The FABLE project aims to characterize nanoparticles in ambient air and indoor environments, synthesize similar nanoparticles in the lab, study how these nanoparticles interact with and potentially affect cells and tissues, and use these findings to inform nanoparticle risk assessments and policies. The project involves various work packages focused on nanoparticle characterization, synthesis, cellular studies, toxicity studies, and risk assessment. Several publications have already resulted from the project's early work, and over the next year the team plans to further engage with policymakers and other groups to communicate their findings.

1. From Airborne exposures
to BiologicaL Effects (FABLE):
the impact of
nanoparticles on health
The University of Birmingham
Juana Maria Delgado-Saborit & Josh Rappoport
on behalf of the FABLE Team
J Ayres, K Chipman, M Viant, J Preece, RM Harrison, J Lead, J Rappoport, JM Delgado-Saborit,
I Chang, N Hodges, J Mazzolini, S Chen, R Weber, S Shei, P Sanderson, A Okam, I Kaur, T Knight

2. FABLE Rationale
• Nanoparticles are being produced in substantial amounts for different reasons/end uses
• These may have significant exposures to both humans and the environment.
• Human exposure to manufactured nanoparticles (NPs - dia 1-100nm) in the environment
may impact on health
• As yet, no quantitative data are available on which to assess risk.
• Hence the need for an inter-disciplinary approach to understand the extent of human
exposures to NPs, the biology of NPs and their potential effects on human health.
• We took as a paradigm metallic NPs produced in the environment from vehicle emissions
aiming to relate real life exposures to biological pathways and toxicity in vitro and in vivo.
• The work addresses 8 of the 17 defined research outcomes in the DEFRA scoping report
on research needs in the nanotechnology area (EMERGNANO, 2009)

3. FABLE Work Packages Framework

4. WP1 Characterisation of ambient NPs
AIMS:
1/ Characterise metallic NPs in outdoor and indoor air
2/ Characterise their physical-chemical structure
3/ Develop models to estimate personal exposures to NPs
DATA COLLECTION ANALYTICAL DETERMINATION DATA ANALYSIS
Outdoor Sampling NPs Ambient
Outdoor Sites Selection Gravimetry Characterization of
Concentrations
Monitoring Outdoor Monitoring Exposure
Sampling ICP-MS Composition
Analysis Bulk Chemistry
Volunteers CeO2 NP Source
Recruitment TEM Morphology Apportionment
Indoor
Monitoring Indoor Monitoring
Sampling Composition
X-EDS Other metal NP Source
Surface Chemistry
Apportionment
UFP Exposure
Measurement (*) Composition
Personal EELS
Oxidative State
Exposure Time/Activity Information for
Modelling Diaries NPs synthesis
PE predicted
GPS data PE Models
exposure
Information for
Toxicological studies
QA/QC

5. WP1 UPDATE
1. DATA COLLECTION
1. Outdoor Monitoring
1. Traffic Roadsides complete – Bham and Newcastle
2. Background & Rural Sites : Very advanced
3. Simultaneous sampling of TR and BS to assess traffic contribution to airborne
NPs
2. Indoor & Personal Monitoring
1. Questionnaires completed and ethical approval obtained
2. Recruitment of subjects started
3. Sampling to start by May 2013
3. Minor modifications
1. Increased the length of sampling to 7 days
2. Reduced the number of planned subjects to accommodate new sampling period
3. Additional sampling equipment recruited to speed up indoor sampling
2. ANALYTICAL DETERMINATION
1. Gravimetry - Size fraction distributions of NPs
2. Microscope (TEM/EDX) –physical-chemical characteristics of some NPs
3. Chemical Analysis (GFAA & ICPMS)– extraction method final testing

6. WP2 Synthesis of Nanoparticles
AIM: To generate NPs of CeO2, V2O5 and ZnO and compare these to
those obtained from ambient sampling.
Synthesis of Nanoparticles/Nanorods
using Physical Routes

7. Synthesis of Metal Oxide Nanoparticles/Nanorods
using Wet Chemical Methods 10 nm
<<CeO2 NPs/NRs, Sol-Gel>> CeO2 NPs Dispersion (in pure water, 18MΩ)
XRD: Cubic CeO2 TEM Bham Aldrich NanoAmor
Nano “Duck”
NPs
d = 8 ± 1.6 nm ζ~52 mV ζ~28 mV ζ~10 mV
(stable) (instable) (Instable)
Found by Sean Chen
1+ month 1 week 1 week and Kenton Arkill
Sample prepared by
Sean Chen
STEM operated by
Kenton Arkill
FABLE, University of
Birmingham
http://polkaperson.tripod.com/id5.html
d = 14 ± 2.5 nm d = 25 ± 3.1 nm Size Variation of CeO2 NPs in Serum Media
(L~μm) (L ~ μm)
<<ZnO NRs, Hydrothermal>> Serum-free medium
XRD: Hexagonal ZnO Preliminary results  NPs aggregate
SEM
Serum-containing medium
 Serum stablized NPs

8. WP3 Cellular entry of manufactured NPs
AIM: To use the data on dosage from WP1 and physico-chemical
characterisation from WPs 1 and 2 to study cellular entry/trafficking of
relevant concentrations of generated metallic NPs in culture and tissues,
imaging their intra-cellular location over time
CeO2 (in water) standard curve
2.5
2
Abs 260/280
1.5
1
0.5
0
0 200 400 600 800 1000
Ceria nanoparticle concentration (µg/ml)
NanoDrop analysis

9. WP3 Cellular entry of manufactured NPs
AIM: To use the data on dosage from WP1 and physico-chemical
characterisation from WPs 1 and 2 to study cellular entry/trafficking of
relevant concentrations of generated metallic NPs in culture and
tissues, imaging their intra-cellular location over time
0.14 0.10
0.12
0.08
0.10
DO (260/280)
DO (260/280)
0.06
0.08
0.06 0.04
0.04
0.02
0.02
0.00
0.00
0 min 30 min 60 min 120 min
0 10 100 1000
CeO2 concentration in µg/ml Time of cell incubation with CeO2

10. WP3 Cellular entry of manufactured NPs
Confocal reflectance microscopy
Pulmonary epithelial cells Macrophages
No NPs CeO2 CeO2
Nucleus Nucleus/NPs Nucleus/NPs
10 m 10 m 10 m

11. WP3 Cellular entry of manufactured NPs
Endocytosis of CeO2 by A549 Cells
Control Endocytosis Inhibitor 6
Mean of fluorescence
5
Reflectance
4
3
2
Brightfield
1
0
Control Endocytosis
Inhibitor
Transmission Electron Microscopy
Vesicle
Plasma membrane Endosomal compartment

12. WP4 Toxicology and new challenges from NPs
AIM: To use the data from WPs 1 to 3 to study the toxicity of these NPs on
respiratory epithelial cells and nerve cells in culture providing information
which will allow an understanding of mechanisms of action and the
development of dose response functions.
- To study the toxicity and biological Effects of NP on cell culture models
and Nasal Epithelial Cells (Aberdeen)
- Cytotoxicity assays
- Oxidative stress
- Genotoxic stress
- Transcriptomics
- Metabolomics
- Subsequently to use these data to inform design of in vivo challenge in
rats at HPA (Chilton)

13. WP5 Policy Relevance and Risk Assessment
AIM: Initial risk assessments to be made from these data: this will involve
regular meetings with relevant policy makers/regulators in relevant
Government departments over the course of the project.
Final Goal of the Research is to use the findings to enhance the responsible
development of nanotechnology:
• Influencing the optimisation of production of “safe” materials based on
mechanistic understanding of features related to potential toxicity
•Clarification of appropriate safety assessment methods
•Optimisation of risk assessment

14. FABLE
Points to highlight for EEHI/ESEI Meeting
Major achievements to date
•Publications from each WP are in various stages of preparation (according to their
date of starting work)
•WP3 – Publication in 2012 (JR , JM and others)
•WP1 – Invitation to present at the World Congress of Nanoscience & Technology
(JMS)
•Invitation to ‘Partnership for Progress’ – An initial discussion between Government
and Toxicologists. Attended in February 2013 (TLK)
Key activities over next 6-12months
•Plans to nurture our links with Policy (DEFRA, BIS, DoT etc)
- Meet again - key personnel of policy-influencing
organisations with all FABLE researchers.
•To nurture our links with Imperial College (first met Oct 2012)
- a follow up meeting in 2013, with plans for collaboration.
Significant changes to original plan None to date