الفهرس 
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Abstract
According to current statistics, cancer is Cancer is second-leading cause of mortality throughout the world. OS is an exceptionally uncommon type of bone malignancy, while being the most prevalent type of malignant bone tumor in youngsters and having a lower incidence in the elderly. OS is a malignant bone tumor with an unclear cause. It contains a variety of subtypes that reveal the presence of pleomorphic cancerous cells that are thought to be the source of osteoid. The main causes of cancer recurrence are cancer cell migration and metastasis. Recurrence occurs in one-third of instances of OS, most commonly as osteolytic bone metastases.
To avoid the adverse effects of surgery, chemotherapeutic drugs, and radiation therapy in the treatment of osteosarcoma, current research has preferred to employ natural products, herbal plant extracts and phytoconstituents with anticancer characteristics have recently been applied to the treatment of OS.
The aim of the present study was to investigate the effect of vitamin C, green tea and their combination on Mg-63 OS cell line after 72 hours.
In this study, we used various assays to confirm the impacts of vitamin C, green tea, and their combination on (Mg-63) cell line cells. The following was the order of these assays: SRB, FC, cytological and morphological analyses, wound healing assay, anti-oxidant test, and statistical analysis all included.
According to the SRB cytotoxic assay, the combination had a greater cytotoxic effect than vitamin C, and the least effect was green tea-treated cells after 72 hours. The FC assay demonstrated that all treated cells in various groups were in cell cycle arrest at Pre-G1 in comparison to the control group. Green tea treated cells caused arrest cell cycle at G2 /M.
The combination had the greatest impact on the cell cycle, followed by vitamin C and finally green tea-treated cells when compared to control cells, the percentages of cells in pre-G1 are increased in all three drugs, with a greater effect in the combination treated cells than in vitamin C and green tea.
Apoptosis assessment explained that all Mg-63 cells treated with vitamin C, green tea and their combination at IC50 after 72 hours induced apoptosis and necrosis but at different levels. The apoptosis assessment revealed that the combination caused an increase in total percentage necrosis, followed by vitamin C and finally green tea green tea treated cells.The percentage of early apoptotic cells was increased with combination more than vitamin C and green tea. The total percentage of apoptosis and necrosis increased with vitamin C treated cells than combination and finally green tea. In the autophagy cell death test, vitamin C and combination increased autophagy cell death with the highest rate to vitamin C more than combination, while autophagy cell death was decreased with green tea treated cells. All these results indicated that combination treated cells were more effective in treating Mg-63 cells followed by vitamin C and finally green tea treated cells by apoptosis and necrosis induction. Autophagy cell death increased more in vitamin C-treated cells than in combination and decreased with green tea treated cells with IC50 after 72 hours.
With wound healing assay, vitamin C, green tea and their combination treated cells inhibited the migration of Mg-63 treated cells after 72 hours but to varying degrees in relation to control groups.
An antioxidant DPPH experiment showed that cells treated with vitamin C were significantly more efficient than cells treated with combinations and green tea. Green tea was the least effective when compared to other previous drugs. This demonstrated that vitamin C alone had a stronger antioxidant capacity than both the mixture and green tea. Vitamin C has a greater antioxidant capacity than trolox.
In our study, we determined and confirmed that vitamin C, green tea and their combination had cytotoxic effects on Mg-63 cells, as well as impacted on cell cycle distribution, apoptosis, necrosis, and autophagy. Cell migration assay had been applied to malignant Mg-63cell line cells which had already been treated with our drugs, and it was discovered that all three natural compounds had their impacts, but to varying degrees. An anti-oxidant test on three drugs indicated that vitamin C has a higher anti-oxidant capability than green tea and a combination. Statistical analysis was compatible with our verified results.