Volume 28, Issue 9 (12-2021)                   RJMS 2021, 28(9): 31-44 | Back to browse issues page

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Ethics code: EE.97.243.93365/scu.ac.ir
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hoveizi E, abdolali nejad P. Evaluation of Graphene Quantum Dots Cytotoxicity on Breast Cancer MCF-7 Cells. RJMS 2021; 28 (9) :31-44
URL: http://rjms.iums.ac.ir/article-1-6364-en.html
Shahid Chamran University of Ahvaz , e.hoveizi@yahoo.com
Abstract:   (657 Views)
Background & Aims: Breast cancer is one of the most common cancers today and how to deal with it is one of the challenges for oncologists today (1). One of the new treatment strategies is the use of nanotechnology in the field of cancer (1, 2). Nanoparticles are the most common elements in nanoscience and technology, whose interesting properties have led to a wide variety of applications (5). In this regard, graphene quantum dots (GQDs) can be used as an excellent option for nanomedical fields such as the release of anti-cancer drugs, cell culture, and tissue engineering (7, 8). Considering the role of nanoparticles as well as the antitumor effects of graphene compounds, it seems the use of these nanoparticles in 3D culture can be one of the useful strategies in suppressing cancer cells with fewer side effects (9). Therefore, the aim of this project was to compare the apoptotic effects of graphene quantum dots on the MCF-7 breast cancer cell line cultured in hydrogel scaffolds.
Methods: In this study, the MCF-7 breast cancer cell line was used as  cancer cells. For cell culture in fibrin hydrogel scaffold, M199 medium containing 10% FBS serum and 1% penicillin/streptomycin and fibrinogen powder with a concentration of 3 mg/ml was used and it was added into each plate well of 24 wells and then cell suspension was added. Then, for 500 μl of scaffolding and cells, 15 μl of thrombin with a concentration of 120 u/ml was added in each well and it was immediately gently aspirated until the culture medium became gelatinous and the cell culture plate was transferred to the incubator. After 2 hours of cell incubation in the incubator (Sina, Iran), On the gelatinous scaffolding of each 24 well plates, 500 μl of M199 medium containing 10% FBS serum and 1% antibiotic was added and transferred to the incubator again. Cell treatment was performed by first removing the culture medium from each well and concentrations of 1, 5, 7, and 10 mg/ml of graphene quantum dots were added. But only the culture medium was added to the control group. The plate was transferred to the incubator and treated for 1, 3, and 5 days. In this study, MCF-7 cells were cultured on a fibrin hydrogel scaffold for SEM imaging and after 24 hours of incubation, the medium was gently removed on each well. 300 μl of PBS was added to each 24-well plate and wash three times and 300 μl of 2.5% glutaraldehyde solution was added to each well and maintained at room temperature for 2 hours. Dehydration was done with increasing concentration of alcohol (30, 50, 70, 80, 90, 100, 100%) and then close the door of the plate and maintained in the freezer for 2 hours and freeze it to dry and dehydrate the fibrin hydrogel. MTT method and acridine orange and DAPI stains were used to measure cell survival and morphology. One-way ANOVA and t-test were used for statistical analysis. The chart was drawn in Excel 2016. P<0.05 was considered a significant difference for samples.
Results: The results of scaffold morphology study using SEM electron microscope, showing the desired mechanical properties and suitable porosity for the scaffold. Therefore, the presence, growth, proliferation, and proper adhesion of cells to the scaffold and the natural morphology of the cells were confirmed. The results of the MTT test showed that the viability and survival of MCF-7 cells after exposure to the mentioned doses of graphene quantum dots were recorded (98, 81, 50, and 31%) respectively. This indicates that the viability of MCF-7 cells is significantly dependent on the concentration of graphene quantum dots. IC50 concentration of GQDs affecting MCF-7 cells was determined to be 7 mg/ml and after 1, 3, and 5 days decreased in a time-dependent manner so that after 24 hours, the average cell survival decreased to 50%, after 3 days to 35% and after 5 days to 22%. These data also showed the cytotoxic effect of IC50 concentration of graphene quantum dots on cells in a time-dependent manner. Also, the results of acridine and DAPI staining in GQDs-treated groups showed compact, fragmented, single, and orange nuclei, which indicated the onset of apoptosis in MCF-7 cells being treated. While in the control group cells that did not receive GQDs, the nuclei were clear, round, large, and in the form of light green clusters, which was a sign that the control group cells were alive.
Conclusion: In this study, the effects of graphene quantum dots on MCF-7 cancer cells cultured on fibrin hydrogel were investigated. The results showed that GODs had a cytotoxic effect on MCF-7 cancer cells, so these effects were dependent on the time and dose in the experiment. In fact, with increasing concentration and time, the toxicity of graphene quantum dots increased. In this study, the IC50 for GODs was calculated to be 7 mg/ml by MTT test. Examination of morphological changes with an inverted microscope and also, qualitative studies of the nucleus characteristics of control and treatment cells confirmed the induction of apoptosis in the treated cells. In addition, the results of acridine orange staining were another confirmation of these findings. As mentioned, graphene and its many derivatives, as well as graphene quantum dots, which is one of the newest products of graphene, have been used for valuable research. According to the results of this study, GQDs have a toxicity effect and induce dose-dependent and time-dependent apoptosis in cultured breast cancer cells on fibrin hydrogel. However, more studies are needed to determine the molecular mechanism involved. In addition, it is suggested that the effects of GQDs on other cancer cells and normal cell lines be investigated and compared.
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Type of Study: Research | Subject: Biology

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