Thinking Small - Nanoparticles in Cosmetic Preparations - A Consumer or Occupational Health Risk ~ ecopa.eu

1. 11
THINKING SMALL: NANOPARTICLES INTHINKING SMALL: NANOPARTICLES IN
COSMETIC PREPARATIONS - A CONSUMERCOSMETIC PREPARATIONS - A CONSUMER
OR OCCUPATIONAL HEALTH RISK?OR OCCUPATIONAL HEALTH RISK?
Gerhard J. Nohynek, M.Sc.,Gerhard J. Nohynek, M.Sc., Ph.D.Ph.D., D.A.B.T., D.A.B.T.
LL’’OREAL GLOBAL SAFETY EVALUATIONOREAL GLOBAL SAFETY EVALUATION
gnohynecgnohynec@@rd.loreal.comrd.loreal.com
ECOPA WORKSHOPECOPA WORKSHOP
DecemberDecember 17, 200517, 2005

2. 22
THE CHALLENGE OF UNDERSTANDINGTHE CHALLENGE OF UNDERSTANDING
NANOSCALE ISNANOSCALE IS SIZESIZE……
NanomaterialsNanomaterials havehave atat leastleast oneone dimensiondimension atat <100 nm<100 nm
1 dimension:1 dimension: layerslayers (films,(films, coatingscoatings))
2 dimensions:2 dimensions: nanotubesnanotubes,, nanowiresnanowires
3 dimensions: nanoparticles (3 dimensions: nanoparticles (NPsNPs))
100 m 10-1 m 10-2 m 10-3 m 10-4 m 10-5 m 10-6 m 10-7 m 10-8 m 10-9 m 10-10 m
(1 m) (1 mm) (1 m) (1 nm)(100 nm)
hair (80 m) RBC (7 m)Football (30 cm) Flea (1 mm) Herpes virus (100 nm)
Fullerene, C60, buckyball
(0.7 nm)
GN/ED: 10/2005
Salicylic acid
(1 nm)
TiO2 nanoparticles (40
to 200 nm

3. 33
SOURCES OF NANOMATERIALSSOURCES OF NANOMATERIALS
BiologicalBiological::
FerritinFerritin
ATPATP synthasesynthase
VirusesViruses,, nanobacteriananobacteria
NaturalNatural::
VulcanicVulcanic ashash
SeaspraySeaspray
ForestForest firesfires
ErosionErosion
Man-madeMan-made::
DieselDiesel exhaustexhaust
FiresFires / toasters/ toasters
ManufacturedManufactured nanoparticlesnanoparticles
OneOne cmcm33 ofof urbanurban airair containscontains
>10.000 nanoparticles>10.000 nanoparticles

4. 44
WORLD-WIDE PRODUCTION OFWORLD-WIDE PRODUCTION OF NPsNPs ((mtmt//yearyear) *) *
* Source: ECETOC, 2005
10103 -3 - 101044
1010221010NanofiltrationNanofiltration, membranes, membranesEnvironmentalEnvironmental
101011<1<1
NanocompositesNanocomposites andand encapsulatesencapsulates,,
targetedtargeted drugdrug deliverydelivery, diagnostic, diagnostic
markers,markers, biosensorsbiosensors
BiotechnologyBiotechnology
>10>1033101022
1010
NanoelectronicNanoelectronic andand optoelectronicoptoelectronic
materialsmaterials,, organicorganic lightlight emittersemitters,,
nanophosphorsnanophosphors
Information,Information,
communicationcommunication
technologiestechnologies
101033101033101033MetalMetal oxidesoxides (ZnO, TiO(ZnO, TiO22))Skin careSkin care
101044 - 10- 1055
1010441010
CeramicsCeramics,, catalystscatalysts, films,, films,
coatingscoatings,, metalsmetals
StructuralStructural
20202020201020102003/042003/04MATERIAL / DEVICEMATERIAL / DEVICEAPPLICATIONAPPLICATION
GN/ED: 10/200
Woldwide research spending on NMs has tripled between
1997 and 2002 (700 to >2000 M$)

5. 55
TWO PRINCIPAL FEATURES MAY AFFECT PHYSICAL ANDTWO PRINCIPAL FEATURES MAY AFFECT PHYSICAL AND
TOXICOLOGICAL PROPERTIES OFTOXICOLOGICAL PROPERTIES OF NPsNPs//NMsNMs
QuantumQuantum effectseffects
importantimportant atat thethe lowlow endend ofof
nanoscalenanoscale
maymay produceproduce changes inchanges in opticaloptical,,
magneticmagnetic, thermal or, thermal or
conductivityconductivity propertiesproperties
IncreasedIncreased surface areasurface area perper
unitunit massmass
1 mL1 mL ofof nanoparticles (2.5 nm; 5nanoparticles (2.5 nm; 5
g/cmg/cm33
)) hashas a surfacea surface ofof 240 m240 m
SurfaceSurface maymay affect dissolutionaffect dissolution
kineticskinetics // bioavailabilitybioavailability oror
increasedincreased surfacesurface activityactivity
GN/ED: 10/200
Supergel: nano-SiOnano-SiO22 ++
waterwater

6. 66
NANO-TOXICOLOGY:NANO-TOXICOLOGY: TYPICAL TOXICOLOGICALTYPICAL TOXICOLOGICAL
PROPERTIES OFPROPERTIES OF NPsNPs DO NOT EXISTDO NOT EXIST
NOTE:NOTE: ToxicologicalToxicological profilesprofiles ofof substances,substances, bulkbulk, micro, nano,, micro, nano,
vapourvapour or solution, tend toor solution, tend to bebe thethe samesame oror similarsimilar
ParticleParticle sizesize ofof a substancea substance hashas littlelittle impact onimpact on itsits toxicologicaltoxicological profile,profile,
UNLESSUNLESS::
SizeSize effecteffect: absorption or absorption: absorption or absorption kineticskinetics maymay bebe affectedaffected byby particleparticle
sizesize
SurfaceSurface effectseffects: surface: surface activityactivity playsplays aa rolerole (relative surface(relative surface increasesincreases
withwith particleparticle sizesize))
EfectEfect onon externalexternal exposureexposure:: smallersmaller particlesparticles longerlonger timetime ofof
settlementsettlement increasedincreased inhalationinhalation exposureexposure
RelativeRelative particleparticle surfacesurface:: mm-particlesmm-particles <<<< microparticlesmicroparticles
<< nanoparticles << solutions or<< nanoparticles << solutions or vapoursvapours ((individualindividual moleculesmolecules))
GN/ED: 10/200

7. 77
INHALATION EXPOSURE:INHALATION EXPOSURE: NPsNPs TEND TO AGGREGATE /TEND TO AGGREGATE /
AGGLOMERATE IN THE AIR *AGGLOMERATE IN THE AIR *
GN/ED: 11/200* A. Maynard, NIOSH, June 2005
NB.: particle size increases by a factor of 104 within 1 minute!
(Major technical challenge for inhalation toxicity studies)
ZnO

8. 88
HUMAN RESPIRATORY DEPOSITION OF PARTICLESHUMAN RESPIRATORY DEPOSITION OF PARTICLES
DepositionDeposition mainlymainly
dependsdepends onon particleparticle sizesize
NanoparticleNanoparticle depositiondeposition
primarilyprimarily by diffusionby diffusion
EffectEffect ofof shapeshape andand
agglomerationagglomeration:: unclearunclear
5-305-30 mm:: extrathoracicextrathoracic
regionregion::
impactionimpaction
1-5 m: thoracic region
bronchial, bronchiolar:
sedimentation,
11 mm:: aveolaraveolar regionregion::
diffusiondiffusion
GN/ED: 11/200

9. 99
INHALATION EXPOSURE TO SMALLINHALATION EXPOSURE TO SMALL
PARTICLES: PARTICLE SIZE DETERMINESPARTICLES: PARTICLE SIZE DETERMINES
REGION OF DEPOSITION *REGION OF DEPOSITION *
* ECETOC, 2005
GN/ED: 10/2005
0
10
20
30
40
50
60
70
80
1 10 100 1000
Diameter, nm
Deposition,%
ET
Bb
AI
Mouth breathing
Nose breathing
NB: at <100 nm, ET (nose, upper airways) deposition increases!
Regional deposition of inhaled NP with
diameters between 1 nm and 1000 nm for
nose and for mouth breathing in the
extrathoracic airways (ET), the bronchial
airways (Bb) and the alveolar region (AI)
during breathing at rest, as predicted by ICRP
66 model (ICRP, 1994)
Silicosis

10. 1010
PHYSIOPATHOLOGY OF LUNG OVERLOADPHYSIOPATHOLOGY OF LUNG OVERLOAD
((InertInert, insoluble, insoluble particlesparticles: TiO: TiO22 andand CB)CB)
MacrophageMacrophage overloadoverload persistent inflammationpersistent inflammation cellcell injuryinjury necrosisnecrosis,, cellcell proliferationproliferation
MechanismMechanism ((rodsentrodsent): inflammation): inflammation cellcell proliferationproliferation fibrosisfibrosis,, andand eventuallyeventually lunglung
tumourstumours
WhenWhen particlesparticles cannotcannot bebe removedremoved by macrophagesby macrophages (fibres)(fibres) increasedincreased raterate ofof cellcell deathdeath,,
inflammationinflammation andand adverseadverse effectseffects fibrosisfibrosis tumourstumours ((asbestosasbestos))
Rats areRats are thethe mostmost sensitivesensitive speciesspecies forfor lunglung overloadoverload ((highhigh breathingbreathing rate /rate / reducedreduced particleparticle
clearance)clearance)
GN/ED: 11/200
Alveolar macrophagemacrophage and
asbestos fiber
Phagocyte migration
Macrophage
phagocytosis and
migration – Fe NPs
(Dr. D. Warheit,
Nov. 2004)

11. 1111
EXAMPLE 1: MICROFINE (EXAMPLE 1: MICROFINE (mfmf) AND ULTRAFINE () AND ULTRAFINE (ufuf) TiO) TiO22::
COMPARATIVE INHALATION TOXICITY *COMPARATIVE INHALATION TOXICITY *
No qualitative differences between uf- and mf-TiO2 toxicity ((nono newnew
hazardshazards withwith uf-TiOuf-TiO2))
EffectsEffects typicaltypical forfor particleparticle overloadoverload withwith associatedassociated changeschanges
SpeciesSpecies sensitivitysensitivity: r>m>h: r>m>h
SimilarSimilar pulmonarypulmonary effectseffects atat 50 mg/m50 mg/m33 mf-TiOmf-TiO2 andand 10 mg/m10 mg/m33 uf-uf-
TiO2TiO2
PulmonaryPulmonary effectseffects//lunglung burdenburden correlatecorrelate bestbest withwith surface areasurface area ofof
totaltotal particleparticle exposureexposure
ComparisonComparison ofof lung-associatedlung-associated lymphlymph nodenode burdenburden suggestssuggests sinimalsinimal
translocationtranslocation ofof ufuf-- andand mf-TiO2mf-TiO2 underunder non-overloadnon-overload conditionsconditions
CONCLUSION:CONCLUSION: ufuf TiOTiO22 maymay requirerequire aa somewhatsomewhat lowerlower TLV valueTLV value thanthan
presentpresent inertinert dustdust value (5-timesvalue (5-times lowerlower?)?)
GN/ED: 10/200
* CIIT, 2003/2004

12. 1212
EXAMPLE 2: INHALATION OF SIOEXAMPLE 2: INHALATION OF SIO22 DUSTS PRODUCESDUSTS PRODUCES
FIBROSIS:FIBROSIS: NPsNPs APPEAR LESS POTENT THAN SIOAPPEAR LESS POTENT THAN SIO22 MPsMPs
Intra-trachealIntra-tracheal (IT) instillation wih 20 mg SiO(IT) instillation wih 20 mg SiO22,, nano-sizenano-size
(10(10±5 nm) or micro-size (1-5 m)±5 nm) or micro-size (1-5 m)
FibroticFibrotic grade of rat lungsgrade of rat lungs
+ = cellular nodules; ++ =+ = cellular nodules; ++ = fibroticfibrotic cellular nodules, +++ = large cellularcellular nodules, +++ = large cellular fibroticfibrotic nodulesnodules
GN/ED: 10/200
** ChenChen etet alal..,, ToxTox IndInd HealthHealth (2005)(2005)
++++NanoNano
SiOSiO22
++/+++++/++++/+++/++MicroMicro
SiOSiO22
0000SalineSaline
22 monthsmonths11 monthmonth

13. 1313
EXAMPLE 3: SINGLE-WALL CARBON NANOTUBESEXAMPLE 3: SINGLE-WALL CARBON NANOTUBES
PRODUCED GRANULOMA FOLLOWING INTRATRACHEALPRODUCED GRANULOMA FOLLOWING INTRATRACHEAL
INSTILLATION IN RODENTS *INSTILLATION IN RODENTS *
Intra-trachealIntra-tracheal instillationinstillation ofof differentdifferent
gradesgrades ofof CNTsCNTs vs.vs. carboncarbon blackblack andand quartzquartz
positive control groupspositive control groups
NoNo effectseffects in CB groups,in CB groups, inflammatoryinflammatory reactionsreactions //
cytotoxicitycytotoxicity followingfollowing quartzquartz exposureexposure ((expectedexpected))
DeathsDeaths ++ interstitialinterstitial epitheloidepitheloid granulomasgranulomas ((foreignforeign
bodybody reactionreaction) in) in CNT-groupsCNT-groups
DeathsDeaths relatedrelated to «to «bolusbolus» administration» administration
(IT instillation)(IT instillation)
New inhalationNew inhalation toxicitytoxicity studiesstudies ongoingongoing (US(US
NTP)NTP)
GN/ED: 10/200
** WarheitWarheit etet alal.,., LamLam etet alal., Inhalation., Inhalation ToxicologyToxicology,, 20042004

14. 1414
LUNG GRANULOMA (RAT) 1 MONTH AFTER SWCNT IT-LUNG GRANULOMA (RAT) 1 MONTH AFTER SWCNT IT-
INSTILLATION: FIBER-, BUT NOT NANO-RELATEDINSTILLATION: FIBER-, BUT NOT NANO-RELATED
TOXICITY *TOXICITY *
* Slide by the courtesy of Dr. D. Warheit / DuPont, USA
ReactionReaction ofof thethe lunglung toto foreignforeign bodiesbodies thatthat cancan notnot bebe removedremoved
((typicaltypical for fibres,for fibres, nano-unrelatednano-unrelated))

15. 1515
GN/ED: 11/200
TransportTransport ofof inhaledinhaled NPsNPs toto
thethe olfactoryolfactory bulbbulb
cerebrumcerebrum cerebellumcerebellum.. (?).. (?)
needsneeds toto bebe confirmedconfirmed
((1414C-studyC-study ongoingongoing))
EXAMPLE 4: TRANSLOCATION OF INHALED 13C NP INTO
THE OLFACTORY BULB AND THE BRAIN? (Oberdörster,
2003)

16. 1616
OCCUPATIONAL EXPOSURE DURING MANUFACTURINGOCCUPATIONAL EXPOSURE DURING MANUFACTURING
OF SWCNT OR CARBON BLACKOF SWCNT OR CARBON BLACK
Concentrations during handling materialConcentrations during handling material
were very low:were very low:
A. Maynard (US NIOSH): always < 53A. Maynard (US NIOSH): always < 53 μμ
g/mg/m33
DuPont (US)DuPont (US) studystudy:: exposureexposure levelslevels
belowbelow limitlimit ofof detectiondetection
EuropeanEuropean studiesstudies in CBin CB manufacturingmanufacturing
plants:plants: veryvery lowlow exposureexposure,, mainlymainly duedue
toto externalexternal sources,sources, suchsuch asas forkfork lifts,lifts,
gasgas heatingheating oror neighbouringneighbouring traffictraffic
((KuhlbuschKuhlbusch etet alal., 2004)., 2004)
InternalInternal cosmeticcosmetic industryindustry datadata suggestsuggest
negligiblenegligible inhalationinhalation riskrisk fromfrom or nor n
TiOTiO22 (<10%(<10% ofof maxmax dustdust value +value +
respiratoryrespiratory protection)protection)
GN/ED: 11/2005
Raw SWCNTs during
handling (DuPont, US)
Carbon Black Manufacture
(DEGUSSA, D)

17. 1717
HUMAN ORAL EXPOSURE TOHUMAN ORAL EXPOSURE TO NPsNPs: PLASMA KINETICS OF: PLASMA KINETICS OF
A POORLY SOLUBLE ORAL DRUG (PHENACETIN): ROLEA POORLY SOLUBLE ORAL DRUG (PHENACETIN): ROLE
OF PARTICLE SIZEOF PARTICLE SIZE
GN/ED: 10/200

18. 1818
HUMAN ORAL EXPOSURE TO NANOPARTICLESHUMAN ORAL EXPOSURE TO NANOPARTICLES
10101212 to 10to 101414 micro or nanoparticles (0.1 to 3micro or nanoparticles (0.1 to 3 mm)) ingestedingested dailydaily
((mainlymainly silicatessilicates andand titaniumtitanium dioxidedioxide).). UnclearUnclear whetherwhether man-man-
mademade NPNP presentpresent anan additionaladditional burdenburden..
SystemicSystemic absorptionabsorption studiesstudies onon NPsNPs hadhad mixedmixed resultsresults::
SomeSome studiesstudies reportreport slightslight systemicsystemic exposureexposure ((liverliver, spleen,, spleen, lymphlymph
nodesnodes,, bloodblood,, GI-mucosaGI-mucosa))
OtherOther studiesstudies reportedreported nono systemicsystemic uptakeuptake byby thethe GI-systemGI-system
NoNo evidenceevidence forfor targettarget organorgan toxicitytoxicity
CONCLUSIONCONCLUSION:: atat presentpresent,, humanhuman systemicsystemic exposureexposure toto
nanoparticlesnanoparticles afterafter oraloral uptakeuptake isis unclearunclear;; nono evidenceevidence forfor
adverseadverse effectseffects or NPor NP storagestorage inin thethe humanhuman bodybody
* ECETOC, 2005; ** Böckmann et al., 2000; *** Jani et
al., 1992; Jani et al., 1994; **** Kanapilly and Diel,
1980; Kreyling et al., 2002
GN/ED: 11/200

19. 1919
HUMAN DERMAL EXPOSURE TOHUMAN DERMAL EXPOSURE TO NPsNPs::
COSMETICSCOSMETICS
GN/ED: 10/200
Use / exposure: NPs used in
cosmetics consist mainly of ZnO or
TiO2 (sunscreens)
Systemic exposure: penetration of
NPs into / through the skin, systemic
exposure?
Hazard: does nano-size increase the
reactivity / toxicity of cosmetic NPs,
such as ZnO or TiO2?
Risk management: can a chemical /
photo-chemical / biological activity of
ZnO or TiO2 be modified (coating)?

20. 2020
PUBLISHED IN VIVO STUDIES ON DERMAL ABSORPTIONPUBLISHED IN VIVO STUDIES ON DERMAL ABSORPTION
OF NANOPARTICLES SHOW NO PENETRATIONOF NANOPARTICLES SHOW NO PENETRATION
NoNo penetrationpenetration intointo epidermisepidermis // dermisdermis inin
vivo,vivo, manman
FluorescentFluorescent polymericpolymeric NPsNPsHoward, P, 2005Howard, P, 2005
((SoTSoT Meeting, 2005)Meeting, 2005)
NoNo evidenceevidence forfor penetrationpenetration intointo livingliving
skin (skin (preliminarypreliminary data, ECETOC, 11/2005)data, ECETOC, 11/2005)
FluorescentFluorescent particlesparticlesEUEU NanodermNanoderm projectproject
((TilmannTilmann ButzButz))
NoNo penetrationpenetration intointo epidermisepidermis // dermisdermis,,
accumulation inaccumulation in thethe folliclefollicle orifice, butorifice, but nono
penetrationpenetration ibtoibto living skin (living skin (pigpig))
PolystyrenePolystyrene NPsNPs, 20, 20 andand
200 nm200 nm
Alvarez-RomanAlvarez-Roman etet alal.,.,
20042004
TiOTiO2 ,2 , 1010 andand 100 nm100 nmPflückerPflücker etet alal., 2001., 2001
NoNo penetrationpenetration intointo epidermisepidermis // dermisdermis inin
vivo,vivo, manman
Microfine TiOMicrofine TiO22LademannLademann, 1999, 1999
Microfine TiOMicrofine TiO22Tan etTan et alal., 1996., 1996
RESULTSRESULTSMATERIALMATERIALSTUDYSTUDY
CONCLUSION:CONCLUSION: NO EVIDENCE THAT TOPICALLY APPLIEDNO EVIDENCE THAT TOPICALLY APPLIED
NANOPARTICLES PENETRATE INTO NORMAL SKINNANOPARTICLES PENETRATE INTO NORMAL SKIN
((NPsNPs willwill alwaysalways penetratepenetrate lessless thanthan a compound in solution!)a compound in solution!) GN/ED: 10/200

21. 2121
IN VIVO PERCUTANEOUS ABSORPTION OF TiOIN VIVO PERCUTANEOUS ABSORPTION OF TiO22
NANOPARTICLES: RECOVERY FROM THE STRATUMNANOPARTICLES: RECOVERY FROM THE STRATUM
CORNEUM OF HUMAN SKIN *CORNEUM OF HUMAN SKIN *
RELATIVE
HORNY
LAYER
THICKNESS
[%]
0
100
TITANIUM CONCENTRATION [ g/square centimetre tape]
5
4
14
NUMBER
OF
TAPE
STRIPPING
1
5
10
15
40
50
60
78
0,05
0,2
0,4
0,4
0,3
1
2
3
Conclusion: some absorption into the upper layers of the stratum corneum,
but no penetration
GN/ED: 10/200* Lademann et al., 1999

22. 2222
PERCUTANEOUS PENETRATION (PIG SKIN) OF ZnO ANDPERCUTANEOUS PENETRATION (PIG SKIN) OF ZnO AND
TiOTiO22––CONTAINING SUNSCREEN-GRADECONTAINING SUNSCREEN-GRADE NPsNPs ININ
COSMETIC FORMULATIONS *COSMETIC FORMULATIONS *
* Gamer et al., Toxicol. In Vitro, 2005, BASF GN/ED: 10/200
ZnO (10%), mean particle size: 80 nm TiOTiO22 (10%),(10%), meanmean particleparticle sizesize: 30-60 x: 30-60 x
10 nm10 nm

23. 2323
IN VITRO PERCUTANEOUS PENETRATION OF TiOIN VITRO PERCUTANEOUS PENETRATION OF TiO22 ANDAND
ZnO IN PIG SKIN (4 mg/cm , 24-hrs): ABSORBED DOSEZnO IN PIG SKIN (4 mg/cm , 24-hrs): ABSORBED DOSE
= ZERO *= ZERO *
0.00.0
0.00.0
0.00.0
ABSORBEDABSORBED
DOSEDOSE
(%)(%)
0.8 to 1.4 **0.8 to 1.4 **
0.00.0
0.00.0
RECEPT.RECEPT.
FLUIDFLUID
(%)(%)
102.3 to102.3 to
106.8106.8
1.4 to 1.5 **1.4 to 1.5 **
98.6 to98.6 to
102.3102.3
NPNP
ZnO, 80 nm, O/WZnO, 80 nm, O/W
emulsionemulsion (10.3%)(10.3%)
86.1 to86.1 to
93.093.0
0.1 to 0.50.1 to 0.50.0 to 0.30.0 to 0.385.4 to 92.985.4 to 92.9
TiOTiO22, 30-60 x 10 nm,, 30-60 x 10 nm,
methicone-coatedmethicone-coated O/WO/W
emulsionemulsion (10%)(10%)
98.2 to98.2 to
100.4100.4
0.1 to 0.30.1 to 0.30.1 to 0.20.1 to 0.297.7 to 100.297.7 to 100.2
TiOTiO22, 30-60 x 10 nm,, 30-60 x 10 nm, silicasilica
// methicone-coatedmethicone-coated O/WO/W
emulsionemulsion (10%)(10%)
RECOV.RECOV.
(%)(%)
SKINSKIN
(%)(%)
TAPETAPE
STRIPSSTRIPS
(%)(%)
SKINSKIN
WASHWASH
(%)(%)
TEST MATERIALTEST MATERIAL
Gamer et al., Toxicol. In Vitro, BASF, 2005; ** values at or below background levels
GN/ED: 10/2005
CONCLUSION:CONCLUSION: NO EVIDENCE FOR PERCUTANEOUS PENENTRATIONNO EVIDENCE FOR PERCUTANEOUS PENENTRATION

24. 2424
Liposomes
100-300 nm
(Caffeine)
Water
Lipid
Nanocapsule
100-600 nm
(Vitamin A, E)
Polymer
Nanoemulsion
50 nm
(transparent)
Oleosomes
150-500 nm
Examples of nanomaterials
NANO-SIZED COSMETIC FORMULATIONSNANO-SIZED COSMETIC FORMULATIONS
Oil

25. 2525
A complete review on liposomes in cosmetics and drugs concluded that liposomes do
not penetrate through the intact stratum cormeum (SC) or enhance penetration of active
ingredients (1995)
Joke Bouwstra (University of Leiden) published more than 30 articles on the
percutaneous penetration of liposomes or similar formulations using 14C-labelled capsule
membranes.
It was concluded that lipids from soft capsules penetrate into the deep layers of the
SC, but were absent in the living skin. Lipids form hard capsules were found only in / on
the superficial leyers of the SC.
Intact capsules – hard or soft - were only found on the surface of the SC
1. Imbert D and Wickett R: Topical delivery with liposomes. Cosmetics and Toiletries magazine. 1995; 111:32-45.
2. Honeywell-Nguyen P et al.: Quantitative assessment of the transport of elastic and rigid vesicle components and a model drug from these vesicle formulations
into human skin in vivo. Journal of Investigative Dermatology. 2004; 123(5):902-10.
3. Van den Bergh B et al.: Interactions of elastic and rigid vesicles with human skin in vitro: electron microscopy and two-photon excitation microscopy.
Biochimica and Biophysica Acta. 1999; 1461:155-173.
NANO-SIZED COSMETIC FORMULATIONS DO NOT
PRODUCE PENETRATION INTO OR THROUGH THE LIVING
SKIN

26. 2626
SKIN PENETRATION OF SMALL MOLECULES INSKIN PENETRATION OF SMALL MOLECULES IN
SOLUTION VS. INSOLUBLESOLUTION VS. INSOLUBLE NPsNPs**
DIFFUSION OF MOLECULES INTO THE
SKIN IS LIKE A BREAKING DAM
* Prof. T. Butz, Chairman Nanoderm Task Force, 11/2005
NPs MOVE BY MECHANICAL FORCE:
WHY SHOULD A ROCK MOVE ONLY
IN ONE DIRECTION? (NO
MECHANISM TO DRIVE ACTIVE
PENETRATION)
?
?

27. 2727
GENOTOXICITY / PHOTO-GENOTOXICITY / PHOTO-
GENOTOXICITY OF TiOGENOTOXICITY OF TiO22 AND ZnOAND ZnO
NANOPARTICLESNANOPARTICLES
GN/ED: 11/200

28. 2828
PHOTO-GENOTOXICITY OF 10 TiOPHOTO-GENOTOXICITY OF 10 TiO22-POWDERS-POWDERS
USED IN COSMETICS *USED IN COSMETICS *
UponUpon requestrequest ofof thethe SCCNFP (1999), anSCCNFP (1999), an industryindustry
consortium (TiOconsortium (TiO22 producersproducers andand cosmeticcosmetic
industryindustry)) investigatedinvestigated
CytotoxicityCytotoxicity
GenotoxicityGenotoxicity andand photo-genotoxicityphoto-genotoxicity ((AmesAmes, CHO), CHO)
TestTest materialmaterial: TiO: TiO22
NPsNPs andand microparticlesmicroparticles
RutileRutile andand anatase (anatase (photo-activephoto-active) TiO) TiO22-crystalline-crystalline formsforms
CoatedCoated andand non-coatednon-coated particlesparticles
ProgramProgram performedperformed by COVANCE / UKby COVANCE / UK
GN/ED: 10/200
* SCCNFP, 24 October, 2000

29. 2929
GENO- AND PHOTO-GENOTOXICITY OF TiOGENO- AND PHOTO-GENOTOXICITY OF TiO22 PARTICLESPARTICLES
(AMES TEST, CHO CELLS)(AMES TEST, CHO CELLS)
negativenegativenegativenegativeAlAl22OO33//StearicStearic acidacid1515RutileRutile88
negativenegative
negativenegative
negativenegative
negativenegative
negativenegative
negativenegative
negativenegative
negativenegative
negativenegative
AmesAmes // Photo-Photo-
AmesAmes
11-2811-28
1515
2020
1717
2020
200.000200.000
6060
6060
1414
MEANMEAN
PARTICLE SIZEPARTICLE SIZE
(nm)(nm)
negativenegativeAlAl22OO33/SiO/SiO22RutileRutile1010
negativenegativeUncoatedUncoatedRutileRutile99
negativenegativeUncoatedUncoatedRutileRutile77
negativenegativeAlAl22OO33//StearicStearic acidacidRutileRutile66
negativenegativeAlAl22OO33//DimethiconeDimethiconeRutileRutile55
negativenegative **UncoatedUncoatedAnataseAnatase44
negativenegativeUncoatedUncoatedAnataseAnatase33
negativenegativeAlAl22OO33/SiO/SiO22AnataseAnatase22
negativenegativeAlAl22OO33//DimethiconeDimethiconeRutileRutile11
CHO /CHO /
Photo-CHOPhoto-CHO
COATINGCOATINGCRYSTALLINECRYSTALLINE
FORMFORM
NAMENAME
* Some inconclusive / positive results at high cytotoxic levels; overall rated negative
GN/ED: 10/200

30. 3030
CYTOTOXICITY AND PHOTO-GENOTOXICITY OF TiOCYTOTOXICITY AND PHOTO-GENOTOXICITY OF TiO22
(NANO/MICRO/COATED/UNCOATED/RUTILE/ANATASE)(NANO/MICRO/COATED/UNCOATED/RUTILE/ANATASE)
PARTICLES *PARTICLES *
AllAll TiOTiO22 materialsmaterials showedshowed minimal cytotoxicity inminimal cytotoxicity in thethe absenceabsence andand
presencepresence ofof UVUV
AllAll TiOTiO22 materialsmaterials werewere negativenegative inin thethe AmesAmes-,-, photo-Amesphoto-Ames andand CHO-,CHO-,
andand photo-CHOphoto-CHO teststests
UV,UV, particleparticle sizesize oror crystallinecrystalline formform hadhad nono effecteffect onon cytoxiccytoxic or genotoxicor genotoxic
potentialpotential ofof TiOTiO22 materialsmaterials
InIn vivovivo ((hairlesshairless micemice), TiO), TiO22 protectsprotects againstagainst genotoxicgenotoxic andand carcinogeniccarcinogenic
activityactivity ofof UV lightUV light
Conclusion:Conclusion: finefine particleparticle TiOTiO22 isis notnot expectedexpected toto presentpresent a genotoxic ora genotoxic or
photo-genotoxicphoto-genotoxic riskrisk forfor humanshumans underunder normal conditionsnormal conditions ofof useuse
* Expert report, Dr. David* Expert report, Dr. David KirklandKirkland, COVANCE, 16, COVANCE, 16 SeptemberSeptember, 1999;, 1999; alsoalso seesee opnionopnion ofof thethe SCCNFP, 24SCCNFP, 24 OctoberOctober 2000,2000,
http://http://europa.eu.inteuropa.eu.int//commcomm//healthhealth//ph_riskph_risk//committeescommittees//sccpsccp//sccp_opinions_en.htmsccp_opinions_en.htm
GN/ED: 10/200GN/ED: 10/200

31. 3131
GENOTOXICITY / PHOTO-GENOTOXICITY OF ZINC OXIDEGENOTOXICITY / PHOTO-GENOTOXICITY OF ZINC OXIDE
(ZnO)(ZnO) NPsNPs: RESULTS OF GLP STUDIES: RESULTS OF GLP STUDIES
Photo-clastogenicPhoto-clastogenic??PositivePositive atat 195195
g/mLg/mL
CHOCHOPhoto-Photo-
clastogenesisclastogenesis **
Photo-clastogenicPhoto-clastogenic??PositivePositive atat 2.5 or 3.02.5 or 3.0
g/mLg/mL
V-79V-79Photo-Photo-
clastogenesisclastogenesis **
EquivocalEquivocalNegativeNegative ((kckc),),
slightlyslightly positive (V-positive (V-
79)79)
HumanHuman
keratinocytes, V-keratinocytes, V-
7979 cellscells
CometComet test /test / photo-photo-
cometcomet testtest
ClastogenicClastogenicPositivePositive atat 10.0 or10.0 or
20.0 g/mL20.0 g/mL
V-79V-79InIn vitrovitro
clastogenesisclastogenesis
ClastogenicClastogenicPositivePositive atat 814814
g/mLg/mL
CHOCHOInIn vitrovitro
clastogenesisclastogenesis
Non-photo-genotoxicNon-photo-genotoxicNegativeNegativeS. typhimuriumS. typhimurium
TA98, 100, 1537TA98, 100, 1537
Photo-AmesPhoto-Ames testtest
Non-photo-genotoxicNon-photo-genotoxicNegativeNegativeS. typhimuriumS. typhimuriumAmesAmes testtest
COMMENTCOMMENTRESULTRESULTTEST ORGANISMTEST ORGANISMTESTTEST
GN/ED: 10/200
4-FOLD INCREASE IN CLASTOGENIC POTENCY: « PHOTO-CLASTOGENIC »?

32. 3232
«« PHOTO-GENOTOXIC RISK » OF ZnOPHOTO-GENOTOXIC RISK » OF ZnO NPsNPs
Microfine ZnOMicrofine ZnO doesdoes notnot penetratepenetrate intointo oror
throughthrough thethe living skinliving skin
ZnOZnO isis clastogenic (chromosome breaks)clastogenic (chromosome breaks)
in vitroin vitro –– nono evidenceevidence for clastogenicityfor clastogenicity
in vivoin vivo
ZnOZnO isis non-photo-reactivenon-photo-reactive,, non-photo-non-photo-
toxictoxic andand non-photo-sensitisingnon-photo-sensitising
InIn vivovivo, ZnO, ZnO preventedprevented photo-damagephoto-damage inin
thethe skinskin ofof hairlesshairless micemice
ZnZn isis an essentialan essential elementelement for DNAfor DNA
polymerasespolymerases andand DNADNA stabilitystability ––
genotoxic?genotoxic?
New dataNew data suggestsuggest thatthat ZnOZnO isis non-photo-non-photo-
genotoxicgenotoxic
GN/ED: 10/200

33. 3333
Most insolubleMost insoluble NPsNPs dodo notnot penetratepenetrate intointo thethe
interiorinterior ofof mammalianmammalian cellscells **
Fluorescent (Sodium Green, Texas Red) polyacrylamide NPs incubated with
CHO cells
GN/ED: 10/200
* Pictures by curtesy of the University of Jena, DE,
2005

34. 3434
NANO-HYPENANO-HYPE andand NANO-FACTSNANO-FACTS
SWCNTsSWCNTs areare -sized-sized, insoluble fibres, insoluble fibres andand produceproduce
toxicitytoxicity typicaltypical for fibresfor fibres
InhalationInhalation ofof NPsNPs producesproduces asbestos-asbestos-
likelike pulmonarypulmonary toxicitytoxicity
SomeSome evidenceevidence for syst. absorption (for syst. absorption (lunglung overloadoverload),),
nono evidenceevidence for adversefor adverse systemicsystemic effectseffects..
Translocation toTranslocation to thethe brainbrain needsneeds toto bebe confirmedconfirmed..
InhaledInhaled NPsNPs areare systemicallysystemically absorbedabsorbed
andand affectaffect thethe CVCV systemsystem andand thethe
brainbrain..
NoNo evidenceevidence forfor penetrationpenetration intointo oror throughthrough thethe
living skin.living skin. DamagedDamaged skinskin needsneeds confirmation, butconfirmation, but
nono mechanismmechanism suggestingsuggesting activeactive penetrationpenetration..
NPsNPs penetratepenetrate throughthrough thethe skinskin andand
produceproduce systemicsystemic exposureexposure
ConflictingConflicting evidenceevidence: possible: possible minorminor syst.syst. exposureexposure
afterafter oraloral uptakeuptake ofof somesome NPsNPs. No. No evidenceevidence forfor
adverseadverse effectseffects. Adverse. Adverse effectseffects on CV or CNSon CV or CNS
systemssystems isis aa hypothesishypothesis onlyonly..
OralOral uptakeuptake ofof NPsNPs producesproduces systemicsystemic
exposureexposure ofof thethe organismorganism andand,,
possiblypossibly, adverse, adverse effectseffects onon thethe
cardiovascularcardiovascular oror CNSsCNSs
SomeSome NPsNPs (TiO(TiO22, CB), CB) werewere somewhatsomewhat moremore toxictoxic
thanthan MPsMPs,, othersothers (SiO(SiO22, ZnO), ZnO) equallyequally oror lessless toxictoxic
InhalationInhalation ofof NPsNPs producesproduces newnew
toxicitiestoxicities.. NPsNPs moremore toxictoxic thanthan MPsMPs
InertInert NPsNPs (TiO(TiO22, CB), CB) werewere carcinogeniccarcinogenic in ratsin rats afterafter
chronicchronic lunglung overloadoverload:: irrelevantirrelevant forfor manman underunder
normalnormal exposureexposure conditionsconditions
NPsNPs areare carcinogeniccarcinogenic afterafter inhalationinhalation
FACTFACTHYPEHYPE

35. 3535
USE OF NANOPARTICLES IN COSMETICS - ISUSE OF NANOPARTICLES IN COSMETICS - IS
THERE A HEALTH RISK: CONCLUSIONTHERE A HEALTH RISK: CONCLUSION
AvailableAvailable datadata suggestsuggest thatthat useuse ofof NPsNPs inin cosmeticcosmetic preparationspreparations
posesposes nono healthhealth riskrisk toto thethe consumerconsumer
ThisThis viewview isis consistentconsistent withwith thethe conclusionconclusion ofof thethe recentrecent ECETOCECETOC
ConferenceConference, 7-9 Nov., 2005 (Chairman Prof. Helmut Greim):, 7-9 Nov., 2005 (Chairman Prof. Helmut Greim):
ConcernConcern levellevel = Inhalation > oral= Inhalation > oral uptakeuptake >>>> dermaldermal exposureexposure
NB:NB: insufficientinsufficient bioavailabilitybioavailability isis thethe major obstacle formajor obstacle for thethe failurefailure ofof newnew drugsdrugs ––
thethe pharmaceuticalpharmaceutical industryindustry wouldwould paypay billionsbillions forfor NPsNPs thatthat areare systemicallysystemically
availableavailable afterafter inhalation, oral orinhalation, oral or topicaltopical adminsitrationadminsitration –– BUTBUT todaytoday,, wewe hardlyhardly havehave
intravenousintravenous NPNP drugdrug formulations.formulations.

36. 3636
UseUse ofof alternativealternative methodsmethods forfor determinationdetermination ofof humanhuman healthhealth risksrisks
ofof NPsNPs
GeneralGeneral principlesprinciples::
Tests mustTests must includeinclude a standard substance ina standard substance in orderorder toto distiguishdistiguish substance-substance-
relatedrelated (ZnO)(ZnO) fromfrom particle-size-relatedparticle-size-related effectseffects ((egeg TiOTiO22 vs nTiOvs nTiO22))
NOTE:NOTE:hazardhazard studiesstudies (in vitro / in vivo(in vitro / in vivo toxicologytoxicology)) onlyonly,, whenwhen penetrationpenetration
intointo living tissue has beenliving tissue has been shownshown
InhalationInhalation
TierTier 1:1: intra-trachealintra-tracheal instillation, rodentinstillation, rodent
TierTier 2: inhalation2: inhalation studystudy, body distribution, rodent, body distribution, rodent
No alternativeNo alternative methodmethod availableavailable
OralOral exposureexposure
Body distribution, rodentBody distribution, rodent
PenetrationPenetration ofof NPsNPs intointo oror throughthrough GI tractGI tract epithelialepithelial cellscells ((screenscreen))
InIn vivovivo toxicitytoxicity studiesstudies ifif bioavailabilitybioavailability ofof NPsNPs >> standard substance?>> standard substance?
DermalDermal exposureexposure
StudiesStudies inin humanhuman subjectssubjects (EM / skin biopsies)(EM / skin biopsies)
HumanHuman oror pigpig skin in vitro (skin in vitro (penetrationpenetration ofof insolubleinsoluble NPsNPs unlikelyunlikely –– FranzFranz cellcell maymay bebe
unsuitedunsuited forfor measuringmeasuring NPNP penetrationpenetration (T.(T. ButzButz))
MicroscopicalMicroscopical methodsmethods (EU(EU NanodermNanoderm projectproject))
CellCell cultures? (cultures? (problematicproblematic, due to absent or, due to absent or compromisedcompromised stratum corneum)stratum corneum)

37. 3737
Barber saucers Hair jacks
GN/ED: 11/200
COSMETIC APPLICATIONS OF NANOBOTSCOSMETIC APPLICATIONS OF NANOBOTS……
T
H
THANK YOU FOR YOUR ATTENTION!THANK YOU FOR YOUR ATTENTION!