The preparation of <100 particles per trial having the same mole fraction of 12 inorganic compounds at diameters of 6.8, 3.8, or 2.6 μm followed by their deposition onto human lung cells (A549) with measurement of the relative downstream differential expression of ICAM-1

The characterization of particulate matter suspended in the troposphere (PM10) based on size is an important basis for assessing the extent of their adverse effects on human health. The relevance of such assessments is anticipated to be significantly improved through the continued development of tools that can identify the chemical components within individual ambient particles, and the injury that they cause. We use recently reported methodology to create mimics of ambient particle types of known size and chemical composition that are levitated within an ac trap. The ac trap uses electric fields to levitate the particles that have a given mass and net elementary charge, and as such the ac trap is a mass-to-charge filter. The ac trap was used to levitate populations of particles where the size of particles in any given population could be altered. The levitated particles are delivered direct from the ac trap to human lung cells (A549), in vitro, with downstream measurement of differential expression of intercellular adhesion molecule (ICAM)-1 and counting of the number of particles actually delivered to the culture using an optical microscope. In this study, the chemical composition of the ambient particle mimics was restricted to inorganic compounds whose relative abundance was purposely designed to mimic the average abundance in Environmental Health Center-93 (EHC-93) particles. The sizes of the multilelement particle types prepared were 6.8 +/- 10.5, 3.8 +/- 0.3, 2.6 +/- 0.2 (mean +/- S.D.). Particles of either elemental carbon, or elemental carbon containing glycerol were used as control particle types. In any given experiment, a known number of particles, but always < 100, of a given size, were deposited onto a small region of an A549 cell culture. Following an 18-h incubation period and anti-body labeling of ICAM-1, the fluorescence emission from a 1.07 mm(2) area of the cell culture centered at the site of particle deposition was acquired. The relative differential expression of ICAM-1 was greatest for multielement particle types having diameters of 2.6 +/- 0.2 mu m for which as few as similar to 15 particles deposited onto the culture resulted in maximal ICAM-1 expression, whereas with multielement particle types having diameters of 6.8 +/- 0.5 mu m, it was necessary to deposit > 50 particles in order to effect comparable ICAM-1 expression. This data set indicates that for multi-element particle types comprised of the same mole fraction of inorganic compounds and of sizes within the course fraction of PM10, the 2.6 mu m particle type was the most potent with respect to effecting differential expression of ICAM-1. (c) 2006 Elsevier B.V. All rights reserved.