التخليق الحيوي لجسيمات أوكسيد الخارصين النانوية باستخدام مستخلص أوراق السدر: التشخيص ودراسة تأثيراته في ذكور الجرذان المعرضة للأدنين

Biosynthesis of Zinc Oxide Nanoparticles Ziziphus-spina christi Leaves Extract: Characterization and Their Effects in Male Rats Exposed to Adenine

Abstract

Zinc oxide nanoparticles (ZnO‐NPs) have been widely studied in recent decades due to their nano-medicine applications based on the physiological and biochemical agents often added as a food supplement in animal diets. This study aimed to synthesis and characterization of (ZnO‐NPs) by the biological method using the plant extract and then investigated its effects in adenine-exposed male rats. Biosynthesized ZnO‐NPs were synthesized using the crude aqueous extract of Ziziphus-spina christi leaves and zinc nitrate hexahydrate (Zn(NO3)2.6H2O). The reaction medium was maintained with a temperature of 60 ◦C and a pH of 9. The change of color from yellow to brown indicates the production of ZnO-NPs. The produced ZnO‐NPs were characterized using several techniques. Ultraviolet-visible (UV-Vis) spectrum showed the maximum peak at 362 nm, this was strong evidence for the forming of ZnO-NPs. Fourier Transform-Infra Red (FT-IR) spectrum revealed the appearance of absorption peaks between 400 and 500 cm-1 returned to Zn-O bonding. X-ray diffraction (XRD) analysis revealed that the hexagonal Wurtzite structure for nanoparticles with an average crystallite size was 38.177 nm. Transmission electron microscopy (TEM) showed that nanoparticles had a spherical shape and fell in the range of 35 – 45 nm in diameter, also the images of scanning electron microscope (SEM) showed the semispherical shape and these particles are in a highly agglomerated form with an average particle size was ranged at diameter: D1 (39.74 nm), D2 (42.25 nm), and D3 (48.19 nm). Finally, atomic force microscopy (AFM) displays a uniform surface with cone-like grains covering the surface of ZnO-NPs, and the surface roughness in the range of 42 to 45 nm. Thirty-six male rats were used in this study and divided randomly into six equal groups (six rats each). Group-I received orally dimethyl sulfoxide (5% v/v/day) alone for 30 days and served as the control group, group-II administrated adenine orally (100 mg/Kg. Bw/day) for 30 days, group III received Ziziphus-spina christi leaves extract (10 mg/Kg. Bw/day) for 30 days, group-IV was administrated ZnO‐NPs (10 mg/Kg. Bw/day) for 30 days, group-V was co-administrated adenine (100 mg/Kg. Bw/day) plus Ziziphus-spina christi leaves extract (10 mg/Kg. Bw/day) for 30 days and the last group-VI received the adenine combined with ZnO‐NPs in the same previous doses for 30 days. At the end of the experiment, the kidney function tests (uric acid, urea, and creatinine) and liver function tests (alanine aminotransferase ALT, aspartate aminotransferase AST and alkaline phosphatase ALP) were determined. Antioxidant status (superoxide dismutase SOD, catalase CAT, total antioxidant capacity TAC, and reduced glutathione GSH) and oxidative stress (Malondialdehyde MDA and nitric oxide NO) were performed. Furthermore, histopathological alterations of kidney and liver tissues were investigated. In vivo of this study, the synthesized ZnO-NPs and Ziziphus-spina christi leaves extract at a dose (10 mg/Kg. Bw) were exhibited the potential effects against adenine-exposed male rats, these confirmed by ameliorative growth rats and the significant decreases in kidney and liver function parameters, uric acid, urea, creatinine, ALT, AST, and ALP in comparison to adenine exposed rats (P ≤ 0.05). Significant elevation was also demonstrated in antioxidants levels SOD, CAT, TAC, and GSH associated with depletion in oxidative stress MDA and NO in both treatment groups fifth and sixth when compared to adenine group rats. Moreover, there were improvements in the marked histological changes of the kidney and liver by comparing with the adenine group. In conclusion, Ziziphus-spina christi leaves extract and ZnO-NPs have ameliorative effects against adenine-exposed male rats, implying that they may be used safely against kidney and liver damage, and no significant effects were observed in normal renal and hepatocyte tissues, indicating that they may be powerful antioxidant, anti-inflammatory, and antitoxic agents for biomedical applications.