The High Potency of Green Synthesized Copper Nanoparticles to Prevent the Toxoplasma gondii Infection in Mice.

PURPOSE: Nowadays, due to the lack of an effective vaccine to prevent the toxoplasmosis, chemotherapy with the combination of pyrimethamine and sulfadiazine is considered as the "gold standard" treatment for toxoplasmosis. Recent reports have exhibited that these synthesized chemical drugs are associated with some serious side effects. The present study aims to evaluate the prophylactic effects of copper nanoparticles (CuNPs) green synthesized by Capparis spinosa fruit methanolic extract alone and combined with atovaquone against chronic toxoplasmosis induced by the Tehran strain of Toxoplasma gondii in mice METHODS: Mice were then orally administrated with CuNPs at the doses of 2 and 4 mg/kg/day and in combined with atovaquone 50 mg/kg for 14 days. Male BALB/c mice were divided into two seven groups include C1 (non-treated non-infected); C2 (treated with normal saline); C3 (Infected mice treated with atovaquone 100 mg/kg/day); Ex1 (treated with CuNPs 2 mg/kg/day); Ex2 (treated with CuNPs 4 mg/kg/day); Ex3 (treated with CuNPs 2 mg/kg/day + atovaquone 50 mg/kg/day); Ex3 (treated with CuNPs 4 mg/kg/day + atovaquone 50 mg/kg/day). On the 15th day, the mice were infected with the intraperitoneal inoculation of 20-25 tissue cysts from the Tehran strain of T. gondii. The mean numbers of brain tissue cysts and the mRNA levels of IL-12, IFN-γ, and inducible nitric oxide synthase (iNOS) in mice of each tested group were measured. RESULTS: CuNPs were green synthesized by C. spinosa methanolic extract. Scanning electron microscopy showed that the particle size of CuNPs was 17 and 41 nm with maximum peak at the wavelength of 414 nm. The mean number of T. gondii tissue cysts in mice of tested groups of Ex1, Ex2, Ex3, and Ex4, significantly decreased as a dose-dependent response compared with control group. Moreover, in similar to the control group C3, no T. gondii tissue cysts was observed in mice of experimental group Ex3 and Ex4. The mRNA levels of IFN-γ, IL-12, and iNO was measured in mice of all tested groups. The mRNA levels of IFN-γ, IL-12, and iNO was increased in all mice of experimental groups in comparison with the control group C2; however, a significant enhancement was detected in mRNA level of IFN-γ, IL-12, and iNO in the tested groups of Ex3 and Ex4 when compared with control group C3. CONCLUSION: The obtained results revealed the high potency of CuNPs alone and combined with atovaquone to prevent toxoplasmosis in mice. Although, the prophylactic effects of CuNPs and other properties, such as improved cellular immunity and low toxicity, are positive topics; however, more studies are required to approve these findings especially in clinical settings.

Cytotoxic and Antileishmanial Effects of Various Extracts of Capparis spinosa L.

Objectives: Cutaneous leishmania sis (CL) is considered as one of the most critical infections worldwide, in which the protozoa of the genus Leishmania infects a person. Today, the common and selective drugs for the treatment of CL are antimonial compounds present some limitations to their usage. The objective of this study is to investigate the cytotoxic and antileishmanial effects of various extracts of Capparis spinosa L. on the in vitro model. Materials and Methods: The primary phytochemical analysis of the C. spinosa extracts was performed to assess the presence of tannins, alkaloids, saponins, flavonoids, terpenoids, and glycosides. Furthermore, the in vitro cytotoxic and antileishmanial effects of C. spinosa extracts on Leishmania tropica promastigote were evaluated. Additionally, these effects on the J774-A1 macrophage cells by colorimetric cell viability 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide assay were also assessed. Results: In this study, the findings of primary phytochemical screening of the C. spinosa extracts demonstrated the existence of flavonoids, tannins, terpenoids, glycosides, and alkaloids in this plant. Importantly, the findings indicated that the aqueous and methanolic extracts of C. spinosa exhibit a high potency to inhibit the growth of L. tropica promastigotes with inhibitory concentration 50 values of aqueous and methanolic extracts being 28.5 and 44.6 µg/mL, respectively. Based on the obtained results, C. spinosa extracts did not display a considerable cytotoxicity on the J774-A1 macrophage cells. Conclusion: The obtained findings exhibited remarkable antileishmanial effects of C. spinosa extracts on L. tropica, thereby indicating the ability of C. spinosa as a herbal product to be developed as a new antileishmanial drug. Nevertheless, supplementary investigations will be obligatory to achieve these findings, especially in human subjects.

Therapeutic Potential of Green Synthesized Copper Nanoparticles Alone or Combined with Meglumine Antimoniate (Glucantime®) in Cutaneous Leishmaniasis.

BACKGROUND: In recent years, the focus on nanotechnological methods in medicine, especially in the treatment of microbial infections, has increased rapidly. AIM: The present study aims to evaluate in vitro and in vivo antileishmanial effects of copper nanoparticles (CuNPs) green synthesized by Capparis spinosa fruit extract alone and combined with meglumine antimoniate (MA). METHODS: CuNPs were green synthesized by C. spinosa methanolic extract. The in vitro antileishmanial activity of CuNPs (10-200 µg/mL) or MA alone (10-200 µg/mL), and various concentrations of MA (10-200 µg/mL) along with 20 µg/mL of CuNPs, was assessed against the Leishmania major (MRHO/IR/75/ER) amastigote forms and, then tested on cutaneous leishmaniasis induced in male BALB/c mice by L. major. Moreover, infectivity rate, nitric oxide (NO) production, and cytotoxic effects of CuNPs on J774-A1 cells were evaluated. RESULTS: Scanning electron microscopy showed that the particle size of CuNPs was 17 to 41 nm. The results demonstrated that CuNPs, especially combined with MA, significantly (p < 0.001) inhibited the growth rate of L. major amastigotes and triggered the production of NO (p < 0.05) in a dose-dependent manner. CuNPs also had no significant cytotoxicity in J774 cells. The mean number of parasites was significantly (p < 0.05) reduced in the infected mice treated with CuNPs, especially combined with MA in a dose-dependent response. The mean diameter of the lesions decreased by 43 and 58 mm after the treatment with concentrations of 100 and 200 mg/mL of CuNPs, respectively. CONCLUSION: The findings of the present study demonstrated the high potency and synergistic effect of CuNPs alone and combined with MA in inhibiting the growth of amastigote forms of L. major, as well as recovery and improving cutaneous leishmaniasis (CL) induced by L. major in BALB/c mice. Additionally, supplementary studies, especially in clinical settings, are required.

Green Synthesis and Characterization of Copper Nanoparticles and Their Effects on Liver Function and Hematological Parameters in Mice.

OBJECTIVES: The present investigation is based on the green synthesis of copper nanoparticles (CuNPs) from aqueous extract of Capparis spinosa L. fruit. Their effects on liver function and hematological parameters in mice were evaluated. MATERIALS AND METHODS: The green synthesis of CuNPs by means of C. spinosa extract was achieved. Ultraviolet-visible spectroscopy, fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy were used to identify the synthesized nanoparticles. BALB/c mice were orally administrated CuNPs at doses of 1000, 2000, and 5000 µg/kg for 2 weeks. Later, the effects of CuNPs on liver function in the treated mice were evaluated by measuring the serum levels of enzymes such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and bilirubin as well as hematological parameters including hemoglobin, hematocrit, white blood cell, red blood cell, and platelet counts. RESULTS: A maximum peak at wavelength 414 nm confirmed the biosynthesis of CuNPs. FTIR spectrum analysis revealed that the factor groups shaped a coating extract on the surface of the nanoparticles. SEM images demonstrated a particle size between 17 and 41 nm. Although some liver enzymes and hematological parameters increased with increasing dose of extract, there was no significant difference (p>0.05) between oral administrations of CuNPs at doses of 1000, 2000, and 5000 µg/kg and the control group. CONCLUSION: The findings revealed that CuNPs biosynthesized from aqueous extract of C. spinosa fruit have no toxic effects on the liver functions and hematological parameters of mice. However, more studies are needed for evaluation of the hepatoprotective effects of CuNPs.

Antinociceptive effects of green synthesized copper nanoparticles alone or in combination with morphine.

OBJECTIVE: The aim of this study was to evaluate the antinociceptive effect of biosynthetic copper nanoparticles from aqueous extract of Capparis spinosa fruit. METHODS: In this study, green synthesis of copper nanoparticles (CuNPs) was performed using C. spinosa extract according to the method described previously. The synthesized CuNPs were characterized using the UV-vis spectroscopy, Fourier transforms of infrared (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray (EDX). The antinociceptive effect of CuNPs was evaluated by tail-flick, hot-plate, and rotarod tests following the oral administration of mice with CuNPs at the concentrations of 25, 50, and 75 mg/kg for two weeks. RESULTS: The obtained maximum peak at the wavelength of 414 nm demonstrated the biosynthesis of the copper nanoparticles. SEM approved the particle size of CuNPs between 17 and 41 nm. The statistical analyses of the data of hot plate and tail-flick tests showed the potent analgesic effect of biosynthetic CuNPs. In this regard, the antinociceptive effect of at the doses of 75 mg/kg and 25 mg/kg plus morphine was significantly higher in comparison with the control group receiving morphine alone (P < 0.05). No significant (p > 0.05) difference was observed after the administration of CuNPs at the doses of 25, 50, and 75 mg/kg in the sensory-motor test. CONCLUSION: The present investigation demonstrated the analgesic effects of CuNPs especially in combination with morphine. These findings can provide a new strategy for producing new antinociceptive medications in the future.