Objectives: Nanoparticles (NPs) are currently used in medical felid such as many diseases diagnosis and treatment, including cancer and autoimmune diseases. A. squamosa L. (family: Annonaceae) is a medicinal plant commonly known as the sugar apple, sweetsop, or sugar-pineapple, Annonaceae family comprising approximately 135 genera and 2300 species. This investigation the side effects of Nano extract of plant seeds by using different tests in vitro by using some normal cell lines and in vivo by using different biomarkers, tests and parameters as well as by the detection of histological changes by using experimental animals. Materials and Methods: The present study has been conducted in the animal house of the of the College of Education for Girls/University of Kufa, during the period extended from Feb, 2023 to June, 100 adult male Swiss albino mice Mus Musculus, weighing 20–30 g, of 8 to 12 weeks of age, will be used in the study. Animals have been divided into ten groups; Group (1) mice will be injected with normal saline 0.5 ml (as control) for two months (once weekly). Group (2) mice male inducing cancer only. Groups (3-4-5-6) mice will be injected with well dispersed Nano extract of plant seeds (40-80-120- 200mg/kg) after that inducing cancer for two months (once weekly). Groups (7-8-9-10) mice will be injected with well dispersed Nano extract of plant seeds (40-80- 120-200mg/kg) for two months (once weekly). After the end of second month of experiment, the mice will all be sacrificed for detection of Nano extract of plant seeds effects on the Hematologic parameters in mice. Males were anesthetized (by injection of 0.3 ml ketamine + 0.1 ml of xylazine/ kg b.w. ip), dissected, and blood samples were taken from the abdominal vein in non-heparinized tubes at the conclusion of each treatment and control. Each rat has a liver and kidney removed for histological analysis. Results: The results of the current study showed that the methanolic extract was obtained in an amount of (6 gm). The result of dissolving (215 gm) of A. squamosa seeds powder in methanol. After adding zinc nitrate Zn(NO₃)₂, there was a change in the color of the seed extract A. squamosa from dark to light in color, indicating that zinc oxide particles Ultrafine ZnO NPs have been formed. FESEM test is used to confirm the shape and size of nanoparticles which shows the results of the scanning electron microscope (SEM) of zinc oxide particles. The composition of ZnO NPs was measured by energy dispersive X-ray spectroscopy analysis (EDS) by determining the presence of elements on the surface of the samples and observing the optical absorption peaks for ZnO NPs. Gave an AFM analysis of the nanoparticles that were synthesized from the seeds of the sour cream plant Information about the shape of the nanoparticles. The XRD study confirms the Wurtzite structure of ZnO nanoparticles and peak formation The narrow Bragg angle indicates the spherical nature of ZnO nanoparticles. At the corner of Bragg, the face-centered, spherical nature of ZnO nanoparticles indicates. Nanoparticle stabilization occurs by some bridging agent which is emphasized by the sharp peaks. An X-ray diffraction pattern is observed ZnONP and no other phase was observed. The FTIR spectra of ZnO nanoparticles, FTIR spectroscopy estimates infrared radiation by the sample plotted against a specific wavelength. The results of the current study showed that there was a significant increase in the body weight of the mice used in the current study before and after the experiment, except for the second group, in which there was a decrease in body weight. The results of the current study also showed that there was no significant difference increase at (P≤0.05) in live weight. The results of the current study also showed that there was a significant difference increase at (P≤0.05) in kidney weight. The results showed a significant difference at (P≤0.05) in concentration of liver enzymes (AST, ALT and ALP) between experimental groups. The results showed a significant difference at (P≤0.05) in concentration of Blood Urea between experimental groups, also the results showed a significant difference at (P≤0.05) in concentration of Creatinine between experimental groups, The results showed a significant difference at (P≤0.05) in concentration of Uric acid between experimental groups, The results showed that there was a significant difference increase at (P≤0.05) in concentration of total protein. The results showed a significant difference at (P≤0.05) in concentration of Potassium between experimental groups, The results showed no significant differences at (P≤0.05) in concentration of Sodium and Calcium between experimental groups, The results showed that there was a significant difference increase at (P≤0.05) in concentration of Chlorine. And in histological changes in liver and kidney its effect by induced groups cancer and other groups of experimental. Conclusions: We conclude from the results of our current study that nano extract of A. squamosa seeds has a clear effect on the studied parameters, especially the liver and kidneys, in the various doses given and according to the time period of the study.
