Positive Correlation Between the Level of Interferon-Gamma and the Severity of Periodontitis

Abstract Objective: To examine the IFN-γ levels in patients with periodontitis and determine the difference in the levels of IFN-γ with the severity of the disease. Material and Methods: The study design was cross-sectional, and the sample consisted of 31 patients, aged between 18 and 64 years. Plaque index (PlI), calculus index (CI), and papillary bleeding index (PBI) were measured. Pocket depth (PD), recession, and clinical attachment loss (CAL) (mm) were measured at six sites per teeth. For mild/moderate periodontitis, pocket depth ≥4 mm in 1-3 sites was required, while the essential criteria for severe periodontitis were pocket depth ≥5 mm, clinical attachment loss >3 mm in more than 3 sites ≥2 quadrants. The IFN-γ levels were measured by performing enzyme-linked immunosorbent assay with the gingival crevicular fluid (GCF) samples. The measurements were made in two different sites, and the severity of periodontitis was categorized based on the pocket depth, attachment loss, and the remaining natural teeth. Kruskal-Wallis test, Mann-Whitney test, and Spearman's rank correlation coefficient were Results: The levels of IFN-γ (pg/mL) were correlated with the severity of the periodontal status, with p <0.05. Clinical parameters of periodontitis also correlated with the level of IFN-γ (pg/mL) Conclusion: Subjects with periodontitis presented greater levels of IFN-γ (pg/mL) in GCF than the periodontal healthy individuals. This result showed the role of IFN-γ in the inflammation.

IFN-γR2 is strongly expressed on endothelial cells of gingival tissues from patients with chronic periodontitis

Abstract Chronic periodontitis (CP) is characterized by gingival inflammation and bone destruction. It has been reported that interferon-gamma (IFN-γ) levels are high in CP patients; however, the IFN-γ receptor (IFN-γR) has not been studied in gingival tissue from these patients. Objective: To evaluate IFN-γ levels and IFN-γR expression in gingival tissue biopsies from chronic periodontitis patients compared with healthy subjects (HS). Material and Methods: Gingival tissues were obtained from all study subjects, CP (n = 18) and healthy subjects (HS) (n = 12). A tissue section of each study subject was embedded in paraffin blocks to determine the expression of IFN-γ R (IFN-γR1 and IFN-γR2) through immunohistochemistry. Another section of the tissue was homogenized and IFN-γ was measured by the ELISA technique. Results: No significant differences were found in the IFN-γR1 expression within the cell layers of the gingival tissue of the study groups. When analyzing the IFN-γR2 expression it was found that IFN-γR2 is strongly expressed in the endothelial cells of CP patients when compared to HS (p<0.05). IFN-γ concentrations in the gingival tissue were significantly higher in CP patients than in HS. No significant correlation between IFN-γ levels and the expression of IFN-γR1 and IFN-γR2 was found. However, a positive correlation between IFN-γ levels and clinical parameters [probing depth (PD) and clinical attachment level (CAL)] was found. Conclusion: The study of IFN-γR expression in gingival tissue samples from patients with CP showed an increase only in the IFN-γR2 chain in endothelial cells when compared to HS.

Periodontal disease and inflammatory blood cytokines in patients with stable coronary artery disease

ABSTRACT Periodontal disease has been associated with elevations of blood cytokines involved in atherosclerosis in systemically healthy individuals, but little is known about this association in stable cardiovascular patients. The aim of this study was to assess the association between periodontal disease (exposure) and blood cytokine levels (outcomes) in a target population of patients with stable coronary artery disease (CAD). Material and Methods This cross-sectional study included 91 patients with stable CAD who had been under optimized cardiovascular care. Blood levels of IL-1β, IL-6, IL-8, IL-10, IFN-γ, and TNF-α were measured by Luminex technology. A full-mouth periodontal examination was conducted to record probing depth (PD) and clinical attachment (CA) loss. Multiple linear regression models, adjusting for gender, body mass index, oral hypoglycemic drugs, smoking, and occurre:nce of acute myocardial infarction were applied. Results CAD patients that experienced major events had higher concentrations of IFN-γ (median: 5.05 pg/mL vs. 3.01 pg/mL; p=0.01), IL-10 (median: 2.33 pg/mL vs. 1.01 pg/mL; p=0.03), and TNF-α (median: 9.17 pg/mL vs. 7.47 pg/mL; p=0.02). Higher numbers of teeth with at least 6 mm of CA loss (R2=0.07) and PD (R2=0.06) were significantly associated with higher IFN-γ log concentrations. Mean CA loss (R2=0.05) and PD (R2=0.06) were significantly related to IL-10 concentrations. Elevated concentrations of TNF-α were associated with higher mean CA loss (R2=0.07). Conclusion Periodontal disease is associated with increased systemic inflammation in stable cardiovascular patients. These findings provide additional evidence supporting the idea that periodontal disease can be a prognostic factor in cardiovascular patients.

Lactobacillus rhamnosus could inhibit Porphyromonas gingivalis derived CXCL8 attenuation

ABSTRACT An increasing body of evidence suggests that the use of probiotic bacteria is a promising intervention approach for the treatment of inflammatory diseases with a polymicrobial etiology. P. gingivalis has been noted to have a different way of interacting with the innate immune response of the host compared to other pathogenic bacteria, which is a recognized feature that inhibits CXCL8 expression. Objective The aim of the study was to determine if P. gingivalis infection modulates the inflammatory response of gingival stromal stem cells (G-MSSCs), including the release of CXCL8, and the expression of TLRs and if immunomodulatory L. rhamnosus ATCC9595 could prevent CXCL8 inhibition in experimental inflammation. Material and Methods G-MSSCs were pretreated with L. rhamnosus ATCC9595 and then stimulated with P. gingivalis ATCC33277. CXCL8 and IL-10 levels were investigated with ELISA and the TLR-4 and 2 were determined through flow cytometer analysis. Results CXCL8 was suppressed by P. gingivalis and L. rhamnosus ATCC9595, whereas incubation with both strains did not abolish CXCL8. L. rhamnosus ATCC9595 scaled down the expression of TLR4 and induced TLR2 expression when exposed to P. gingivalis stimulation (p<0.01). Conclusions These findings provide evidence that L. rhamnosus ATCC9595 can modulate the inflammatory signals and could introduce P. gingivalis to immune systems by inducing CXCL8 secretion.

Expressão do interferon-γ em biópsias gengivais de pacientes com periodontite crônica severa / Expression of interferon-γ in gingival biopsies from patients with severe chronic periodontitis

O objetivo desse estudo foi analisar a expressão do interferon gama (INF-γ) em biópsias gengivais de sítios rasos e profundos de pacientes com periodontite crônica severa, e correlacionar a expressão do INF- γ no fluido gengival. Foram coletadas biópsias de 22 sítios profundos, 18 sítios rasos e 14 sítios controles. O fluido gengival foi coletado de 12 sítios profundos e 8 sítios rasos. Profundidade de bolsa à sondagem (PB); nível de inserção clínica (NIC); índice de placa visível (IPV) e índice de sangramento gengival (ISG) foram avaliados. As análises morfológica e imuno-histoquímica dos tecidos removidos foram realizadas. A intensidade da marcação do INF- γ nos tecidos foi avaliada semiquantitativamente. Houve uma tendência a um padrão de marcação similar nos sítios rasos e profundos, com predomínio de marcação fraca nos sítios profundos. Não houve diferenças estatisticamente significantes entre os três grupos. Não houve correlação significante (Spearman) com a expressão do INF- γ nos tecidos epiteliais e conjuntivos com os dados clínicos e o fluido gengival. Concluímos que não foi possível observar diferenças na expressão do INF- γ em biópsias gengivais nos sítios rasos e profundos de pacientes com periodontite quando comparados à indivíduos saudáveis, o que pode ser atribuído ao caráter bifásico do INF- γ.

The aim of this study was to analyze the expression of interferon-gamma (INF-γ) in gingival biopsies from shallow and deep sites in patients with severe chronic periodontitis and correlate the expression of INF-γ in the gingival crevicular fluid (GCF). Biopsies were collected from 22 deep sites, 18 shallow sites and 14 control sites. The GCF samples were collected from 12 deep and 8 shallow sites. Probing pocket in the epithelial and connective tissues with the clinical data and with the GCF. We conclude that there were no differences in the expression of INF-γ in gingival biopsies in shallow and deep sites of pacients with periodontitis compared to healthy subjects, which may be attributed to the biphasic character of INF-γ.

Expressão do interferon-γ em biópsias gengivais de pacientes com periodontite crônica severa / Expression of interferon-γ in gingival biopsies from patients with severe chronic periodontitis

O objetivo desse estudo foi analisar a expressão do interferon-gamma (INF-) em biópsias gengivais de sítios rasos e profundos de pacientes com periodontite crônica severa. O objetivo secundário foi correlacionar a expressão do INF- no fluido gengival com os sítios onde foram coletadas as biópsias gengivais. Foram coletadas biópsias de 22 pacientes portadores de periodontite crônica generalizada ou localizada severas (idade média 45,5  DP 8,9 anos), sendo 22 sítios profundos e 18 sítios rasos. O grupo controle foi composto por 14 pacientes clinicamente saudáveis (idade média 39,35  DP 16,5 anos). No total, foram 54 biópsias coletadas de 36 pacientes. As amostras do fluido gengival foram coletadas de alguns dos mesmos sítios de onde foram realizadas as biópsias, totalizando 12 sítios profundos, 8 sítios rasos e 4 sítios controle. Foram utilizados os parâmetros clínicos de avaliação de profundidade de bolsa à sondagem (PB); nível de inserção clínica (NIC); índice de placa visível (IPV) e índice de sangramento gengival (ISG). O tecido foi removido com punch de 2 mm de diâmetro, na área cirúrgica (grupo controle) ou na consulta para raspagem subgengival com ou sem acesso cirúrgico (grupo teste) e armazenados em Eppendorffs com 1 ml de solução de formaldeído a 10% para posterior análise morfológica e imuno-histoquímica. A intensidade da marcação do INF- foi avaliada semiquantitativamente nas células epiteliais, plasmócitos, macrófagos, fibroblastos e células endoteliais, considerando-se marcação forte (escore 2), marcação fraca (escore 1) ou ausência de marcação (escore 0). O teste de Kruskal-Wallis foi utilizado para comparar a expressão do INF- nos tecidos epitelial e conjuntivo, entre os três grupos (sítios profundos e rasos da periodontite e sítios controle). Observamos uma tendência a um padrão de marcação similar nos sítios rasos e profundos, com predomínio de marcação fraca nos sítios profundos. Nos sítios controle a marcação do...

The aim of this study was to analyze the expression of interferon-gamma (INF-γ) in gingival biopsies from shallow and deep sites in patients with severe chronic periodontitis. The secondary objective was to correlate the expression of INF-γ in the gingival crevicular fluid (GCF) with sites where the gingival biopsies were collected. Biopsies from 22 patients with chronic generalized or localized severe periodontitis (mean age 45.5 ± SD 8.9 years), 22 deep and 18 shallow sites were collected. The control group consisted of 14 patients clinically healthy (mean age 39.35 ± SD 16.5 years). In total, 54 biopsies were collected from 36 patients. The GCF samples were collected from some of the same sites where biopsies were performed, totaling 12 deep sites, 8 shallow sites and 4 control sites. Clinical evaluation parameters including probing pocket depth (PPD), clinical attachment level (CAL), visible plaque index (VPI) and gingival bleeding index (GBI) were used. The tissue was removed with a 2 mm of diameter punch, in the surgery area (control group) or in the subgingival scaling with or without surgical access (test group) and stored in an Eppendorff with 1 ml of 10% formaldehyde for subsequent morphological and immunohistochemical analysis. The intensity of staining was evaluated semi - quantitatively in the epithelial cells, plasma cells, macrophages, fibroblasts and endothelial cells, considering strong staining (score 2), weak staining (score 1) or absence of staining (score 0). The Kruskal-Wallis test was used to compare the expression of INF-γ in epithelial and connective tissues between the three groups (shallow and deep sites of periodontitis and control sites). We observed a trend for a similar pattern of staining in the shallow and deep sites, with a predominance of weak staining in deep sites. In control sites the staining of the epithelium was predominant. However, we could not demonstrate statistically significant differences between...

Avaliação do padrão de metilação em genes relacionados à resposta imune em polpa dental humana / Evaluation of pattern of mehylation in genes related to the immune response in human dental pulp

Receptores de reconhecimento padrão (PRRs), como o Toll-like receptor 2 (TLR-2) e o TLR4, participam da ativação do sistema imune por microrganismos quando a polpa dental está inflamada. Supões-se que seus níveis de expressão em tecido pulpar humano podem ser modulados por fatores epigenéticos, especialmente a metilação do DNA. O objetivo deste estudo foi avaliar o padrão de metilação dos genes TLR-2 e CD14 - um correceptor do TLR4 - em polpas dentais saudáveis e inflamadas. Adicionalmente, verificou-se a associação entre o padrão de metilação do gene interferon-gama (IFN-y), previamente por nós identificado, com sua expressão gênica em polpas dentárias...

Efeito do tratamento periodontal não cirúrgico em pacientes com periodontite crônica e agressiva: achados microbiológicos e imunológicos / Effect of non surgical periodontal treatment in chronic and aggressive subjects: microbiological and immunological findings

O objetivo do presente estudo foi comparar a expressão de IL-1β, IL-4, IL-8, interferon-γ, atividade de elastase e a composição do perfil microbiano subgengival antes e depois do tratamento periodontal não cirúrgico em pacientes com doença peridontal crônica generalizada (PC) e agressiva generalizada (PA). Vinte pacientes com PC e quatorze com PA foram avaliados. Dados clínicos, fluido gengival e biofilme subgengival foram analisados na visita inicial (VI) e 3 meses (3M) após o tratamento periodontal não cirúrgico. Amostras de fluido gengival (FG) foram coletadas com tiras de papel e os níveis de: IL-1β, IL-4, IL-8 e INF-γ foram medidos, utilizando um tipo de imunoensaio multiplexado (Luminex). Atividade da elastase foi avaliada por um ensaio enzimático. Amostras de placa subgengival foram analisadas através do checkerboard DNA-DNA hybridization. Na avaliação de 3 meses após terapia periodontal foi encontrado melhora significativa para todos os parâmetros clínicos em ambos os grupos. Foram encontradas reduções significativas na atividade de elastase nos sítios rasos e profundos dos pacientes do grupo PA e nos sítios profundos do grupo PC, também foi achado um aumento significativo de INF-γ nos sítios rasos do grupo PA. Os dados microbiológicos, mostraram reduções significativas para os níveis dos membros do complexo vermelho (P. gingivalis, T. forsythia, T.denticola), e para as espécies E.nodatum e P.micra no grupo PC. No grupo PA, ocorreram reduções significativas no níveis de P. gingivalis, T. forsythia, Fusobacterium nucleatum ss polymorphum e Fusobacterium periodonticum. Quando as respostas clínica e imunológica 3M após terapia foram comparadas entre os grupos, apenas diferenças sutis foram observadas. Nenhuma diferença microbiológica foi encontrada entre os grupos após a terapia. Em conclusão, os achados suportam nossa hipótese de que as periodontites cronica e agressiva respondem de forma semelhante ao tratamento periodontal nao cirúrgico.

Our aim was to compare the expression of IL1β, IL-4, IL-8, INF-γ, elastase activity and the composition of the subgingival microbiota profile before and after non-surgical periodontal treatment in patients with untreated generalized chronic (GCP) and aggressive periodontitis (GAgP). Twenty patients with GCP and 14 with GAgP were evaluated. Clinical data, GCF and plaque were analyzed at baseline and 3 months after non-surgical periodontal treatment. IL-1β, IL-4, IL-8 and INFγ were analyzed in a luminex assay. Elastase activity was assessed by an enzymatic assay. Subgingival plaque samples were analyzed using checkerboard DNA-DNA hybridization. Three months after periodontal therapy significant improvement for all clinical parameters in both groups were found. We have found significant reductions in the elastase activity in shallow and deep sites from GAgP and in deep sites from GCP group. A significant increase in INF-γ in the shallow sites from GAgP group were also found. Microbiological data showed significant reductions in the levels of members of the red complex (P. gingivalis, T. forsythia, T. denticola), and for the species E.nodatum and P.micra on GCP group. In the GAgP group, there were significant reductions in levels of P. gingivalis, T. forsythia, Fusobacterium nucleatum and Fusobacterium polymorphum ss periodonticum. When the clinical and immunological responses after therapy were compared between groups, only slight differences were found. No microbiological difference was found between groups after therapy. In conclusion, the findings support our hypothesis that the chronic and aggressive periodontitis respond equally well to non-surgical periodontal treatment.

Análise dos parâmetros clínicos periodontais e expressão genética de interferons alfa, gama e genes relacionados em indivíduos portadores de Síndrome de Down com doença periodontal / Periodontal clinical parameters analysis and genetics expression of Interferon-alpha, Interferon-gamma and genes related in individuais with Down Syndrome and periodontal disease

A doença periodontal (DP) em indivíduos com Síndrome de Down (SD) se desenvolve com alta prevalência, precocemente, de modo rápido e generalizado em comparação com indivíduos não-sindrômicos. Foi demonstrado que portadores da SD apresentam resposta imune diminuída em relação aos cromossomicamente normais. O objetivo desta pesquisa foi investigar diferenças nos parâmetros clínicos periodontais e níveis de expressão dos genes Interferon-gama (IFNG), Interferon-gama receptor 1 (IFNGR1), Interferon-gama receptor 2 (IFNGR2), Interferon-alfa (IFNA), Interferon-alfa receptor 1 (IFNAR1), Interferon-alfa receptor 2 (IFNAR2), Janus-quinase 1 (JAK1), Transdutor de sinal e ativador da transcrição 1 (STAT1) e Fator de regulação de interferon 1 (IRF1) em indivíduos com SD que apresentam ou não DP e em indivíduos cromossomicamente normais. Fizeram parte deste estudo 80 indivíduos entre 7 e 57 anos de idade subdivididos em 4 grupos: SD com DP (A); indivíduos com SD sem DP (B); indivíduos não-sindrômicos (Controle) com DP (C) e indivíduos Controle sem DP (D). A expressão gênica foi investigada por meio de quantificação relativa utilizando a técnica da Reação em Cadeia da Polimerase (PCR) em Tempo Real. Para o índice sangramento à sondagem (SS) não houve diferença entre os grupos A e C. A periodontite crônica localizada foi o tipo prevalente tanto entre indivíduos com SD como Controle. Considerando os parâmetros clínicos, não foram encontradas diferenças na periodontite crônica localizada entre os indivíduos com SD e Controle, assim como para a periodontite crônica generalizada. Com relação à análise genética, observou-se que indivíduos dos grupos com SD em relação aos grupos cromossomicamente normais (A+B-C+D) tiveram uma expressão de IFNG semelhante ao observado entre indivíduos do grupo Controle com DP (CD). No entanto, quando a DP acomete indivíduos com SD (A-B), o IFNG não apresenta uma resposta imune tão eficiente quanto para os indivíduos cromossomicamente normais (C-D). Na expressão do gene IFNA, indivíduos com SD (A+B-C+D) têm uma expressão consideravelmente menor que os indivíduos cromossomicamente normais com DP (C-D), indicando uma imunodeficiência em um importante mecanismo de controle da inflamação. Indivíduos com SD apresentaram expressão significativamente maior dos genes IFNGR2 e IFNAR1, os quais residem no cromossomo 21. A expressão de JAK1 nos indivíduos com SD independente de DP (A+B-C+D) foi acentuadamente menor, enquanto que a expressão do gene IRF1 foi significativamente maior nesses indivíduos em comparação aos indivíduos cromossomicamente normais (C-D). Portanto, a SD provavelmente pode interferir na cascata da via de sinalização do IFNG e IFNA contribuindo para uma menor eficiência do sistema imune de indivíduos frente à um estímulo inflamatório como a DP

Periodontal disease (PD) in individuals with Down Syndrome (DS) has an early, quickly and widespread onset and high prevalence when compared with individuals without the Syndrome. Only poor oral hygiene does not explain the severe periodontal destruction seen in DS patients. It has been shown that DS patients have a weaker immune response than people with normal number of chromosomes. The aim of this study was to investigate differences in periodontal clinical parameters and the expression levels of the genes Interferon-gamma (IFNG), Interferongamma receptor 1 (IFNGR1), Interferon-gamma receptor 2 (IFNGR2), interferon-alpha (IFNA), interferon-alpha receptor 1 (IFNAR1), Interferonalpha receptor 2 (IFNAR2), Janus-kinase 1 (JAK1), Signal transducers and activators of transcription 1 (STAT1) and Interferon regulatory factor 1 (IRF1) in DS patients with and without periodontal disease in comparison with chromossomically normal individuals. A total of 80 individuals aged 7 to 57 years participated in this study and were divided into 4 groups: DS with PD (A); DS without PD (B); individuals without DS (control) with PD (C) and individuals without DS (control) and without PD (D). A quantitative RT-qPCR was used to investigate gene expression. There was no difference between groups A and C regarding the bleeding on probing (BOP) index. The most prevalent type of periodontitis seen in this study was the localized chronic periodontitis, both in individuals with and without DS. Considering the clinical parameters, localized and generalized chronic periodontitis did not differ between individuals with and without DS. Regarding genetic analysis, individuals of the groups with DS in relation to the groups without DS (A+B-C+D) showed an IFNG expression similar to that seen among the individuals of groups control with PD (C-D). However, individuals with DS (A+B-C+D), the IFNG immune response is not as efficient as that of individuals without DS (C-D). Individuals of groups with DS (A+B) have a considerably smaller expression of the IFNA gene than those of groups without DS (C+D), indicating an immune deficiency in an important inflammation control mechanism. Individuals with DS presented a significantly greater expression of the genes IFNGR2 and IFNAR1, both located in chromosome 21. JAK1 expression in individuals with DS regardless of PD (A+B-C+D) was markedly smaller, while the expression of the IRF1 gene was significantly greater when compared with individuals without DS (C+D). Therefore, DS can probably interfere in the IFNG and IFNA signaling pathways, making the response of the immune system of individuals with DS to inflammatory stimuli, such as PD, less efficient

Avaliação da citotoxicidade de materiais obturadores de canais radiculares: influência na liberação de fator de necrose tumoral-alfa e interferon-y e óxido nítrico em cultura de células murinas / Evaluation of the citotoxicity of endodontic sealers: influency on liberation of tumor necrotic factor-alpha, interferon gamma and nitric oxide in murine cells culture

Os macrófagos constituem uma população celular do sistema imune. Estas células podem ser ativadas por uma variedade de estímulos e suas principais funções incluem a fagocitose de partículas estranhas, apresentação de antígenos, produção de citocinas e compostos intermediários do nitrogênio (NO) e do oxigênio (H2O2). Os cimentos endodônticos são capazes de promover uma estimulação do sistema imune. Neste estudo, foram analisados os níveis de quantificação das citocinas, além do mediador óxido nítrico, como uma medida de estimulação de macrófagos peritoneais de camundongos. Através de análise estatística dos dados, foram observados os níveis de citotoxicidade dos macrófagos de camundongos estimulados pelos diferentes cimentos endodônticos, meio RPMI-1640 (grupo controle) e LPS (grupo controle +). Os diferentes cimentos testados apresentaram concentrações com diferentes citotoxicidades: Sealapex 35 ug/ml, Polímero de Mamona 8,75 ug/ml, do Epiphany 17,5 ug/mle do AH Plus 70 ug/ml. Após a adequação das concentrações viáveis dos cimentos testados conclui-se que o material que mais estimulou a liberação de NO foi Primer...