RESUMEN

BACKGROUND: In the present study, we examined the effects of Fe3O4@bio-MOF nanoparticle (Nano-FO) plus artemisinin (Art) and glucantime (Glu) or shark cartilage extract (ShCE) on Leishmania major in vitro and in vivo. METHODS: This experimental study was conducted at the laboratory of Department of Parasitology, Tarbiat Modares University, Tehran, Iran during 2016-2017. The promastigote and amastigote assays were performed were conducted at the presence of 3.12-400 µg/mL of the drug combinations. According to in vitro IC50 results, the combinations of 12.5µg/mL Nano-FO with 25 µg/mL Art as well as 200 µg/mL Glu and 0.5 mL of 20 mg/kg of ShCE were used to treat BALB/c mice. During and at the end of the treatment, the lesion sizes were measured. Parasite loads, cytokine levels were evaluated at the end of the treatment. RESULTS: The IC50 of Fe3O4@bio-MOF-Artemisinin (Nano-FO/Art), Fe3O4@bio-MOF-Glucantime (Nano-FO/Glu), and Fe3O4@bio-MOF-Shark cartilage extract (Nano-FO/ShCE) on promasitigotes were 12.58±0.12, 235±0.17, and 18.54±0.15, respectively. These results on amastigotes were 10.32±0.01, 187±0.03, and 338±0.07 µg/mL, respectively. The apoptosis percentage of these combinations were 32.54%, 20.59%, and 15.68% in promastigotes and 15.68%, 12.84%, and 3.51% in infected macrophages, respectively with no toxicity on uninfected macrophages. In vivo results showed that the size of lesions significantly decreased against all drugs combinations, but Nano-FO/Art combination with Selectivity Index of 23.62 value was safe, and more effective on healing of lesions than other drugs combinations (P=0.003). CONCLUSION: This study suggested that Nano-FO/Art combination can be considered as an anti-leishmania combination therapy in CL induced by L. major.

RESUMEN

In this study we examined enhancement effects of Artemisinin plus Glucantime and shark cartilage extract on promastigotes and amastigotes of L.infantum in in-vitro condition.The toxicity of artemisinin, glucantime, and shark cartilage extract on the L. infantum promastigotes and amastigote-infected macrophages was evaluated using MTT assay. The role of these drugs inducing apoptosis in promastigotes, un- infected, and parasite- infected macrophages was also studied. Using promastigote assay, IC50 values of artemisinin and glucantime as standalone drugs as well as in combination were obtained to be 50, 400, and 100µg/mL respectively. The flow cytometry analysis of apoptotic promastigotes stained with Annexin-V FITC staining showed that artemisinin, glucantime, artemisinin plus glucantime, artemisinin plus shark cartilage extract, and shark cartilage extract alone applied at their IC50 concentrations resulted in 53.5%, 73.92%, 64.46%, 49.9%, and 47.34% apoptosis respectively. The results of MTT assay indicated that cytotoxicity of artemisinin, glucantime, artemisinin plus glucantime, shark cartilage plus artemisinin, and shark cartilage in infected macrophages after 72h was 75%, 84%, 82%, 30%, and 3% respectively. In un- infected macrophages, cytotoxicity of Artemisinin, Glucantime, Artemisinin plus Glucantime and shark cartilage was 15%, 31%, 21%, 2%, and 0% respectively.This study suggests that artemisinin, glucantime, artemisinin plus glucantime, and shark cartilage extract have significant killing effects on promastigotes and amastigotes. Also, it proved that artimisinin alone and in combination with glucantime and shark cartilage extract has little toxic effect on macrophages, but could induce apoptosis in L.infantum promastigotes and amastigote-infected macrophages. Thus, these chemicals can be used as alternative drugs for in-vivo studies.

RESUMEN

OBJECTIVES: Because leishmaniasis is related to the impaired functioning of T-cells, the use of an immunomodulator can increase the efficacy of antileishmanial therapy in visceral leishmaniasis. In this study, we used shark cartilage extract with artemisinin and glucantime against visceral leishmaniasis in BALB/c mice, and evaluated the synergistic therapeutic effect. MATERIALS AND METHODS: The culturing method and quantitative real-time PCR by using the kDNA gene was used to detect parasite loads in the spleen and liver. INF-γ and IL-4 cytokine levels and survival rates were assayed. RESULTS: The drug therapy with target drugs reduced parasite burden in the spleen and liver significantly. Although parasite burden was lower in the artemisinin treated group than in the glucantime treated group (P<0.05). The mice survival rate records, throughout the experimental period, showed highly significant survival rates in the test groups compared to the control group (P<0.001). The results of cytokine assay in mice treated with glucantime-shark cartilage extract combination indicated significant increases of IFNγ and IL-4 (P<0.05). Although the increase of IFNγ was more notable than IL-4. The synergistic therapeutic effect is shown in all groups except in the group treated with shark cartilage extract-artemisinin combination. The IFN-γ in glucantime-shark cartilage extract combination treated group was higher than in other groups (P<0.05). The survival rate in this group was more than in other groups too (P<0.05). CONCLUSION: Combination therapy with shark cartilage extract as an immunomodulator can increase antileishmanial effects of antimony drugs in VL treatment.

RESUMEN

In this work, a magnetic bio-metal-organic framework (MBMOF) nanocomposite with porous-layer open morphology is synthesized through a simple sonochemical approach and its effects on Leishmania major (MRHO/IR/75/ER) under both in vitro and in vivo conditions are investigated. The effects of sonication time, initial concentration of reagents and sonication power on size and morphology of MBMOF nanocomposites have been investigated and optimized. A comparison was then made between the structural information of the nanostructures and that of the bio-metal-organic framework crystals. Using the powder X-ray diffraction (PXRD), field emission scanning electron microscope (FE-SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), energy dispersive analysis of X-ray (EDAX), vibrating sample magnetometer (VSM), thermogravimetric analysis (TGA), and Brunauer-Emmet-Teller (BET) techniques, the prepared MBMOF nanocomposites were characterized. The mean numbers of promastigotes (cell/ml) in different MBMOF concentrations (3.12, 6.25, 12.5, 25, 50, 100, 200 and 400 µg mL-1) were determined by direct counting after 24, 48 and 72 h. Using MTT assays, the cytotoxic impacts of the MBMOF nanocomposites on promastigotes, intracellular amastigotes, and J774 macrophages were estimated. In order to investigate their therapeutic effects, the prepared MBMOF nanocomposites (25 and 12.5 µg mL-1) were used as ointment three times a week to treat Leishmania major in BALB/c mice. The lesion size and weight of mice were assessed before and during the treatment. The parasitic loads were measured in spleen and liver through the culture. After 72 h, the INF-γ and IL-4 cytokines levels in the supernatant of the spleen culture were measured. To the best of the authors' knowledge, this study is the first to attempt to synthesize the bio-MOFs through an in-situ sonosynthesis route under ultrasound irradiation and examine their cytotoxicity effects on Leishmania major under in vitro and in vivo conditions.

Asunto(s)

Leishmania major/efectos de los fármacos , Nanopartículas de Magnetita/química , Compuestos Orgánicos/química , Ondas Ultrasónicas , Animales , Línea Celular , Modelos Animales de Enfermedad , Femenino , Interferón gamma/metabolismo , Interleucina-4/metabolismo , Leishmaniasis Cutánea/tratamiento farmacológico , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/parasitología , Macrófagos/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Compuestos Orgánicos/farmacología , Compuestos Orgánicos/uso terapéutico , Difracción de Polvo , Espectrometría por Rayos X , Espectroscopía Infrarroja por Transformada de Fourier , Bazo/efectos de los fármacos , Bazo/metabolismo , Bazo/parasitología , Termogravimetría

RESUMEN

The early diagnosis of visceral leishmaniasis (VL) using Direct Agglutination Tests (DAT) and its treatment and control are essential actions taken in rural health centers in endemic foci of the infection based on the national protocols set by the Iranian Ministry of Health and Medical Education. Eleven clinically confirmed VL patients with typical VL symptoms and negative results of DAT, admitted to the pediatrics department of Valiasr Hospital in Meshkinshahr underwent parasitological tests. 7 of the total of 11 patients had positive result of bone marrow puncture smears and all of them had negative results of DAT. Thus factors that had led to false negative DAT results were examined. The patients' blood samples were collected in microhematocrit tubes using the finger prick technique, centrifuged and their plasma then separated. The complete medical history of the patients was taken using a questionnaire. The laboratory staff therefore checked the quality of anti-leishmania antigen, materials and equipment used. The patients' medical history showed that they had all been administered corticosteroid medications such as dexamethasone or hydrocortisone prior to visiting the laboratory. The DAT was repeated in these patients 2-3 weeks after their last administration of corticosteroids. The antibody titers were positive this time. A total of 3 of the collected specimens (27.3 %) showed a titer of 1.3200, 5 (46 %) showed a titer of 1.1600 and 3 (27.3 %) a titer of 1.800. Due to the effects of some medications, particularly corticosteroids, on serological tests, the patients' full medical history should be taken prior to performing this test and physicians working at endemic regions of this infection should be notified about these drug interactions.