Comparative in vitro study of single and four layer graphene oxide nanoflakes — Cytotoxicity and cellular uptake

Perużyńska, Magdalena; Cendrowski, Krzysztof; Barylak, Martyna; Tkacz, Marta; Piotrowska, Katarzyna; Kurzawski, Mateusz; Mijowska, Ewa; Droździk, Marek

doi:10.1016/j.tiv.2017.03.005

Cited by 26 publications

(28 citation statements)

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“…Functionalization of nanoparticles with biocompatible polymers increases their stability under physiological conditions. It also minimizes their interactions with other biomolecules and reduce immunological responses [ 8 , 18 ]. The effects of GONPs on RAW cells, under LPS stimulated conditions, were not evaluated in the current study.…”

Section: Discussionmentioning

confidence: 99%

“…Graphene oxide induced oxidative stress and immunotoxicity in vivo in zebrafish [ 1 ]. Upon in vitroexposure to GONPs, decrease in cell viability, DNA damage, increased reactive oxygen species (ROS) production and induction of inflammatory factors were recorded [ 4 , 8 , 9 ]. The cytokines selected in this in vitro study (i.e., IL-6, IL-10 and IFNγ) are indicative of the inflammatory, humoral and cell mediated immune systems respectively.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Graphene Oxide Nanoparticles on the Immune System Biomarkers Produced by RAW 264.7 and Human Whole Blood Cell Cultures

et al. 2018

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Graphene oxide nanoparticles (GONPs) have attracted a lot of attention due to their many applications. These applications include batteries, super capacitors, drug delivery and biosensing. However, few studies have investigated the effects of these nanoparticles on the immune system. In this study, the in vitro effects of GONPs on the immune system was evaluated by exposing murine macrophages, RAW 264.7 cells and human whole blood cell cultures (to GONPs. The effects of GONPs on RAW cells were monitored under basal conditions. The whole blood cell cultures were exposed to GONPs in the presence or absence of the mitogens lipopolysaccharide (LPS) and phytohaemmagglutinin (PHA). A number of parameters were monitored for both RAW and whole blood cell cultures, these included cytotoxicity, inflammatory biomarkers, cytokines of the acquired immune system and a proteome profile analysis. The GONPs were cytotoxic to both RAW and whole blood cell cultures at 500 μg/mL. In the absence of LPS, GONPs elicited an inflammatory response from the murine macrophage, RAW and whole blood cell cultures at 15.6 and 5 μg/mL respectively. This activation was further corroborated by proteome profile analysis of both experimental cultures. GONPs inhibited LPS induced interleukin 6 (IL-6) synthesis and PHA induced interferon gamma (IFNγ) synthesis by whole blood cell cultures in a dose dependent manner. In the absence of mitogens, GONPs stimulated IL-10 synthesis by whole blood cell cultures. The current study shows that GONPs modulate immune system biomarkers and that these may pose a health risk to individuals exposed to this type of nanoparticle.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Effects of Graphene Oxide Nanoparticles on the Immune System Biomarkers Produced by RAW 264.7 and Human Whole Blood Cell Cultures

et al. 2018

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“…The cytotoxicity induced by graphene and GO are entirely different because of their difference in physiochemical properties of these two nanomaterials. Hence, most of the researchers have concentrated on the use of GO because of its high solubility and dispersibility in comparison to other graphene-based materials (Bianco, 2013;Peruzynska et al, 2017). glutathione in bluegill sunfish cells (BF-2) exposed to graphene oxide at 0, 40, 60, 80 and 100 μg ml -1 .…”

Section: Discussionmentioning

confidence: 99%

Graphene oxide induces cytotoxicity and oxidative stress in bluegill sunfish cells

Srikanth

Sundar

Pereira

et al. 2017

J of Applied Toxicology

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Graphene oxide (GO) is considered a promising material for biological application due to its unique properties. However, the potential toxicity of GO to aquatic organism particularly bluegill sun fish cells (BF-2) is unexplored or remains poorly understood. GO-induced cytotoxicity and oxidative stress in BF-2 cells were assessed using a battery of biomarkers. Two different biological assays (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red uptake were used to evaluate the cytotoxicity of GO on BF-2 cells. It was found that GO induced dose- and time-dependent cytotoxicity on BF-2 cells. BF-2 cells exposed to lower concentration of GO (40 μg ml ) for 24 induced morphological changes when compared to their respective controls. As evidence for oxidative stress lipid peroxidation, superoxide dismutase, catalase, reactive oxygen species and 8-hydroxy-2'-deoxyguanosine levels were increased and glutathione levels were found to decline in BF-2 cells after treatment with GO. Our findings demonstrate that GO when exposed to BF-2 fish cells cause oxidative stress.

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“…toxicity of GBMs such as graphene oxide (GO). [9][10][11][12][13] However, there are few if any studies in which the long-term impact is examined. Here, we addressed the impact of GO sheets with varying lateral dimensions on human lung cells by applying RNA sequencing coupled with computational analysis of the transcriptomics data.…”

Section: Introductionmentioning

confidence: 99%

Next‐Generation Sequencing Reveals Differential Responses to Acute versus Long‐Term Exposures to Graphene Oxide in Human Lung Cells

Mukherjee

Gupta

Klöditz

et al. 2020

Small

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Numerous studies have addressed the biological impact of graphenebased materials including graphene oxide (GO), yet few have focused on long-term effects. Here, RNA sequencing is utilized to unearth responses of human lung cells to GO. To this end, the BEAS-2B cell line derived from normal human bronchial epithelium is subjected to repeated, low-dose exposures of GO (1 or 5 µg mL −1 ) for 28 days or to the equivalent, cumulative amount of GO for 48 h. Then, samples are analyzed by using the NovaSeq 6000 sequencing system followed by pathway analysis and gene ontology enrichment analysis of the differentially expressed genes. Significant differences are seen between the low-dose, long-term exposures and the high-dose, short-term exposures. Hence, exposure to GO for 48 h results in mitochondrial dysfunction. In contrast, exposure to GO for 28 days is characterized by engagement of apoptosis pathways with downregulation of genes belonging to the inhibitor of apoptosis protein (IAP) family. Validation experiments confirm that long-term exposure to GO affects the apoptosis threshold in lung cells, accompanied by a loss of IAPs. These studies reveal the sensitivity of RNA-sequencing approaches and show that acute exposure to GO is not a good predictor of the long-term effects of GO.The ORCID identification number(s) for the author(s) of this article can be found under https://doi.www.small-journal.com relationships of graphene-based materials (GBMs). [8] Previous studies have shown that the lateral dimensions as well as the number of layers play important roles in the acute toxicity of GBMs such as graphene oxide (GO). [9][10][11][12][13] However, there are few if any studies in which the long-term impact is examined. Here, we addressed the impact of GO sheets with varying lateral dimensions on human lung cells by applying RNA sequencing coupled with computational analysis of the transcriptomics data. Specifically, we asked whether transcriptomics approaches could be used to distinguish short-term and long-term exposures to GO and if so, which biological processes were involved. We then applied cell-based assays in order to validate the results. www.small-journal.comSmall 2020, 16, 1907686 Figure 8. Long-term exposure to GO affects the susceptibility of lung cells to apoptosis. To validate the RNA-sequencing-based prediction regarding apoptosis, we exposed BEAS-2B cells to GO-US, GO-S, and GO-L twice weekly for 28 days at 1 and 5 µg mL −1 , and harvested cells at weekly intervals A-D) for analysis using the DNA content assay. Untreated control cells maintained in culture under the same conditions were included for comparison. The results showed a dose-and time-dependent increase of apoptosis following low-dose exposure to GO while control cells were unaffected. 12 of 15) www.advancedsciencenews.com (

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Comparative in vitro study of single and four layer graphene oxide nanoflakes — Cytotoxicity and cellular uptake

Cited by 26 publications

References 40 publications

Effects of Graphene Oxide Nanoparticles on the Immune System Biomarkers Produced by RAW 264.7 and Human Whole Blood Cell Cultures

Effects of Graphene Oxide Nanoparticles on the Immune System Biomarkers Produced by RAW 264.7 and Human Whole Blood Cell Cultures

Graphene oxide induces cytotoxicity and oxidative stress in bluegill sunfish cells

Next‐Generation Sequencing Reveals Differential Responses to Acute versus Long‐Term Exposures to Graphene Oxide in Human Lung Cells

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