Mechanism of neutrophil activation and toxicity elicited by engineered nanomaterials.

Helinor Johnston, David M Brown, Nilesh Kanase, Matthew Euston, Birgit K Gaiser, Calum T Robb, Elisabeth Dyrynda, Adriano G Rossi, Euan R Brown, Vicki Stone
Author Information
  1. Helinor Johnston: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom. Electronic address: h.johnston@hw.ac.uk.
  2. David M Brown: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.
  3. Nilesh Kanase: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.
  4. Matthew Euston: Institute of Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.
  5. Birgit K Gaiser: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.
  6. Calum T Robb: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom; MRC Centre for Inflammation Research, Queen's Medical Research Institute, University of Edinburgh, Edinburgh EH16 4TJ, United Kingdom.
  7. Elisabeth Dyrynda: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.
  8. Adriano G Rossi: MRC Centre for Inflammation Research, Queen's Medical Research Institute, University of Edinburgh, Edinburgh EH16 4TJ, United Kingdom.
  9. Euan R Brown: Institute of Biological Chemistry, Biophysics and Bioengineering, School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.
  10. Vicki Stone: School of Life Sciences, Heriot-Watt University, Edinburgh EH14 4AS, United Kingdom.

Abstract

The effects of nanomaterials (NMs) on biological systems, especially their ability to stimulate inflammatory responses requires urgent investigation. We evaluated the response of the human differentiated HL60 neutrophil-like cell line to NMs. It was hypothesised that NM physico-chemical characteristics would influence cell responsiveness by altering intracellular Ca2+ concentration [Ca2+]i and reactive oxygen species production. Cells were exposed (1.95-125 μg/ml, 24 h) to silver (Ag), zinc oxide (ZnO), titanium dioxide (TiO2), multi-walled carbon nanotubes (MWCNTs) or ultrafine carbon black (ufCB) and cytotoxicity assessed (alamar blue assay). Relatively low (TiO2, MWCNTs, ufCB) or high (Ag, ZnO) cytotoxicity NMs were identified. Sub-lethal impacts of NMs on cell function were investigated for selected NMs only, namely TiO2, Ag and ufCB. Only Ag stimulated cell activation. Within minutes, Ag stimulated an increase in [Ca2+]i (in Fura-2 loaded cells), and a prominent inward ion current (assessed by electrophysiology). Within 2-4 h, Ag increased superoxide anion release and stimulated cytokine production (MCP-1, IL-8) that was diminished by Ca2+ inhibitors or trolox. Light microscopy demonstrated that cells had an activated phenotype. In conclusion NM toxicity was ranked; Ag>ufCB>TiO2, and the battery of tests used provided insight into the mechanism of action of NM toxicity to guide future testing strategies.

Keywords

Grants

  1. G0601481/Medical Research Council
  2. MR/K013386/1/Medical Research Council

MeSH Term

Calcium
Cell Survival
Cytokines
HL-60 Cells
Humans
Nanostructures
Nanotubes, Carbon
Neutrophil Activation
Silver
Soot
Superoxides
Titanium
Zinc Oxide

Chemicals

Cytokines
Nanotubes, Carbon
Soot
Superoxides
titanium dioxide
Silver
Titanium
Zinc Oxide
Calcium

Word Cloud

Created with Highcharts 10.0.0AgNMscellNMTiO2ufCBstimulatedtoxicitynanomaterialsCa2+[Ca2+]iproductionhZnOcarbonMWCNTscytotoxicityassessedactivationWithincellsMechanismeffectsbiologicalsystemsespeciallyabilitystimulateinflammatoryresponsesrequiresurgentinvestigationevaluatedresponsehumandifferentiatedHL60neutrophil-likelinehypothesisedphysico-chemicalcharacteristicsinfluenceresponsivenessalteringintracellularconcentrationreactiveoxygenspeciesCellsexposed195-125μg/ml24silverzincoxidetitaniumdioxidemulti-wallednanotubesultrafineblackalamarblueassayRelativelylowhighidentifiedSub-lethalimpactsfunctioninvestigatedselectednamelyminutesincreaseFura-2loadedprominentinwardioncurrentelectrophysiology2-4increasedsuperoxideanionreleasecytokineMCP-1IL-8diminishedinhibitorstroloxLightmicroscopydemonstratedactivatedphenotypeconclusionrankedAg>ufCB>TiO2batterytestsusedprovidedinsightmechanismactionguidefuturetestingstrategiesneutrophilelicitedengineeredCa2+NanomaterialNeutrophilToxicity

Similar Articles

Cited By