Establishment of an in vivo simulating co‐culture assay platform for genotoxicity of multi‐walled carbon nanotubes

Fukai, Emi; Sato, Haruna; Watanabe, Masatoshi; Nakae, Dai; Totsuka, Yukari

doi:10.1111/cas.13534

Cited by 22 publications

(21 citation statements)

References 44 publications

(73 reference statements)

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“…The authors reported a stronger mutation frequency in co-cultured GDL1 cells and RAW 264.7 macrophages over monocultured GDL1 cells noting a greater IL-1α and IL-1β production when macrophages were present. The authors also reported the presence of the free-radical-induced DNA lesion, 8-hydroxydeoxyguanosine (8-OHdG), in co-cultured GLD1 cells was greater than monocultured cells indicative of secondary mechanisms [ 11 ]. In the recent work of Evans and colleagues investigating SPION toxicity in co-cultures of 16HBE14o − and d.THP-1 macrophages, the authors reported secondary genotoxicity at concentrations of 10, 50 and 100 µg/ml.…”

Section: Discussionmentioning

confidence: 99%

Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro

et al. 2021

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Background Toxicological evaluation of engineered nanomaterials (ENMs) is essential for occupational health and safety, particularly where bulk manufactured ENMs such as few-layer graphene (FLG) are concerned. Additionally, there is a necessity to develop advanced in vitro models when testing ENMs to provide a physiologically relevant alternative to invasive animal experimentation. The aim of this study was to determine the genotoxicity of non-functionalised (neutral), amine- and carboxyl-functionalised FLG upon both human-transformed type-I (TT1) alveolar epithelial cell monocultures, as well as co-cultures of TT1 and differentiated THP-1 monocytes (d.THP-1 (macrophages)). Results In monocultures, TT1 and d.THP-1 macrophages showed a statistically significant (p < 0.05) cytotoxic response with each ENM following 24-h exposures. Monoculture genotoxicity measured by the in vitro cytokinesis blocked micronucleus (CBMN) assay revealed significant (p < 0.05) micronuclei induction at 8 µg/ml for amine- and carboxyl-FLG. Transmission electron microscopy (TEM) revealed ENMs were internalised by TT1 cells within membrane-bound vesicles. In the co-cultures, ENMs induced genotoxicity in the absence of cytotoxic effects. Co-cultures pre-exposed to 1.5 mM N-acetylcysteine (NAC), showed baseline levels of micronuclei induction, indicating that the genotoxicity observed was driven by oxidative stress. Conclusions Therefore, FLG genotoxicity when examined in monocultures, results in primary-indirect DNA damage; whereas co-cultured cells reveal secondary mechanisms of DNA damage.

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Section: Discussionmentioning

confidence: 99%

Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro

et al. 2021

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“…The authors reported a stronger mutation frequency in co-cultured GDL1 cells and RAW 264.7 macrophages over monocultured GDL1 cells noting a greater IL-1α and IL-1β production when macrophages were present. The authors also reported the presence of the free-radical-induced DNA lesion, 8-hydroxydeoxyguanosine (8-OHdG), in cocultured GLD1 cells was greater than monocultured cells indicative of secondary mechanisms (Fukai et al, 2018). In the recent work of Evans and colleagues investigating SPION toxicity in co-cultures of 16HBE14o − and d.THP-1 macrophages, the authors reported secondary genotoxicity at concentrations of 10, 50 and 100 µg/ml.…”

Section: Discussionmentioning

confidence: 96%

Few-Layer Graphene Induces Both Primary and Secondary Genotoxicity in in Vitro Alveolar Barrier Models

Burgum

Evans

et al. 2020

Preprint

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Background: Toxicological evaluation of engineered nanomaterials (ENMs) is essential for occupational health and safety, particularly where bulk manufactured ENMs such as few-layer graphene (FLG) are concerned. Additionally, there is a necessity to develop advanced in vitro models when testing ENMs to provide a physiologically relevant alternative to invasive animal experimentation. The aim of this study was to determine the genotoxicity of non-functionalised (neutral), amine- and carboxyl-functionalised FLG upon both human-transformed type-I (TT1) alveolar epithelial cell monocultures, as well as co-cultures of TT1 and differentiated THP-1 monocytes (d.THP-1 (macrophages)). Results: In monocultures, TT1 and d.THP-1 macrophages showed a statistically significant (p<0.05) cytotoxic response with each ENM following 24-hour exposures. Monoculture genotoxicity measured by the in vitro cytokinesis blocked micronucleus (CBMN) assay revealed significant (p<0.05) micronuclei induction at 8µg/ml for amine- and carboxyl-FLG. Transmission electron microscopy (TEM) revealed ENMs were internalised by TT1 cells within membrane-bound vesicles. In the co-cultures, ENMs induced genotoxicity in the absence of cytotoxic effects. Co-cultures pre-exposed to 1.5mM N-acetylcysteine (NAC), showed baseline levels of micronuclei induction, indicating that the genotoxicity observed was driven by oxidative stress. Conclusions: Therefore, FLG genotoxicity when examined in monocultures, results in primary-indirect DNA damage; whereas co-cultured cells reveal secondary mechanisms of DNA damage.

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“…On the other hand, enhanced cytokines IL-10, and TGF-β, restricted tumor metastasis. 36 , 41 Indeed, the role of tumor destruction of CNTs did not rule out. Another study on breast cancer was performed with CNTs, which confirmed that CNTs could reduce tumor size.…”

Section: Discussionmentioning

confidence: 99%

“…The presence of MWCNTs in vivo resulted in cytokines production such as TNF‐α and IL‐1β from immune cells which involved in creating the toxicity. 36 In a 90-day study by the intravenous injection of MWCNT, no changes in mice weight were observed, while the toxic effects were observed, and also the mice survive, despite the toxicity. 37 In another study on Sprague–Dawley rats with injected intratracheally of 0.5, 2, and 5 mg MWCNTs, TNF-α discharge was increased from macrophages.…”

Section: The Pulmonary Toxicity Assessment Of Carbon Nanotubesmentioning

confidence: 98%

<p>The Applications of Carbon Nanotubes in the Diagnosis and Treatment of Lung Cancer: A Critical Review</p>

Sheikhpour¹,

Naghinejad²,

Kasaeian³

et al. 2020

IJN

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The importance of timely diagnosis and the complete treatment of lung cancer for many people with this deadly disease daily increases due to its high mortality. Diagnosis and treatment with helping the nanoparticles are useful, although they have reasonable harms. This article points out that the side effects of using carbon nanotube (CNT) in this disease treatment process such as inflammation, fibrosis, and carcinogenesis are very problematic. Toxicity can reduce to some extent using the techniques such as functionalizing to proper dimensions as a longer length, more width, and greater curvature. The targeted CNT sensors can be connected to various modified vapors. In this regard, with helping this method, screening makes non-invasive diagnosis possible. Researchers have also found that nanoparticles such as CNTs could be used as carriers to direct drug delivery, especially with chemotherapy drugs. Most of these carriers were multi-wall carbon nanotubes (MWCNT) used for cancerous cell targeting. The results of laboratory and animal researches in the field of diagnosis and treatment became very desirable and hopeful. The collection of researches summarized has highlighted the requirement for a detailed assessment which includes CNT dose, duration, method of induction, etc., to achieve the most controlled conditions for animal and human studies. In the discussion section, 4 contradictory issues are discussed which are invited researchers to do more research to get clearer results.

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Establishment of an in vivo simulating co‐culture assay platform for genotoxicity of multi‐walled carbon nanotubes

Cited by 22 publications

References 44 publications

Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro

Few-layer graphene induces both primary and secondary genotoxicity in epithelial barrier models in vitro

Few-Layer Graphene Induces Both Primary and Secondary Genotoxicity in in Vitro Alveolar Barrier Models

<p>The Applications of Carbon Nanotubes in the Diagnosis and Treatment of Lung Cancer: A Critical Review</p>

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