Comparison of the Biological
Effects of Sudbury Particulate
Matter (SPaM) with Other
Particulate Types: Acute and
Chronic Studies
Sarah M. White
Stacey Ritz, PhD.
Health Effects of Air
Pollution
• Mortality estimates:
• 6 000 annually in Ontario (OMA)
• 21 000 annually in Canada (CMA)
• Compare to breast cancer (5 300), prostate
cancer (4 300), influenza & pneumonia (8 000),
lung cancer (20 000) (StatsCan & Canadian
Cancer Society)
• 500 000 annually worldwide (WHO)
Health Effects of Air
Pollution
 Asthma exacerbations
 Trigger for cardiovascular events in those
with pre-existing disease
 Exposure may facilitate the development
of other disorders
 Allergy
 Atherosclerosis
 Autoimmunity (?)
Mechanisms of Health
Effects
 Oxidative stress:
 reactive organic compounds and transition
metals involved in production of reactive oxygen
species (ROS)
 Inflammation:
 Release of cytokines by airway epithelial
cells and alveolar macrophages
Particulate Matter (PM)
 The PM component of air pollution is
known to have adverse health effects
 Smaller particles are more toxic
 Experimental models of PM:
 Diesel exhaust particles (DEPs): from a
diesel engine, standardized
 Ambient particles: collected from ambient
air, reflects local environment
DEP
(Diesel exhaust Particles)
 carbon core with organic
compounds
 polyaromatic hydrocarbons
(PAH), quinones, aldehydes,
nitroaromatic hydrocarbons
and heterocyclics.
 DEP makes up a
significantly large portion of
PM



Depends on local contributors to PM
Mostly fine and ultrafine in size
Standardized DEP is widely used
in experimental settings (cell
culture, animal studies, human
studies)
Transmission electron
microscopy image of DEP
http://www.transportation.anl.gov
/engines/diesel_structure.html
EHC-93
 Ambient urban sample from Ottawa ON
 Bronchial epithelial cells exposed to
EHC-93 upregulate expression of
inflammatory cytokines
 GM-CSF, IL-8 and IL-1 (Fujii et al, 2001)
 Used in numerous cell culture and animal
models
Sudbury Particulate
Matter (SPaM)
 Collected by Kati
McCartney of the Ritz
lab in 2006
 Reflects local
environment in Sudbury,
which is impacted by
local industrial activities
Image of the particulate collected
from one filter (McCartney, 2009)
Transition Metal Content
of PM Samples
(McCartney ,2009)
Hypothesis
• Ambient PM samples will elicit greater
inflammatory effects than DEPs
• Transition metal content
• Tested in 3 experimental settings:
• Cell cultures (bronchial epithelial cells)
• Acute exposure in mice
• Chronic exposure in mice
Cell Culture: ALI
 Previous work by McCartney et al indicated that
exposure to SPaM elicited enhanced pro-inflammatory
cytokine production (IL-8) in cells grown under
standard conditions
 BEAS2B cells were grown in ALI (Air Liquid Interface)
cell culture to better model in vivo.
http://www.mattek.com/pages/in_vitro_basics/ali-schematic-600.gif
IL-8 production by airway epithelial
cells after PM exposure
IL-8 concentrations in Particulate Matter treated
BEASE2B Cells in ALI Culture
50
IL-8 Concentration in (pg/mL)
45
40
35
30
25
20
15
10
5
0
Control
DEP
EHC-93
Particulate Matter treatment (50ug)
(ANOVA on Ranks P=0.017)
SPaM
Acute Study
 Balb/c mice exposed to 250ug
of PM via intranasal instillation
 Background exposure to PM
reduced by specialized caging
system
 Mice euthanized at 12h postexposure
 Airway inflammation
 Acute phase response
Airway Neutrophilia after
acute exposure to PM
(ANOVA P=0.006)
Control
DEP
EHC-93
SPaM
Chronic Study
 Balb/c mice exposed to aerosolized
suspensions of PM in chambers
 15 min a day, five days a week
 Concentration of aerosol in chamber is 400ug/m3
 estimated dose of 24 ug/kg/week
 Comparable to ambient exposure in typical urban area
Neutrophilic Airway
Inflammation During Chronic
Exposure
Percentage of Neutrophils found in BAL from PM Treated Balb/c mice
9
Percentage of Neutrophils
8
7
6
2 weeks
4 weeks
6 weeks
5
4
3
2
1
0
Control
DEP
EHC
Treatment Group
SPaM
Implications
 The acute study reinforces the need for
occupational health and safety for workers who
can be exposed to large doses of PM.
 There was signs of inflammation in all PM treated
groups
 The chronic study shows some evidence of
physiological changes in the respiratory
epithelium
Acknowledgements
 Special thanks to all the members of Team Ritz:
 Kati McCartney, Cathy Brummer, Jane Bulloch, Nya Fraleigh,
Sandhya Khurana PhD.
 Especially Stacey Ritz PhD.
 To all the NOSM Research lab members especially Joe Eibl and
Heather Peltch for their support.
Questions