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

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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!

0

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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

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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

0

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25 mg/L 10 mg/L 5 mg/L

Gro

wth

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ucti

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[%

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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

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[%

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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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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

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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 d.hoefer@hohenstein.de

www.hohenstein.de