RESUMO
We focused on apoptotic blebs from Leishmania major-infected macrophages as a vaccine for cutaneous leishmaniasis. Apoptosis was induced in L. major-infected J774A.1 cells in order to prepare apoptotic blebs. Test groups of BALB/c mice were immunized with these at doses of 1 × 106, 5 × 106 or 1 × 107 blebs. An immunization control group received Leishmania lysate antigens. The results showed that as the number of apoptotic bodies increased, the lymphocyte proliferation index increased, and this was proportional to IFN-γ level in the test groups. Additionally, the difference of IFN-γ, IL-4, IFN-γ/IL-4 ratio, or total IgG (p < 0.0001) in all groups was statistically significant compared to the negative control group. The highest IFN-γ (514.0 ± 40.92 pg/mL) and IFN-γ/IL-4 ratio (2.94 ± 0.22) were observed in the group that received 1 × 107 apoptotic blebs. The highest levels of IL-4 (244.6 ± 38.8 pg/mL) and total IgG (5626 ± 377 µg/mL) were observed in the immunization control group. Reflecting these data, no lesions were observed in any of the groups vaccinated with apoptotic blebs after 12 weeks. In summary, the use of apoptotic blebs from L. major-infected macrophages is protective against the challenge with L. major in this animal model.
Assuntos
Leishmania major , Leishmaniose Cutânea , Leishmaniose , Vacinação , Animais , Camundongos , Antígenos de Protozoários , Citocinas , Leishmania major/patogenicidade , Leishmaniose Cutânea/prevenção & controle , Macrófagos , Camundongos Endogâmicos BALB CRESUMO
INTRODUCTION: This in vitro study aimed at comparing the effect of agitating the final irrigant solutions of root canal by ultrasonic or using 808 nm Diode laser on the apical seal of canal. METHODS AND MATERIALS: A total of 90 extracted human maxillary central incisors were prepared up to size #45 and were randomly assigned to 4 experimental groups (n=20) and two control groups (n=5) respectively, as follows: I): 3 mL of 5.25% NaOCl was agitated as final irrigant solution with ultrasonic for 30 sec. The ultrasonic tip was 1 mm shorter than the working length, II): 3 mL of 5.25% NaOCl was agitated as final irrigant with 808 nm Diode laser for 30 sec. Fiber tip, placed in 1 mm shorter from working length was spirally moved coronally, III): 3 mL of 17% EDTA was agitated as final irrigant with 808 nm Diode laser for 30 sec and was applied similar to group II, IV): 3 mL of 17% EDTA was stimulated as final irrigant with ultrasonic for 30 sec and was applied similar to I. Apical seal was assessed by Dual Chamber technique using Bovine Serum Albumin protein. Kruskal-Wallis and Mann Whitney tests were used with significance level lower than 0.05% for statistical analysis. RESULTS: The average leakage in the negative control, positive control, and groups I, II, III, IV were: 0.00, 13.5±5.1, 1.72±2.9, 5.12±5.6, 3.36±3.7, 2.4±4.2, respectively. Statistical analysis showed significant difference between groups (P<0.05). There was a significant difference between groups 1 and 2 in terms of protein leakage. CONCLUSION: Agitating 5.25% sodium hypochlorite solution as the final irrigant with ultrasonic is more effective in apical leakage reduction compared to other groups.