Assessing the cytotoxicity of ambient particulate matter (PM) using Chinese hamster ovary (CHO) cells and its relationship with the PM chemical composition and oxidative potential

Yixiang Wang, Michael Jacob Plewa, Ujjal Kumar Mukherjee, Vishal Verma

Research output: Contribution to journalArticle

Abstract

We assessed mammalian cell cytotoxicity of ambient PM 2.5 and investigated its association with the oxidative potential (OP) and chemical composition of the particles. Sixteen PM samples spanning in various seasons (fall, winter, spring and summer) were collected from an urban site in central Illinois. Cytotoxicity (LC 50 ) in terms of the volume of air that kills 50% of the cells were calculated, which varied from 4.3 to 7.2 m 3 of air. The OP was measured by two assays – the dithiothreitol (DTT) and the surrogate lung fluid (SLF) assay. In DTT assay, we measured two endpoints – hydroxyl radicals ( OH) generation and DTT consumption (the conventionally measured endpoint), while only OH generation was measured in the SLF assay. Although, all three endpoints in the OP assays correlated significantly (P ≤ 0.05) with LC 50 , the correlation of reactive oxygen species (ROS) generation in DTT and SLF assays was much higher (r > 0.80 for OH generation versus LC 50 ) than the DTT consumption (r = 0.58). To further understand the components in PM that drive cytotoxicity and OP, concentration of water-soluble metals (Fe, Cu, Co, Cr, Mn, Ni, Pb, V, Hg, and Zn), organic carbon (OC), water soluble organic carbon (WSOC), and elemental carbon (EC) were measured. Among all the chemical components, Fe, Cu and WSOC correlated most (r > 0.70; P ≤ 0.01) with the cytotoxicity. DTT consumption correlated only with OC and WSOC (r > 0.80; P ≤ 0.01), while OH generation in DTT and SLF assay correlated with both WSOC (r > 0.70; P ≤ 0.01) and metals (i.e. Fe and Cu; r > 0.75; P ≤ 0.01). Our results suggest a strong link between the PM 2.5 OP and its cytotoxicity. Furthermore, the synergistic interactions among the organic compounds (i.e. WSOC) and metals (Fe and Cu) to enhance the ROS generation, which are more effectively captured in OH generation endpoints in DTT and SLF assay than the DTT consumption, appear to be largely responsible for the observed mammalian cell cytotoxicity of PM 2.5 .

Original languageEnglish (US)
Pages (from-to)132-141
Number of pages10
JournalAtmospheric Environment
Volume179
DOIs
StatePublished - Apr 2018

Fingerprint

particulate matter
chemical composition
assay
organic carbon
fluid
water
metal
cytotoxicity
urban site
air
hydroxyl radical
organic compound
consumption
winter
carbon
summer

Keywords

  • Chinese hamster ovary (CHO) cells
  • Cytotoxicity
  • Dithiothreitol
  • Reactive oxygen species (ROS) generation
  • Surrogate lung fluid

ASJC Scopus subject areas

  • Environmental Science(all)
  • Atmospheric Science

Cite this

@article{1951816b58c74c4cb7ed928cf800f141,
title = "Assessing the cytotoxicity of ambient particulate matter (PM) using Chinese hamster ovary (CHO) cells and its relationship with the PM chemical composition and oxidative potential",
abstract = "We assessed mammalian cell cytotoxicity of ambient PM 2.5 and investigated its association with the oxidative potential (OP) and chemical composition of the particles. Sixteen PM samples spanning in various seasons (fall, winter, spring and summer) were collected from an urban site in central Illinois. Cytotoxicity (LC 50 ) in terms of the volume of air that kills 50{\%} of the cells were calculated, which varied from 4.3 to 7.2 m 3 of air. The OP was measured by two assays – the dithiothreitol (DTT) and the surrogate lung fluid (SLF) assay. In DTT assay, we measured two endpoints – hydroxyl radicals ( • OH) generation and DTT consumption (the conventionally measured endpoint), while only • OH generation was measured in the SLF assay. Although, all three endpoints in the OP assays correlated significantly (P ≤ 0.05) with LC 50 , the correlation of reactive oxygen species (ROS) generation in DTT and SLF assays was much higher (r > 0.80 for • OH generation versus LC 50 ) than the DTT consumption (r = 0.58). To further understand the components in PM that drive cytotoxicity and OP, concentration of water-soluble metals (Fe, Cu, Co, Cr, Mn, Ni, Pb, V, Hg, and Zn), organic carbon (OC), water soluble organic carbon (WSOC), and elemental carbon (EC) were measured. Among all the chemical components, Fe, Cu and WSOC correlated most (r > 0.70; P ≤ 0.01) with the cytotoxicity. DTT consumption correlated only with OC and WSOC (r > 0.80; P ≤ 0.01), while • OH generation in DTT and SLF assay correlated with both WSOC (r > 0.70; P ≤ 0.01) and metals (i.e. Fe and Cu; r > 0.75; P ≤ 0.01). Our results suggest a strong link between the PM 2.5 OP and its cytotoxicity. Furthermore, the synergistic interactions among the organic compounds (i.e. WSOC) and metals (Fe and Cu) to enhance the ROS generation, which are more effectively captured in • OH generation endpoints in DTT and SLF assay than the DTT consumption, appear to be largely responsible for the observed mammalian cell cytotoxicity of PM 2.5 .",
keywords = "Chinese hamster ovary (CHO) cells, Cytotoxicity, Dithiothreitol, Reactive oxygen species (ROS) generation, Surrogate lung fluid",
author = "Yixiang Wang and Plewa, {Michael Jacob} and Mukherjee, {Ujjal Kumar} and Vishal Verma",
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T1 - Assessing the cytotoxicity of ambient particulate matter (PM) using Chinese hamster ovary (CHO) cells and its relationship with the PM chemical composition and oxidative potential

AU - Wang, Yixiang

AU - Plewa, Michael Jacob

AU - Mukherjee, Ujjal Kumar

AU - Verma, Vishal

PY - 2018/4

Y1 - 2018/4

N2 - We assessed mammalian cell cytotoxicity of ambient PM 2.5 and investigated its association with the oxidative potential (OP) and chemical composition of the particles. Sixteen PM samples spanning in various seasons (fall, winter, spring and summer) were collected from an urban site in central Illinois. Cytotoxicity (LC 50 ) in terms of the volume of air that kills 50% of the cells were calculated, which varied from 4.3 to 7.2 m 3 of air. The OP was measured by two assays – the dithiothreitol (DTT) and the surrogate lung fluid (SLF) assay. In DTT assay, we measured two endpoints – hydroxyl radicals ( • OH) generation and DTT consumption (the conventionally measured endpoint), while only • OH generation was measured in the SLF assay. Although, all three endpoints in the OP assays correlated significantly (P ≤ 0.05) with LC 50 , the correlation of reactive oxygen species (ROS) generation in DTT and SLF assays was much higher (r > 0.80 for • OH generation versus LC 50 ) than the DTT consumption (r = 0.58). To further understand the components in PM that drive cytotoxicity and OP, concentration of water-soluble metals (Fe, Cu, Co, Cr, Mn, Ni, Pb, V, Hg, and Zn), organic carbon (OC), water soluble organic carbon (WSOC), and elemental carbon (EC) were measured. Among all the chemical components, Fe, Cu and WSOC correlated most (r > 0.70; P ≤ 0.01) with the cytotoxicity. DTT consumption correlated only with OC and WSOC (r > 0.80; P ≤ 0.01), while • OH generation in DTT and SLF assay correlated with both WSOC (r > 0.70; P ≤ 0.01) and metals (i.e. Fe and Cu; r > 0.75; P ≤ 0.01). Our results suggest a strong link between the PM 2.5 OP and its cytotoxicity. Furthermore, the synergistic interactions among the organic compounds (i.e. WSOC) and metals (Fe and Cu) to enhance the ROS generation, which are more effectively captured in • OH generation endpoints in DTT and SLF assay than the DTT consumption, appear to be largely responsible for the observed mammalian cell cytotoxicity of PM 2.5 .

AB - We assessed mammalian cell cytotoxicity of ambient PM 2.5 and investigated its association with the oxidative potential (OP) and chemical composition of the particles. Sixteen PM samples spanning in various seasons (fall, winter, spring and summer) were collected from an urban site in central Illinois. Cytotoxicity (LC 50 ) in terms of the volume of air that kills 50% of the cells were calculated, which varied from 4.3 to 7.2 m 3 of air. The OP was measured by two assays – the dithiothreitol (DTT) and the surrogate lung fluid (SLF) assay. In DTT assay, we measured two endpoints – hydroxyl radicals ( • OH) generation and DTT consumption (the conventionally measured endpoint), while only • OH generation was measured in the SLF assay. Although, all three endpoints in the OP assays correlated significantly (P ≤ 0.05) with LC 50 , the correlation of reactive oxygen species (ROS) generation in DTT and SLF assays was much higher (r > 0.80 for • OH generation versus LC 50 ) than the DTT consumption (r = 0.58). To further understand the components in PM that drive cytotoxicity and OP, concentration of water-soluble metals (Fe, Cu, Co, Cr, Mn, Ni, Pb, V, Hg, and Zn), organic carbon (OC), water soluble organic carbon (WSOC), and elemental carbon (EC) were measured. Among all the chemical components, Fe, Cu and WSOC correlated most (r > 0.70; P ≤ 0.01) with the cytotoxicity. DTT consumption correlated only with OC and WSOC (r > 0.80; P ≤ 0.01), while • OH generation in DTT and SLF assay correlated with both WSOC (r > 0.70; P ≤ 0.01) and metals (i.e. Fe and Cu; r > 0.75; P ≤ 0.01). Our results suggest a strong link between the PM 2.5 OP and its cytotoxicity. Furthermore, the synergistic interactions among the organic compounds (i.e. WSOC) and metals (Fe and Cu) to enhance the ROS generation, which are more effectively captured in • OH generation endpoints in DTT and SLF assay than the DTT consumption, appear to be largely responsible for the observed mammalian cell cytotoxicity of PM 2.5 .

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KW - Reactive oxygen species (ROS) generation

KW - Surrogate lung fluid

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