Apparatus for preparing mimics of suspended particles in the troposphere and their controlled deposition onto individual lung cells in culture with measurement of downstream biological response.
Inhalation exposure to particles <10 microm in size that are suspended in the troposphere (PM10) is a factor in respiratory and cardiovascular diseases. The extent of the injury, local to systemic inflammation, is dependent on the number, size, and composition of the particles to which an individual is exposed. The physical properties of and compounds on PM10 that are responsible for these adverse effects on human health are the subject of intense investigation. Here, we report a laboratory method that involved the creation of 1-120 particles per trial that were of known size and composition, followed by deposition of them directly onto individual human lung cells within a cell culture, and after an incubation period, a downstream biological response was measured. To illustrate this methodology, particles that each contained 50 pg of lipopolysaccharide were created and deposited onto individual cells over a region <0.36 mm2 within a genetically modified A549 cell culture. The biological readout was the relative expression of intercellular cell adhesion molecule 1 after 24 h of incubation using an immunocytochemistry assay. The apparatus and methodology introduced here enables studies at the interface between the relevant but diverse areas of atmospheric particle chemistry and lung cell biology to identify the chemical and physical factors of PM10 that cause/exacerbate respiratory and cardiovascular diseases by triggering various biological pathways.
Haddrell
Allen E
AE
Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6.
Ishii
Hiroshi
H
van Eeden
Stephan F
SF
Agnes
George R
GR
D001272:Atmosphere / Q000737:chemistry*; D001273:Atmosphere Exposure Chambers; D002460:Cell Line; D002478:Cells, Cultured; D004867:Equipment Design; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000032:analysis*; D008168:Lung / Q000166:cytology* / Q000378:metabolism*; D010316:Particle Size; D052638:Particulate Matter / Q000032:analysis* / Q000493:pharmacokinetics*; D012680:Sensitivity and Specificity
D008070:Lipopolysaccharides; D052638:Particulate Matter; D018799:Intercellular Adhesion Molecule-1