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Back to Journal »International Journal of Nanomedicine» Volume 16
The role of oxidative stress in the cytotoxicity and apoptosis of CHANG and HuH-7 cells induced by La2O3 nanoparticles
Author Almukhlafi H, Ali D, Almutairi B, Yaseen KN, Alyami N, Almeer R, Alkahtani S, Alarifi S
Published on May 20, 2021, the 2021 volume: 16 pages 3487-3496
DOI https://doi.org/10.2147/IJN.S302478
Single anonymous peer review
Editor approved for publication: Prof. Dr. Anderson Oliveira Lobo
Hanouf Almukhlafi, Daoud Ali, Bader Almutairi, Khadijah N Yaseen, Nouf Alyami, Rafa Almeer, Saad Alkahtani, Saud Alarifi Department of Zoology, King Saud University Faculty of Science, Riyadh, Saudi Arabia Communications: Saud Alarifi Department of Zoology, Faculty of Science, King Saud University, BOX 2455, Riyadh, 11451, Saudi Arabia Tel 966 11 4679816 Fax 966 11 4678514 Email [email protected] Introduction: Nanoparticles are widely used in pharmaceuticals, agriculture, food processing industry and many other fields. At present In experiments, we have determined the toxicity mechanism of lanthanum oxide nanoparticles (La2O3 NPs) to human liver cell lines. Methods: Before the study, we used dynamic light scattering (DLS) and transmission electron microscopy (TEM) to characterize the size and shape of La2O3 NPs. La2O3 NPs have an average size of 32 ± 1.6 nm and are in the form of flakes. Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays to determine La2O3 NPs 24 hours to CHANG and Cytotoxicity of HuH-7 cells. Results: Cytotoxicity was observed in both cells in a concentration-dependent manner, but the toxicity of NPs to HuH-7 was greater than that of CHANG cells. The fluorescent dye 2',7'-dichlorofluorescein diacetate (DCFDA) was used to measure the production of reactive oxygen species (ROS), and a higher green fluorescence was observed in HuH-7 cells than in CHANG cells. Compared with CHANG cells, glutathione (GSH) and other biomarkers of oxidative stress were reduced in HuH-7 cells, and the antioxidant enzyme superoxide dismutase (SOD) was increased, but the level of SOD was decreased. The apoptotic cells were determined by using fluorescence activated cell sorting (FACS) analysis. The largest percentage of apoptotic cells observed in HuH-7 cells was 300 μg/mL. DNA double-strand breaks were observed by comet test. Within 24 hours of 300 μg/mL La2O3 NPs, CHANG cells had the most DNA damage than HuH-7 cells. Conclusion: Therefore, this study shows that La2O3 NPs are toxic to human hepatocytes and are more toxic in HuH-7 cells than CHANG cells. Keywords: La2O3 NPs, apoptosis, oxidative stress, CHANG and HuH-7 cells
Bangladesh, India, China, Pakistan and other Asian countries have abundant rare earth metal resources. These rare earth metals enter water bodies and the environment during smelting and mining, causing pollution. Nowadays, different nanotechnology technologies are used to manufacture various types of nanomaterials. In particular, rare earth elements such as lanthanum are widely used in micro-fertilizers and micro-feeds in agriculture, animal husbandry and aquaculture. 1-3 Lanthanum accumulates in the liver of normal and uremic rats. 4 Lacour et al. 5 Exposure to lanthanum carbonate can accumulate in the body and induce potential liver toxicity. Producing eco-friendly nanomaterials is a huge challenge, and this challenge has made nanotechnology one of the most researched and well-funded fields in the past few decades. Metal nanomaterials are used as basic materials for cosmetics and skin care products. At present, rare earth metals are mainly used in animal husbandry, agriculture, and health industries. 6 Due to the wide application of rare earth metals in various fields, it enters the environment, animal and nutrition levels, and enters the human body. Chen et al.7 reported that the chronic bioaccumulation of heavy metals or rare earth metals in the human body can lead to teratogenicity and reproductive toxicity. Brabu et al. 8 reported the in vitro and in vivo biocompatibility of La2O3NPs. Lanthanum chloride and samarium trinitrate can reduce sperm count, fertility and sperm deformity. 9 Rare earth metals can reduce superoxide dismutase, glutathione and glutathione peroxidase, and increase the level of malondialdehyde and apoptosis activity during spermatogenesis. 10 Excessive ROS production in cells can induce toxic cells. Schumacker et al.11 reported that mitochondrial dysfunction occurred due to excessive ROS. The generation of ROS and apoptosis are the potential mechanisms of NPs toxicity. 12 Zhuang et al. 13 reported that due to excessive ROS production, different enzyme activities and induction of lipid peroxidation were observed in cells. Studies have shown that oxidative stress is usually related to the induction of inflammation. As far as I know, no studies have confirmed the harmful effects of La2O3 NPs on human liver cells. The purpose of this study is to study the toxic effects of La2O3 NPs on human normal cells and cancer cells.
Lanthanum oxide (La2O3) nanoparticles (La2O3NPs, 99.99%, 10-100 nm, Stock#: US3265) were purchased from US Research Inc. Houston TX USA. MTT, H2-DCFH-DA), DMSO and Annexin V FITC were purchased from Sigma-Aldrich (St. Louis, Missouri, USA). DMEM and fetal bovine serum were purchased from Gibco, USA.
Using scanning electron/transmission electron microscope (SEM/TEM) (JEOL Inc., Tokyo, Japan) and X-ray diffraction (XRD) (Rigaku, Tokyo, Japan) operating at 200 kV at 9 kW and plus Smart Lab Guidance software (Smart Lab Studio II software package). As mentioned, the average hydrodynamic size and zeta potential of dH2O and La2O3NPs in the culture medium were observed by DLS (Nano-Zeta Sizer-HT, Malvern, UK). 14 We used a 300 μg/mL La2O3NPs suspension for DLS measurement, which is the maximum exposure concentration used to assess cell viability.
CHANG and HuH-7 cells were purchased from the American Type Culture Collection (ATCC). These cells were subcultured in DMEM containing 10% FBS and 10,000 U/mL antibiotics in a 5% CO2 incubator at 37°C.
CHANG and HuH-7 cells were subcultured overnight before being exposed to La2O3NPs. The stock solution of La2O3 NPs was prepared in double-distilled water at a ratio of 1 mg La2O3 NPs/mL DDW and diluted according to the experimental dose (0–400 µg/mL). Control cells were not exposed to NPs and were regarded as controls for each experiment.
After 24 hours of incubation with La2O3NPs in CHANG and Huh-7 cells, mitochondrial activity was determined by MTT test. Mix 15 MTT solution (100 µL) into each well at a final concentration of 0.5 mg/mL, and then continue to incubate for an additional 3.5 hours. The obtained formazan was diluted in dimethyl sulfoxide (DMSO), and the absorbance was measured at 570 nm using a BioTek Epoch plate reader (BioTek Instruments, Winooski, VT, USA) and Gen5 software (version 1.09).
The lactate dehydrogenase (LDH) technology is used as described. 16 The cells were treated with different concentrations of La2O3NPs for one day. After treatment, the culture plate was centrifuged at 1200 rpm for 10 minutes. Then remove the supernatant (100 µL) from the culture plate. LDH activity was detected in 1.0 mL reaction mixture containing 100 µL pyruvate (2.5 mg/mL phosphate buffer) and 100 µL reduced nicotinamide adenine dinucleotide (NADH; 2.5 mg/mL phosphate buffer), AndThe remaining volume is adjusted with phosphate buffer (0.1 mol/L, pH 7.4). The rate of NADH oxidation was determined by using a spectrophotometer (Varian-Cary 300 Bio) at 25°C to track the decrease in absorbance at 340 nm at 3 minute intervals. The amount of LDH released is expressed in terms of LDH activity (IU/L) in the culture medium.
In order to study the ability of La2O3NPs to penetrate into CHANG and HuH-7 cells within 24 hours, analysis was carried out by transmission electron microscopy (TEM). The cells (3×104) were seeded in a cell culture dish (35mm) for 24 hours. The suspension of La2O3 NPs was exposed for 24 hours. Shang et al.17 reported that the higher the exposure concentration of nanoparticles, the less absorption of nanoparticles. Therefore, we chose a medium concentration (100 µg/mL) of La2O3 NPs for the absorption test. CHANG and HuH-7 cells were incubated with La2O3 NPs under the same environmental conditions for 24 hours.
After exposure, we fixed the cells in 2.7% glutaraldehyde for 60 minutes and prepared cell slices according to the method of Ciorîță et al. 18 Place the slice on a copper net and check with TEM Jeol JEM 1010 (JEOL, Tokyo, Japan), run at 80 kV, and use it with a Mega View III digital camera. Confirm whether the dense accumulation of electrons seen in the cell is La2O3 NPs.
After 24 hours of exposure to La2O3 NPs (0, 10, 20, 50, 100, and 300 µg/mL), the production of intracellular ROS in CHANG and HuH-7 cells was measured as described. 19
After 24 hours of NPs treatment, the cells were collected with 0.25% trypsin and disrupted by ultrasound. After that, cell lysates were used to detect oxidative stress parameters such as reduced glutathione (GSH) and superoxide (SOD) according to the following methods. Using BSA as the standard, the amount of protein was determined using the method described by Bradford20.
The GSH level was measured using the method of Alarifi et al. 21 The assay mixture contains phosphate buffer, 5,5-dithiobis-(2-nitrobenzoic acid) and cell extracts. Monitor the reaction at 412 nm and express the amount of GSH as GSH mM/mg protein.
Perform SOD testing according to this method. 21 The assay mixture contains 50 mM Na2CO3, 1.6 mM NBT, Triton x-100 (10%) and 100 mM hydroxylamine-HCl and cell lysates, and the optical density is determined to be at 560 nm.
After exposing La2O3 NPs (0, 10, 20, 50, 100 and 300 µg/mL) in CHANG and HuH-7 cells using a confocal microscope (ZEISS LSM), AO and EtBr staining were used to detect apoptotic and necrotic cells 900) .
La2O3 NP (0, 10, 20, 50, 100 and 300 µg/mL) was exposed to CHANG and HuH-7 for 24 hours. The cells were washed with cold PBS and collected in Eppendorf tubes and centrifuged at 1000 rpm for 5 minutes. The cell pellet is mixed with 5 µL Annexin-V FITC and 10 µL PI in binding buffer (485 µL). The cell suspension was incubated at room temperature in the dark for 30 minutes. After incubation, the cell suspension was detected by flow cytometry (Becton-Dickinson Immunocytometry Systems, Sunnyvale, CA, USA). The fluorescence of FITC bound to Annexin-V and propidium iodide bound to DNA was detected as red fluorescence in each event. The results were analyzed by FACS Diva 6.1.2 software.
Use comet assay to determine DNA damage in CHANG and HuH-7 cells. 22,23
The data was analyzed using SPSS 26.0 software (IBM) and expressed as mean ± standard error (SE). The statistical difference between the control group and the exposed group was determined by one-way analysis of variance with the least significant difference test. * A p value of <0.05 is considered statistically significant.
La2O3NP was purchased from American Research Nanomaterials Corporation (Houston, Texas, USA) and has been characterized by SEM, TEM, XRD and DLS methods. The nanoparticles are inhomogeneous sheet-like structures in shape, and show the aggregation mode determined by TEM. Figures 1A and B show the SEM and TEM images of La2O3 nanoparticles, respectively. The shape of La2O3NPs is a sheet-like structure (Figure 1A and B). The average size of NP is 32 ± 1.6 nm (Figure 1C). Figure 1D shows the XRD spectrum of La2O3 NPs with a purity of 99.5%. After La2O3 NPs were suspended in Milli Q water and DMEM, their size was measured using Zetasizer. The size and zeta potential were 296±6.0 nm and ~10.7 ±3.7 mV and 161±9.9 nm and ~11.5 ±4.9 mV, respectively (Table 1) . Table 1 Physical characterization of La2O3 nanoparticles using dynamic light scattering. The data represents the mean ± standard error (± SE) of three independent experiments. Figure 1 (A) SEM image of La2O3NPs (B). TEM image of La2O3NPs (C). Distribution of La2O3NPs in aqueous suspension (D). XRD spectrum of La2O3NPs.
Table 1 Physical characterization of La2O3 nanoparticles using dynamic light scattering. Data represents the average of three independent experiments ± standard error (± SE)
Figure 1 (A) SEM image of La2O3NPs (B). TEM image of La2O3NPs (C). Distribution of La2O3NPs in aqueous suspension (D). XRD spectrum of La2O3NPs.
We have observed the internalization of La2O3 NPs in CHANG, HUH-7 cells were treated with 100 μg/mL for 24 hours, and untreated cells were used as a control using TEM. La2O3 NPs enter the cell due to their small size and form clusters of NPs inside the cell (Figure 2A-D). Figure 2 TEM micrograph taken by La2O3 NPs cells (A). CHANG control (B). CHANG was maintained at 100 μg/mL La2O3 NPs for 24 hours (C). HUH-7 control (D). HUH-7 was treated with 100 μg/mL La2O3 NPs for 24 hours. Black arrow = small NP polymer.
Figure 2 TEM micrograph taken by La2O3 NPs cells (A). CHANG control (B). CHANG was maintained at 100 μg/mL La2O3 NPs for 24 hours (C). HUH-7 control (D). HUH-7 was treated with 100 μg/mL La2O3 NPs for 24 hours. Black arrow = small NP polymer.
We measured the cell viability of CHANG and HuH-7 cells after being exposed to La2O3 NP (0, 10, 20, 50, 100, and 300 µg/mL) for 24 hours. The cytotoxicity results are shown in Figure 3A and B. La2O3NPs induced cytotoxicity in CHANG and HuH-7 cells in a dose-dependent manner. At a higher dose of 300 µg/mL, the toxic effects of La2O3 NPs observed in CHANG cells were stronger than in HuH-7 cells (Figure 3A). Figure 3 24-hour cytotoxicity of La2O3NPs to CHANG and HuH-7 cells, as assessed in (A). MTT (B). LDH detection. Each value represents the mean ± SE of three experiments. n = 3, *p <0.05 compared with control.
Figure 3 24-hour cytotoxicity of La2O3NPs to CHANG and HuH-7 cells, as assessed in (A). MTT (B). LDH detection. Each value represents the mean ± SE of three experiments. n = 3, *p <0.05 compared with control.
The effect of La2O3 NPs on the plasma membrane of two types of cells was observed by using the LDH test. As the concentration of NPs increases, the leakage of LDH enzyme increases. Therefore, it was confirmed that La2O3NPs deteriorated the cell membrane, resulting in cell apoptosis and cytotoxicity (Figure 3B). The cytotoxicity patterns in both cells showed irregular patterns (Figure 3A and B).
Oxidative stress was determined by measuring ROS, GSH and SOD enzymes in CHANG and HuH-7 cells. The formation of ROS in cells is determined by capturing DCF fluorescence. Compared with HuH-7 cells (Figure 4A and K), in CHANG cells (Figure 4A and E), the green fluorescence intensity observed in 100 µg/mL La2O3 NPs is higher. The intensity of DCF fluorescence (green) increases and decreases irregularly (Figure 4A-K). GSH and SOD were measured and statistically analyzed with control cells. GSH levels decreased after exposure to La2O3NPs, and the maximum deflection angle was found to be 300 µg/mL La2O3NPs in CHANG cells (Figure 5A). The SOD levels in the two cells increased irregularly (Figure 5B). Figure 4 ROS production in CHANG and HuH-7 cells 24 hours after La2O3NPs exposure (A). 24 hours after exposure to La2O3NPs, the percentage of DCF fluorescence intensity and green fluorescence in CHANG and HuH-7 (B). Control CHANG cells (C). CHANG cells at a concentration of 20 μg/mL (D). CHANG cells at a concentration of 50 μg/mL (E). CHANG cells are at 100 μg/mL (F). CHANG cells, the concentration is 300 μg/mL (G). Control HuH-7 cells (H). HuH-7 cells at a concentration of 20 μg/mL (I). HuH-7 cells at a concentration of 50 μg/mL (J). HuH-7 cells at a concentration of 100 μg/mL (K). HuH-7 cells, the concentration is 300 μg/mL. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control. Figure 5 La2O3NPs 24 hours after exposure to CHANG and HuH-7 (A). The level of GSH (B). SOD in the cell. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Figure 4 ROS production in CHANG and HuH-7 cells 24 hours after La2O3NPs exposure (A). 24 hours after exposure to La2O3NPs, the percentage of DCF fluorescence intensity and green fluorescence in CHANG and HuH-7 (B). Control CHANG cells (C). CHANG cells at a concentration of 20 μg/mL (D). CHANG cells at a concentration of 50 μg/mL (E). CHANG cells are at 100 μg/mL (F). CHANG cells, the concentration is 300 μg/mL (G). Control HuH-7 cells (H). HuH-7 cells at a concentration of 20 μg/mL (I). HuH-7 cells at a concentration of 50 μg/mL (J). HuH-7 cells at a concentration of 100 μg/mL (K). HuH-7 cells, the concentration is 300 μg/mL. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Figure 5 La2O3NPs 24 hours after exposure to CHANG and HuH-7 (A). The level of GSH (B). SOD in the cell. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Using FACS to evaluate the effects of NPs on the apoptosis and necrosis of CHANG and HuH-7, we found that under 300 µg/mL La2O3 NPs, 49% of CHANG cells and 32% of HuH-7 cells Of apoptotic cells (Figure 6K). We have observed the necrotic effect of NPs in 100 and 300 µg/mL La2O3 NPs in CHANG cells (Figure 6K). We have shown in Figure 6A-J. The statistical analysis of early apoptotic and necrotic cells is determined by (FACS), as shown in Figure 6K. Figure 6 24 hours scatter plot of apoptosis and necrosis of CHANG and HuH-7 after La2O3NPs exposure (A). Control the CHANG cell (B). CHANG cells at a concentration of 20 μg/mL (C). CHANG cells at a concentration of 50 μg/mL (D). CHANG cells at 100 μg/mL (E). CHANG cells are at 300 μg/mL (F). Control HuH-7 cells (G). HuH-7 cells at a concentration of 20 μg/mL (H). HuH-7 cells at a concentration of 50 μg/mL (I). HuH-7 cells at a concentration of 100 μg/mL (J). HuH-7 cells, the concentration is 300 μg/mL. (K) The percentages of CHANG and HuH-7 of apoptosis and necrosis 24 hours after La2O3NPs exposure. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Figure 6 24 hours scatter plot of apoptosis and necrosis of CHANG and HuH-7 after La2O3NPs exposure (A). Control the CHANG cell (B). CHANG cells at a concentration of 20 μg/mL (C). CHANG cells at a concentration of 50 μg/mL (D). CHANG cells at 100 μg/mL (E). CHANG cells are at 300 μg/mL (F). Control HuH-7 cells (G). HuH-7 cells at a concentration of 20 μg/mL (H). HuH-7 cells at a concentration of 50 μg/mL (I). HuH-7 cells at a concentration of 100 μg/mL (J). HuH-7 cells, the concentration is 300 μg/mL. (K) The percentages of CHANG and HuH-7 of apoptosis and necrosis 24 hours after La2O3NPs exposure. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Figures 7A and B show the apoptotic and necrotic cells in CHANG and HuH-7 by staining AO/EtBr, and the images were captured by a confocal microscope. The percentages of apoptotic and necrotic cells are shown in Figure 7C. The largest apoptotic cells were observed in HuH-7 cells at 300 g/mL (Figure 7C). Figure 7 After exposure to La2O3NPs, apoptosis and necrosis cells were induced in CHANG and HuH-7 for 24 hours (A). 24 hours after exposure to La2O3NPs, AO/Etbr stained fluorescence in CHANG cells (B). 24 hours after exposure to La2O3NPs (C), AO/Etbr stained fluorescent HuH-7 cells. The percentage of apoptotic cells in CHANG and HuH-7 cells after 24 hours of exposure to La2O3NPs. Each value represents the mean ± SE of three experiments. * p <0.05 compared with control cells. Arrows indicate apoptotic cells.
Figure 7 After exposure to La2O3NPs, apoptosis and necrosis cells were induced in CHANG and HuH-7 for 24 hours (A). 24 hours after exposure to La2O3NPs, AO/Etbr stained fluorescence in CHANG cells (B). 24 hours after exposure to La2O3NPs (C), AO/Etbr stained fluorescent HuH-7 cells. The percentage of apoptotic cells in CHANG and HuH-7 cells after 24 hours of exposure to La2O3NPs. Each value represents the mean ± SE of three experiments. * p <0.05 compared with control cells. Arrows indicate apoptotic cells.
DNA fragmentation was observed in CHANG and HuH-7 cells (Figure 8A-K). DNA fragmentation was found in both cells in a dose-dependent manner. The largest DNA damage was found in HuH-7 cells at a concentration of 300 µg/mL La2O3 NP (Figure 8J and K). Under the conditions of 20 and 50 µg/mL La2O3 NPs, the incidence of DNA damage in CHANG cells was higher than that in HuH-7 cells. At 100 µg/mL La2O3 NPs, the DNA damage of the two cells is slightly equal (Figure 8D, I, and K). Compared with the control group, the DNA damage of La2O3 NPs was significantly increased in CHANG and HuH-7 cells (p <0.05) (Figure 8A-K). Figure 8 DNA damage in CHANG and HuH-7 after exposure to La2O3NPs for 24 hours (A). Control the CHANG cell (B). CHANG cells at a concentration of 20 μg/mL (C). CHANG cells at a concentration of 50 μg/mL (D). CHANG cells at 100 μg/mL (E). CHANG cells are at 300 μg/mL (F). Control HuH-7 cells (G). HuH-7 cells at a concentration of 20 μg/mL (H). HuH-7 cells at a concentration of 50 μg/mL (I). HuH-7 cells at a concentration of 100 μg/mL (J). HuH-7 cells, the concentration is 300 μg/mL. (K) The percentage of DNA damage in the tail of CHANG and HuH-7 within 24 hours after exposure to La2O3NPs. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Figure 8 DNA damage in CHANG and HuH-7 after exposure to La2O3NPs for 24 hours (A). Control the CHANG cell (B). CHANG cells at a concentration of 20 μg/mL (C). CHANG cells at a concentration of 50 μg/mL (D). CHANG cells at 100 μg/mL (E). CHANG cells are at 300 μg/mL (F). Control HuH-7 cells (G). HuH-7 cells at a concentration of 20 μg/mL (H). HuH-7 cells at a concentration of 50 μg/mL (I). HuH-7 cells at a concentration of 100 μg/mL (J). HuH-7 cells, the concentration is 300 μg/mL. (K) The percentage of DNA damage in the tail of CHANG and HuH-7 within 24 hours after exposure to La2O3NPs. Each value represents the mean ± SE of three experiments. * p <0.05 vs. control.
Due to advanced technology, nanomaterials have now become an important part of our daily lives, and they are both good and bad for human health. With the increasing application of La2O3 NPs in daily-use products in recent decades, they are inevitably discharged into the natural environment. 25 In this experiment, we observed the toxicity potential of La2O3 NPs on CHANG and HuH-7 cells. Before La2O3 NPs were exposed to cells, we have used TEM, XRD and Zetasizer to characterize the nature and size of the nanoparticles. TEM analysis found that the average size of NP was 32 nm, with a sheet-like structure shape (Figure 1A and B). In this study, ROS played a major role in inducing the toxicity and apoptosis of CHANG and HuH-7 cells. We observed the biodistribution and internalization of toxic La2O3 NPs in CHANG (Figure 2B) and HuH-7 cells (Figure 2C). Su et al. 26 reported that La3 ion induces apoptosis in cancer cells. We have observed the toxic effects of La2O3 NPs at all concentrations in the two cells, but the maximum toxicity in CHANG cells is greater than that in HuH-7 cells at 300 µg/mL for 24 hours. The current findings are consistent with the findings of Sambale et al.27 regarding the toxicity of silver nanoparticles in mammalian cell lines. Therefore, La2O3 NPs exposure outperformed in reducing the activity and leakage of lactate dehydrogenase in CHANG and HuH-7 cells. Some researchers have reported the phytotoxicity of La2O3 NPs to Cucumis sativus L.28. In this experiment, we studied the production of intracellular reactive oxygen species, which increased in a dose-dependent manner, followed by a higher ROS production in CHANG cells compared to HuH-7 cells. As we discovered in our experiments, the effect of ROS may be due to a pathway mediated by mitochondria. Victor et al. 29 reported that in the extrinsic pathway of apoptosis, ROS has not been approved as an activator due to lanthanum NP. Due to nanoparticles, cell apoptosis takes place in an intrinsic way. 30 NP produces free radicals and degrades cells by producing reactive oxygen species. We have used flow cytometry to confirm the apoptotic response of La2O3 NPs. Apoptosis and necrotic cells are induced at higher concentrations of NPs.
In summary, we observed that the toxicity potential of La2O3 NPs depends on the cell type of human hepatocytes. La2O3 NPs exhibit cytotoxicity and apoptosis, which is probably due to their size effect and the induction of ROS and oxidative stress. The most important finding of this study is that La2O3 NPs passively internalize into these hepatocytes and induce toxicity. Based on our findings, we observed that HuH-7 cells are more sensitive to La2O3 NPs than CHANG cells. In my further study, I will further study the toxicity of La2O3 NPs in vivo model.
The author would like to thank the Dean of the Institute of Science at King Saud University for funding this work through the No (RG-1441-180) research group.
The authors report no conflicts of interest in this work.
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