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彗星试验:评价纳米药物递送系统遗传毒性的方法。

Comet assay: a method to evaluate genotoxicity of nano-drug delivery system.

机构信息

Research Center for Pharmaceutical Nanotechnology, Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Bioimpacts. 2011;1(2):87-97. doi: 10.5681/bi.2011.012. Epub 2011 Aug 6.

Abstract

INTRODUCTION

Drug delivery systems could induce cellular toxicity as side effect of nanomaterials. The mechanism of toxicity usually involves DNA damage. The comet assay or single cell gel electrophoresis (SCGE) is a sensitive method for detecting strand damages in the DNA of a cell with applications in genotoxicity testing and molecular epidemiology as well as fundamental research in DNA damage and repair.

METHODS

In the current study, we reviewed recent drug delivery researches related to SCGE.

RESULTS

We found that one preference for choosing the assay is that comet images may result from apoptosis-mediated nuclear fragmentation. This method has been widely used over the last decade in several different areas. Overall cells, such as cultured cells are embedded in agarose on a microscope slide, lysed with detergent, and treated with high salt. Nucleoids are supercoiled DNA form. When the slide is faced to alkaline electrophoresis any breakages present in the DNA cause the supercoiling to relax locally and loops of DNA extend toward the anode as a ''comet tail''.

CONCLUSION

This article provides a relatively comprehensive review upon potentiality of the comet assay for assessment of DNA damage and accordingly it can be used as an informative platform in genotoxicity studies of drug delivery systems.

摘要

简介

药物输送系统可能会引发细胞毒性作为纳米材料的副作用。毒性的机制通常涉及 DNA 损伤。彗星试验或单细胞凝胶电泳(SCGE)是一种灵敏的方法,用于检测细胞 DNA 中的链损伤,可应用于遗传毒性测试和分子流行病学以及 DNA 损伤和修复的基础研究。

方法

在目前的研究中,我们回顾了与 SCGE 相关的最近的药物输送研究。

结果

我们发现,选择该测定法的一个偏好是彗星图像可能是由凋亡介导的核碎片引起的。该方法在过去十年中已在几个不同领域得到广泛应用。总体而言,细胞(例如培养细胞)在显微镜载玻片上的琼脂糖中嵌入,用去污剂裂解,并用高盐处理。核小体是超螺旋 DNA 形式。当幻灯片面临碱性电泳时,DNA 中存在的任何断裂都会导致局部超螺旋松弛,并且 DNA 环会向阳极延伸形成“彗星尾巴”。

结论

本文对彗星试验在评估 DNA 损伤方面的潜力进行了相对全面的综述,因此它可以作为药物输送系统遗传毒性研究的信息平台。

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