Yi Jinghai, Chen Bean T, Schwegler-Berry Diane, Frazer Dave, Castranova Vince, McBride Carroll, Knuckles Travis L, Stapleton Phoebe A, Minarchick Valerie C, Nurkiewicz Timothy R
Center for Cardiovascular and Respiratory Sciences, West Virginia University, USA.
J Vis Exp. 2013 May 7(75):e50263. doi: 10.3791/50263.
Inhalation is the most likely exposure route for individuals working with aerosolizable engineered nano-materials (ENM). To properly perform nanoparticle inhalation toxicology studies, the aerosols in a chamber housing the experimental animals must have: 1) a steady concentration maintained at a desired level for the entire exposure period; 2) a homogenous composition free of contaminants; and 3) a stable size distribution with a geometric mean diameter < 200 nm and a geometric standard deviation σg < 2.5 (5). The generation of aerosols containing nanoparticles is quite challenging because nanoparticles easily agglomerate. This is largely due to very strong inter-particle forces and the formation of large fractal structures in tens or hundreds of microns in size (6), which are difficult to be broken up. Several common aerosol generators, including nebulizers, fluidized beds, Venturi aspirators and the Wright dust feed, were tested; however, none were able to produce nanoparticle aerosols which satisfy all criteria (5). A whole-body nanoparticle aerosol inhalation exposure system was fabricated, validated and utilized for nano-TiO2 inhalation toxicology studies. Critical components: 1) novel nano-TiO2 aerosol generator; 2) 0.5 m(3) whole-body inhalation exposure chamber; and 3) monitor and control system. Nano-TiO2 aerosols generated from bulk dry nano-TiO2 powders (primary diameter of 21 nm, bulk density of 3.8 g/cm(3)) were delivered into the exposure chamber at a flow rate of 90 LPM (10.8 air changes/hr). Particle size distribution and mass concentration profiles were measured continuously with a scanning mobility particle sizer (SMPS), and an electric low pressure impactor (ELPI). The aerosol mass concentration (C) was verified gravimetrically (mg/m(3)). The mass (M) of the collected particles was determined as M = (Mpost-Mpre), where Mpre and Mpost are masses of the filter before and after sampling (mg). The mass concentration was calculated as C = M/(Q*t), where Q is sampling flowrate (m(3)/min), and t is the sampling time (minute). The chamber pressure, temperature, relative humidity (RH), O2 and CO2 concentrations were monitored and controlled continuously. Nano-TiO2 aerosols collected on Nuclepore filters were analyzed with a scanning electron microscope (SEM) and energy dispersive X-ray (EDX) analysis. In summary, we report that the nano-particle aerosols generated and delivered to our exposure chamber have: 1) steady mass concentration; 2) homogenous composition free of contaminants; 3) stable particle size distributions with a count-median aerodynamic diameter of 157 nm during aerosol generation. This system reliably and repeatedly creates test atmospheres that simulate occupational, environmental or domestic ENM aerosol exposures.
对于从事可雾化工程纳米材料(ENM)工作的个人来说,吸入是最有可能的接触途径。为了正确进行纳米颗粒吸入毒理学研究,容纳实验动物的实验室内的气溶胶必须具备以下条件:1)在整个暴露期间将浓度稳定维持在所需水平;2)成分均匀且无污染物;3)具有稳定的粒径分布,几何平均直径<200 nm,几何标准偏差σg<2.5(5)。生成包含纳米颗粒的气溶胶极具挑战性,因为纳米颗粒很容易团聚。这主要是由于颗粒间作用力非常强,会形成尺寸达数十或数百微米的大型分形结构(6),难以分解。对几种常见的气溶胶发生器进行了测试,包括雾化器、流化床、文丘里吸气器和赖特粉尘进料器;然而,没有一种能够产生满足所有标准的纳米颗粒气溶胶(5)。制造、验证并使用了一种全身纳米颗粒气溶胶吸入暴露系统用于纳米TiO₂吸入毒理学研究。关键组件包括:1)新型纳米TiO₂气溶胶发生器;2)0.5 m³全身吸入暴露室;3)监测与控制系统。由大量干燥纳米TiO₂粉末(初级直径为21 nm,堆积密度为3.8 g/cm³)产生的纳米TiO₂气溶胶以90 LPM(10.8次空气交换/小时)的流速输送到暴露室。使用扫描迁移率粒径谱仪(SMPS)和低压冲击器(ELPI)连续测量粒径分布和质量浓度曲线。通过重量法验证气溶胶质量浓度(C)(mg/m³)。收集颗粒的质量(M)确定为M =(Mpost - Mpre),其中Mpre和Mpost分别是采样前后过滤器的质量(mg)。质量浓度计算为C = M/(Q*t),其中Q是采样流速(m³/分钟),t是采样时间(分钟)。持续监测并控制室内压力、温度、相对湿度(RH)、O₂和CO₂浓度。对收集在核孔滤膜上的纳米TiO₂气溶胶进行扫描电子显微镜(SEM)和能量色散X射线(EDX)分析。总之,我们报告称,生成并输送到我们暴露室的纳米颗粒气溶胶具有:1)稳定的质量浓度;2)成分均匀且无污染物;3)在气溶胶生成期间,粒径分布稳定,计数中值空气动力学直径为157 nm。该系统可靠且可重复地创建模拟职业、环境或家庭ENM气溶胶暴露的测试环境。