• Mohamed A. Hashem Clinical Pathology Department, Faculty of Veterinary Medicine, 44511, Zagazig University, Zagazig, Sharkia Governorate, Egypt, Corresponding author, E-mail: mhashem.vet@gmail.com
  • Asmaa Z. M. Zidan Belkas Veterinary Administration, 35631, Dakahlia Governorate, Egypt
  • Shefaa Ali El-Mandrawy Clinical Pathology Department, Faculty of Veterinary Medicine, 44511, Zagazig University, Zagazig, Sharkia Governorate, Egypt




AgNPs, cisplatin, hepatocellular carcinoma, hematology, biochemical, TNFα, IL6


Silver nanoparticles (AgNPs) have various applications such as their use in the medical field. As they have been reported to show antimicrobial and anti-tumor effects. The main purpose of the current study was to explore the anti-cancer effects of AgNPs administration alone or in a combination with cisplatin (CP) against hepatocellular carcinoma (HCC) in rats. Seventy-five rats (Sprague Dawley albino rats) were used in the present study. Rats were assigned to 5 groups. Group 1 served as normal control. Group 2 was injected intraperitoneally (IP) with a single dose (200 ml/kg) of diethylnitrosamine (DEN), then one week later carbon tetrachloride (CCl4) was injected IP (0.2 ml/kg) two times weekly for 14 successive weeks for the induction of HCC. Group 3 was treated with AgNPs (4 mg/kg) daily. Group 4 was treated with CP (6 mg/kg) once a week. Group 5 was treated with a combination of AgNPs (4 mg/kg/daily) and CP (2.5 mg/kg/weekly). Groups 3, 4, and 5 were treated for 3 successive weeks after induction of HCC. Hematological, biochemical, antioxidant activities, proinflammatory cytokines, and apoptotic genes were evaluated in the current study. Hematological results denoted normocytic normochromic anemia in all examined groups except group 2 which showed macrocytic hypochromic anemia, with thrombocytopenia and leukocytosis in all groups. Significant hypoproteinemia, hypoalbuminemia, and hypoglobulinemia were detected in all groups, with no significant changes (P < 0.05) in globulin in group 5. Significantly decreases in GSH and SOD were recorded in all groups. While the serum AST, ALT activities, and levels of total bilirubin, urea, creatinine, IL-6, TNF-α, AFP, VEGF, BAX, and caspase-3 were markedly elevated. The results revealed a remarkable improvement in group 4  than groups 3, and 5. The obtained results were supported by immunohistochemical and histopathological investigations of the liver tissue.


● 1. Jeevanandam J, Barhoum A, Chan YS, et al. Review on nanoparticles and nanostructured materials: history, sources, toxicity, and regulations. Beilstein J Nanotechnol 2018; 9: 1050–74.

● 2. Colvin VL, Schlamp MC, Alivisatos A. Light-emitting diodes made fromcadmium selenide nanocrystals and a semiconducting polymer. Nature 1994; 370:354–7.

● 3. Zhang X-F, Liu Z-G, et al. Silver nanoparticles: synthesis, characterization, properties, applications, and therapeutic approaches. Inter J Mol Sci 2016; 17: 1534.

● 4. Vlăsceanu GM, Marin Ş, Ţiplea RE, et al. Silver nanoparticles in cancer therapy. Nanobiomat Cancer Ther 2016; 29–56.

● 5. El-Serag HB. Hepatocellular carcinoma. New England J Med 2011; 365: 1118–27.

● 6. Papatheodoridis GV, Lampertico P, Manolakopoulos S, et al. Incidence of hepatocellular carcinoma in chronic hepatitis B patients receiving nucleos(t)ide therapy: a systematic review. J Hepatol 2010; 53: 348–56.

● 7. Roncalli M, Terracciano L, Di Tommaso L, et al. Liver precancerous lesions and hepatocellular carcinoma: the histology report. Digest Liver Dis 2011; 43: S361–72.

● 8. Emadi A, Jones RJ, Brodsky RA. Cyclophosphamide and cancer: golden anniversary. Nat Rev Clin Oncol 2009; 6: 63.

● 9. Nelius T, Klatte T, de Riese W, et al. Clinical outcome of patients with docetaxel-resistant hormone-refractory prostate cancer treated with second-line cyclophosphamide-based metronomic chemotherapy. Med Oncol 2010; 27: 363-–7.

● 10. Herrera-Pérez Z, Gretz N, Dweep HA Comprehensive Review on the Genetic Regulation of Cisplatin-induced Nephrotoxicity. Curr Genomics 2016; 17: 279–93.

● 11. Miller RP, Tadagavadi RK, Ramesh G, et al. Mechanisms of Cisplatin nephrotoxicity. Toxins (Basel) 2010; 2: 2490–518.

● 12. Abou Zaid OAR, El- Senousy YA, Badawy Abdel Fattah M, et al. Biochemical studies on evaluation of new composite against hepatocellularcarcinoma-induced in rats: recent therapeutic approaches. Benha Vet Med J 2016; 1: 124–36.

● 13. Shaloam D, Paul BT. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 2014; 364–78.

● 14. Motawi TK, El-Boghdady NA, El-Sayed AM, et al. Comparative study of the effects of PEGylated interferon-α2a versus 5-fluorouracil on cancer stem cells in a rat model of hepatocellular carcinoma. Tumour Biol 2016; 37: 1617–25.

● 15. Singh D, Singh M, Yadav E, et al. Attenuation of diethylnitrosamine (DEN) - Induced hepatic cancer in experimental model of Wistar rats by Carissa carandas embedded silver nanoparticles. Biomed Pharmacother 2018; 108: 757–65.

● 16. Uehara T, Pogribny IP, Rusyn I. The DEN and CCl4 -induced mouse model of fibrosis and inflammation-associated hepatocellular carcinoma. Curr Protoc 2021;1 (8): e211.

● 17. Guoqiang Z, Wenying W. Synthesis of silver nanoparticles and their antiproliferation against human lung cancer cells in vitro. Orient J Chem 2012; 28: 651–5.

● 18. Tiwari DK, Jin T, Behari J. Dose-dependent in vivo toxicity assessment of silver nanoparticle in Wistar rats. Toxicol Mech Methods 2011; 21: 13–24.

● 19. Zayed MN, Aly HF, Moneim El-Mezayen HA, et al. Effect of co-administration of Bee honey and some chemotherapeutic drugs on the dissemination of hepatocellular carcinoma in rats. Toxicol Rep 2019; 6: 875–88.

● 20. Parasuraman S, Raveendran R, Kesavan R. Blood sample collection in small laboratory animals. J Pharmacol Pharmacother 2010; 1: 87–9.

● 21. Beutler E, Duron O, Kelly MBJ. Lab Clin Med 1963; 61: 882

● 22. Nishikimi M, Roa NA, Yogi K. Biochem Bioph Res Common 1972; 46: 849–54.

● 23. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) Method. Methods 2001; 25: 402–8.

● 24. Suvarna KS, Layton C, Bancroft JD. Bancroft’s theory and practice of histological techniques e-book. Elsevier Health Sciences; Amsterdam, The Netherlands: 2019.

● 25. Pedrycz A, Czerny K. Immunohistochemical study of proteins linked to apoptosis in rat fetal kidney cells following prepregnancy adriamycin administration in the mother. Acta Histochem 2008; 110: 519–23.

● 26. Asif IHR. Haematological Profile In. Hepatocellular Carcinoma before and after Chemotherapy. ACTA Pharmaceutica Turcica 1999; 4: 149–52.

● 27. Culling CF, Allison RT, Barr WT. cellular pathology technique. 4th ed. New York: Butterworth. 642. 1985.

● 28. Hauck S, Kapczinski F, Roesler R, et al. Serum brain-derived neurotrophic factor in patients with trauma psychopathology. Prog Neuropsychopharmacol Biol Psychiatry 2010; 34 (3): 459–62.

● 29. Rajendoran V, Krishnaswamy K. Effect of Solanum villosum extract and its silver nanoparticles on the hematopoietic system of diethylnitrosamine-induced hepatocellular carcinoma in rats. Inno J of Health Sci 2016; 5: 13–16.

● 30. Reddy MV. Statistical methods in psychiatry research and SPSS. Oakville: Apple Academic Press; 2019.

● 31. Hassan I, Chibber S, Naseem I. Ameliorative effect of riboflavin on the cisplatin-induced nephrotoxicity and hepatotoxicity. Food Chem Toxicol 2010; 48: 2052–8.

● 32. Marković SD, Djačić DS, Cvetković DM, et al. Effects of acute in vivo cisplatin and selenium treatment on hematological and oxidative stress parameters in red blood cells of rats. Biol Trace Elem Res 2011; 142: 660-70.

● 33. Hassen MT, Ahmed NJ, Mohamed HK, Therapeutic effect of silver nanoparticles against diethyl nitrosamin and carbon tettrachloride-induced hepatocellular carcinoma in rats. Inter J Pharm Pharmaceut Sci 2020; 1–8.

● 34. Adeyemi OS, Akanji MA. Psidium guajava leaf extract: effects on rat serum homeostasis and tissue morphology. Comp Clin Pathol 2012; 21: 401–7.

● 35. Abou Zaid OAR, El-Senousy YA, Badawy AFM, et al. Biochemical studies on evaluation of new composite against hepatocellular carcinoma-induced in rats: recent therapeutic approaches. Benha Vet Med J 2016; 30: 124–36.

● 36. Singh D, Chaudhary D, Kumar V, Verma A. Amelioration of diethylnitrosamine (DEN) induced renal oxidative stress and inflammation by Carissa carandas embedded silver nanoparticles in rodents. Toxicol Rep 2021; 8: 636–45

● 37. Hassan HA, Edrees GM, El-Gamel EM, et al. Amelioration of cisplatin-induced nephrotoxicity by grape seed extract and fish oil is mediated by lowering oxidative stress and DNA damage. Cytotechnology 2014; 66: 419–29.

● 38. Jiang M, Dong Z. Regulation and pathological role of p53 in cisplatin nephrotoxicity. J Pharmacol Exp Ther 2008; 327: 300-7.

● 39. Abeer MB. The possible protective effect of ambroxol on cisplatin-induced hepato and nephrotoxicity in rat, Faculty of Pharmacy, Cairo University, Master thesis 2019.

● 40. Medhat A, Mansour S, El-Sonbaty S, et al. Evaluation of the antitumor activity of platinum nanoparticles in the treatment of hepatocellular carcinoma induced in rats. Tumour Biol 2017; 39 (7): 1010428317717259.

● 41. Kadasa NM, Abdallah H, Afifi M, Gowayed S. Hepatoprotective effects of curcumin against diethyl nitrosamine induced hepatotoxicity in albino rats. Asian Pac J Cancer Prev 2015; 16: 103–8.

● 42. Yuan YG, Zhang S, Hwang JY, et al. Silver nanoparticles potentiates cytotoxicity and apoptotic potential of camptothecin in human cervical cancer cells. Oxid Med Cell Longev 2018; 2018: 6121328

● 43. Arora S, Jain J, Rajwade JM, Paknikar KM. Cellular responses induced by silver nanoparticles: In vitro studies. Toxicol Lett 2008; 179 (2): 93–100.

● 44. Farkas J, Christian P, Gallego-Urrea JA, et al. Uptake and effects of manufactured silver nanoparticles in rainbow trout (Oncorhynchus mykiss) gill cells. Aquat Toxicol 2011;101: 117-2.

● 45. Jayaprakash R, Ramesh V, Sridhar MP, et al. Antioxidant activity of ethanolic extract of Tinospora cordifolia on N-nitrosodiethylamine (diethylnitrosamine) induced liver cancer in male Wister albino rats. J Pharm Bioallied Sci 2015; 7: S40–S45.

● 46. Hassan HA, Serag HM, Abdel-Hamid NM, et al. Synergistic curative effect of chicory extract and cisplatin against thioacetamide-induced hepatocellular carcinoma. Hepatoma Res 2015; 1: 147–54.

● 47. Aggarwal V, Tuli HS, Varol A, et al. Role of reactive oxygen species in cancer progression: molecular mechanisms and recent advancements. Biomolecules 2019; 9: 735.

● 48. Hübner G, Brauchle M, Smola H, et al. Differential regulation of pro-inflammatory cytokines during wound healing in normal and glucocorticoid-treated mice. Cytokine 1996; https://doi.org/10.1006/cyto.1996.0074.

● 49. Fehaid A, Fujii R, Sato T, et al. Silver nanoparticles affect the inflammatory response in a lung epithelial cell line. Open Biotechnol J 2020; 14 (1): 113–23.

● 50. Zhang B, Ramesh G, Uematsu S, et al. TLR4 signaling mediates inflammation and tissue injury in nephrotoxicity. J Amer Soc Nephrol 2008; 19 (5): 923–32.

● 51. Shin SH, Ye MK, Kim HS, et al. The effects of nano-silver on the proliferation and cytokine expression by peripheral blood mononuclear cells. Inter Immunopharmacol 2007; 7: 1813–18.

● 52. Abdel-Hamid NM, Nazmy MH, Nazmy WH. Vitamin C and diallyl sulfide as chemosensitizers to cisplatin in treating hepatocellular carcinoma. J Solid Tumors 2011; 90–102.

● 53. Zhang X, Lu Y, Rong C, et al. Role of superoxide dismutase in hepatitis B virus-related hepatocellular carcinoma. J Res Med Sci 2016; 21: 94.

● 54. Taha H, Elfar N, Haffez H, et al. Raptinal silver nanoparticles: new therapeutic advances in hepatocellular carcinoma mouse model. Naunyn Schmiedebergs Arch Pharmacol 2021; 394: 279–89.

● 55. Matsui D, Nagai H, Mukozu T, et al. VEGF in patients with advanced hepatocellular carcinoma receiving intra-arterial chemotherapy. Anticancer Res 2015; 35: 2205–10.

● 56. Kemp MM, Kumar A, Mousa S, et al. Gold and silver nanoparticles conjugated with heparin derivative possess anti-angiogenesis properties. Nanotechnol 2009; 20: 455104.

● 57. Li Z, Liu H, Lang J, et al. Effects of cisplatin on surgically induced endometriosis in a rat model. Oncol Lett 2018; 16: 5282–90.




How to Cite




Veterinary Medicine and The One Health Concept