Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA

Abstract Objective The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC). Material and Methods Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and » serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05. Results In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC. Conclusion mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.

Demetallization of Enterococcus faecalis biofilm: a preliminary study

Abstract Objectives To determine the concentration of calcium, iron, manganese and zinc ions after the application of chelator to Enterococcus faecalis biofilms. Material and Methods Fifty bovine maxillary central incisors were prepared and inoculated with E. faecalis for 60 days. The following were used as irrigation solutions: 17% EDTA (pH 3, 7 and 10), 2.5% sodium hypochlorite (NaOCl) combined with 17% EDTA (pH 3, 7 and 10), distilled water (pH 3, 7 and 10), and 2.5% NaOCl. Each solution was kept in the root canal for five minutes. Fifteen uncontaminated root canals were irrigated with 17% EDTA (pH 3, 7 and 10). Six teeth were used as bacterial control. The number of calcium, iron, manganese and zinc ions was determined using flame atomic absorption spectrometry. Mean ± standard deviation (SD) values were used for descriptive statistics. Results Calcium chelation using 17% EDTA at pH 7 was higher than at pH 3 and 10, regardless of whether bacterial biofilm was present. The highest concentration of iron occurred at pH 3 in the presence of bacterial biofilm. The highest concentration of manganese found was 2.5% NaOCl and 17% EDTA at pH 7 in the presence of bacterial biofilm. Zinc levels were not detectable. Conclusions The pH of chelating agents affected the removal of calcium, iron, and manganese ions. The concentration of iron ions in root canals with bacterial biofilm was higher after the use of 17% EDTA at pH 3 than after the use of the other solutions at all pH levels.

Avaliação da atividade antibacteriana de diferentes materiais restauradores provisórios / Evaluation of the antibacterial activity of different temporary restorative materials

O objetivo deste estudo foi avaliar a propriedade antimicrobiana de diferentes materiais restauradores provisórios: Óxido de zinco e eugenol, óxido de zinco pronto para uso [Coltosoll, resinoso (Bioplic) e guta-percha. Foram utilizadas 10 amostras de cada material acondicionados em poços de m-enterococcus ágar contaminado com Enterococcus faecolis. Poços vazios cons­tituíram o grupo controle. As placas foram encubadas a 37°C por 24 horas, e os halos de inibição foram analisados. Os dados obtidos foram submetidos ao teste de Kruskal-Wallis complemen­tado pelo teste de Dunn (p<0,05). Os resultados apresentaram que o óxido de zinco e eugenol apresentou maior halo de inibição quando comparado à guta-percha e o resinoso, porém não obteve diferença significante ao óxido de zinco pronto para uso que foi diferente somente em relação ao grupo controle de crescimento bacteriano zero. Pode-se concluir que na observação após 24 h de inserção, o Óxido de zinco e eugenol demonstrou maior efeito antibacteriano que o resinoso e a guta-percha, porém semelhante ao óxido de zinco pronto para uso.

The aim of this study was to evaluate the antibacterial activity of different temporary res­torative materiais: Zinc oxide and eugenol, ready-to-use zinc oxide, resin-based (Bioplic) and Gutta-percha. Ten samples of each material were placed into wells done on m-enterococcus agar contaminated with Enterococcus faecalis. Empty wells were the control group. The pla­tes were incubated at 3rC for 24 hours, and the inhibition zone around the materiais was analyzed. Data were submitted to Kruskal-Wallis complemented by Dunn's test (p <0.05) The results showed that the zinc oxide and eugenol showed greater inhibition zone compared to gutta-percha and resin-based material, no significant difference was found between them and ready-to-use zinc oxide. Ready-to-use zinc oxide was significant difference to control group of growth inhibition zero. It can be concluded that after 24 h, the zinc oxide and eugenol has more antibacterial effect than resin-based material and gutta-percha, but it showed similarity to ready-to-use zinc oxide.

Effects of lysed Enterococcus faecalis FK-23 on allergen-induced peritoneal accumulation of eosinophils and serum total IgE concentration in inbred mice.

Recent clinical trials have shown the possibility of probiotics in prevention and treatment of allergic diseases. The purpose of this experimental study was to assess the influence of lysed Enterococcus faecalis FK-23 (LFK) on allergic responses in different mouse strains. We performed a comparative study on the effects of LFK for allergen-induced peritoneal accumulation of eosinophils and serum total IgE concentration by using BALB/c, C57BL/6, C3H/HeN and C3H/HeJ mice. There was no significant difference in total number of peritoneal accumulated cells induced by cedar pollen allergen between the control and LFK groups in any strain of mice (p > 0.05); however, the ratio of eosinophils to total accumulated cells was significantly decreased in LFK-treated mice of BALB/c (p = 0.016), C3H/HeN (p = 0.010) and C3H/HeJ (p = 0.004), but not C57BL/6 (p > 0.05). No significant difference in serum total IgE concentration was found between the control and LFK groups of different mouse strains (p > 0.05). These results reveal a different effect of LFK on suppressing allergen-induced local eosinophila in inbred strains of mice, suggesting the effectiveness of probiotics on limiting allergy might be under the influence of individual genetic background.

Detection and characterization of enterocins from Enterococcus sp.

Examination of 90 isolates of Enterococcus sp. revealed production of enterocin by two isolates of E. faecalis which was inhibitory to Listeria monocytogenes. Two isolates of E. gallinarum produced enterocin active against Staphylococcus aureus. None of the isolates antagonized Salmonella enteritidis. The enterocins of E. faecalis isolates were inactivated by alpha-chymotrypsin but not by trypsin and papain, while those of E. gallinarum were resistant to all the three enzymes. Enterocins produced by all 4 strains were resistant to heating at 60 degrees C for 30 min and 80 degrees C for 10 min, but sensitive to 121 degrees C for 15 min. At 100 degrees C for 10 min, two enterocins, one each of E. faecalis and E. gallinarum were inactivated, while the remaining two retained the bactericidal activity.

Tissue reactions to a component of root canal system bacteria: lipoteichoic acid

Lipoteichoic acid (LTA), present in Gram-positive microorganisms, has physiochemical characteristics that allow it to act as an immunogen. Due to polymicrobial characteristics of root canal infections, LTA can participate in the development of periapical disease. The reaction of the rat subcutaneous tissue to Teflon tube implants, filled with Fibrinol soaked in lipoteichoic acid (concentration of 150 æg/ml), was observed. Lipoteichoic acid provoked an inflammatory tissue reaction