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银纳米粒子:对人骨髓间充质干细胞的 DNA 损伤、毒性和功能障碍的评估。

Silver nanoparticles: evaluation of DNA damage, toxicity and functional impairment in human mesenchymal stem cells.

机构信息

Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany.

出版信息

Toxicol Lett. 2011 Feb 25;201(1):27-33. doi: 10.1016/j.toxlet.2010.12.001. Epub 2010 Dec 8.

Abstract

Silver nanoparticles (Ag-NPs) are the most frequent commercialized nanomaterial currently. Due to a distinct lack of information on hazardous properties of Ag-NPs in human cells, a study was conducted to evaluate Ag-NP induced DNA damage, cell death and functional impairment in human mesenchymal stem cells (hMSCs). Initially, Ag-NPs and their cellular distribution were characterized by transmission electron microscopy (TEM). Ag-NPs were exposed to hMSCs for 1, 3 and 24h. Cytotoxicity was measured by the trypan blue exclusion test and the fluorescein-diacetate test, DNA damage was evaluated by the comet assay and chromosomal aberration test. Cytokine release of IL-6, IL-8 and VEGF was observed using the ELISA technique. Additionally, hMSC migration capability was tested in a transwell system. TEM revealed a Ag-NP distribution to cytoplasm and nucleus. Cytotoxic effects were seen at concentrations of 10 μg/ml for all test exposure periods. Both, comet assay and chromosomal aberration test showed DNA damage after 1, 3, and 24h at 0.1 μg/ml. A significant increase of IL-6, IL-8 and VEGF release indicates hMSC activation. Migration ability was not impaired at subtoxic concentrations. In conclusion, we demonstrated cyto- and genotoxic potential of Ag-NPs in hMSCs at significantly higher concentrations as compared to antimicrobial effective levels.

摘要

银纳米颗粒(Ag-NPs)是目前商业化最广泛的纳米材料。由于缺乏关于 Ag-NPs 在人类细胞中危害特性的信息,因此进行了一项研究,以评估 Ag-NP 对人骨髓间充质干细胞(hMSCs)的 DNA 损伤、细胞死亡和功能障碍的诱导作用。首先,通过透射电子显微镜(TEM)对 Ag-NPs 及其细胞分布进行了表征。将 Ag-NPs 暴露于 hMSCs 中 1、3 和 24h。通过台盼蓝排斥试验和荧光二乙酸酯试验测量细胞毒性,通过彗星试验和染色体畸变试验评估 DNA 损伤。使用 ELISA 技术观察白细胞介素 6(IL-6)、白细胞介素 8(IL-8)和血管内皮生长因子(VEGF)的细胞因子释放。此外,在 Transwell 系统中测试了 hMSC 的迁移能力。TEM 显示 Ag-NP 分布于细胞质和细胞核。在所有测试暴露期,浓度为 10μg/ml 时均观察到细胞毒性作用。彗星试验和染色体畸变试验均显示,在 0.1μg/ml 时,1、3 和 24h 后均发生 DNA 损伤。IL-6、IL-8 和 VEGF 释放的显著增加表明 hMSC 被激活。在亚毒性浓度下,迁移能力未受损。总之,我们在明显高于抗菌有效浓度的水平下,证明了 Ag-NPs 在 hMSCs 中的细胞毒性和遗传毒性。

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