Cellular and Ecotoxicological Analysis of ... · COMPETENCE IN TEXTILES Cellular and Ecotoxicological Analysis of Nanofunctionalized Textiles (Project TECHNOTOX) Prof. Dr. Dirk Höfer

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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90

100

TiO2 ZnO AgPURE

Gro

wth

red

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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

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

0

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60

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90

100

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

0

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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

0

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80

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

20

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40

50

60

70

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

0

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80

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100

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