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Reducing X-Ray Induced Oxidative Damages in Fibroblasts with Graphene Oxide.

Qiao Y, Zhang P, Wang C, Ma L, Su M - Nanomaterials (Basel) (2014)

Bottom Line: A major issue of X-ray radiation therapy is that normal cells can be damaged, limiting the amount of X-rays that can be safely delivered to a tumor.A variety of techniques such as cytotoxicity, genotoxicity, oxidative assay, apoptosis, γ-H2AX expression, and micro-nucleus assay have been used to assess the protective effect of GO in cultured fibroblast cells.Thus, low concentration GO can be used as an effective radio-protective agent in occupational and therapeutic settings.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Engineering, Worcester Polytechnic Institute, Worcester, MA 01609, USA.

ABSTRACT

A major issue of X-ray radiation therapy is that normal cells can be damaged, limiting the amount of X-rays that can be safely delivered to a tumor. This paper describes a new method based on graphene oxide (GO) to protect normal cells from oxidative damage by removing free radicals generated by X-ray radiation using grapheme oxide (GO). A variety of techniques such as cytotoxicity, genotoxicity, oxidative assay, apoptosis, γ-H2AX expression, and micro-nucleus assay have been used to assess the protective effect of GO in cultured fibroblast cells. It is found that although GO at higher concentration (100 and 500 μg/mL) can cause cell death and DNA damage, it can effectively remove oxygen free radicals at a lower concentration of 10 μg/mL. The level of DNA damage and cell death is reduced by 48%, and 39%, respectively. Thus, low concentration GO can be used as an effective radio-protective agent in occupational and therapeutic settings.

No MeSH data available.


Related in: MedlinePlus

Oxidative stress induced by GO and X-ray. Fluorescent images of cells (A); cells treated with 10 μg/mL GO (B); cells exposed to 1.25 Gy X-ray (C); and cells treated with 10 μg/mL GO and then exposed to 1.25 Gy X-ray (D); Flow cytometry results of cells after different treatment: cells (E); cells treated with GO (F); cells exposed to 1.25 Gy X-ray (G); and cells treated with GO and exposed to X-ray (H).
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Figure 4: Oxidative stress induced by GO and X-ray. Fluorescent images of cells (A); cells treated with 10 μg/mL GO (B); cells exposed to 1.25 Gy X-ray (C); and cells treated with 10 μg/mL GO and then exposed to 1.25 Gy X-ray (D); Flow cytometry results of cells after different treatment: cells (E); cells treated with GO (F); cells exposed to 1.25 Gy X-ray (G); and cells treated with GO and exposed to X-ray (H).

Mentions: X-ray radiation can generate ROS [33], which can cause alteration of membrane lipids, proteins, and nucleic acids. The effect of GO in reducing ROS generation is tested as follows. Fibroblasts are treated with 10 μg/mL GO and exposed to 1.25 Gy X-ray radiation, followed by H2DCFDA staining and flow cytometry. In addition, DNA is stained with Hoechst 33342 (Life Technologies, Eugene, OR, USA) for observation of nucleus. Figure 4A,B are fluorescence images of untreated cells and cells treated with 10 μg/mL GO alone, where DNA in nucleus (blue color) can be seen clearly; while carboxy-DCF (green color) cannot be seen due to small amount of ROS. In comparison, cells treated with 1.25 Gy X-ray show strong green fluorescence (Figure 4C). Most importantly, there is weak green fluorescence from cells that are treated with 10 μg/mL GO before exposing to 1.25 Gy X-ray (Figure 4D). Flow cytometry results are shown in Figure 4E–H, where the peaks shift toward the right is proportional to the level of ROS. Compared to untreated cells (4E) and cells treated with GO (4F), cells exposed to X-ray (4G) show a significant shift towards right, suggesting a large population cells have high level of ROS. X-ray irradiation of GO treated cells does not shift the peak much compared to cells alone, or cells treated with 10 μg/mL GO (4H). Taking cells without GO treatment and without X-ray exposure as standard (0.5% ROS), cells treated with GO show 2.5% ROS; cells exposed to X-ray show 99.7% ROS, and cells treated with GO and then exposed to X-ray show 21.2% ROS. These results confirm that GO can significantly reduce X-ray induced ROS generation.


Reducing X-Ray Induced Oxidative Damages in Fibroblasts with Graphene Oxide.

Qiao Y, Zhang P, Wang C, Ma L, Su M - Nanomaterials (Basel) (2014)

Oxidative stress induced by GO and X-ray. Fluorescent images of cells (A); cells treated with 10 μg/mL GO (B); cells exposed to 1.25 Gy X-ray (C); and cells treated with 10 μg/mL GO and then exposed to 1.25 Gy X-ray (D); Flow cytometry results of cells after different treatment: cells (E); cells treated with GO (F); cells exposed to 1.25 Gy X-ray (G); and cells treated with GO and exposed to X-ray (H).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 4: Oxidative stress induced by GO and X-ray. Fluorescent images of cells (A); cells treated with 10 μg/mL GO (B); cells exposed to 1.25 Gy X-ray (C); and cells treated with 10 μg/mL GO and then exposed to 1.25 Gy X-ray (D); Flow cytometry results of cells after different treatment: cells (E); cells treated with GO (F); cells exposed to 1.25 Gy X-ray (G); and cells treated with GO and exposed to X-ray (H).
Mentions: X-ray radiation can generate ROS [33], which can cause alteration of membrane lipids, proteins, and nucleic acids. The effect of GO in reducing ROS generation is tested as follows. Fibroblasts are treated with 10 μg/mL GO and exposed to 1.25 Gy X-ray radiation, followed by H2DCFDA staining and flow cytometry. In addition, DNA is stained with Hoechst 33342 (Life Technologies, Eugene, OR, USA) for observation of nucleus. Figure 4A,B are fluorescence images of untreated cells and cells treated with 10 μg/mL GO alone, where DNA in nucleus (blue color) can be seen clearly; while carboxy-DCF (green color) cannot be seen due to small amount of ROS. In comparison, cells treated with 1.25 Gy X-ray show strong green fluorescence (Figure 4C). Most importantly, there is weak green fluorescence from cells that are treated with 10 μg/mL GO before exposing to 1.25 Gy X-ray (Figure 4D). Flow cytometry results are shown in Figure 4E–H, where the peaks shift toward the right is proportional to the level of ROS. Compared to untreated cells (4E) and cells treated with GO (4F), cells exposed to X-ray (4G) show a significant shift towards right, suggesting a large population cells have high level of ROS. X-ray irradiation of GO treated cells does not shift the peak much compared to cells alone, or cells treated with 10 μg/mL GO (4H). Taking cells without GO treatment and without X-ray exposure as standard (0.5% ROS), cells treated with GO show 2.5% ROS; cells exposed to X-ray show 99.7% ROS, and cells treated with GO and then exposed to X-ray show 21.2% ROS. These results confirm that GO can significantly reduce X-ray induced ROS generation.

Bottom Line: A major issue of X-ray radiation therapy is that normal cells can be damaged, limiting the amount of X-rays that can be safely delivered to a tumor.A variety of techniques such as cytotoxicity, genotoxicity, oxidative assay, apoptosis, γ-H2AX expression, and micro-nucleus assay have been used to assess the protective effect of GO in cultured fibroblast cells.Thus, low concentration GO can be used as an effective radio-protective agent in occupational and therapeutic settings.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Engineering, Worcester Polytechnic Institute, Worcester, MA 01609, USA.

ABSTRACT

A major issue of X-ray radiation therapy is that normal cells can be damaged, limiting the amount of X-rays that can be safely delivered to a tumor. This paper describes a new method based on graphene oxide (GO) to protect normal cells from oxidative damage by removing free radicals generated by X-ray radiation using grapheme oxide (GO). A variety of techniques such as cytotoxicity, genotoxicity, oxidative assay, apoptosis, γ-H2AX expression, and micro-nucleus assay have been used to assess the protective effect of GO in cultured fibroblast cells. It is found that although GO at higher concentration (100 and 500 μg/mL) can cause cell death and DNA damage, it can effectively remove oxygen free radicals at a lower concentration of 10 μg/mL. The level of DNA damage and cell death is reduced by 48%, and 39%, respectively. Thus, low concentration GO can be used as an effective radio-protective agent in occupational and therapeutic settings.

No MeSH data available.


Related in: MedlinePlus