Comparison of the Biological Comparison of the Biological Effects of Sudbury Particulate Effects of Sudbury Particulate Matter (SPaM) with Other Matter (SPaM) with Other Particulate Types: Acute and Particulate Types: Acute and Chronic StudiesChronic Studies

Sarah M. WhiteSarah M. White

Stacey Ritz, PhD.Stacey Ritz, PhD.

Health Effects of Air Health Effects of Air PollutionPollution

• 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)

Asthma exacerbationsAsthma exacerbations Trigger for cardiovascular events in those Trigger for cardiovascular events in those

with pre-existing diseasewith pre-existing disease Exposure may facilitate the development Exposure may facilitate the development

of other disordersof other disorders Allergy Allergy AtherosclerosisAtherosclerosis Autoimmunity (?)Autoimmunity (?)

Health Effects of Air Health Effects of Air PollutionPollution

Mechanisms of Health Mechanisms of Health EffectsEffects

Oxidative stress:Oxidative stress: reactive organic compounds and transition reactive organic compounds and transition

metals involved in production of reactive oxygen metals involved in production of reactive oxygen species (ROS)species (ROS)

Inflammation:Inflammation: Release of cytokines by airway epithelial Release of cytokines by airway epithelial

cells and alveolar macrophagescells and alveolar macrophages

Particulate Matter (PM)Particulate Matter (PM)

The PM component of air pollution is The PM component of air pollution is known to have adverse health effectsknown to have adverse health effects Smaller particles are more toxicSmaller particles are more toxic

Experimental models of PM:Experimental models of PM: Diesel exhaust particles (DEPs): from a Diesel exhaust particles (DEPs): from a

diesel engine, standardizeddiesel engine, standardized Ambient particles: collected from ambient Ambient particles: collected from ambient

air, reflects local environmentair, reflects local environment

DEP DEP (Diesel exhaust Particles)(Diesel exhaust Particles)

carbon core with organic carbon core with organic compounds compounds polyaromatic hydrocarbons polyaromatic hydrocarbons

(PAH), quinones, aldehydes, (PAH), quinones, aldehydes, nitroaromatic hydrocarbons nitroaromatic hydrocarbons and heterocyclics.and heterocyclics.

DEP makes up a DEP makes up a significantly large portion of significantly large portion of PMPM Depends on local contributors to PMDepends on local contributors to PM Mostly fine and ultrafine in sizeMostly fine and ultrafine in size

Standardized DEP is widely used Standardized DEP is widely used in experimental settings (cell in experimental settings (cell culture, animal studies, human culture, animal studies, human studies)studies)

Transmission electron microscopy image of DEP http://www.transportation.anl.gov/engines/diesel_structure.html

EHC-93EHC-93

Ambient urban sample from Ottawa ONAmbient urban sample from Ottawa ON Bronchial epithelial cells exposed to Bronchial epithelial cells exposed to

EHC-93 upregulate expression of EHC-93 upregulate expression of inflammatory cytokinesinflammatory cytokines GM-CSF, IL-8 and IL-1 (Fujii et al, 2001)GM-CSF, IL-8 and IL-1 (Fujii et al, 2001)

Used in numerous cell culture and animal Used in numerous cell culture and animal modelsmodels

Sudbury Particulate Sudbury Particulate Matter (SPaM)Matter (SPaM)

Collected by Kati Collected by Kati McCartney of the Ritz McCartney of the Ritz lab in 2006 lab in 2006

Reflects local Reflects local environment in Sudbury, environment in Sudbury, which is impacted by which is impacted by local industrial activitieslocal industrial activities

Image of the particulate collected from one filter (McCartney, 2009)

Transition Metal Content Transition Metal Content of PM Samplesof PM Samples

(McCartney ,2009)(McCartney ,2009)

Hypothesis 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: ALICell Culture: ALI Previous work by McCartney et al indicated that Previous work by McCartney et al indicated that

exposure to SPaM elicited enhanced pro-inflammatory exposure to SPaM elicited enhanced pro-inflammatory cytokine production (IL-8) in cells grown under cytokine production (IL-8) in cells grown under standard conditionsstandard conditions

BEAS2B cells were grown in ALI (Air Liquid Interface) BEAS2B cells were grown in ALI (Air Liquid Interface) cell culture to better model cell culture to better model in vivoin vivo..

http://www.mattek.com/pages/in_vitro_basics/ali-schematic-600.gif

IL-8 production by airway epithelial IL-8 production by airway epithelial cells after PM exposurecells after PM exposure

IL-8 concentrations in Particulate Matter treated BEASE2B Cells in ALI Culture

0

5

10

15

20

25

30

35

40

45

50

Control DEP EHC-93 SPaM

Particulate Matter treatment (50ug)

IL-8

Co

ncen

trati

on

in

(p

g/m

L)

(ANOVA on Ranks P=0.017)

Acute StudyAcute Study

Balb/c mice exposed to 250ug of Balb/c mice exposed to 250ug of PM via intranasal instillationPM via intranasal instillation Background exposure to PM Background exposure to PM

reduced by specialized caging reduced by specialized caging systemsystem

Mice euthanized at 12h post-Mice euthanized at 12h post-exposureexposure Airway inflammationAirway inflammation Acute phase responseAcute phase response

Airway Neutrophilia after Airway Neutrophilia after acute exposure to PMacute exposure to PM

(ANOVA P=0.006)

Control

DEP

EHC-93

SPaM

Chronic StudyChronic Study Balb/c mice exposed to aerosolized Balb/c mice exposed to aerosolized

suspensions of PM in chamberssuspensions of PM in chambers 15 min a day, five days a week15 min a day, five days a week Concentration of aerosol in chamber is 400ug/mConcentration of aerosol in chamber is 400ug/m33 estimated dose of 24 ug/kg/week estimated dose of 24 ug/kg/week

Comparable to ambient exposure in typical urban areaComparable to ambient exposure in typical urban area

Neutrophilic Airway Neutrophilic Airway Inflammation During Chronic Inflammation During Chronic ExposureExposure

Percentage of Neutrophils found in BAL from PM Treated Balb/c mice

0

1

2

3

4

5

6

7

8

9

Control DEP EHC SPaM

Treatment Group

Per

cen

tag

e o

f N

eutr

op

hils

2 weeks4 weeks6 weeks

Implications Implications

The acute study reinforces the need for The acute study reinforces the need for occupational health and safety for workers who occupational health and safety for workers who can be exposed to large doses of PM. can be exposed to large doses of PM. There was signs of inflammation in all PM treated There was signs of inflammation in all PM treated

groups groups

The chronic study shows some evidence of The chronic study shows some evidence of physiological changes in the respiratory epithelium physiological changes in the respiratory epithelium

AcknowledgementsAcknowledgements

Special thanks to all the members of Team Ritz:Special thanks to all the members of Team Ritz: Kati McCartney, Cathy Brummer, Jane Bulloch, Nya Fraleigh, Kati McCartney, Cathy Brummer, Jane Bulloch, Nya Fraleigh,

Sandhya Khurana PhD. Sandhya Khurana PhD. Especially Stacey Ritz PhD.Especially Stacey Ritz PhD.

To all the NOSM Research lab members especially Joe Eibl and To all the NOSM Research lab members especially Joe Eibl and

Heather Peltch for their support.Heather Peltch for their support.

QuestionsQuestions