COMPETENCE IN TEXTILES
Cellular and Ecotoxicological Analysis of
Nanofunctionalized Textiles (Project TECHNOTOX)
Prof. Dr. Dirk Höfer
BfR, Berlin, February 9th 2012
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No. 2
Hohenstein InstituteDepartment Hygiene, Environment and Medicine
Hohenstein is a public and independent textile research institute.
� Research & Development using public funding
Department Hygiene, Environment & Medicine
� Life science studies on functionalized textiles
� Efficacy and safety of products along the textile chain
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No. 3
Interaction Textile │ Human
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No. 4
Interaction Textile │ Environment
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No. 5
Silver and the human skin microflora
• Field study with 60 subjects (21 – 65 years)
• microbiological and dermatological supervision
• Halved shirts (verum/placebo) were worn
for 8 h/d over a period of 5 weeks
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No. 6
Human skin microflora was not affected
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No. 7
Skin physiology experiments
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No. 8
Research project TECHNOTOX (2011 – 2013)
acc. to Krug & Wick (2011), Angew. Chem. 123(6): 1294-1314
Exposureassessment
Risk-characterization
Risk communication
Risk reduction
Risk prevention
Biologicalhazard
in vitro & in vivobioassays
physico-chemicalmodels
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No. 9
Nanomaterials in textiles
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No. 10
Use of organic biocides and silver in textiles
• 100 t/a organic biocides
• 28 t/a silver
• <0.2 t/a metallic nanosilver (mainly 50-500 nm AgCl particles)
Source: Burkhardt et al. (2011), Forschungsbericht HSR, UMTEC, Rapperswil (CH)
Biocide concentrations in textilesSilver use worldwide
coins
3.1%investment
1.9%
industrial use38.2%
biocide
0.2-0.5%
photography
23.8%
jewellery32.5%
µ-compositAg
AgCl
Ag-zeolite
Triclosan
Quat
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No. 11
Fibre functionalization with nanosilver
Surface modification
Material modification
� Different abrasion behaviour � Exposition
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No. 12
Textile abrasion
Mechanical stress (worst case scenario) Detection and collection
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No. 13
Distribution sites
Human
Eco
Source: PLoS Source: www.rad-zep.deSource: wikipedia
Food chain
skin alimentary tract Lung
bacteria DaphniaZebra fish
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No. 14
The lung as ‚main entrance‘ for nanoparticles
Size matters
from Krug & Wick (2011), Angew. Chem. 123(6): 1294-1314
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No. 15
TECHNOTOX lung model
37°C heating plate
Aerosol generator Particle detectorflow chamber = „Lung“
30 min; 2,2 l/min
10
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No. 16
Lung model
Lung cell vitality after 30 min exposure to NP-aerosol
30% = Thresholdacc. to DIN EN ISO 10993-5
Zytotoxizität der Lungenzellen nach Benebelung im Aerosolgenerator
0
10
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60
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100
TiO2 ZnO AgPURE
Gro
wth
red
ucti
on
[%
]
Cell growth reduction after NP-aerosol exposure
TiO2 ZnO nAg (OECD NM300)
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No. 17
Cell toxicity studies– according to DIN EN ISO 10993-5
Different cell types vary in their susceptibility!
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PK
Sol 1
1:2
Sol 1
1:4
Sol 1
1:1
0
aus
Luft
gewasc
hene
Ag P
artik
el 1
:2
aus
Luft
gewasc
hene
Ag P
artik
el 1
:4
aus
Luft
gewasc
hene
Ag P
artik
el 1
:10
10 m
g/L
AgPU
RE
7,5
mg/
L AgP
UR
E 5
mg/L
AgP
URE
0,5
mg/
L AgP
UR
E
0,05
mg/
L AgPU
RE
50 m
g/L
iSyS
SU
N
500
mg/
L FLE
XO
FIX
50 m
g/L
FLEXO
FIX
Gro
wth
red
ucti
on
[%
]
Leber
Bindegewebe
Lunge
Makrophage / Immun
Liver
Connective tissue
Lung
Macrophages
POS Si-Sol1:2
Si-Sol1:4
Si-Sol1:10
nAg wash-out (air)
1:2 1:4 1:10
nAg (OECD NM300) product
[mg/l]
0.510 7.5 5 0.05 TiO2
product50 mg/l
SiO2
product500 mg/l
SiO2
product50 mg/l
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No. 18
nAg versus AgNO3 – lung cells
Silver ions are toxic for cells
Lung cells A549
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60
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25 mg/L 10 mg/L 5 mg/L
Gro
wth
red
ucti
on
[%
]
AgPURE
AgNO3
nAg
AgNO3
25 mg/l 10 mg/l 5 mg/l
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No. 19
nAg versus AgNO3 – liver cells
Dose-response relation
Liver cells HepG2
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25 mg/L 10 mg/L 5 mg/L 2mg/L 1mg/L
Gro
wth
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ucti
on
[%
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AgPURE
AgNO3
nAg
AgNO3
25 mg/l 10 mg/l 5 mg/l 2 mg/l 1 mg/l
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No. 20
Effects of nAg on Daphnia magna
• acc. to DIN EN ISO 6341:2010
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100
120
Pos
itivk
ontro
lle
Nega
tivko
ntro
lle 0,1
0,25 0,
5 1
2,5 5 10
nAg (NM300) product [mg/L]
To
te D
ap
hn
ien
[%
]
24h
48h
72h
POS NEG 0.1 0.25 0.5 1 2.5 5 10
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No. 21
Effects of aerosols and textile abrasion samples
• Luminescent bacteria test acc. to DIN EN ISO 11483-1
Sol 2 b) Abrieb
Luft- Wasser
Wasser
Referenz (Nr.21)
30 min
-75
-50
-25
0
25
50
75
100
Zinksulfat (pos.
Kontrolle)
Ag-Pure aus Luft
(Nr.15)
Hem
mun
g [%
]
POS nAg (NM300) Sol (SiO2) H2O(ZnSO4) wash-out (air) abrasion
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No. 22
Effects of textile abrasion samples
Daphnia magna lethality after 48 h exposition
Lethality of Daphnia magna
0
10
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60
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80
90
100
0,974 mg/L AgPURE 8,1 mg/L SiO2 Sol 2b) Abrieb Positivkontrolle
Leth
ali
ty [
%]
nAg (NM300) SiO2 Sol (SiO2) POS wash-out (air) wash-out (air) abrasion
final conc. 0.1 mg/l final conc. 8.1 mg/l
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No. 23
Zebrafish early larval stage test
acc. to DIN EN ISO 15988
Lethality of Danio rerio
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neg NM300 600 ppm NM300 1200 ppm
Leth
ali
ty [
%]
neg NM300 product600 ppm 1200 ppm
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No. 24
Outlook TECHNOTOX project
• More realistic exposition scenarios
• Testing further textile abrasion samples
� genotoxic potential
� zebrafish early larval stage test
• Human 3D skin model
• Mucosa penetration (mouth, gut)
• Resorption into the blood
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No. 25
Outlook TECHNOTOX dialog
• Scientific publications
• Public presentation in web
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No. 26
Conclusion
• Safety of products and processes is assessed with
standardized biological test systems. These should be
reliable, robust, sensitive and predictive.
• Realistic working concentrations must be used. No
overload experiments.
• Textile products should be evaluated seperately
depending on the exposition scenario.
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No. 27
Thanks for your attention !
Prof. Dr. Dirk Höfer
Hohenstein Institute
Schloss Hohenstein
74357 Bönnigheim
Telefon +49 7143 271 432
Fax +49 7143 271 94432
E-Mail [email protected]
www.hohenstein.de