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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

Immunostaining images of cells (A); cells treated with GO (B); cells exposed to X-ray (C); and cells treated with GO and then exposed to X-ray (D); flow cytometry results of cells after different treatments (E–H).
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Figure 3: Immunostaining images of cells (A); cells treated with GO (B); cells exposed to X-ray (C); and cells treated with GO and then exposed to X-ray (D); flow cytometry results of cells after different treatments (E–H).

Mentions: Once damaged, a group of DNA repair proteins accumulate around damaged sites. γ-H2AX is a protein that is related to repairing DNA double strand breaks [31,32]. Detecting γ-H2AX is one way to quantify double strand breaks of DNA. Four samples are studied: (1) cells; (2) cells treated with 10 μg/mL GO; (3) cells exposed to 1.25 Gy X-ray and (4) cells treated with 10 μg/mL GO and then exposed to 1.25 Gy X-ray. DNAs are stained with DAPI for observation of nuclei. Figure 3 shows the immunostaining and flow cytometry results of γ-H2AX expression, where γ-H2AX is shown in green and DNA is shown in blue. Figure 3A,B are fluorescence images of cells and cells treated with 10 μg/mL GO alone, where DNA (blue color) can be seen clearly; while γ-H2AX (green color) cannot be seen due to small amount of γ-H2AX. In comparison, cells treated with 1.25 Gy X-ray has shown strong green fluorescence (3C). There is weak green fluorescence from cells that are treated with 10 μg/mL GO before exposing to 1.25 Gy X-ray (3D). Figure 3E–H shows the flow cytometry results, where the shift of the peak towards the right is proportional to the level of γ-H2AX. Comparing cells alone (3E) and cells treated with GO (3F), cells exposed to X-ray (3G) show a significant shift towards the right, suggesting a large population of cells have high levels of γ-H2AX, as well as double DNS strand breaks. X-ray radiation of GO treated cells does not shift the peak much compared to cells alone, or cells treated with 10 μg/mL GO (3H). Taking cells without GO treatment and without X-ray exposure as standard (0.6% γ-H2AX), cells treated with GO show 0.5% γ-H2AX, cells exposed to X-ray show 94.6% γ-H2AX, and cells treated with GO and then exposed to X-ray show 13.1% γ-H2AX. These results confirm GO can significantly reduce X-ray induced DNA double strand breaks.


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

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

Immunostaining images of cells (A); cells treated with GO (B); cells exposed to X-ray (C); and cells treated with GO and then exposed to X-ray (D); flow cytometry results of cells after different treatments (E–H).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4269382&req=5

Figure 3: Immunostaining images of cells (A); cells treated with GO (B); cells exposed to X-ray (C); and cells treated with GO and then exposed to X-ray (D); flow cytometry results of cells after different treatments (E–H).
Mentions: Once damaged, a group of DNA repair proteins accumulate around damaged sites. γ-H2AX is a protein that is related to repairing DNA double strand breaks [31,32]. Detecting γ-H2AX is one way to quantify double strand breaks of DNA. Four samples are studied: (1) cells; (2) cells treated with 10 μg/mL GO; (3) cells exposed to 1.25 Gy X-ray and (4) cells treated with 10 μg/mL GO and then exposed to 1.25 Gy X-ray. DNAs are stained with DAPI for observation of nuclei. Figure 3 shows the immunostaining and flow cytometry results of γ-H2AX expression, where γ-H2AX is shown in green and DNA is shown in blue. Figure 3A,B are fluorescence images of cells and cells treated with 10 μg/mL GO alone, where DNA (blue color) can be seen clearly; while γ-H2AX (green color) cannot be seen due to small amount of γ-H2AX. In comparison, cells treated with 1.25 Gy X-ray has shown strong green fluorescence (3C). There is weak green fluorescence from cells that are treated with 10 μg/mL GO before exposing to 1.25 Gy X-ray (3D). Figure 3E–H shows the flow cytometry results, where the shift of the peak towards the right is proportional to the level of γ-H2AX. Comparing cells alone (3E) and cells treated with GO (3F), cells exposed to X-ray (3G) show a significant shift towards the right, suggesting a large population of cells have high levels of γ-H2AX, as well as double DNS strand breaks. X-ray radiation of GO treated cells does not shift the peak much compared to cells alone, or cells treated with 10 μg/mL GO (3H). Taking cells without GO treatment and without X-ray exposure as standard (0.6% γ-H2AX), cells treated with GO show 0.5% γ-H2AX, cells exposed to X-ray show 94.6% γ-H2AX, and cells treated with GO and then exposed to X-ray show 13.1% γ-H2AX. These results confirm GO can significantly reduce X-ray induced DNA double strand breaks.

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