Chitosan nanoparticles from Artemia salina inhibit progression of hepatocellular carcinoma in vitro and in vivo

  • Mai M. Elkeiy
  • Abeer A. Khamis
  • Mona M. El-Gamal
  • Maha M. Abo Gazia
  • Zeinb A. Zalat
  • Mohammed A. El-MagdEmail author
Nanotechnology, Nanopollution, Nanotoxicology and Nanomedicine (NNNN)


This study was conducted to evaluate the effect of chitosan nanoparticles (CNPs) isolated from Artemia salina against hepatocellular carcinoma (HCC) both in vitro (HepG2) and in vivo (diethylnitrosamine-induced HCC in rats) and to investigate the involved underlying mechanisms. Administration of CNPs decreased HCC progression as evidenced by (1) induced HepG2 cell death as detected by MTT assay; (2) induced necrosis as indicated by acridine orange/propidium iodide (AO/PI) red staining, annexin V/7-AAD positive staining (detected by flow cytometry), and upregulated expression of necrosis markers (PARP1 and its downstream target, RIP1 genes), but no effect on apoptosis as revealed by insignificant changes in caspase 3 activity and mRNA levels of Bax and AIF; (3) increased intracellular ROS and decreased mitochondrial membrane potential in HepG2; (4) decreased liver relative weight, serum levels of liver enzymes (ALT, AST, and ALP), total bilirubin, and cancer markers (AFP and GGT), number and area of GST-P positive tumor nodules; and (5) reduced oxidative stress (decrease in MDA levels) and increased activities of SOD, CAT, and GPx enzymes in rat liver. The preventive (pre-treatment) effect of CNPs was better than the therapeutic (post-treatment) effect. Collectively, administration of CNPs inhibited HCC progression in vitro and in vivo, possibly through induction of necrosis, rather than apoptosis, and induction of antioxidant enzyme activities in vivo, but with stimulation of ROS production in vitro. Thus, CNPs could be used as a promise agent for treating HCC after application of further confirmatory clinical trials.


Chitosan nanoparticles Artemia salina Hepatocellular carcinoma Necrosis Oxidative stress 



Apoptosis-inducing factor


Alkaline phosphatase


Alanine transaminase


Analysis of variance


Aspartate transaminase




Diethyl nitosoamine


γ-Glutamyl transferase


Glutathione peroxidase


Hepatocellular carcinoma;


Mitochondrial permeability transition


Mitochondrial membrane potential


Poly(ADP-ribose) polymerase 1


Serine/threonine kinases receptor interacting protein 1


Superoxide dismutase


Tukey’s honestly significant difference


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biochemistry Division, Chemistry Department, Faculty of ScienceTanta UniversityTantaEgypt
  2. 2.Zoology Department, Faculty of ScienceTanta UniversityTantaEgypt
  3. 3.Department of Histology, Faculty of MedicineKafrelsheikh UniversityKafr El SheikhEgypt
  4. 4.Clinical Pharmacy Department, Faculty of PharmacyAlazhar UniversityCairoEgypt
  5. 5.Anatomy Department, Faculty Veterinary MedicineKafrelsheikh UniversityKafr El SheikhEgypt

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