G-3669

Evaluation of anti-leishmanial effect of Copper Oxide (CuO) nanoparticles doped with AgNo3 (CuO-Ag) synthesis to Ferula szowitsiana on Leishmania major promastigotes In-Vitro

 Pegah Babaali ¹ ℗, Fateme Habibi ¹, Mahmoodreza Behravan ² ©, Samira Eghbali ³   

 
Abstract

Introduction: Cutaneous leishmaniasis is one of the most important health-threatening diseases in Iran and around the world. Glucantime is currently used for treatment, but due to its side effects and high resistance, the use of alternative therapeutic methods, especially nanoparticles and also metal nanoparticles with plant extracts, has attracted the attention of researchers. The aim of this study was to evaluate the anti-leishmanial effect of Copper Oxide (CuO) nanoparticles doped with AgNo3 (CuO-Ag) synthesized using Ferula szowitsiana on Leishmania major promastigotes In-Vitro. Methods and Materials: In this experimental study, the anti-leishmanial activity of Copper Oxide nanoparticles doped with AgNo3 (CuO-Ag) synthesized using F.szowitsiana at concentrations of 0, 0.75, 1.5, 3, 6, 12, 48, and 96 μg/mL was investigated at exposure times of 24, 48, and 72 hours on Leishmania major promastigotes of the standard strain prepared from the School of Public Health, Tehran University of Medical Sciences. F.szowitsiana was collected, extracted, and prepared at different concentrations. Then, silver-doped CuO nanoparticles were synthesized along with F.szowitsiana extract, and X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) tests were used to identify and examine their structure. Next, the number of live parasites (percentage of viability) was counted by Trypan Blue dye using hemocytometer and light microscope for each of the nanoparticles. Glucantime and distilled water were considered as positive and negative controls, respectively. The IC50 of all nanoparticles was also calculated after 24, 48, and 72 hours. All reactions were performed three times. A t-test was used to compare the data obtained from the test and control samples. This study was approved by the Ethics Code IR.BUMS.REC.1402.356. Result: The synthesis of CuO-Ag nanoparticles using F.szowitsiana, was confirmed by color change, XRD (crystalline phases), TEM (spherical/irregular shape, agglomeration), and SEM (22–47 nm size). The results of this study showed that all concentrations of CuO nanoparticles, CuO-Ag nanoparticles, and CuO-Ag nanoparticles with F.szowitsiana extract have anti-leishmanial activity. CuO-Ag with F.szowitsiana at 96 μg/mL (72 h) had the highest effect (62%) with the lowest IC50 (2.4 μg/mL) compared to others. The highest IC50 was related to copper nanoparticles at 24 hours (150 μg/mL) and the lowest IC50 was related to copper oxide nanoparticles doped with silver with F.szowitsiana extract at 72 hours (2.4 μg/mL). Conclusion and Discussion: The results showed that different concentrations of nanoparticles had a major inhibitory effect on the growth of Leishmania. Silver-doped CuO nanoparticles with F.szowitsiana extract at higher concentrations on Leishmania major promastigotes in vitro after 72 hours showed a better anti-leishmanial effect than other nanoparticles. Combining nanoparticles with plant extracts can develop new effective treatments for leishmaniasis. Further studies are recommended to determine the effect of these nanoparticles on amastigote forms and in vivo experiments.

Keywords: Nanoparticle, Leishmania major, Copper Oxide, AgNo3, Ferula szowitsiana, In-Vitro