Nanotoxicity of Occupational Dust Generated in Granite Stone Saw Mill

I. Ahmad, M. Khan, Govil Patil
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引用次数: 13

Abstract

The risk of occupational exposure to particles of granite exists during quarrying, saw milling and construction work environment. These particles are known to have predominantly micro-size as well as nanosize. Moreover, toxicological profiles of these particles are not known. An attempt was therefore made to study size-dependent cytotoxicity of granite particles using lung epithelial cell line A549. Occupational dust settled around the saw mill was collected and analyzed for their particle size in microscale and found to be 49%, 31% and 20% of 30-50 μm, 10-30μm and 5-10 μm, respectively. With the help of ball mill occupational dust was grinded to particle size 5-8 μm observed under optical micrometry which was further nanotised to 80-130 nm as characterized by DLS. For experimentation occupational dust (OD), microdust 5-8 μm (MD) and nanodust 83±31nm (ND) were exposed to A549 for 72 hr to study cytotoxicity, oxidative stress and inflammatory bioactivity. Detailed cytotoxicity studies showed LC50 values as 326 μg/ml, 269 μg/ml and 127 μg/ml for OD, MD and ND, respectively. Effects of LC50 concentrations of OD, MD and ND on cellular release of LDH, content of GSH, LPO and ROS production were helpful to assess the differential toxicity caused by microand nanoparticles of granite. Granite particle mediated release of LDH in the incubation media , in comparison with control were 1.3-fold, 2.1 -fold and 3.2-fold in the case of OD, MD and ND, respectively. Exposure of these particles caused significant depletion in cellular GSH i.e. 19.8% and 33.9% due to MD and ND exposure, respectively. Particle mediated ROS production increased by 1.4-fold, 2.3-fold and 3.4fold, by OD, MD and ND, respectively. These particles also enhanced LPO profiles significantly by 1.4-fold, 4.0-fold and 6.5fold due to OD, MD and ND, respectively. Inflammatory bioactivities of these particles were explored acknowledging TNF-α, IL-1β and IL-6 as markers of inflammation. Expressions of these cytokines were significantly enhanced by ND in comparision with MD and OD. Present study suggests that nanosizing of granite particle from microsize enhances cytotoxic and inflammatory bioactivities of granite particle. Particle sizedependent toxicological profiles of granite would help extrapolating health effects of populations exposed to different work environment risk right from quarrying, stone saw milling and civil installation. It is concluded that different safety standards are required depending upon the work environment prevalent with microor nano-size granite particles. Keywords— Granite, Nanoparticles, Cytotoxicity, ROS, TNF-α,
花岗岩锯石厂职业粉尘的纳米毒性研究
在采石、锯磨和建筑工作环境中存在职业接触花岗岩颗粒的风险。众所周知,这些颗粒主要具有微尺寸和纳米尺寸。此外,这些颗粒的毒理学特征尚不清楚。因此,我们尝试用肺上皮细胞系A549研究花岗岩颗粒的大小依赖性细胞毒性。对锯木厂周围职业性粉尘进行微尺度粒度分析,发现30-50 μm、10-30μm和5-10 μm的粉尘粒径分别为49%、31%和20%。利用球磨机将职业性粉尘磨成5 ~ 8 μm的粒径,经光学显微镜观察,经DLS表征,进一步纳米化至80 ~ 130 nm。实验采用职业性粉尘(OD)、5 ~ 8 μm微尘(MD)和83±31nm纳米粉尘(ND)暴露于A549 72小时,研究A549的细胞毒性、氧化应激和炎症生物活性。细胞毒性实验表明,OD、MD和ND的LC50分别为326、269和127 μg/ml。OD、MD和ND的LC50浓度对LDH细胞释放、GSH含量、LPO和ROS生成的影响有助于评估花岗岩微颗粒和纳米颗粒的毒性差异。花岗岩颗粒介导的LDH在OD、MD和ND培养液中的释放量分别是对照的1.3倍、2.1倍和3.2倍。暴露于这些颗粒导致细胞谷胱甘肽显著减少,分别为19.8%和33.9%,原因是暴露于MD和ND。OD、MD和ND分别使颗粒介导的ROS产量增加1.4倍、2.3倍和3.4倍。OD、MD和ND对LPO剖面的影响分别为1.4倍、4.0倍和6.5倍。研究了这些颗粒的炎症生物活性,确认TNF-α、IL-1β和IL-6是炎症的标志物。与MD和OD相比,ND显著增强了这些细胞因子的表达。目前的研究表明,花岗岩颗粒的纳米化可以增强花岗岩颗粒的细胞毒性和炎症生物活性。花岗岩颗粒大小相关的毒理学概况将有助于推断暴露于采石、石锯磨和民用安装等不同工作环境风险的人群的健康影响。结果表明,微纳米级花岗岩颗粒普遍存在于不同的工作环境中,需要不同的安全标准。关键词:花岗岩,纳米颗粒,细胞毒性,ROS, TNF-α,
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