Chitosan and Grifola Frondosa nanoparticles insulate liver dysfunction in EAC-bearing mice

Aliaa M Radwan; Doaa T Gebreel; Sahar Allam; Afaf El-Atrash; Ehab Tousson

doi:10.1093/toxres/tfae050

. 2024 Mar 28;13(2):tfae050. doi: 10.1093/toxres/tfae050

Chitosan and Grifola Frondosa nanoparticles insulate liver dysfunction in EAC-bearing mice

Aliaa M Radwan ^1,^✉, Doaa T Gebreel ², Sahar Allam ³, Afaf El-Atrash ⁴, Ehab Tousson ⁵

PMCID: PMC10980792 PMID: 38559757

Abstract

Background

Ehrlich ascites carcinoma (EAC) is a rapidly growing and undifferentiated tumor that can prompt oxidative stress and liver toxicity, whereas chitosan and Grifola Frondosa have widely recognized biological qualities. Therefore, our study designed to assess the potential ameliorative ability of chitosan nanoparticles (CS NPs) and Grifola Frondosa nanoparticles (GF-loaded casein NPs) on EAC-induced hepatic injury in mice.

Methods

A total of 60 female albino mice were segregated into 6 groups (10 mice each), G1, control group; G2, CS NPs group; G3, GF-loaded casein NPs group; G4, EAC group; G5, EAC treated with CS NPs; G6, EAC treated with GF-loaded casein NPs.

Results

According to the findings, EAC considerably increased serum activities of ALT, AST, ALP as well as LDL, cholesterol, and triglycerides levels coincided with marked decrease in albumin and total protein content in liver tissue. At the same time, it drastically lowered GSH levels and catalase activity while significantly elevating MDA levels. In addition, EAC caused DNA damage and apoptosis by decreasing Bcl-2 while increasing p53 expressions. However, either CS NPs or GF-loaded casein NPs therapy improved liver architecture and functioning, increased antioxidant parameters, and prevented hepatocyte death in EAC mice.

Conclusions

Our findings concluded that CS NPs and GF-loaded casein NPs have insulating functions against EAC-induced hepatic damage in mice.

Keywords: Ehrlich ascites carcinoma, chitosan NPs, Grifola Frondosa NPs, apoptosis, DNA damage

Introduction

Cancer is a prevalent and serious health concern that threatens both industrialized and developing countries' daily lives as a consequence of external as well as internal influences.¹ Among women, breast cancer is the most common type and the second prominent cause of death. Ehrlich ascites carcinoma (EAC) is a rapidly proliferating undifferentiated mammary adenocarcinoma with a short prognosis and 100% malignancy.²^,³ EAC is also highly transplantable through consecutive intraperitoneal (i/p) passages from one animal to another.⁴ EAC is a tumor model that mimics human carcinogenesis and is extremely susceptible to chemotherapy, making it broadly utilized in cancer and chemotherapeutic research.⁵ However, it has been reported that EAC causes severe degeneration and pathological lesions in the liver. The mechanisms of such hepatotoxicity were discovered to be caused by the building up of haemorrhagic ascitic fluid inside the peritoneal cavity, where cells multiply and migrate to invade the internal organs.⁶ These tumor cells can disturb the antioxidant machinery in the liver tissues of tumor host, resulting in a lack of cellular redox hemostasis.⁷ Furthermore, they induce the flux of oxy-radicals and hydrogen peroxide generation, causing oxidative harm by attacking the cellular DNA strands and prompting site-specific mutation, culminating in cytotoxicity.^8–10

Chitosan is a naturally occurring polymer generated from chitin that can be present in several forms of seafood, including shrimp and crayfish, with a high concentration in their shell.¹¹ Chitosan’s nontoxicity, biocompatibility, and biodegradability properties have led to a wide range of uses and increased interest.¹² Chitosan exhibits a variety of biological qualities, particularly antibacterial, anticancer, and hemostatic effects.¹³ Furthermore, chitosan’s positive charge, which allows for non-covalent binding with biological tissues, has been employed in medication administration.¹⁴ Chitosan may be effective in cancer treatment because it inhibits tumor growth, angiogenesis, and metastasis.¹¹^,¹⁵ Chitosan-based nanoparticles have chitosan properties as well as nanoparticle qualities such as surface and interface effects, small size, and quantum size effects.¹⁶ The unique size-dependent physicochemical characteristics of nanoparticles frequently encourage their use in various products; nevertheless, these same unique properties also contribute to unique physiological reactions in biological systems when these materials interact with them. Because nanoparticles can move more easily than bulkier molecules, they can be more hazardous than larger particles.¹⁷ Numerous investigations have found that chitosan and chitosan NPs are effective anticancer, antioxidant, anti-inflammatory, and hepatoprotective agents.^18–21

Anti-inflammatory, antiviral, and hypoglycemic effects of several mushrooms have been demonstrated. Further, multiple investigations have indicated the protective effects of mushroom extracts against chemically induced liver damage by lowering inflammation and oxidative stress.²²^,²³Grifola frondosa, popularly recognized as maitake, is a prominent edible mushroom that has also been consumed as medicine for thousands of years in Japan and China.²⁴Grifola frondosa's main active ingredients are polysaccharides, which are responsible for its anticancer, antioxidant, antidiabetic, and hepatoprotective properties.²⁵^,²⁶ Hence, this work aimed to investigate the ameliorative and protective roles of chitosan nanoparticles (CS NPs) and Grifola frondosa-loaded casein nanoparticles (GF-loaded casein NPs) against Ehrlich ascites carcinoma-induced hepatotoxicity in mice.

Materials and methods

Animals and experimental design

Sixty female Swiss albino mice weighing 20–25 g and aged 10–12 weeks were procured from the National Research Center’s animal house (Dokki, Giza, Egypt). The mice were accommodated in adequate plastic cages and fed regular food with free access to water.

Animal care and treatment were carried out in compliance with the Tanta University Faculty of Science’s animal care guidelines, as authorized by the Institutional Animal Care and Use Committee (IACUC-SCI-TU-0166). Chitosan nanoparticles (CS NPs, average particle size ranged from 10 to 60 nm) were purchased from NANOFAB TECHNOLOGY (Giza, Egypt). Grifola Frondosa nanoparticles used throughout this work were prepared and characterized according to.²⁷ Six groups of ten mice were randomly formed, the experimental groups and treatments employed in this work are presented in Table 1.

Table 1.

The experimental groups used in this work.

Group	Dose (mg/Kg)	Treatments
G1	-	Saline
G2	100	CS NPs²⁸
G3	100	GF-loaded casein NPs²⁹
G4	-	EAC cells³⁰
G5	100	EAC + CS NPs
G6	100	EAC + GF-loaded casein NPs

Open in a new tab

G1, control group; G2 and G3, mice received CS NPs and GF-loaded casein NPs daily by oral gavage for 2 weeks respectively; G4, mice injected once with 2.5×10⁶ EAC cells intraperitoneally; G5 and G6, mice injected with EAC cells on day 0 then on the next day treated with CS NPs and GF-loaded casein NPs for 2 weeks respectively.

Blood collection and tissue sampling

At the end of the experiment, mice were subjected to fasting overnight before being euthanized with sodium pentobarbital through intravenous injection and necropsied. Blood was drawn from each mouse’s inferior vena cava and placed in non-heparinized glass tubes. The serum was then centrifuged at 2,250 × g for 20 min and stored at −20 °C for subsequent biochemical assessment. Liver tissues were isolated, cleaned and rinsed with the cold normal saline. Then divided into three parts: the tissue homogenate was prepared in the first part and the supernatant was collected by centrifugation at 10,000 × g for 20 min then kept at −80 °C for further antioxidant parameters evaluation. The second part was kept at −80 °C for DNA fragmentation analysis. The third part was immersed in 10% neutral buffered formalin for histological and immunohistochemical investigation.

Biochemical parameters

Liver functions including aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), albumin, and total protein were determined in serum of different experimental groups as previously illustrated.³¹^,³² Moreover, serum total cholesterol and triglycerides were estimated as previously reported³³^,³⁴ respectively. Serum HDL-cholesterol was evaluated according to the method of³⁵ and LDL-cholesterol was calculated using the previously reported method.³⁶

Oxidative stress and antioxidant parameters

Hepatic oxidative stress was assessed colorimetrically by detecting thiobarbituric acid reactive substances (TBARS) and the extent of lipid peroxidation was quantified as MDA equivalents.³⁷ Reduced glutathione (GSH) and catalase activity (CAT) were evaluated as previously established³⁸ and³⁹ respectively.

DNA fragmentation analysis

QIAamp DNA Mini Kit was used to isolate and purify DNA from hepatic tissues, and DNA concentration was quantified at 260 nm using Nano Volume Spectrometer Scan drop 200 (Analytik Jena). The DNA samples were then separated on 1.8% agarose gel containing ethidium bromide.⁴⁰^,⁴¹

Histopathological assessment

Hepatic tissue samples were preserved for 48 h in 10% formalin solution before being prepared for paraffin sectioning. The sections were then dyed with hematoxylin and eosin to investigate the typical histologic structure.⁴²

Immunohistochemical detection of p53 and Bcl-2 expression

The expression of tumor suppressor (p53), and antiapoptotic (Bcl-2) proteins was conducted using avidin-biotin complex method according to aforementioned studies.⁴³^,⁴⁴

Statistical assessment

Data were presented as mean ± SEM. The significance of variance between experimental groups was assessed using one-way ANOVA followed by Tukey’s test in GraphPad Prism software. When P value was <0.05, the significance was considered acceptable.

Results

CS NPs and GF-loaded casein NPs improve liver functions in EAC-bearing mice

Despite the albumin level being considerably raised in G2 group, a non-remarkable difference in liver function biomarkers were observed in CS NPs (G2) and GF-loaded casein NPs (G3) groups compared to normal G1 group (Fig. 1). Conversely, there was a significant increase (P ˂ 0.0001) in hepatic ALT, AST, and ALP activities with a marked decrease (P ˂ 0.0001) in total protein and albumin concentration in EAC group (G4). The treatment of EAC-bearing mice either with CS NPs (G5 group) or GF-loaded casein NPs (G6 group) demonstrated a remarkable decline in hepatic enzymes with an apparent improvement in albumin and total protein content compared to untreated EAC mice (G4 group).

Fig. 1 — Effect of CS NPs and GF-loaded casein NPs on liver function tests in EAC-bearing mice. Values were presented as mean ± SEM, n = 10. Small (a,b) letters differed significantly from G1 and G4 respectively at P ≤ 0.05. ALT: Alanine aminotransferase; AST: aspartate aminotransferase; ALP: alkaline phosphatase.

CS NPs and GF-loaded casein NPs reduce LDL, cholesterol, and triglycerides level in EAC-bearing mice

The levels of HDL, LDL, cholesterol, and triglycerides in the CS NPs treated (G2) group were not statistically different from those in the normal (G1) group. In GF-loaded casein NPs (G3) group, triglycerides concentration increased markedly (P ˂ 0.0001) but other lipid profile parameters remained unchanged. Untreated EAC mice (G4) have higher LDL, cholesterol, and triglycerides concentration than control (G1) group. The administration of CS NPs (G5) or GF-loaded casein NPs (G6) in EAC mice caused a substantial reduction (P ˂ 0.0001) in LDL, cholesterol, and triglycerides levels as shown in Fig. 2.

Fig. 2 — Effect of CS NPs and GF-loaded casein NPs on lipid profile in EAC-bearing mice. Values were presented as mean ± SEM, n = 10. Small (a,b) letters differed significantly from G1 and G4 respectively at P ≤ 0.05.

CS NPs and GF-loaded casein NPs attenuate oxidative stress and enhance antioxidant system in liver tissues of EAC-bearing mice

As illustrated by Table 2, there was a considerable increase (P ˂ 0.05) in MDA levels and a substantial suppression (P ˂ 0.0001) of GSH content and CAT activity in the liver of the EAC-bearing group (G4) as compared to the G1 control group. Supplementation with CS NPs (G5) and GF-loaded casein NPs (G6), on the other hand, returned the decrease in GSH content and enhanced CAT activity in liver tissues to normal levels. Furthermore, the administration of GF-loaded casein NPs (G6) to EAC-bearing mice considerably lowered (P ˂ 0.05) the increased levels of MDA compared to untreated EAC (G4) group.

Table 2.

Hepatic oxidative stress and antioxidant parameters in EAC-bearing mice after treatment with CS NPs and GF-loaded casein NPs.

Groups	MDA (μmol/g tissue)	GSH (μmol/g tissue)	CAT (μmol/min/mg protein)
G1	117.60 ± 7.95	1115.00 ± 50.83	1093.00 ± 38.00
G2	93.59 ± 8.74	1395.00^a ± 63.44	1260.00^a ± 49.94
G3	90.77 ± 12.80	1240.00 ± 77.04	1126.00 ± 38.82
G4	237.90^a ± 12.07	422.50^a ± 27.58	454.40^a ± 29.66
G5	139.70 ± 9.15	1041.00^b ± 30.39	888.00^ab ± 50.03
G6	120.00^b ± 5.28	913.40^ab ± 58.73	773.80^ab ± 50.14

Open in a new tab

Values were presented as mean ± SEM. Small (a,b) letters differed significantly from G1 and G4 respectively at P ≤ 0.05.

CS NPs and GF-loaded casein NPs reduce DNA fragmentation in liver tissues of EAC-bearing mice

Figure 3A represents the agarose gel electrophoresis of total genomic DNA isolated from liver tissues of all experimental groups. As demonstrated in this image, CS NPs (G2) and GF-loaded casein NPs (G3) groups did not exhibit DNA fragmentation as compared to G1 control group. Unlike the control group, EAC-bearing mice (G4) demonstrated smeared DNA fragmentation. In addition, the treatment of EAC-bearing mice with CS NPs extensively (P ˂ 0.01) reduced DNA fragmentation, as evidenced by lower optical density of extracted DNA (Fig. 3B).

Fig. 3 — DNA fragmentation analysis of DNA extracted from hepatic tissues of different experimental groups. A) Descriptive photograph of DNA resolved on 1.8% agarose gel stained with ethidium bromide. B) Data were presented as mean ± SEM. Small (a,b) letters differed significantly from G1 and G4 respectively at P ≤ 0.05.

CS NPs and GF-loaded casein NPs improve hepatic structure in EAC-bearing mice

Figure 4 depicts the histological alterations in the liver sections from the various groups. Hepatocytes with large round nuclei, eosinophilic cytoplasm, and few spaced hepatic sinusoids placed in-between the hepatic cords with a fine arrangement of Kupffer cells were seen in liver sections from the G1, G2, and G3 groups. Liver sections in G4 (EAC) group, on the other hand, revealed severe tissue injury with marked degeneration, apoptosis, and inflammation of hepatic tissue. EAC mice treated with CS NPs in G5 group exhibited clear improvement of all changes in hepatic cells compared to untreated EAC (G4) group. Further, mild to moderate damage, mild apoptosis, cellular infiltrations, and cytoplasmic vacuolization of hepatocytes was observed in EAC group treated with GF-loaded casein NPs (G6).

Fig. 4 — Liver sections photomicrographs stained with haematoxylin and eosin. A–C) Normal liver structure with hepatocytes (Hp) with prominent round nuclei, eosinophilic cytoplasm, and central veins (Cv) in G1-G3. D) Severe tissue injury with marked degeneration, apoptosis (arrow heads) and inflammation (arrows) of hepatic tissue were observed in liver of EAC (G4). E) Treated EAC mice with CS NPs group (G5) exhibited mild cellular inflammation (arrows) of hepatic tissue. F) Mild to moderate damage, mild apoptosis (arrow heads), cellular infiltrations (arrows), and cytoplasmic vacuolization of hepatocytes was observed in EAC group treated with GF-loaded casein NPs (G6).

CS NPs and GF-loaded casein NPs upregulate Bcl-2 and downregulate p53 expression in liver tissues of EAC-bearing mice

Figure 5 showed that Bcl-2 expression in hepatocytes was strongly positive in liver sections from the control (G1), CS NPs (G2), and GF-loaded casein NPs (G3) groups. Negative to mild reaction for Bcl-2 expression was shown in the hepatocytes of EAC (G4) group (Fig. 5D). On the contrary, Bcl-2 demonstrated mild to moderate expressions in liver of EAC mice treated with either CS NPs (G5 group) or GF-loaded casein NPs (G6 group) as seen in Fig. 5E and F.

Fig. 5 — Liver sections photomicrographs stained with Bcl2 expression (denoted by arrows). A–C) Control (G1), CS NPs (G2) and GF-loaded casein NPs (G3) groups showed strong positive reaction for Bcl-2 expressions in hepatocytes. D) Negative to mild reaction for Bcl-2 expression was shown in the hepatocytes of EAC (G4) group. E and F) Mild to moderate expressions for Bcl-2 expressions in hepatocytes in liver sections in EAC mice treated with either CS NPs (G5) or GF-loaded casein NPs (G6) groups respectively.

Furthermore, the liver sections in control (G1), CS NPs (G2) and GF-loaded casein NPs (G3) groups showed faint to mild positive reaction for P53 expressions in hepatocytes (Fig. 6A–C). On the contrary, strong positive reactivity of p53 expressions was seen in the hepatocytes of mice inoculated with EAC (G4) (Fig. 6D). The treatment of EAC mice with CS NPs (G5 group) revealed mild positive reaction for P53 expressions in hepatocytes. Moreover, Moderate to strong positive reactivity of p53 expression in hepatocytes of EAC mice treated with GF-loaded casein NPs as compared to EAC (G4) group (Fig. 6E and F).

Fig. 6 — Liver sections photomicrographs stained with P53 expression (denoted by arrows). A–C) Liver sections in control (G1), CS NPs (G2) and GF-loaded casein NPs (G3) groups showed faint to mild positive reaction for P53 expressions in hepatocytes. D) Strong positive reactivity of p53 expressions in hepatocytes of mice inoculated with EAC (G4). E) Mild positive reaction for P53 expressions in hepatocytes in treated EAC mice with CS NPs (G5). F) Moderate to strong positive reaction for P53 expressions in hepatocytes in treated EAC mice with GF-loaded casein NPs.

Discussion

Ehrlich ascites carcinoma has characteristics comparable to human tumor as it grows rapidly and is undifferentiated.⁴⁵ Tumor growth has been attributed to genomic instability, which causes DNA damage and gene alterations, leading to internal organ injuries.⁴⁶ It has been revealed that EAC cells can infiltrate vital organs in the animal body such as the liver and kidney, causing inflammatory cell aggregation and mitochondrial degradation.⁸^,⁴⁷ Thus, the present work was implemented to consider the ameliorative role of chitosan and Grifola Frondosa nanoparticles against EAC-induced liver toxicity in mice.

The current study found that EAC inoculation caused liver malfunction, as seen by an increase in serum ALT, AST, and ALP activity and a decrease in total protein and albumin levels as well as elevation of lipid profile markers, including TG, cholesterol, and LDL. Our findings are in line with earlier studies.⁶^,⁴⁸^,⁴⁹ The increased levels of liver enzymes are an indicator of hepatic metabolism disruption caused by cancer proliferation, resulting in increase in cell permeability and hepatocytes disintegration, causing their enzymes to be released into the plasma.⁵⁰ The treatment of EAC mice either with CS NPs or GF-loaded casein NPs improved liver functions, indicating that these substances can protect hepatic cellular membrane from damage. These findings corroborate those of Al-Baqami and his colleagues, and Elsonbaty and his colleagues who reported that the administration of chitosan nanoparticles alleviate the hepatotoxic effect induced by D-galactose and diethylnitrosamine in rat respectively.⁵¹^,⁵² In addition, a previous study demonstrated that water extract of Grifola frondosa ameliorate carbon tetrachloride-induced liver damage in albino rats.⁵³

The observed decline in liver marker levels following administration of CS NPs or GF-loaded casein NPs may be attributable, in part, to the potential of these compounds to decline oxidative stress and boost the antioxidant defense status.⁵⁴ In the current work, EAC inoculation caused a considerable elevation in hepatic MDA level while decreasing GSH content and CAT activity. This could be because EAC inoculation induces an increase in lipid peroxidation and a decrease in antioxidant potential, which causes liver damage. Our results are consistent with earlier research.^55–57 Treatment of EAC animals with either CS NPs or GF-loaded casein NPs significantly recovered redox equilibrium and mitigated the effect of EAC cells by raising GSH content and CAT activity, while suppressing the EAC-mediated increase in MDA levels. Previous studies elucidated the protective effect of CS NPs against oxidative damage persuaded by H₂O₂ in murine macrophage cells and by D-galactose in liver of male rats which is considerably proved the current study’s findings and confirmed the antioxidant activity of CS NPs.⁵⁸^,⁵⁹ The particle size of chitosan nanoparticles considered the key factor in regulating its absorption, distribution, and accumulation, consequently, influence its effectiveness. Small-sized NPs are helpful for extended circulation and distribution in a variety of tissues. Further, the particle size should not be as small as feasible, as this will result in greater instability of the nanosystem under physiological settings and will not have the desired effect.⁶⁰ Previous studies reported that chitosan nanoparticles of 110–390 nm have similar cytotoxicity against lung cancer cells.⁶¹^,⁶² On the other hand, another study demonstrated that human gastric carcinoma cells are very sensitive to chitosan nanoparticles of 65 nm.⁶³ This may be attributable to the correlation between the size of nanoparticles, their cellular uptake and cytotoxicity.⁶² The current study used chitosan NPs ranging in size from 10 to 60 nm, which assisted in NP uptake by absorptive cells via endocytosis. Moreover, Li et al reported that Grifola Frondosa water extract attenuated the oxidative stress and hepatic fibrosis provoked by CCl4 in experimental rats.⁶⁴ The antioxidant activity of GF may be attributed to their ability to chelate metals and function as free radical scavengers.⁶⁵

Various studies found that EAC promoted oxidative stress in experimental animals due to ROS overproduction, which was linked to liver injury, by disrupting several physiological functions and triggering cell apoptosis via inducing DNA damage.^66–68 The current work reported that EAC- induced liver damage was accompanied by hepatic tissue DNA damage, as shown by the appearance of smeared DNA fragmentation and necrosis. Our results agree with Tousson et al and Hashem et al.⁶^,⁶⁹ The treatment of EAC-bearing mice with CS NPs resulted in a significant increase in DNA optical density compared to untreated EAC mice. EAC mice treated with GF-loaded casein NPs, on the other hand, revealed non-significant change in DNA optical density. These findings confirm the findings of Saleh and El-Shorbagy, who discovered that chitosan is an effective preventive treatment for liver ischemia reperfusion by reducing DNA fragmentation and suppressing apoptosis.⁷⁰

Tumor suppressor protein (P53) is a transcription factor implicated in cell cycle regulation, apoptosis, and cancer repression.⁷¹ Further, Bcl-2, a key anti-apoptotic protein that belongs to the Bcl-2 family, blocks the release of pro-apoptotic molecules and cytochrome C from the mitochondria, hence preventing apoptosis and allowing cancer cells to persist.⁷² Immunohistochemical results indicated a high expression of apoptotic proteins, P53, with low expression of Bcl-2 in the livers of EAC-bearing mice, revealing that EAC tumors proliferate rapidly. These findings were consistent with those of Tousson and his colleagues, who reported that EAC increased P53 protein expression in mouse hepatic tissue as compared to a normal control group.⁶ When CS NPs or GF-loaded casein NPs were given to EAC mice, they lowered the immunoreactivity of p53 expression while increasing the expression of Bcl-2 protein compared to the untreated EAC mice. Our results are coherent with prior study which found that CS NPs treatment reduced p53 immunoreactivity in rats injected with the hepatocarcinogen 2-nitropropane.²¹

Conclusion

The data of the current work collectively demonstrated that CS NPs and GF-loaded casein NPs have an ameliorative role against EAC-induced liver injury in mice by enhancing liver functions, balancing the oxidative/antioxidant status, and inhibiting apoptosis.

Acknowledgments

Not applicable.

Contributor Information

Aliaa M Radwan, Biochemistry Division, Chemistry Department, Faculty of Science, Tanta University, El Geish street, Tanta, Gharbia Governorate 31527, Egypt.

Doaa T Gebreel, Medical Equipment Department, Faculty of Allied Medical Sciences, Pharos University, Canal El Mahmoudia Street, beside, Green Plaza 21648, Alexandria, Egypt.

Sahar Allam, Zoology Department, Faculty of Science, Tanta University, El Geish street, Tanta, Gharbia Governorate 31527, Egypt.

Afaf El-Atrash, Zoology Department, Faculty of Science, Tanta University, El Geish street, Tanta, Gharbia Governorate 31527, Egypt.

Ehab Tousson, Zoology Department, Faculty of Science, Tanta University, El Geish street, Tanta, Gharbia Governorate 31527, Egypt.

Author contributions

E.T. and A.E. conceptualized and designed the study. A.M.R., S.A. and D.T.G. carried out the initial analysis. A.M.R. wrote the draft of the manuscript. ET reviewed and revised the final version of the manuscript.

Funding

Not applicable.

Conflict of interest statement. There are no conflicts of interest to declare.

Data availability

The data utilized to maintain the outcomes of this study are available from the corresponding author upon request.

Ethical approval

The Institutional Animal Care and Use Committee (IACUC-SCI-TU-0166) had the capability to underwrite this study which was guided by the rules provided by the Moral Committee of the Faculty of Science at Tanta University.

References

1. Abd-Elghany AA, Ahmed SM, Masoud MA, Atia T, Waggiallah HA, El-Sakhawy MA, Mohamad EA. Annona squamosa L. extract-loaded niosome and its anti-ehrlich ascites’ carcinoma activity. ACS Omega. 2022:7(43):38436–38447. [DOI] [PMC free article] [PubMed] [Google Scholar]
2. Oshiba RT, Touson E, Elsherbini YM, Abdraboh ME. Melatonin: a regulator of the interplay between FoxO1, miR96, and miR215 signaling to diminish the growth, survival, and metastasis of murine adenocarcinoma. Biofactors. 2021:47(5):740–753. [DOI] [PubMed] [Google Scholar]
3. Hasan AF, Alankooshi AA, Alankooshi AA, Abbood AS, Dulimi AG, Mohammed al-Khuzaay H, Elsaedy EA, Tousson E. Impact of B-Glucan against Ehrlich ascites carcinoma induced renal toxicity in mice. OnLine J Biol Sci. 2023:23(1):103–108. [Google Scholar]
4. Frajacomo FTT, de Souza Padilha C, Marinello PC, Guarnier FA, Cecchini R, Duarte JAR, Deminice R. Solid Ehrlich carcinoma reproduces functional and biological characteristics of cancer cachexia. Life Sci. 2016:162:47–53. [DOI] [PubMed] [Google Scholar]
5. Saleh N, Allam T, Korany RM, Abdelfattah AM, Omran AM, Abd Eldaim MA, Hassan AM, el-Borai NB. Protective and therapeutic efficacy of hesperidin versus cisplatin against ehrlich ascites carcinoma-induced renal damage in mice. Pharmaceuticals. 2022:15(3):294. [DOI] [PMC free article] [PubMed] [Google Scholar]
6. Tousson E, Hafez E, Abo Gazia MM, Salem SB, Mutar TF. Hepatic ameliorative role of vitamin B17 against Ehrlich ascites carcinoma-induced liver toxicity. Environ Sci Pollut Res. 2020:27(9):9236–9246. [DOI] [PubMed] [Google Scholar]
7. Areida SK, Abd El-Azim AO, Amer ME. Protective and curative effect of Thymoquinone on Ehrlich solid carcinoma inoculated mice. Egypt J Hosp Med. 2015:58(1):129–142. [Google Scholar]
8. Abd Eldaim MA, Tousson E, el Sayed IET, Abd Elmaksoud AZ, Ahmed AA. Ameliorative effects of 9-diaminoacridine derivative against Ehrlich ascites carcinoma-induced hepatorenal injury in mice. Environ Sci Pollut Res. 2021:28(17):21835–21850. [DOI] [PubMed] [Google Scholar]
9. Bhattacharyya A, Mandal DP, Lahiry L, Bhattacharyya S, Chattopadhyay S, Ghosh UK, Sa G, Das T. Black tea-induced amelioration of hepatic oxidative stress through antioxidative activity in EAC-bearing mice. J Environ Pathol Toxicol Oncol. 2007:26(4):245–254. [DOI] [PubMed] [Google Scholar]
10. Alotaibi B, Tousson E, El-Masry TA, Altwaijry N, Saleh A. Ehrlich ascites carcinoma as model for studying the cardiac protective effects of curcumin nanoparticles against cardiac damage in female mice. Environ Toxicol. 2021:36(1):105–113. [DOI] [PubMed] [Google Scholar]
11. Ding J, Guo Y. Recent advances in chitosan and its derivatives in cancer treatment. Front Pharmacol. 2022:13:888740. [DOI] [PMC free article] [PubMed] [Google Scholar]
12. Sultan MH, Moni SS, Madkhali OA, Bakkari MA, Alshahrani S, Alqahtani SS, Alhakamy NA, Mohan S, Ghazwani M, Bukhary HA, et al. Characterization of cisplatin-loaded chitosan nanoparticles and rituximab-linked surfaces as target-specific injectable nano-formulations for combating cancer. Sci Rep. 2022:12(1):468. [DOI] [PMC free article] [PubMed] [Google Scholar]
13. Mahgoob AAE, Tousson E, Abd Eldaim MA, Ullah S, Al-Sehemi AG, Algarni H, El Sayed IET. Ameliorative role of chitosan nanoparticles against silver nanoparticle-induced reproductive toxicity in male albino rats. Environ Sci Pollut Res. 2023:30(7):17374–17383. [DOI] [PubMed] [Google Scholar]
14. Tang L, Li J, Zhao Q, Pan T, Zhong H, Wang W. Advanced and innovative nano-systems for anticancer targeted drug delivery. Pharmaceutics. 2021:13(8):1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
15. Lima BV, Oliveira MJ, Barbosa MA, Goncalves RM, Castro F. Immunomodulatory potential of chitosan-based materials for cancer therapy: a systematic review of in vitro, in vivo and clinical studies. Biomater Sci. 2021:9(9):3209–3227. [DOI] [PubMed] [Google Scholar]
16. Divya K, Jisha M. Chitosan nanoparticles preparation and applications. Environ Chem Lett. 2018:16(1):101–112. [Google Scholar]
17. Gupta R, Xie H. Nanoparticles in daily life: applications, toxicity and regulations. J Environ Pathol Toxicol Oncol. 2018:37(3):209–230. [DOI] [PMC free article] [PubMed] [Google Scholar]
18. Dawoud SF, Al-Akra TM, Zedan AM. Hepatoprotective effects of chitosan and chitosan nanoparticles against biochemical, genetic, and histological disorders induced by the toxicity of emamectin benzoate. Rep Biochem Mol Biol. 2021:10(3):506–514. [DOI] [PMC free article] [PubMed] [Google Scholar]
19. Gebreel DT, Elatrash AM, Atta FA, Tousson E, Allam S. Roles of chitosan nanoparticles on Ehrlich ascites carcinoma induced changes in some blood parameters. Asian Oncol Res J. 2021:3(4):71–77. [Google Scholar]
20. Yao H-T, Luo M-N, Li C-C. Chitosan oligosaccharides reduce acetaminophen-induced hepatotoxicity by suppressing CYP-mediated bioactivation. J Funct Foods. 2015:12:262–270. [Google Scholar]
21. Shaheen S, Arafah MM, Alshanwani AR, Fadda LM, Alhusaini AM, Ali HM, Hasan IH, Hagar H, Alharbi FM, AlHarthii A. Chitosan nanoparticles as a promising candidate for liver injury induced by 2-nitropropane: implications of P53, iNOS, VEGF, PCNA, and CD68 pathways. Sci Prog. 2021:104(2):003685042110118. [DOI] [PMC free article] [PubMed] [Google Scholar]
22. Soares AA, Sá-Nakanishi AB, Bracht A, Costa SM, Koehnlein EA, Souza CG, Peralta RM. Hepatoprotective effects of mushrooms. Molecules. 2013:18(7):7609–7630. [DOI] [PMC free article] [PubMed] [Google Scholar]
23. Gallego P, Luque-Sierra A, Falcon G, Carbonero P, Grande L, Bautista JD, Martín F, Del Campo JA. White button mushroom extracts modulate hepatic fibrosis progression, inflammation, and oxidative stress in vitro and in LDLR−/− mice. Food Secur. 2021:10(8):1788. [DOI] [PMC free article] [PubMed] [Google Scholar]
24. Ma X, Li C, Qi W, Li X, Wang S, Cao X, Wang C. Protective effect of extracellular polysaccharides from Grifola frondosa mycelium on CCl4-injured liver in vitro. Bioact Carbohydr Diet Fibre. 2015:6(1):7–14. [Google Scholar]
25. Zhang W, Jiang X, Zhao S, Zheng X, Lan J, Wang H, Ng TB. A polysaccharide-peptide with mercury clearance activity from dried fruiting bodies of maitake mushroom Grifola frondosa. Sci Rep. 2018:8(1):17630. [DOI] [PMC free article] [PubMed] [Google Scholar]
26. Chen Y, Liu D, Wang D, Lai S, Zhong R, Liu Y, Yang C, Liu B, Sarker MR, Zhao C. Hypoglycemic activity and gut microbiota regulation of a novel polysaccharide from Grifola frondosa in type 2 diabetic mice. Food Chem Toxicol. 2019:126:295–302. [DOI] [PubMed] [Google Scholar]
27. Abosharaf HA, Gebreel DT, Allam S, El-Atrash A, Tousson E. Ehrlich ascites carcinoma provokes renal toxicity and DNA injury in mice: therapeutic impact of chitosan and maitake nanoparticles. Basic Clin Pharmacol Toxicol. 2024:134(4):472–484. [DOI] [PubMed] [Google Scholar]
28. Xu Y, Wen Z, Xu Z. Chitosan nanoparticles inhibit the growth of human hepatocellular carcinoma xenografts through an antiangiogenic mechanism. Anticancer Res. 2009:29(12):5103–5109. [PubMed] [Google Scholar]
29. Zhao J, He R, Zhong H, Liu S, Hussain M, Sun P. Synergistic antitumor effect of Grifola frondose polysaccharide-protein complex in combination with cyclophosphamide in H22 tumor-bearing mice. Molecules. 2023:28(7):2954. [DOI] [PMC free article] [PubMed] [Google Scholar]
30. Mutar TF, Tousson E, Hafez E, Abo Gazia M, Salem SB. Ameliorative effects of vitamin B17 on the kidney against Ehrlich ascites carcinoma induced renal toxicity in mice. Environ Toxicol. 2020:35(4):528–537. [DOI] [PubMed] [Google Scholar]
31. Rashid N, Siddique W, Zaheer Z. Evaluation of effect of Argyrolobiumroseum aqueous extract on carbon tetrachloride induced liver injury in rabbits. Pak J Med Health Sci. 2018:12(1):161–165. [Google Scholar]
32. Tietz NW, Berger S. Fundamentals of clinical chemistry. Medicine. 1976:3(1). [Google Scholar]
33. Allain CC, Poon LS, Chan CS, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clin Chem. 1974:20(4):470–475. [PubMed] [Google Scholar]
34. Fossati P, Principe L. Enzymatic colorimetric method to determination triglycerides. Clin Chem. 1982:28(2077):1330.6176371 [Google Scholar]
35. Lopes-Virella MF, Stone P, Ellis S, Colwell JA. Cholesterol determination in high-density lipoproteins separated by three different methods. Clin Chem. 1977:23(5):882–884. [PubMed] [Google Scholar]
36. Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem. 1972:18(6):499–502. [PubMed] [Google Scholar]
37. Esterbauer H, Cheeseman KH. [42] Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 1990:186:407–421. [DOI] [PubMed] [Google Scholar]
38. Balasubashini MS, Rukkumani R, Viswanathan P, Menon VP. Ferulic acid alleviates lipid peroxidation in diabetic rats. Phytother Res. 2004:18(4):310–314. [DOI] [PubMed] [Google Scholar]
39. Aebi H. [13] Catalase in vitro. Methods Enzymol. 1984:105:121–126. [DOI] [PubMed] [Google Scholar]
40. Abd Eldaim MA, Tousson E, El Sayed IET, Awd WM. Ameliorative effects of Saussurea lappa root aqueous extract against Ethephon-induced reproductive toxicity in male rats. Environ Toxicol. 2019:34(2):150–159. [DOI] [PubMed] [Google Scholar]
41. Morsi DS, el-Nabi SH, Elmaghraby MA, Abu Ali OA, Fayad E, Khalifa SA, el-Seedi HR, el-Garawani IM. Anti-proliferative and immunomodulatory potencies of cinnamon oil on Ehrlich ascites carcinoma bearing mice. Sci Rep. 2022:12(1):11839. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
42. Bancroft JD, Gamble M. Theory and practice of histological techniques. 6th ed. Churchill Livingstone, Elsevier, China; 2008. [Google Scholar]
43. Tousson E, Hafez E, Zaki S, Gad A. P53, Bcl-2 and CD68 expression in response to amethopterin-induced lung injury and ameliorating role of l-carnitine. Biomed Pharmacother. 2014:68(5):631–639. [DOI] [PubMed] [Google Scholar]
44. Tousson E, Hafez E, Zaki S, Gad A. The cardioprotective effects of L-carnitine on rat cardiac injury, apoptosis, and oxidative stress caused by amethopterin. Environ Sci Pollut Res. 2016:23(20):20600–20608. [DOI] [PubMed] [Google Scholar]
45. Kabel AM, Abdel-Rahman MN, El-Sisi AE-DE, Haleem MS, Ezzat NM, El Rashidy MA. Effect of atorvastatin and methotrexate on solid Ehrlich tumor. Eur J Pharmacol. 2013:713(1–3):47–53. [DOI] [PubMed] [Google Scholar]
46. Chakraborty T, Bhuniya D, Chatterjee M, Rahaman M, Singha D, Chatterjee BN, Datta S, Rana A, Samanta K, Srivastawa S, et al. Acanthus ilicifolius plant extract prevents DNA alterations in a transplantable Ehrlich ascites carcinoma-bearing murine model. World J Gastroenterol. 2007:13(48):6538–6548. [DOI] [PMC free article] [PubMed] [Google Scholar]
47. Lawal R, Özaslan M. Effect of medicinal plants on ehrlich ascites carninoma cells (a review). Zeugma Biol Sci. 2021:2(2):19–27. [Google Scholar]
48. Al-Khuzaay HM, Al-Juraisy YH, Alwan AH. Therapeutic effect of β-Glucan on Ehrlich ascites carcinoma that induced liver toxicity. 2023. Preprint, 10.22541/au.167938202.28621136/v1. [DOI]
49. Aldubayan MA, Elgharabawy RM, Ahmed AS, Tousson E. Antineoplastic activity and curative role of avenanthramides against the growth of ehrlich solid tumors in mice. Oxidative Med Cell Longev. 2019:2019:1–12. [DOI] [PMC free article] [PubMed] [Google Scholar]
50. Eissa MM, Gaafar MR, Younis LK, Ismail CA, El Skhawy N. Prophylactic antineoplastic activity of toxoplasma gondii RH derived antigen against ehrlich solid carcinoma with evidence of shared antigens by comparative immunoblotting. Infect Agent Cancer. 2023:18(1):1–13. [DOI] [PMC free article] [PubMed] [Google Scholar]
51. Al-Baqami N, Hamza RZ. Synergistic antioxidant capacities of vanillin and chitosan nanoparticles against reactive oxygen species, hepatotoxicity, and genotoxicity induced by aging in male Wistar rats. Hum Exp Toxicol. 2021:40(1):183–202. [DOI] [PubMed] [Google Scholar]
52. Elsonbaty S, Moawad F, Abdelghaffar M. Antioxidants and hepatoprotective effects of chitosan nanoparticles against hepatotoxicity induced in rats. Benha Vet Med J. 2019:36(1):252–261. [Google Scholar]
53. Lee J-S, Kim H-S, Lee Y-J, Yong C-S, Choi H-G, Han G-D, Kim J-A, Lee J-S. Hepatoprotective effect of Grifola frondosa water extract on carbon tetrachloride-induced liver injury in rats. Food Sci Biotechnol. 2008:17(1):203–207. [Google Scholar]
54. Sandri G, Rossi S, Bonferoni MC, Ferrari F, Zambito Y, Di Colo G, Caramella C. Buccal penetration enhancement properties of N-trimethyl chitosan: influence of quaternization degree on absorption of a high molecular weight molecule. Int J Pharm. 2005:297(1–2):146–155. [DOI] [PubMed] [Google Scholar]
55. Elsadek MF, Essam el-Din MM, Ahmed BM. Evaluation of anticarcinogenic and antioxidant properties of Eruca sativa extracts versus Ehrlich ascites carcinoma in mice. J King Saud Univ-Sci. 2021:33(4):101435. [Google Scholar]
56. Attia AA, Salama AF, Eldiasty JG, Mosallam SAE-R, El-Naggar SA, El-Magd MA, Nasser HM, Elmetwalli A. Amygdalin potentiates the anti-cancer effect of Sorafenib on Ehrlich ascites carcinoma and ameliorates the associated liver damage. Sci Rep. 2022:12(1):6494. [DOI] [PMC free article] [PubMed] [Google Scholar]
57. Mohammed FZ, Tousson E, Yassin F. Anticancer activity and antioxidant potential of crocus Sativus. L (saffron) aqueous extract against Ehrlich ascites carcinoma cells in Swiss albino mice. Biochem Lett. 2020:16(1):20–49. [Google Scholar]
58. Wen Z-S, Liu L-J, Qu Y-L, OuYang X-K, Yang L-Y, Xu Z-R. Chitosan nanoparticles attenuate hydrogen peroxide-induced stress injury in mouse macrophage RAW264. 7 cells. Mar Drugs. 2013:11(10):3582–3600. [DOI] [PMC free article] [PubMed] [Google Scholar]
59. Al-Eisa RA. Synergistic antioxidant capacity of chitosan nanoparticles and lycopene against aging hepatotoxicity induced by D-galactose in male rats. Int J Pharm. 2018:14(6):811–825. [Google Scholar]
60. Jafernik K, Ładniak A, Blicharska E, Czarnek K, Ekiert H, Wiącek AE, Szopa A. Chitosan-based nanoparticles as effective drug delivery systems—a review. Molecules (Basel, Switzerland). 2023:28(4):1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
61. Ma Z, Lim L-Y. Uptake of chitosan and associated insulin in Caco-2 cell monolayers: a comparison between chitosan molecules and chitosan nanoparticles. Pharm Res. 2003:20(11):1812–1819. [DOI] [PubMed] [Google Scholar]
62. Huang M, Khor E, Lim L-Y. Uptake and cytotoxicity of chitosan molecules and nanoparticles: effects of molecular weight and degree of deacetylation. Pharm Res. 2004:21(2):344–353. [DOI] [PubMed] [Google Scholar]
63. Qi L-F, Xu Z-R, Li Y, Jiang X, Han X-Y. In vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells. World J Gastroenterol. 2005:11(33):5136–5141. [DOI] [PMC free article] [PubMed] [Google Scholar]
64. Li C, Meng M, Guo M, Wang M, Ju A, Wang C. The polysaccharides from Grifola frondosa attenuate CCl 4-induced hepatic fibrosis in rats via the TGF-β/Smad signaling pathway. RSC Adv. 2019:9(58):33684–33692. [DOI] [PMC free article] [PubMed] [Google Scholar]
65. Yeh J-Y, Hsieh L-H, Wu K-T, Tsai C-F. Antioxidant properties and antioxidant compounds of various extracts from the edible basidiomycete Grifola frondosa (Maitake). Molecules. 2011:16(4):3197–3211. [DOI] [PMC free article] [PubMed] [Google Scholar]
66. Bhattacharya S, Prasanna A, Majumdar P, Kumar RS, Haldar PK. Antitumor efficacy and amelioration of oxidative stress by Trichosanthes dioica root against Ehrlich ascites carcinoma in mice. Pharm Biol. 2011:49(9):927–935. [DOI] [PubMed] [Google Scholar]
67. Patra S, Muthuraman MS, Prabhu A, Priyadharshini RR, Parthiban S. Evaluation of antitumor and antioxidant activity of Sargassum tenerrimum against Ehrlich ascites carcinoma in mice. Asian Pac J Cancer Prev. 2015:16(3):915–921. [DOI] [PubMed] [Google Scholar]
68. El Sayed RAA, Hanafy ZEM, Abd El Fattah HF, Mohamed AK. Possible antioxidant and anticancer effects of plant extracts from Anastatica hierochuntica, Lepidium sativum and Carica papaya against Ehrlich ascites carcinoma cells. Cancer Biol Ther. 2020:10(1):1–16. [Google Scholar]
69. Hashem MA, Mahmoud EA, Abd-Allah NA. Alterations in hematological and biochemical parameters and DNA status in mice bearing Ehrlich ascites carcinoma cells and treated with cisplatin and cyclophosphamide. Comp Clin Pathol. 2020:29(2):517–524. [Google Scholar]
70. Saleh H, El-Shorbagy HM. Chitosan protects liver against ischemia-reperfusion injury via regulating Bcl-2/Bax, TNF-α and TGF-β expression. Int J Biol Macromol. 2020:164:1565–1574. [DOI] [PubMed] [Google Scholar]
71. DeVita V, Lawrence T, Rosenberg S. Chapter 8. Ljungman M, editor. In: Cancer: Principles & Practice of Oncology. 10th ed. Wolters Kluwer Health; 2015. [Google Scholar]
72. Lucken-Ardjomande S, Martinou J-C. Regulation of Bcl-2 proteins and of the permeability of the outer mitochondrial membrane. C R Biol. 2005:328(7):616–631. [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

The data utilized to maintain the outcomes of this study are available from the corresponding author upon request.

[ref1] 1. Abd-Elghany AA, Ahmed SM, Masoud MA, Atia T, Waggiallah HA, El-Sakhawy MA, Mohamad EA. Annona squamosa L. extract-loaded niosome and its anti-ehrlich ascites’ carcinoma activity. ACS Omega. 2022:7(43):38436–38447. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref2] 2. Oshiba RT, Touson E, Elsherbini YM, Abdraboh ME. Melatonin: a regulator of the interplay between FoxO1, miR96, and miR215 signaling to diminish the growth, survival, and metastasis of murine adenocarcinoma. Biofactors. 2021:47(5):740–753. [DOI] [PubMed] [Google Scholar]

[ref3] 3. Hasan AF, Alankooshi AA, Alankooshi AA, Abbood AS, Dulimi AG, Mohammed al-Khuzaay H, Elsaedy EA, Tousson E. Impact of B-Glucan against Ehrlich ascites carcinoma induced renal toxicity in mice. OnLine J Biol Sci. 2023:23(1):103–108. [Google Scholar]

[ref4] 4. Frajacomo FTT, de Souza Padilha C, Marinello PC, Guarnier FA, Cecchini R, Duarte JAR, Deminice R. Solid Ehrlich carcinoma reproduces functional and biological characteristics of cancer cachexia. Life Sci. 2016:162:47–53. [DOI] [PubMed] [Google Scholar]

[ref5] 5. Saleh N, Allam T, Korany RM, Abdelfattah AM, Omran AM, Abd Eldaim MA, Hassan AM, el-Borai NB. Protective and therapeutic efficacy of hesperidin versus cisplatin against ehrlich ascites carcinoma-induced renal damage in mice. Pharmaceuticals. 2022:15(3):294. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref6] 6. Tousson E, Hafez E, Abo Gazia MM, Salem SB, Mutar TF. Hepatic ameliorative role of vitamin B17 against Ehrlich ascites carcinoma-induced liver toxicity. Environ Sci Pollut Res. 2020:27(9):9236–9246. [DOI] [PubMed] [Google Scholar]

[ref7] 7. Areida SK, Abd El-Azim AO, Amer ME. Protective and curative effect of Thymoquinone on Ehrlich solid carcinoma inoculated mice. Egypt J Hosp Med. 2015:58(1):129–142. [Google Scholar]

[ref8] 8. Abd Eldaim MA, Tousson E, el Sayed IET, Abd Elmaksoud AZ, Ahmed AA. Ameliorative effects of 9-diaminoacridine derivative against Ehrlich ascites carcinoma-induced hepatorenal injury in mice. Environ Sci Pollut Res. 2021:28(17):21835–21850. [DOI] [PubMed] [Google Scholar]

[ref9] 9. Bhattacharyya A, Mandal DP, Lahiry L, Bhattacharyya S, Chattopadhyay S, Ghosh UK, Sa G, Das T. Black tea-induced amelioration of hepatic oxidative stress through antioxidative activity in EAC-bearing mice. J Environ Pathol Toxicol Oncol. 2007:26(4):245–254. [DOI] [PubMed] [Google Scholar]

[ref10] 10. Alotaibi B, Tousson E, El-Masry TA, Altwaijry N, Saleh A. Ehrlich ascites carcinoma as model for studying the cardiac protective effects of curcumin nanoparticles against cardiac damage in female mice. Environ Toxicol. 2021:36(1):105–113. [DOI] [PubMed] [Google Scholar]

[ref11] 11. Ding J, Guo Y. Recent advances in chitosan and its derivatives in cancer treatment. Front Pharmacol. 2022:13:888740. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref12] 12. Sultan MH, Moni SS, Madkhali OA, Bakkari MA, Alshahrani S, Alqahtani SS, Alhakamy NA, Mohan S, Ghazwani M, Bukhary HA, et al. Characterization of cisplatin-loaded chitosan nanoparticles and rituximab-linked surfaces as target-specific injectable nano-formulations for combating cancer. Sci Rep. 2022:12(1):468. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref13] 13. Mahgoob AAE, Tousson E, Abd Eldaim MA, Ullah S, Al-Sehemi AG, Algarni H, El Sayed IET. Ameliorative role of chitosan nanoparticles against silver nanoparticle-induced reproductive toxicity in male albino rats. Environ Sci Pollut Res. 2023:30(7):17374–17383. [DOI] [PubMed] [Google Scholar]

[ref14] 14. Tang L, Li J, Zhao Q, Pan T, Zhong H, Wang W. Advanced and innovative nano-systems for anticancer targeted drug delivery. Pharmaceutics. 2021:13(8):1151. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref15] 15. Lima BV, Oliveira MJ, Barbosa MA, Goncalves RM, Castro F. Immunomodulatory potential of chitosan-based materials for cancer therapy: a systematic review of in vitro, in vivo and clinical studies. Biomater Sci. 2021:9(9):3209–3227. [DOI] [PubMed] [Google Scholar]

[ref16] 16. Divya K, Jisha M. Chitosan nanoparticles preparation and applications. Environ Chem Lett. 2018:16(1):101–112. [Google Scholar]

[ref17] 17. Gupta R, Xie H. Nanoparticles in daily life: applications, toxicity and regulations. J Environ Pathol Toxicol Oncol. 2018:37(3):209–230. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref18] 18. Dawoud SF, Al-Akra TM, Zedan AM. Hepatoprotective effects of chitosan and chitosan nanoparticles against biochemical, genetic, and histological disorders induced by the toxicity of emamectin benzoate. Rep Biochem Mol Biol. 2021:10(3):506–514. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref19] 19. Gebreel DT, Elatrash AM, Atta FA, Tousson E, Allam S. Roles of chitosan nanoparticles on Ehrlich ascites carcinoma induced changes in some blood parameters. Asian Oncol Res J. 2021:3(4):71–77. [Google Scholar]

[ref20] 20. Yao H-T, Luo M-N, Li C-C. Chitosan oligosaccharides reduce acetaminophen-induced hepatotoxicity by suppressing CYP-mediated bioactivation. J Funct Foods. 2015:12:262–270. [Google Scholar]

[ref21] 21. Shaheen S, Arafah MM, Alshanwani AR, Fadda LM, Alhusaini AM, Ali HM, Hasan IH, Hagar H, Alharbi FM, AlHarthii A. Chitosan nanoparticles as a promising candidate for liver injury induced by 2-nitropropane: implications of P53, iNOS, VEGF, PCNA, and CD68 pathways. Sci Prog. 2021:104(2):003685042110118. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref22] 22. Soares AA, Sá-Nakanishi AB, Bracht A, Costa SM, Koehnlein EA, Souza CG, Peralta RM. Hepatoprotective effects of mushrooms. Molecules. 2013:18(7):7609–7630. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref23] 23. Gallego P, Luque-Sierra A, Falcon G, Carbonero P, Grande L, Bautista JD, Martín F, Del Campo JA. White button mushroom extracts modulate hepatic fibrosis progression, inflammation, and oxidative stress in vitro and in LDLR−/− mice. Food Secur. 2021:10(8):1788. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref24] 24. Ma X, Li C, Qi W, Li X, Wang S, Cao X, Wang C. Protective effect of extracellular polysaccharides from Grifola frondosa mycelium on CCl4-injured liver in vitro. Bioact Carbohydr Diet Fibre. 2015:6(1):7–14. [Google Scholar]

[ref25] 25. Zhang W, Jiang X, Zhao S, Zheng X, Lan J, Wang H, Ng TB. A polysaccharide-peptide with mercury clearance activity from dried fruiting bodies of maitake mushroom Grifola frondosa. Sci Rep. 2018:8(1):17630. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref26] 26. Chen Y, Liu D, Wang D, Lai S, Zhong R, Liu Y, Yang C, Liu B, Sarker MR, Zhao C. Hypoglycemic activity and gut microbiota regulation of a novel polysaccharide from Grifola frondosa in type 2 diabetic mice. Food Chem Toxicol. 2019:126:295–302. [DOI] [PubMed] [Google Scholar]

[ref27] 27. Abosharaf HA, Gebreel DT, Allam S, El-Atrash A, Tousson E. Ehrlich ascites carcinoma provokes renal toxicity and DNA injury in mice: therapeutic impact of chitosan and maitake nanoparticles. Basic Clin Pharmacol Toxicol. 2024:134(4):472–484. [DOI] [PubMed] [Google Scholar]

[ref28] 28. Xu Y, Wen Z, Xu Z. Chitosan nanoparticles inhibit the growth of human hepatocellular carcinoma xenografts through an antiangiogenic mechanism. Anticancer Res. 2009:29(12):5103–5109. [PubMed] [Google Scholar]

[ref29] 29. Zhao J, He R, Zhong H, Liu S, Hussain M, Sun P. Synergistic antitumor effect of Grifola frondose polysaccharide-protein complex in combination with cyclophosphamide in H22 tumor-bearing mice. Molecules. 2023:28(7):2954. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref30] 30. Mutar TF, Tousson E, Hafez E, Abo Gazia M, Salem SB. Ameliorative effects of vitamin B17 on the kidney against Ehrlich ascites carcinoma induced renal toxicity in mice. Environ Toxicol. 2020:35(4):528–537. [DOI] [PubMed] [Google Scholar]

[ref31] 31. Rashid N, Siddique W, Zaheer Z. Evaluation of effect of Argyrolobiumroseum aqueous extract on carbon tetrachloride induced liver injury in rabbits. Pak J Med Health Sci. 2018:12(1):161–165. [Google Scholar]

[ref32] 32. Tietz NW, Berger S. Fundamentals of clinical chemistry. Medicine. 1976:3(1). [Google Scholar]

[ref33] 33. Allain CC, Poon LS, Chan CS, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clin Chem. 1974:20(4):470–475. [PubMed] [Google Scholar]

[ref34] 34. Fossati P, Principe L. Enzymatic colorimetric method to determination triglycerides. Clin Chem. 1982:28(2077):1330.6176371 [Google Scholar]

[ref35] 35. Lopes-Virella MF, Stone P, Ellis S, Colwell JA. Cholesterol determination in high-density lipoproteins separated by three different methods. Clin Chem. 1977:23(5):882–884. [PubMed] [Google Scholar]

[ref36] 36. Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem. 1972:18(6):499–502. [PubMed] [Google Scholar]

[ref37] 37. Esterbauer H, Cheeseman KH. [42] Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 1990:186:407–421. [DOI] [PubMed] [Google Scholar]

[ref38] 38. Balasubashini MS, Rukkumani R, Viswanathan P, Menon VP. Ferulic acid alleviates lipid peroxidation in diabetic rats. Phytother Res. 2004:18(4):310–314. [DOI] [PubMed] [Google Scholar]

[ref39] 39. Aebi H. [13] Catalase in vitro. Methods Enzymol. 1984:105:121–126. [DOI] [PubMed] [Google Scholar]

[ref40] 40. Abd Eldaim MA, Tousson E, El Sayed IET, Awd WM. Ameliorative effects of Saussurea lappa root aqueous extract against Ethephon-induced reproductive toxicity in male rats. Environ Toxicol. 2019:34(2):150–159. [DOI] [PubMed] [Google Scholar]

[ref41] 41. Morsi DS, el-Nabi SH, Elmaghraby MA, Abu Ali OA, Fayad E, Khalifa SA, el-Seedi HR, el-Garawani IM. Anti-proliferative and immunomodulatory potencies of cinnamon oil on Ehrlich ascites carcinoma bearing mice. Sci Rep. 2022:12(1):11839. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]

[ref42] 42. Bancroft JD, Gamble M. Theory and practice of histological techniques. 6th ed. Churchill Livingstone, Elsevier, China; 2008. [Google Scholar]

[ref43] 43. Tousson E, Hafez E, Zaki S, Gad A. P53, Bcl-2 and CD68 expression in response to amethopterin-induced lung injury and ameliorating role of l-carnitine. Biomed Pharmacother. 2014:68(5):631–639. [DOI] [PubMed] [Google Scholar]

[ref44] 44. Tousson E, Hafez E, Zaki S, Gad A. The cardioprotective effects of L-carnitine on rat cardiac injury, apoptosis, and oxidative stress caused by amethopterin. Environ Sci Pollut Res. 2016:23(20):20600–20608. [DOI] [PubMed] [Google Scholar]

[ref45] 45. Kabel AM, Abdel-Rahman MN, El-Sisi AE-DE, Haleem MS, Ezzat NM, El Rashidy MA. Effect of atorvastatin and methotrexate on solid Ehrlich tumor. Eur J Pharmacol. 2013:713(1–3):47–53. [DOI] [PubMed] [Google Scholar]

[ref46] 46. Chakraborty T, Bhuniya D, Chatterjee M, Rahaman M, Singha D, Chatterjee BN, Datta S, Rana A, Samanta K, Srivastawa S, et al. Acanthus ilicifolius plant extract prevents DNA alterations in a transplantable Ehrlich ascites carcinoma-bearing murine model. World J Gastroenterol. 2007:13(48):6538–6548. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref47] 47. Lawal R, Özaslan M. Effect of medicinal plants on ehrlich ascites carninoma cells (a review). Zeugma Biol Sci. 2021:2(2):19–27. [Google Scholar]

[ref48] 48. Al-Khuzaay HM, Al-Juraisy YH, Alwan AH. Therapeutic effect of β-Glucan on Ehrlich ascites carcinoma that induced liver toxicity. 2023. Preprint, 10.22541/au.167938202.28621136/v1. [DOI]

[ref49] 49. Aldubayan MA, Elgharabawy RM, Ahmed AS, Tousson E. Antineoplastic activity and curative role of avenanthramides against the growth of ehrlich solid tumors in mice. Oxidative Med Cell Longev. 2019:2019:1–12. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref50] 50. Eissa MM, Gaafar MR, Younis LK, Ismail CA, El Skhawy N. Prophylactic antineoplastic activity of toxoplasma gondii RH derived antigen against ehrlich solid carcinoma with evidence of shared antigens by comparative immunoblotting. Infect Agent Cancer. 2023:18(1):1–13. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref51] 51. Al-Baqami N, Hamza RZ. Synergistic antioxidant capacities of vanillin and chitosan nanoparticles against reactive oxygen species, hepatotoxicity, and genotoxicity induced by aging in male Wistar rats. Hum Exp Toxicol. 2021:40(1):183–202. [DOI] [PubMed] [Google Scholar]

[ref52] 52. Elsonbaty S, Moawad F, Abdelghaffar M. Antioxidants and hepatoprotective effects of chitosan nanoparticles against hepatotoxicity induced in rats. Benha Vet Med J. 2019:36(1):252–261. [Google Scholar]

[ref53] 53. Lee J-S, Kim H-S, Lee Y-J, Yong C-S, Choi H-G, Han G-D, Kim J-A, Lee J-S. Hepatoprotective effect of Grifola frondosa water extract on carbon tetrachloride-induced liver injury in rats. Food Sci Biotechnol. 2008:17(1):203–207. [Google Scholar]

[ref54] 54. Sandri G, Rossi S, Bonferoni MC, Ferrari F, Zambito Y, Di Colo G, Caramella C. Buccal penetration enhancement properties of N-trimethyl chitosan: influence of quaternization degree on absorption of a high molecular weight molecule. Int J Pharm. 2005:297(1–2):146–155. [DOI] [PubMed] [Google Scholar]

[ref55] 55. Elsadek MF, Essam el-Din MM, Ahmed BM. Evaluation of anticarcinogenic and antioxidant properties of Eruca sativa extracts versus Ehrlich ascites carcinoma in mice. J King Saud Univ-Sci. 2021:33(4):101435. [Google Scholar]

[ref56] 56. Attia AA, Salama AF, Eldiasty JG, Mosallam SAE-R, El-Naggar SA, El-Magd MA, Nasser HM, Elmetwalli A. Amygdalin potentiates the anti-cancer effect of Sorafenib on Ehrlich ascites carcinoma and ameliorates the associated liver damage. Sci Rep. 2022:12(1):6494. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref57] 57. Mohammed FZ, Tousson E, Yassin F. Anticancer activity and antioxidant potential of crocus Sativus. L (saffron) aqueous extract against Ehrlich ascites carcinoma cells in Swiss albino mice. Biochem Lett. 2020:16(1):20–49. [Google Scholar]

[ref58] 58. Wen Z-S, Liu L-J, Qu Y-L, OuYang X-K, Yang L-Y, Xu Z-R. Chitosan nanoparticles attenuate hydrogen peroxide-induced stress injury in mouse macrophage RAW264. 7 cells. Mar Drugs. 2013:11(10):3582–3600. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref59] 59. Al-Eisa RA. Synergistic antioxidant capacity of chitosan nanoparticles and lycopene against aging hepatotoxicity induced by D-galactose in male rats. Int J Pharm. 2018:14(6):811–825. [Google Scholar]

[ref60] 60. Jafernik K, Ładniak A, Blicharska E, Czarnek K, Ekiert H, Wiącek AE, Szopa A. Chitosan-based nanoparticles as effective drug delivery systems—a review. Molecules (Basel, Switzerland). 2023:28(4):1963. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref61] 61. Ma Z, Lim L-Y. Uptake of chitosan and associated insulin in Caco-2 cell monolayers: a comparison between chitosan molecules and chitosan nanoparticles. Pharm Res. 2003:20(11):1812–1819. [DOI] [PubMed] [Google Scholar]

[ref62] 62. Huang M, Khor E, Lim L-Y. Uptake and cytotoxicity of chitosan molecules and nanoparticles: effects of molecular weight and degree of deacetylation. Pharm Res. 2004:21(2):344–353. [DOI] [PubMed] [Google Scholar]

[ref63] 63. Qi L-F, Xu Z-R, Li Y, Jiang X, Han X-Y. In vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells. World J Gastroenterol. 2005:11(33):5136–5141. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref64] 64. Li C, Meng M, Guo M, Wang M, Ju A, Wang C. The polysaccharides from Grifola frondosa attenuate CCl 4-induced hepatic fibrosis in rats via the TGF-β/Smad signaling pathway. RSC Adv. 2019:9(58):33684–33692. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref65] 65. Yeh J-Y, Hsieh L-H, Wu K-T, Tsai C-F. Antioxidant properties and antioxidant compounds of various extracts from the edible basidiomycete Grifola frondosa (Maitake). Molecules. 2011:16(4):3197–3211. [DOI] [PMC free article] [PubMed] [Google Scholar]

[ref66] 66. Bhattacharya S, Prasanna A, Majumdar P, Kumar RS, Haldar PK. Antitumor efficacy and amelioration of oxidative stress by Trichosanthes dioica root against Ehrlich ascites carcinoma in mice. Pharm Biol. 2011:49(9):927–935. [DOI] [PubMed] [Google Scholar]

[ref67] 67. Patra S, Muthuraman MS, Prabhu A, Priyadharshini RR, Parthiban S. Evaluation of antitumor and antioxidant activity of Sargassum tenerrimum against Ehrlich ascites carcinoma in mice. Asian Pac J Cancer Prev. 2015:16(3):915–921. [DOI] [PubMed] [Google Scholar]

[ref68] 68. El Sayed RAA, Hanafy ZEM, Abd El Fattah HF, Mohamed AK. Possible antioxidant and anticancer effects of plant extracts from Anastatica hierochuntica, Lepidium sativum and Carica papaya against Ehrlich ascites carcinoma cells. Cancer Biol Ther. 2020:10(1):1–16. [Google Scholar]

[ref69] 69. Hashem MA, Mahmoud EA, Abd-Allah NA. Alterations in hematological and biochemical parameters and DNA status in mice bearing Ehrlich ascites carcinoma cells and treated with cisplatin and cyclophosphamide. Comp Clin Pathol. 2020:29(2):517–524. [Google Scholar]

[ref70] 70. Saleh H, El-Shorbagy HM. Chitosan protects liver against ischemia-reperfusion injury via regulating Bcl-2/Bax, TNF-α and TGF-β expression. Int J Biol Macromol. 2020:164:1565–1574. [DOI] [PubMed] [Google Scholar]

[ref71] 71. DeVita V, Lawrence T, Rosenberg S. Chapter 8. Ljungman M, editor. In: Cancer: Principles & Practice of Oncology. 10th ed. Wolters Kluwer Health; 2015. [Google Scholar]

[ref72] 72. Lucken-Ardjomande S, Martinou J-C. Regulation of Bcl-2 proteins and of the permeability of the outer mitochondrial membrane. C R Biol. 2005:328(7):616–631. [DOI] [PubMed] [Google Scholar]

PERMALINK

Chitosan and Grifola Frondosa nanoparticles insulate liver dysfunction in EAC-bearing mice

Aliaa M Radwan

Doaa T Gebreel

Sahar Allam

Afaf El-Atrash

Ehab Tousson

Abstract

Background

Methods

Results

Conclusions

Introduction

Materials and methods

Animals and experimental design

Table 1.

Blood collection and tissue sampling

Biochemical parameters

Oxidative stress and antioxidant parameters

DNA fragmentation analysis

Histopathological assessment

Immunohistochemical detection of p53 and Bcl-2 expression

Statistical assessment

Results

CS NPs and GF-loaded casein NPs improve liver functions in EAC-bearing mice

Fig. 1.

CS NPs and GF-loaded casein NPs reduce LDL, cholesterol, and triglycerides level in EAC-bearing mice

Fig. 2.

CS NPs and GF-loaded casein NPs attenuate oxidative stress and enhance antioxidant system in liver tissues of EAC-bearing mice

Table 2.

CS NPs and GF-loaded casein NPs reduce DNA fragmentation in liver tissues of EAC-bearing mice

Fig. 3.

CS NPs and GF-loaded casein NPs improve hepatic structure in EAC-bearing mice

Fig. 4.

CS NPs and GF-loaded casein NPs upregulate Bcl-2 and downregulate p53 expression in liver tissues of EAC-bearing mice

Fig. 5.

Fig. 6.

Discussion

Conclusion

Acknowledgments

Contributor Information

Author contributions

Funding

Data availability

Ethical approval

References

Associated Data

Data Availability Statement

ACTIONS

PERMALINK

RESOURCES

Similar articles

Cited by other articles

Links to NCBI Databases