الفهرس 
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Abstract
The present study was conducted to compare the in vitro therapeutic efficacy and in vivo side effects of gemcitabine loaded silver nanoparticles with gemcitabine alone. Synthesis and characterization of AgNPs: AgNPs were characterized by, dynamic light scattering, transmission electron microscopy (TEM), the zeta potential measurement, the Fourier transform infrared spectroscopy. Drug loading: The GEM-AgNPs were subsequently characterized by TEM and FTIR, and their zeta size was measured.
In vitro experiments: Cell culture:. The cells were cultured as a monolayer in T-25 flasks and seeded to attain 70% confluency before seeding, propagation, or treatment. Cell viability assay: At the end of the incubation period, dark purple formazan crystals were produced and solubilized with isopropanol. Optical density (OD) was detected at 570 nm using a BioTek ELx800 microplate reader. The experiment was replicated three times, and the concentration required for 50% inhibition of viability (IC50) was determined. Annexin V-FITC and propidium iodide (PI) staining for apoptosis assay: The cells were incubated with 5 μL Annexin V-FITC for 10 min and 5 μL PI for 5 min, respectively, in the dark at room temperature, after which Annexin V-FITC Apoptosis Detection Kit (BioVision, USA) was used per the manufacturer’s protocol. Cell cycle analysis: Cell cycle perturbations induced by GEM-AgNPs (100 μg/mL) were evaluated by performing PI staining of DNA using Propidium Iodide Flow Cytometry Kit (ab139418) per the manufacturer’s protocol.
The results: 1-Characterization of AgNPs and GEM-AgNPs According to the FTIR analysis, the following could be suggested: GEM molecules were successfully attached to AgNPs surfaces, and O=H and N-H groups of GEM are the active sites for the attachment to AgNPs.
2-In vitro study: On the basis of this information, the current research used GEM-AgNPs (100 μg/mL) for subsequent tests.
Effect of GEM-AgNPs on apoptosis in HepG2: Cell cycle analysis: Cell flow cytometry was performed to evaluate the effect of GEM-AgNPs on cell cycle progression.
3-In vivo study: The number of surviving animals at the completion of the study was as follows: control group, 10/10; GEM group, 8/10; AgNPs group, 10/10; and GEM-AgNP group, 9/10.
I- Effect of AgNPs and/or Gemcitabine on some hematological studies in adult rats: Decrease in RBC count, Hb concentration, PCV, total WBC count.
II- Effect of AgNPs and/or Gemcitabine on the hepatic and kidney oxidative/ antioxidant status in adult rats: Our results revealed that i.p GEM injection in adult male rats caused a marked reduction in CAT, GSH, and GSH-Px levels, as well as a marked elevation in MDA, NO, and MPO concentrations in hepatic and renal tissues.
III-- Effect of AgNPs and/or Gemcitabine on some hepatic and kidney functions of adult rats: Data recorded in this research confirmed renal dysfunction in GEM-treated rats, which evidenced by a marked elevation in urea creatinine, and uric acid levels compared with those in the normal rats. Marked elevation in serum ALP, ALT, AST, total bilirubin, uric acid, urea, creatinine levels recorded in rats injected with GEM (134 mg/kg) i.p..
IV- Histopathological findings: The effects in Livers of AgNPs on GEM-induced hepatopathy in the GEM-AgNP-treated animals were average with a significant reduction in the severity but not the frequency of steatosis, coagulative necrosis, hemorrhages, edema, and inflammatory cell infiltration and a non-significant reduction in the other GEM-induced hepatic lesions. Kidneys of the AgNP-treated rats exhibited normal histological characteristics except.