Elsevier

Toxicology in Vitro

Volume 29, Issue 5, August 2015, Pages 997-1005
Toxicology in Vitro

Particle-induced cell migration assay (PICMA): A new in vitro assay for inflammatory particle effects based on permanent cell lines

https://doi.org/10.1016/j.tiv.2015.04.005Get rights and content
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Highlights

  • In vitro model for particle induced neutrophilic lung inflammation.

  • The assay can differentiate between inflammatory and inert particles.

  • The test is easy to handle and avoids the use of primary cells.

Abstract

Inflammation is a decisive pathophysiologic mechanism of particle toxicity and accumulation of neutrophils in the lung is believed to be a crucial step in this process. This study describes an in vitro model for investigations of the chemotactic attraction of neutrophils in response to particles using permanent cell lines. We challenged NR8383 rat macrophages with particles that were characterized concerning chemical nature, crystallinity, and size distribution in the dry state and in the culture medium. The cell supernatants were used to investigate migration of differentiated human leukemia cells (dHL-60 cells). The dose range for the tests was determined using an impedance-based Real-Time Cell Analyzer. The challenge of NR8383 cells with 32–96 μg cm−2 coarse and nanosized particles resulted in cell supernatants which induced strong and dose-dependent migration of dHL-60 cells. Quartz caused the strongest effects – exceeding the positive control “fetal calf serum” (FCS) several-fold, followed by silica, rutile, carbon black, and anatase. BaSO4 served as inert control and induced no cell migration. Particles caused NR8383 cells to secrete chemotactic compounds. The assay clearly distinguished between the particles of different inflammatory potential in a highly reproducible way. Specificity of the test is suggested by negative results with BaSO4.

Abbreviations

CCL2
chemokine (C–C motif) ligand 2
CD
cluster of differentiation
COPD
chronic obstructive pulmonary disease
CXCL
chemokine (C–X–C motif) ligand
dHL-60 cells
differentiated human leukemia-60 cells
DLS
dynamic light scattering
EDX
energy-dispersive X-ray spectroscopy
FCS
fetal calf serum
IL
interleukin
JNK
c-Jun activating kinase
MCP-1
monocyte chemo-attractant protein-1
MIP-2
macrophage inflammatory protein 2
NF-κB
nuclear factor-κB
RTCA
Real-Time Cell Analyzer
SEM
scanning electron microscopy
XRD
X-ray powder diffraction

Keywords

Particles
Neutrophilic inflammation
Cell migration
NR8383 cells
dHL-60 cells

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