Cytotoxicity of PM(2.5) and PM(2.5--10) ambient air pollutants assessed by the MTT and the Comet assays.
Ambient air particulate matters are classified into two distinct modes in size distribution, namely the coarse and fine particles. Correlation between high particulate concentration and adverse effects on human populations has long been recognized, however, the toxicology of these adverse effects has not been clarified. In the current report, the cytotoxic effects of the solvent-extractable organic compounds (SEOC) from fine particles smaller than 2.5 microm (PM(2.5)) and from coarse particles between 2.5-10 microm (PM(2.5-10)) were studied. Nine 24h consecutive monthly samples were tested to determine the correlation between cytotoxicity and total SEOC in two size fractions of particulate air pollution. Cytotoxicity of SEOC was measured by two micro-scale mammalian cells-based bioassays: the MTT cell proliferation assay, and the Comet assay for the detection of DNA damage. A well-defined mammalian cell line - Rat 6 rodent fibroblast was employed in the study. The SEOC extracts of air particulate matters were sub divided into two equal parts. One part was dissolved in DMSO, the other in KOH/hexane and then conjugated with bovine serum albumin to produce a lipid-soluble fraction for testing. The DMSO fraction would contain mainly the polycyclic aromatic hydrocarbons (PAH), alkanes and alkanols, while the lipid-soluble fraction would be enriched with fatty acids. The results from MTT assay showed that cytotoxicity of the PM(2.5) was much more severe than the PM(2.5-10), suggesting that toxic SEOC were confined to the fine particles. By and large, the DMSO solubles were much more toxic than the lipid solubles. The degree of cytotoxicity of the DMSO soluble samples is positively correlated to the amount of particulates present in the ambient air. For the PM(2.5), the winter samples were significantly more toxic than the summer samples in terms of cell killing, which seemed to be a direct reflection of the total loading of organic matter in the samples. Results from Comet assays showed that SEOC samples of PM(2.5) derived from winter months induced DNA damage at dosages resulting in no obvious cell killing in the MTT assay. Thus, long-term exposure to non-killing dosage of air pollutants may lead to the accumulation of DNA lesions, which may be one of the mechanisms responsible for the chronic adverse health effects of particulate air pollution.
Hsiao
W L
WL
Department of Biology, The Hong Kong University of Science and Technolgy, Clear Water Bay, Kowloon, Hong Kong, PR China. bowhsiao@ust.hk
Mo
Z Y
ZY
Fang
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Shi
X M
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Wang
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D000336:Aerosols; D000393:Air Pollutants / Q000737:chemistry / Q000633:toxicity*; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D004247:DNA / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009153:Mutagens / Q000633:toxicity*; D009930:Organic Chemicals / Q000737:chemistry / Q000633:toxicity; D010316:Particle Size; D051381:Rats; D012621:Seasons; D012995:Solubility; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D000336:Aerosols; D000393:Air Pollutants; D009153:Mutagens; D009930:Organic Chemicals; D013778:Tetrazolium Salts; D013844:Thiazoles; D004247:DNA; C022616:thiazolyl blue