodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of cyclophilin A (CYPA), extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase 7 (MMP-7) in human specimens of clinically healthy gingiva (n=32), biofilm-induced gingivitis (n=28), and chronic periodontitis (n=30). Immunopositivity for CYPA, EMMPRIN and MMP-7 differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (p < 0.001). Immunoexpression of CYPA and MMP-7 was higher in the intense inflammatory infiltrate observed mainly in cases of periodontitis. Analysis of possible correlations between the immunoexpression of EMMPRIN, MMP-7 and CYPA and between the expression of these proteins and clinical parameters (probing depth and clinical attachment loss) showed a positive correlation of CYPA expression with MMP-7 (r = 0.831; p < 0.001) and EMMPRIN (r = 0.289; p = 0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r = 0.726; p < 0.001), EMMPRIN (r = 0.345; p = 0.001), and CYPA (r = 0.803; p < 0.001). These results suggest that CYPA, EMMPRIN and MMP-7 are associated with the pathogenesis and progression of periodontal disease. (AU)

La respuesta inmunológica innata y la destrucción del tejido óseo en la enfermedad periodontal: revisión de la literatura / The innate immune response and tissue destruction in periodontal disease bone. A literatura review

La periodontitis desencadena una respuesta inmunológica del huésped contra los microorganismos de la placa dentobacteriana y sus productos. La activación de los receptores Toll-like (TLR) y Nod-like (NLR) en presencia de antígenos bacterianos inducen la respuesta inflamatoria y celular, con la consecuente expresión de citocinas inflamatorias como interleucina 1 (IL-1), interleucina 6 (IL-6), interleucina-8 (IL-8), interleucina 11 (IL-11), interleucina 18 (IL-18) y el factor de necrosis tumoral alfa (TNF-?), así como moléculas antibacterianas como b-defensinas, catelicidina y calprotectina. La IL-1 se asocia con la activación severa de las metaloproteinasas de la matriz extracelular (MMPs) que promueve la perdida de los tejidos de sostén del diente. La IL-6 estimula la diferenciación de los osteoclastos y estimula la síntesis de IL-1. El TNF-? induce la diferenciación de osteoclastos, la resorción ósea y tiene actividad sinérgica con IL-1?. Los neutrófilos forman una barrera entre el epitelio de unión y la placa dentobacteriana cuya función sinérgica es la actividad secretora de especies de oxigeno reactivas y proteínas bactericidas con el mecanismo fagocítico cuya actividad afecta la integridad de los tejidos del periodonto. Los derivados lipídicos del ácido araquidónico como Prostanglandina E2 (PGE2), han sido estrechamente relacionados con la pérdida del hueso alveolar por estimulación de los osteoclastos. Todo esto sugiere que la persistencia de las bacterias, una excesiva respuesta inflamatoria y/o una inadecuada resolución de la inflamación pueden afectar la estructura del tejido óseo durante la enfermedad periodontal.

Periodontitis triggers a host immune response against plaque microorganisms and their products. Activation of Toll-like receptors (TLRs) and Nod-like (NLR) by bacterial antigens triggers an inflammatory response and cell, with the expression of inflammatory cytokines such as interleukin 1 (IL-1), interleukin 6 (IL-6) , interleukin-8 (IL-8), interleukin 11 (IL-11), interleukin 18 (IL-18) and tumor necrosis factor alpha (TNF-?) and antibacterial molecules as b-defensins, cathelicidin and calprotectin . Among the cytokines IL-1? is associated with severe activation of extracellular matrix metalloproteinases (MMPs) that promotes the loss of the tooth-supporting tissues. IL-6 stimulates osteoclast differentiation and stimulates the synthesis of IL-1. TNF-? induces the differentiation of osteoclasts, bone resorption and has synergistic activity with IL-1?. In the cellular response, the neutrophils form a barrier between the junctional epithelium and plaque and secrete reactive oxygen species and bactericidal proteins with the phagocytic mechanism whose activity affects the integrity of the periodontal tissues. Also an arachidonic acid lipid derivative as prostaglandin E2 (PGE2) has been closely related to the loss of alveolar bone by stimulating osteoclast. This suggests that the persistence of bacteria, excessive inflammatory response and / or an inadequate resolution of the inflammation can affect the structure of the bone tissue during periodontal disease.

Antimicrobial peptides and nitric oxide production by neutrophils from periodontitis subjects

Neutrophils play an important role in periodontitis by producing nitric oxide (NO) and antimicrobial peptides, molecules with microbicidal activity via oxygen-dependent and -independent mechanisms, respectively. It is unknown whether variation in the production of antimicrobial peptides such as LL-37, human neutrophil peptides (HNP) 1-3, and NO by neutrophils influences the pathogenesis of periodontal diseases. We compared the production of these peptides and NO by lipopolysaccharide (LPS)-stimulated neutrophils isolated from healthy subjects and from patients with periodontitis. Peripheral blood neutrophils were cultured with or without Aggregatibacter actinomycetemcomitans-LPS (Aa-LPS), Porphyromonas gingivalis-LPS (Pg-LPS) and Escherichia coli-LPS (Ec-LPS). qRT-PCR was used to determine quantities of HNP 1-3 and LL-37 mRNA in neutrophils. Amounts of HNP 1-3 and LL-37 proteins in the cell culture supernatants were also determined by ELISA. In addition, NO levels in neutrophil culture supernatants were quantitated by the Griess reaction. Neutrophils from periodontitis patients cultured with Aa-LPS, Pg-LPS and Ec-LPS expressed higher HNP 1-3 mRNA than neutrophils from healthy subjects. LL-37 mRNA expression was higher in neutrophils from patients stimulated with Aa-LPS. Neutrophils from periodontitis patients produced significantly higher LL-37 protein levels than neutrophils from healthy subjects when stimulated with Pg-LPS and Ec-LPS, but no difference was observed in HNP 1-3 production. Neutrophils from periodontitis patients cultured or not with Pg-LPS and Ec-LPS produced significantly lower NO levels than neutrophils from healthy subjects. The significant differences in the production of LL-37 and NO between neutrophils from healthy and periodontitis subjects indicate that production of these molecules might influence individual susceptibility to important periodontal pathogens.

Decreased phagocytic function in neutrophils and monocytes from peripheral blood in periodontal disease

Phagocytosis by neutrophils and monocytes constitutes the main defense mechanism against bacterial challenges in periodontitis. Phagocytosis by neutrophils has already been evaluated, whereas phagocytic function of monocytes has hardly been addressed so far. Objectives: The aim of this study was to assess phagocytosis by neutrophils and monocytes in periodontitis. Material and Methods: The sample included 30 subjects with severe periodontitis and 27 control subjects without periodontal disease. The phagocytic index (PhI) was calculated as the mean number of adhered/ingested Saccharomyces cerevisiae per phagocytozing monocyte or neutrophil multiplied by the percentage of phagocytes involved in phagocytosis. Results: A significant reduction in phagocyte functions was observed in individuals with periodontitis. The median of PhI of neutrophils using nonsensitized S. cerevisiae was 3 for the control group, and 1.5 for the periodontitis group (p=0.01, Mann-Whitney test). The median of PhI of monocytes with non-sensitized S. cerevisiae was 26.13 for the control group, and 13.23 for the periodontitis group (p=0.03, Mann Whitney test). The median of PhI of monocytes assessed with sensitized S. cerevisiae was 97.92 for the control group and 60.1 for the periodontitis group (p=0.005, t-test). Conclusion: The data demonstrated a reduction in the function of phagocytes, suggesting a decrease in immune defenses in periodontitis.

Effect of non-surgical periodontal treatment on the subgingival microbiota of patients with chronic kidney disease

This study investigated the effect of non-surgical periodontal therapy on the composition of subgingival microbiota of patients with chronic kidney disease (CKD). Sixteen CKD pre-dialysis individuals (CKD) and 14 individuals without clinical evidence of kidney disease (C) presenting chronic periodontitis were treated by scaling and root planing. Subgingival samples were collected from each patient and analyzed for their composition by checkerboard at baseline and 3 months post-therapy. Significant differences between groups at baseline were sought by the Mann-Whitney and χ² tests. Changes over time were examined by the Wilcoxon test. At baseline, the CKD group had significantly lower counts of E. faecalis compared to the C group (p < 0.05). After treatment, the levels of a greater number of species were reduced in the C group. Higher levels of A. israelii, C. rectus, F. periodonticum, P. micra, P. nigrescens, T. forsythia, N. mucosa, and S. anginosus (p < 0.05) were found in the CKD group compared to the C group. Also, non-responsive sites in CKD individuals harbored significantly higher levels of pathogenic species (T. forsythia, P. gingivalis, T. denticola, Fusobacterium spp., D. pneumosintes, E. faecalis and S. aureus; p < 0.05) than sites that responded to therapy, as well as non-responsive sites in the C group. The periodontitis-associated subgingival microbiota of CKD and systemically healthy individuals was similar in composition. However, high levels of pathogenic species persisted in the subgingival microbiota of patients with CKD after treatment.

Langerhans cells in periodontal disease of HIV- and HIV+ patients undergoing highly active antiretroviral therapy

The aim of this study was to assess and compare quantitatively the presence of S100+ Langerhans cells (LC) by immunochemistry techniques in HIV+ and HIV- gingivitis and periodontitis subjects. Additionally, it aimed to evaluate the correlation among densities of these cells with CD4+ and CD8+ T cells, and viral load levels in HIV+ subjects, all using Highly Active Antiretroviral Therapy (HAART). The samples were allocated into four groups: 1) 15 subjects with moderate chronic periodontitis (MCP), HIV+; 2) 15 subjects with MCP, HIV-; 3) 10 subjects with gingivitis (G), HIV+; and 4) 10 subjects with G, HIV-. The S100+ cells were assessed in the pocket epithelium, gingival epithelium, and lamina propria. A statistically significant increase of total S100+ cells in HIV+ periodontitis subjects was observed in relation to HIV- periodontitis subjects. No increase of S100+ cells with increased inflammation was observed. No statistically significant correlation among S100+ cells and blood levels of CD4, CD8, and viral load was observed. In conclusion, the use of HAART can aid in achieving viral loads, and it is suggested that it may prevent the destruction of the LC.

E-cadherin and CD1a expression in gingival epithelium in periodontal health, disease and post-treatment.

Background: Epithelial integrity is important for maintenance of periodontal health. It is not fully known if non-surgical periodontal therapy is capable of recreating the epithelial barrier in its functional state. Patients and Methods: Sixty-five patients (31 males and 34 females) were included in the study. They were divided into group A (healthy gingiva 16 patients), group B (gingivitis 17 patients), group C (periodontitis 17 patients), and group D (post-treatment 15 patients). Gingival samples were collected and immunohistochemical study was done using E-cadherin and CD1a antibody. Statistical analysis was done using analysis of variance (ANOVA), followed by Tukey-Kramer multiple comparison test for CD1a and Tukey's highly significant difference (HSD) test for E-cadherin. Result: There was a statistically significant difference (P<0.001) in the expression of E-cadherin between healthy (1.846±0.555), gingivitis (1.100±0.994), and periodontitis group (0.700±0.483). Similarly, there was a statistically significant difference (P<0.001) in the expression of CD1a between healthy (75.70±3.09), gingivitis (42.53±3.09), and periodontitis group (29.07±3.08). However, the expression of E-cadherin (1.242±0.653) and CD1a in post-treatment samples (52.18±2.90) was lower with no statistically significant difference when compared to health. Discussion: The significant reduction in E-cadherin and CD1a levels in periodontal disease when compared to health could possibly be a result of invasion by the periodontopathogens and its subsequent sequel. Although, the post-treatment samples showed significant improvement when compared to disease, the reduction in E-cadherin and CD1a levels when compared to gingival health suggests that the epithelial barrier was not yet fully established in its functional state.

Análise da expressão de citocinas e dos receptores RAGE no periodonto de ratos diabéticos / Cytokine and RAGE receptor expression analyses in periodontium of diabetic rats

As doenças periodontais (DPs) são alterações inflamatórias crônicas que acometem os tecidos de sustentação do órgão dental. A presença do diabetes é refletida em maior severidade e prevalência das DPs tanto em humanos quanto em modelos experimentais. Contudo, os mecanismos biológicos envolvidos no aumento da prevalência e da severidade permanecem pouco conhecidos. Desta forma, o objetivo deste estudo foi avaliar o número de células marcadas por imunohistoquímica para TNF- , IL-1 , IL-6, RANKL, MMP-2, MMP-9 e para os receptores RAGE na doença periodontal experimental decorrente da indução do diabetes em ratos. Inicialmente, os ratos (n=25) foram submetidos à indução do diabetes através de administração endovenosa de aloxana (42mg/kg) e, juntamente com o grupo controle (n=25), acompanhados por 1, 3, 6, 9 e 12 meses. Em seguida, as hemimandíbulas foram coletadas e submetidas aos procedimentos de imunohistoquímica. Os resultados revelam que a presença do diabetes resulta em alterações significativas no número de células imunomarcadas para diferentes mediadores do processo inflamatório. Nos animais diabéticos, foi observado aumento estatisticamente significativo (p<0,05 ANOVA) no número de células imunomarcadas para TNF- (6 e12 meses), IL-1 (12 meses), IL-6 (9 e 12 meses), RANKL (9 meses) e para os receptores RAGE (6, 9 e 12 meses). Não foram observadas diferenças em relação ao número de células imunomarcadas para MMP-2 e MMP-9 entre os grupos controle e experimental, apesar da tendência a aumento no número de células MMP-9+ nos ratos após 12 meses da indução do diabetes (p>0,05 ANOVA). Assim, a desregulação na expressão de citocinas inflamatórias e fatores osteoclastogênicos parece ser um dos mecanismos biológicos envolvidos no aumento da prevalência e da severidade das doenças periodontais em decorrência do diabetes.

Periodontal diseases (PD) are chronic inflammatory diseases leading the destruction of connective tissue and alveolar bone supporting the teeth. The establishment of diabetes increases PD prevalence and severity in humans and experimental model. However, biological mechanisms regarding to increase of prevalence and severity remains poorly known. The aim of this study was to evaluate the number of immuno-staining cells to TNF- IL-1 , IL-6, MMP-2, MMP-9, RANKL and RAGE receptors in experimental periodontal disease in diabetic rats. Diabetes was induced in Wistar rats (n=25) by endovenous administration of 42 mg/kg of alloxan, and together with control animals (n=25), were analyzed at 1, 3, 6, 9 and 12 months after diabetes induction. The animals were sacrificed and the jaws were removed and submitted to immunohistochemistry procedures. Our data demonstrated that diabetes induction and progression resulted in significant alterations in number of immuno-staining cells to different mediators of inflammatory process. In diabetic rats, we observed an increased number of immuno-staining cells to TNF- (6 and 12 months), IL-1 (12 months), IL-6 (9 e 12 months), RANKL (9 months) and RAGE receptors (6, 9 and 12 months) (p<0,05 ANOVA). Regarding to MMP-2+ and MMP-9+ cells, we did not found differences between control and experimental groups. However, we found a trend of towards in MMP-9+ cells in diabetic rats after 12 months of diabetes induction (p>0,05 ANOVA). Then, our data demonstrated that diabetes establishment and progression resulted in an increase of immuno-staining cells to TNF- , IL-1 IL-6, RANKL and RAGE receptors. Taken together, desregulation of inflammatory cytokines and osteoclastogenic factor expression seems to be one of biological mechanisms involved in the increase of periodontal disease prevalence and severity associated with diabetes.

Análise da expressão de citocinas e dos receptores RAGE no periodonto de ratos diabéticos / Cytokine and RAGE receptor expression analyses in periodontium of diabetic rats

As doenças periodontais (DPs) são alterações inflamatórias crônicas que acometem os tecidos de sustentação do órgão dental. A presença do diabetes é refletida em maior severidade e prevalência das DPs tanto em humanos quanto em modelos experimentais. Contudo, os mecanismos biológicos envolvidos no aumento da prevalência e da severidade permanecem pouco conhecidos. Desta forma, o objetivo deste estudo foi avaliar o número de células marcadas por imunohistoquímica para TNF- , IL-1 , IL-6, RANKL, MMP-2, MMP-9 e para os receptores RAGE na doença periodontal experimental decorrente da indução do diabetes em ratos. Inicialmente, os ratos (n=25) foram submetidos à indução do diabetes através de administração endovenosa de aloxana (42mg/kg) e, juntamente com o grupo controle (n=25), acompanhados por 1, 3, 6, 9 e 12 meses. Em seguida, as hemimandíbulas foram coletadas e submetidas aos procedimentos de imunohistoquímica. Os resultados revelam que a presença do diabetes resulta em alterações significativas no número de células imunomarcadas para diferentes mediadores do processo inflamatório. Nos animais diabéticos, foi observado aumento estatisticamente significativo (p<0,05 ANOVA) no número de células imunomarcadas para TNF- (6 e12 meses), IL-1 (12 meses), IL-6 (9 e 12 meses), RANKL (9 meses) e para os receptores RAGE (6, 9 e 12 meses). Não foram observadas diferenças em relação ao número de células imunomarcadas para MMP-2 e MMP-9 entre os grupos controle e experimental, apesar da tendência a aumento no número de células MMP-9+ nos ratos após 12 meses da indução do diabetes (p>0,05 ANOVA). Assim, a desregulação na expressão de citocinas inflamatórias e fatores osteoclastogênicos parece ser um dos mecanismos biológicos envolvidos no aumento da prevalência e da severidade das doenças periodontais em decorrência do diabetes.

Periodontal diseases (PD) are chronic inflammatory diseases leading the destruction of connective tissue and alveolar bone supporting the teeth. The establishment of diabetes increases PD prevalence and severity in humans and experimental model. However, biological mechanisms regarding to increase of prevalence and severity remains poorly known. The aim of this study was to evaluate the number of immuno-staining cells to TNF- IL-1 , IL-6, MMP-2, MMP-9, RANKL and RAGE receptors in experimental periodontal disease in diabetic rats. Diabetes was induced in Wistar rats (n=25) by endovenous administration of 42 mg/kg of alloxan, and together with control animals (n=25), were analyzed at 1, 3, 6, 9 and 12 months after diabetes induction. The animals were sacrificed and the jaws were removed and submitted to immunohistochemistry procedures. Our data demonstrated that diabetes induction and progression resulted in significant alterations in number of immuno-staining cells to different mediators of inflammatory process. In diabetic rats, we observed an increased number of immuno-staining cells to TNF- (6 and 12 months), IL-1 (12 months), IL-6 (9 e 12 months), RANKL (9 months) and RAGE receptors (6, 9 and 12 months) (p<0,05 ANOVA). Regarding to MMP-2+ and MMP-9+ cells, we did not found differences between control and experimental groups. However, we found a trend of towards in MMP-9+ cells in diabetic rats after 12 months of diabetes induction (p>0,05 ANOVA). Then, our data demonstrated that diabetes establishment and progression resulted in an increase of immuno-staining cells to TNF- , IL-1 IL-6, RANKL and RAGE receptors. Taken together, desregulation of inflammatory cytokines and osteoclastogenic factor expression seems to be one of biological mechanisms involved in the increase of periodontal disease prevalence and severity associated with diabetes.

Etiología y terapia de la periodontitis agresiva: estado actual / Etiology and therapy of aggressive periodontitis: an update

La periodontitis agresiva es una patología de etiología multifactorial que se presenta en pacientes sistémicamente sanos, con rápida pérdida de inserción y ósea, acompañada de agregación familiar. Puede ser localizada o generalizada, siendo las bacterias predominantes el actinobacillus actinomycetemcomitans y la porphyromona gingivalis. Ellas desencadenan una respuesta inmuno-inflamatoria, la cual es la mayor responsable de la gran destrucción tisular. Se ha sugerido que la presencia de virus del tipo herpes pueda jugar un rol importante ne este cuadro. La terapia básica y el mantenimienot son, como en toda la patología gingivo-periodontal, la base del tratamiento. La cirugía reconstructiva y la terapia farmacológica antimicrobiana, han mostrado ser altamente efectivas y de utilización frecuente

Immunoglobulin concentration in gingival tissue of type 2 diabetic patients with periodontitis.

BACKGROUND: Diabetes mellitus is considered as a risk factor for the initiation and progression of periodontal disease. The diabetic patients often exhibit decreased immune response and increased susceptibility to infection. In the present study, a quantitative estimation of the gingival tissue immunoglobulin concentrations in diabetic and non diabetic subjects with periodontitis was assessed and compared with that of clinically healthy gingiva. METHOD: 40 gingival tissue samples obtained from 20 diabetic (Type 2) and 20 non-diabetic subjects were subjected to quantitative estimation of immunoglobulins G, A, and M. The data thus obtained were compared to the level of immunoglobulin found in clinically healthy gingiva. RESULTS: The IgG and IgA level in the tissues of both diabetic and non-diabetic subjects with periodontitis were found to be significantly higher than that of healthy subjects. The diabetic group also showed a significantly higher IgG and IgA levels compared to the non-diabetic group with periodontitis. CONCLUSION: These findings support the concept that the humoral immune response plays an important role in the pathogenesis of periodontal disease in diabetics. The significantly higher levels of immunoglobulin in the gingival tissues might be a protective mechanism against the increased bacterial challenge in diabetic subjects.

Análise qualitativa e quantitativa do padrão de expressão imuno-histoquímica das células dendríticas na doença periodontal / Quantitative and qualitative analysis of immunohistochemical expression profile of dendritcs cells in periodontal disease

As células dendríticas agem como células apresentadoras de antígeno (APC) nas respostas imunológicas. Na doença periodontal elas são responsáveis no início e na manutenção da resposta inflamatória. O presente trabalho se propôs a uma análise imuno-histoquímica da quantidade de células dendríticas, através do anticorpo anti- SI00 e da localização das células marcadas em 17 espécimes de gengivite crônica e 25 de periodontite crônica. As células s-100+ foram evidenciadas nas camadas basal e suprabasais do epitélio, bem como, na lâmina própria, especialmente na porção papilar, com morfologia e número variados. A análise estatística não demonstrou diferença significativa no número de células S-100+ entre os casos de gengivite e periodontite crônicas

Parto prematuro e baixo peso ao nascer associados à doença periodontal: aspectos clínicos, microbiológicos e imunológicos / Preterm and low birth weight and the relationship with periodontal disease: clinical, microbiological and immunological aspects

Sendo o peso da criança, ao nascer, o determinante mais importante das chances que o recém-nascido tem de sobreviver, crescer e se desenvolver saudavelmente, a prevenção do parto prematuro toma-se um importante fator de saúde pública. Evidências apontam que as infecções crônicas podem ter um papel fundamental no estabelecimento do trabalho de parto prematuro. A doença periodontal é uma reação imuno-inflamatória decorrente de uma infecção crônica causada por microrganismos gram-positivos e gram-negativos. Ela é capaz de elevar os níveis séricos de mediadores inflamatórios que, em conjunto com bacteremias e endotoxinemias transitórias, poderiam afetar o período gestacional. Assim, vários estudos têm sido dirigidos a fim de verificar o possível papel da doença periodontal como fator de risco ao parto prematuro de neonatos de baixo peso. Este artigo vem mostrar, através de uma revisão analítica da literatura, a associação entre infecção e trabalho de parto prematuro e as evidências encontradas sobre uma possível relação causal entre doença periodontal e nascimento prematuro de baixo peso. Concluiu-se que a doença periodontal é capaz de aumentar os níveis de mediadores inflamatórios associados ao trabalho de parto; que maiores níveis de periodontopatógenos podem estar relacionados ao nascimento prematuro de baixo peso; que o tratamento da doença periodontal nas mulheres grávidas, no início da gestação, parece diminuir o risco de ocorrência de nascimento prematuro de baixo peso; quanto maior a severidade da doença periodontal, maiores são as chances de NPBP

La periodoncia del nuevo milenio / Periodontics of the new millenium

El nuevo paradigma de la biopatología periodontal introduce el concepto de biofilms, en lugar de la placa bacteriana. Estos son necesarios pero insuficientes para determinar el curso de la enfermedad periodontal. En la actualidad, juegan un rol importantísimo los factores de riesgo (genéticos adquiridos o medio ambiente) en el comienzo, progresión y respuesta al tratamiento de la enfermedad periodontal. Además, la periodontitis está considerada como alto riesgo para enfermedades sistémicas tales como enfermedades cardiovasculares, partos prematuros y diabetes

A possible defective estimation of antineutrophil cytoplasmic antibodies in systemic lupus erythematosus due ot the coexistence of periodontitis: preliminary observations

The objectives of this study is to determine if periodontitis-related ANCA hinder the accurate estimation of this kind of autoantibodies in systemic lupus erythematosus (SLE), due to the frequent coexistence of SLE and periodontitis, and the high incidence of antineutrophil cytoplasmic antibodies (ANCA) in this periodontal condition. Thirty SLE, thirty periodontitis lacking systemic involvement patients, and twenty healthy controls were utilized in this study. The periodontal condition and the presence of ANCA in sera of all individuals was carefully evaluated. For ANCA determination an EIA essay was utilized, directed to a neutrophil granular extract and six neutrophil granule proteins. Sixty percent of SLE patients had periodontitis, and sixty-five percent were ANCA positive. Eighty three percent of all ANCA cases were coexisting with periodontitis. A significant association (p > 0.005) between periodontitis and ANCA was found (Chi Square Test). Fifty percent of the patients with periodontitis lacking systemic involvement were ANCA positive. The results obtained in this study suggest that the figures of ANCA previously reported for SLE, might be overestimated due to the inadvertent presence of periodontitis