Enterococcus faecalis Demonstrates Pathogenicity through Increased Attachment in an Ex Vivo Polymicrobial Pulpal Infection.

This study investigated the host response to a polymicrobial pulpal infection consisting of Streptococcus anginosus and Enterococcus faecalis, bacteria commonly implicated in dental abscesses and endodontic failure, using a validated ex vivo rat tooth model. Tooth slices were inoculated with planktonic cultures of S. anginosus or E. faecalis alone or in coculture at S. anginosus/E. faecalis ratios of 50:50 and 90:10. Attachment was semiquantified by measuring the area covered by fluorescently labeled bacteria. Host response was established by viable histological cell counts, and inflammatory response was measured using reverse transcription-quantitative PCR (RT-qPCR) and immunohistochemistry. A significant reduction in cell viability was observed for single and polymicrobial infections, with no significant differences between infection types (∼2,000 cells/mm2 for infected pulps compared to ∼4,000 cells/mm2 for uninfected pulps). E. faecalis demonstrated significantly higher levels of attachment (6.5%) than S. anginosus alone (2.3%) and mixed-species infections (3.4% for 50:50 and 2.3% for 90:10), with a remarkable affinity for the pulpal vasculature. Infections with E. faecalis demonstrated the greatest increase in tumor necrosis factor alpha (TNF-α) (47.1-fold for E. faecalis, 14.6-fold for S. anginosus, 60.1-fold for 50:50, and 25.0-fold for 90:10) and interleukin 1ß (IL-1ß) expression (54.8-fold for E. faecalis, 8.8-fold for S. anginosus, 54.5-fold for 50:50, and 39.9-fold for 90:10) compared to uninfected samples. Immunohistochemistry confirmed this, with the majority of inflammation localized to the pulpal vasculature and odontoblast regions. Interestingly, E. faecalis supernatant and heat-killed E. faecalis treatments were unable to induce the same inflammatory response, suggesting E. faecalis pathogenicity in pulpitis is linked to its greater ability to attach to the pulpal vasculature.

Influência da infecção endodôntica associada ou não à diabetes nos níveis da citocina pró-inflamatória IL-17 nos tecidos periapical, hepático e renal de ratos Wistar / Influence of endodontic infection with or without diabetes in the levels of proinflammatory cytokine IL-17 in periapical, hepatic and renal tissues of Wistar rats

O objetivo deste trabalho foi avaliar a influência da infecção endodôntica associada ou não à diabetes, nos níveis da citocina pró-inflamatória IL-17, nos tecidos periapical, hepático e renal de ratos Wistar. Foram utilizados 40 ratos machos divididos em quatro grupos contendo 10 animais cada grupo: ratos normoglicêmicos (N); ratos normoglicêmicos com periodontite apical (N-PA); ratos com diabetes experimental (DE); ratos com diabetes experimental e com periodontite apical (DE-PA). A diabetes foi induzida por meio da administração de estreptozotocina via veia peniana. Foi realizada avaliação glicêmica, para confirmar o desenvolvimento da doença, no sexto e no último dia do experimento. A infecção endodôntica foi induzida por meio da exposição pulpar do primeiro molar superior direito ao meio bucal. Após 30 dias, foram coletados o fígado e o rim, para a quantificação da IL-17 pelo método ELISA, bem como a hemi-maxila direita para a quantificação da IL-17 pela técnica imunoistoquímica. Os valores obtidos nos testes paramétricos foram tabulados e analisados, estatisticamente, por meio dos testes de análise de variância (ANOVA) e teste de Tukey (p<0,05). Os valores obtidos em scores foram analisados, estatisticamente, por meio dos testes de Kruskal-Wallis e teste de Dun (p<0,05). Os resultados mostraram que os ratos diabéticos (DE e DE-PA) apresentaram maior quantidade de IL-17 nos tecidos hepático e renal quando comparados aos ratos normoglicêmicos (N e N-PA) (p<0,05). A análise histológica mostrou que a periodontite apical de ratos diabéticos (DE-PA) possui maior severidade quando comparada com a dos ratos normoglicêmicos (N-PA) (p<0,05), além disso, houve um aumento significativo dos níveis de IL-17 na periodontite apical dos ratos diabéticos (DE-PA) quando comparados aos normoglicêmicos (N-PA) (p<0,05). Entretanto, a infecção endodôntica induzida em um único elemento dentário não foi capaz de alterar os níveis de IL-17 nos tecidos hepático e renal (p>0,05)...

The aim of this study was to evaluate the influence of endodontic infection with or without diabetes, in the levels of proinflammatory cytokine IL-17 in apical periodontitis and hepatic and renal tissues of Wistar rats. Forty male Wistar rats were divided into four groups of ten animals each: normoglycemic rats (N), normoglycemic rats with apical periodontitis (N-AP), rats with experimental diabetes (ED), rats with experimental diabetes and apical periodontitis (ED-AP). Diabetes was induced by injection of streptozotocin in the penile vein. Glycemic control was performed to confirm the development of the disease at the sixth and last day of the experimental period. The endodontic infection was induced by exposure of the upper right first molar to the oral environment. After 30 days, the hepatic and renal tissues were collected for the IL-17 quantification by ELISA, as well the right hemi-jaw for the IL-17 quantification by immunohistochemistry. The values obtained in parametric tests were tabulated and analyzed statistically by means of analysis of variance tests (ANOVA) and Tukey test (p<0.05). The values obtained for scores were statistically analyzed by Kruskal-Wallis and Dun test (p<0.05). The results showed that diabetic rats (ED and ED-AP) showed a higher IL-17 levels in the hepatic and renal tissues compared to normoglycemic rats (N and N-AP) (p<0.05). The apical periodontitis in diabetic rats (ED-AP) had higher severity when compared to normoglycemic rats (N-PA) (p<0.05), furthermore, there was a large increase in IL-17 levels in apical periodontitis of diabetics rats (ED-AP) when compared with normoglycemic rats (N-AP) (p<0.05). However, the induced endodontic infection did not alter IL-17 levels in hepatic and renal tissues (p>0.05). It may be concluded that diabetes increases IL-17 levels in hepatic and renal tissues, as well as enhances the IL-17 production in apical periodontitis. Diabetes also increases the severity and development of apical...

Influência da infecção endodôntica associada ou não à diabetes nos níveis da citocina pró-inflamatória IL-17 nos tecidos periapical, hepático e renal de ratos Wistar / Influence of endodontic infection with or without diabetes in the levels of proinflammatory cytokine IL-17 in periapical, hepatic and renal tissues of Wistar rats

O objetivo deste trabalho foi avaliar a influência da infecção endodôntica associada ou não à diabetes, nos níveis da citocina pró-inflamatória IL-17, nos tecidos periapical, hepático e renal de ratos Wistar. Foram utilizados 40 ratos machos divididos em quatro grupos contendo 10 animais cada grupo: ratos normoglicêmicos (N); ratos normoglicêmicos com periodontite apical (N-PA); ratos com diabetes experimental (DE); ratos com diabetes experimental e com periodontite apical (DE-PA). A diabetes foi induzida por meio da administração de estreptozotocina via veia peniana. Foi realizada avaliação glicêmica, para confirmar o desenvolvimento da doença, no sexto e no último dia do experimento. A infecção endodôntica foi induzida por meio da exposição pulpar do primeiro molar superior direito ao meio bucal. Após 30 dias, foram coletados o fígado e o rim, para a quantificação da IL-17 pelo método ELISA, bem como a hemi-maxila direita para a quantificação da IL-17 pela técnica imunoistoquímica. Os valores obtidos nos testes paramétricos foram tabulados e analisados, estatisticamente, por meio dos testes de análise de variância (ANOVA) e teste de Tukey (p<0,05). Os valores obtidos em scores foram analisados, estatisticamente, por meio dos testes de Kruskal-Wallis e teste de Dun (p<0,05). Os resultados mostraram que os ratos diabéticos (DE e DE-PA) apresentaram maior quantidade de IL-17 nos tecidos hepático e renal quando comparados aos ratos normoglicêmicos (N e N-PA) (p<0,05). A análise histológica mostrou que a periodontite apical de ratos diabéticos (DE-PA) possui maior severidade quando comparada com a dos ratos normoglicêmicos (N-PA) (p<0,05), além disso, houve um aumento significativo dos níveis de IL-17 na periodontite apical dos ratos diabéticos (DE-PA) quando comparados aos normoglicêmicos (N-PA) (p<0,05). Entretanto, a infecção endodôntica induzida em um único elemento dentário não foi capaz de alterar os níveis de IL-17 nos tecidos hepático e renal (p>0,05)...

The aim of this study was to evaluate the influence of endodontic infection with or without diabetes, in the levels of proinflammatory cytokine IL-17 in apical periodontitis and hepatic and renal tissues of Wistar rats. Forty male Wistar rats were divided into four groups of ten animals each: normoglycemic rats (N), normoglycemic rats with apical periodontitis (N-AP), rats with experimental diabetes (ED), rats with experimental diabetes and apical periodontitis (ED-AP). Diabetes was induced by injection of streptozotocin in the penile vein. Glycemic control was performed to confirm the development of the disease at the sixth and last day of the experimental period. The endodontic infection was induced by exposure of the upper right first molar to the oral environment. After 30 days, the hepatic and renal tissues were collected for the IL-17 quantification by ELISA, as well the right hemi-jaw for the IL-17 quantification by immunohistochemistry. The values obtained in parametric tests were tabulated and analyzed statistically by means of analysis of variance tests (ANOVA) and Tukey test (p<0.05). The values obtained for scores were statistically analyzed by Kruskal-Wallis and Dun test (p<0.05). The results showed that diabetic rats (ED and ED-AP) showed a higher IL-17 levels in the hepatic and renal tissues compared to normoglycemic rats (N and N-AP) (p<0.05). The apical periodontitis in diabetic rats (ED-AP) had higher severity when compared to normoglycemic rats (N-PA) (p<0.05), furthermore, there was a large increase in IL-17 levels in apical periodontitis of diabetics rats (ED-AP) when compared with normoglycemic rats (N-AP) (p<0.05). However, the induced endodontic infection did not alter IL-17 levels in hepatic and renal tissues (p>0.05). It may be concluded that diabetes increases IL-17 levels in hepatic and renal tissues, as well as enhances the IL-17 production in apical periodontitis. Diabetes also increases the severity and development of apical...