j. Cosmet. 51, 103-125 (March/April 2000) sci.

Photodegradation humanhair: An SEM study of

SIGRID B. RUETSCH, Y. KAMATH, and H.-D. WEIGMANN,

TRI/Princeton, P.O. Box 625, Princeton, 08542. NJ

Accepted publication for January3 I, 2000.

Synopsis

This studyusesfield emission scanning electronmicroscopy (FESEM) to monitor the effects UV irraof diation on the physicalnature of hair fibers. Long-term UV irradiation/humidification cycling causes thinningand fusionof the surface cuticlecell, aswell asfusionof the cuticular sheath into a solid,rigid, and brittle unit. While intercellular cohesion within the cuticularsheathis high, possibly due to crosslinking of theproteins the intra- andintercellular in domains, cellsthemselves brittle. A newlyobserved the are fracture patternof long-term UV-exposed fibers suggests fusion the regions of attacked mostseverely UV by light into one rigid and brittle mass, incapable extension of due to lossof all originalelasticproperties. Unlike chemical oxidation, whichresults partialdissolution h H202) andthencomplete in (1 solubilization (4 h H202) of the melaningranules, photochemical oxidation produces entirelydifferentresults. Evenafter long-termUV irradiation/humidification (95% RH) cycling,the melaningranules appear physically intact. Lossof colordoesnot occuras long as the melanin granules intact. are The severity photodegradation of during UV irradiation/humidification cycling becomes apparentupon brief(seconds) contact these of fiberswith alkalinehydrogen peroxide. Suchcontact results instantaneous in disintegration the components of within the cuticlecells.Formationof sac-likestructures (AllwiSrden sacs) occurs due to osmoticpressure within seconds exposure alkaline hydrogenperoxidecaused phoof to by tochemically degraded proteinswithin the surface cuticle cells.The cellsswell until they burst and their contents drain, leavingbehindcuticularmembranes, which may detachor fuseto the fiber surface. UV irradiationhasalsoseverely photodegraded melaningranules the and preconditioned them for accelerated solubilization uponcontact the fiberswith alkalinehydrogen of peroxide.
The effects both relativehumidity and spectral of energydistributionon the photochemical oxidationof the hair fiberarestudied.Results obtained various at relativehumiditiesin two differentfadingunits, namely,

the QUV Accelerated Weathering Testerand the Atlas Weather-Ometer ("AW") are compared. Scale thinning and fusionobserved during UV/humidificationcyclingare greatlyreduced with exposure low at humiditieswithout humidification cycles. Upon post-treatment with water, fibersirradiatedat a constant 10% RH in the QUV showscale thinningandfusionsimilarto that of fibersexposed UV/humidification to cycling.This indicates that photodegradation occurs low humidity as well. Fibersexposed constant at at 20%, 50%, and 70% humidity in the "AW" showonly moderate scale thinning, evenafterpost-treatment with water.The total solarspectrum usedin the "AW" apparentlycauses severe less photodegradation of the proteins than the UV light of the QUV.

INTRODUCTION

When exposed sunlight,hair is knownto undergo to changes morphological, in chemical, and mechanical characteristics (1-4). The lower wavelength rangeof the UV com103

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ponentof sunlight is known to be responsible thesechanges. recentyears,UV for In radiationat lowerwavelengths significantly has increased to the deterioration the due of ozonelayer. While the earth'satmosphere filters out most radiation below 295 nm, depletionof the ozonelayer, and therefore the reductionin the screening effectof the atmosphere, permits lower wavelengthcomponents reachthe earth'ssurface. to These lower wavelengthregionsof the UV radiation receivedby the earth are the most energetic therefore cause and can severe photodegradation. UV rangeof the sunlight The can be divided into three wavelengthregions,namely UV-A (320-400 nm), UV-B (280-320 nm), and UV-C (<280 nm). UV-C radiationis totally filtered out by the atmosphere and is only experienced space. in
We havecarriedout extensive studies, investigating variousmicroscopic by techniques the effectsof photochemical oxidationon specificaspects hair damagesuchas the of appearance the surface of cuticlecell, intercellular cohesion, scale lifting during longitudinal extension,the integrity of the cuticular sheath, the physical nature of the melaningranules, characteristic fracture patterns,and lossin hair color.In otherwords, we haveexamined radiation-induced changes the physical in ratherthanchemical nature of the hair fiber. Hair fibers used in these studieswere exposed UV radiation/ to humidificationcycling in a QUV Accelerated Weathering Tester. Various follow-up treatments theselong-termUV-irradiated fibersillustratethe extent of photodegraof dation inflicted upon the hair proteins.

More recently, havealsoexamined effects both the relativehumidity and the we the of spectral energydistributionin the radiationon the photochemical oxidation the hair of fiber. Comparisons weremadebetween results the obtained various at relativehumidities in two different fading units, namely, the QUV Accelerated Weathering Tester

(290-400 nm; UV-A; kn = 340 nm) andthe AtlasWeather-Ometer "AW" (Xenon ,

solarsimulator,250-800 nm). Using FESEM, we characterized differentapproaches by the extentof photodamage inflicteduponthe physical natureof the hair fiber in general and the cuticulain particular.

EXPERIMENTAL
MATERIALS

Hair type. Rootsections 14-in-long,brownEuropean fibers of hair fromDeMeoBrothers wereused.The majoraxisof thesehair fibersrangedfrom 70 lamto 120 lam.Because
of this variationin size,single hair fibersalsovaried in the depth of shade.Fiberswith the largermajor axiswere more elliptical than fiberswith a smallermajor axis.
UV EXPOSURE CONDITIONS

QUV Accelerated Weathering Tester. QUV simulates The the sunlightin the range of
290-400 nm, with an irradiancemaximum of the fluorescentbulb at 340 nm. The

irradianceintensity factor has been chosento be 1.35 compared 1.0 for regular to

sunlight. energy The density the340-nm at wavelength iskept constant0.97W/m2. at

The totalenergy density theUV light in thewavelength of range 300-400 nm is 5.06 of
mW/cm 2.

Hair fiberswereexposed two sets conditions the QUV: (a) 0, 100, 300, 500, and to of in

PHOTODEGRADATION

OF HUMAN

HAIR

105

700 hours alternating at three-hour cycles humidification 95% RH; 40C)andUV of (at irradiation (50C;RH decreasing from 95% to 10% within 30 min, then remaining at
10% for 2.5 h); and (b) 0, 100, 200, and 300 hours continuous UV irradiation at
constant 10% RH and 50C.

At/asWeather-Ometer , ("AW"). The "AW" hasa spectral distribution rangingfrom UV throughthe visiblerange,simulatingsunlight.Theseexposure conditions correspond to an "average" Miami summersunlight.The energydensityat the 340-nm wave45

length keptconstant 0.3W/mi . Thismeans at thespecific is at that wavelength340 of nm,theenergy density theQUV (0.97W/m2)is approximately times in three greater than thatof the"AW"(0.3W/mP). However, thewavelength of 300-400nm, in range theenergy densities theQUV (300-400nm yield5.06mW/cm andthe "AW" of 2) (300-400nm yield4.46 mW/cm aresimilar, 2) although distribution quite the is
different.The specific spectral distributions the "AW" in the rangeof 250-300 nm in

yield 0.012mW/cm and , 400-800nmyield 36.80mW/cm Thetotal 2. energy density in the "AW" is 41.272mW/cm Specific . exposure conditions the hairfibers of to
UV/visible radiation in the "AW" were 0, 100, 200, and 300 hours at constant20%, 50%, and 70% RH and 50C.

Post-treatment of hairfibers alkalinehydrogen in peroxide. fibersexposed UV irraHair to diation/humidification 95% RH) cyclingwere subjected post-treatment 6% (at to in alkalinehydrogen peroxide fromseconds to two hours. up This wasdoneto illustrateand characterize severe the extentof photodegradation inflicted uponthe fiber during treatment in the QUV.

Post-treatment of samples irradiated lowand intermediate at humidities water. with UV irradiation/humidification 95% RH) cyclinglead to extensive (at "thinning" of the surface cuticlecellsand "fusion" the scale of edges. Differentiationof the surface cuticlecell, so characteristic the untreated of hair fiber, eventually disappears longerexposure at times. We concluded that the presence moisturein the highly swollenfiber during the of humidification cycleis responsible transporting degraded, for the low-molecular-weight proteinfragments of the cuticlecellandpossibly out into the cortex, therebycausing the collapse and thinning of the surface cuticlecell.
Hair fibersirradiated low humidities(from 10% to 70% RH) in the two fadingunits at do not showthis featureof thinning and fusionof the cuticlecell. Apparently,the lack of mobile water in the hair fiber and the lack of swelling of the fiber eliminate the transport photodegraded of fragments of the cuticlecell. To mobilizethe photodeout gradedproteins,hair fibersirradiatedunder low RH conditions both the QUV and in "AW" were subjectedto a water post-treatment.The fibers were immersedfor 60 minutesin lukewarm,deionized water, air-dried, and then examinedlongitudinallyin the SEM. This wasto establish whetherpost-treatment warm water would result in in thinning and fusionof the cuticlecell by diffusingphotodegraded materialsout of the cuticlecell, assuming that photo-oxidation the hair proteinshad occurred all. of at

INVESTIGATIVE

METHOD

Field emission scanning electron microscopy. Longitudinal and cross-sectional segments of untreatedand UV-exposedfiberswere mountedon double-sided tape and coatedwith

approximatelyfkofplatinum. hairfiber 90 The topography interior examined and were

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JOURNAL OF COSMETIC SCIENCE

for photo-oxidative damage a Hitachi S-4500 digital field emission in scanning electron
microscope (FESEM).

RESULTS
BACKGROUND

AND

DISCUSSION

Photochemical degradation hair affects of both hair proteins and hair pigments andhas beenshown occur to primarilyin the wavelength regionof 254 to 400. In recent infrared spectroscopic studies hair exposed weathering, of to Dubief (3) shows decrease lysine a in and histidine,anda drastic increase cysteic in acidcontent tip endsof hair compared in to root ends. This suggests scission the disulfide of crosslinks during the photochemical oxidation process formationof cysteic and acid.This assumption supported Robis by binsandBahl(5), whohaveshown electron by spectroscopy chemical for analysis (ESCA)
that both UV-B (280-320 nm) and UV-A (320-400 nm) oxidize sulfur in hair. Oxi-

dationwasshown occur to primarilyin the fiberperiphery, namely,the cuticular sheath, producing steepgradientto lessoxidizedhair in the fiber core.Theseresults not a are surprising, sincedisulfideis at its highestconcentration the A-layer and the exocuin ticle, where the highest level of photochemical oxidationoccurs.The ESCA spectra suggest that high levels cystine of S-sulfonate cysteic and acidareformedin hairexposed to photochemical oxidationin the wavelength regionof 254-500 nm.

Our research strongly (6) supports results Dubief(3) and Robbins the of andBahl(5).
Using UV microspectrophotometry, haveshown we that high levelsof photodegradation products formedthroughout hair fiber cross are the section during long-termexposure in the 290-400 nm range.UV irradiation-induced photodegradation products the of hair proteins revealed an extension the absorbance are by of plateauand a shift in peaks from 290 nm in untreated hair to 315 nm in UV-exposedhair. There is also the development an absorbance of shoulder the 330-340 nm range,well isolated in fromthe absorbance the bulk of the hair fiber (Figure1). Formation the photo-oxidized of of hair proteins be traced can andmapped, evenquantified, scanning by across fibercross hair sections the wavelengths the absorbance at of shoulder, m -- 330 nm). Thesephoto(}t degradation products especially are pronounced blond(unpigmented) in Piedmont hair, with the highestlevelofphotodegradation occurring the cuticularregion(A-layerand in exocuticle), where cystineis at its highestconcentration (Figure 2).

It is a generallyaccepted concept that the mechanism photochemical for oxidationof cystinefollowsthe C-S scission pathway,wherebyoxidative scission yieldsS-sulfonic acid that is finally degraded light to cysteic by acid (7,8). In contrast the chemical to oxidationthat followsthe S-Sscission pathwayand yieldstwo molesof cysteic acidper mole of reacted disulfide,only onemole of cysteic acid is produced from eachmole of reacted disulfidein the C-S scission pathway.The progressive oxidationpathways the for two typesof scission are:
S-S Scission (chemical oxidation):

R-S-S-R---> R-SO-S-R> R-SO2-S-R- R-SO2-SO-R- R-SO2-SO2R 2R-SO3H - C-S Scission (photochemical oxidation):

R-S-S-R> R-S-S-OH > R-S-SO2H> R-S-SO3H> R-SO3H + H2SO 4

+ R-OH

PHOTODEGRADATION

OF HUMAN

HAIR

107

2.0

1.2

f. o

Untreated

Hair

0.8

0.4

.o

230. o

264. o

298. o

332. o

366. o

400.

COLLAPSE

AND

FUSION

OF THE

SURFACE

CUTICLE

CELL

The conditions havechosen this studymay seemsomewhat we in extreme;however,we felt they werenecessary be able to learnaboutthe behavior the hair fiber and the to of

changes its physical of naturebroughtaboutby severe irradiation. UV Observations in the FESEMhadshown that nearlyall hair fibersexposed long-termalternating to cycles of UV irradiation (290-400 nm; kma = 340 nm) andhumidification x (95% RH; 42C) displayed unusual an topography. The hair fibershavea smoothtopography similar to
that of a man-madefiber, with little of the characteristic differentiationof the cuticle cell of undamaged hair. Figure 3a shows the typical appearance thickness a normal and of

cuticle cell of unaltered,untreatedhair fibers.However,after only 100 hoursof UV irradiationand humidificationin the QUV, a slight thinning of the surface cuticle cell andfusion the scale at edges apparent are (Figure3b). After 300 hoursof UV exposure, a more pronounced collapse the surface of cuticles and fusionof the scaleedges the to underlying cuticlecellsis seen (Figure3c). 700 hoursof UV exposure produced has hair fiberswith a smooth topography, lackingcleardifferentiation the cuticlecells(Figure of 3d) because extreme of cuticular thinningandfusionto the underlying cuticlecells.The overalldecrease thickness the surface in of cuticlecell asa functionof exposure time to UV irradiationwas obtainedin the FESEM using built-in software measuring for distances a nanometer on scalein the axial, radial, and diagonaldirection of fibers. Decreases scalethickness in were measured the samehigh magnificationand conat verted to the appropriate scale.The resultsare shownin Figure 4. Our hypothesis explainingthis photochemical damagephenomenon as follows:this is progressive thinningandfusionof the surface cuticlecell, underthe conditions used, we is mostlikely dueto photochemical degradation the proteinsin the surface of cuticlecell

108
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o.4

0.2

0.0

0
0.8

20

40

60

80

1O0

120

140

UV exposedhair
0.6

0.4

O.2

0.0

20

40

60

80

100

120

140

Distance (mm)
Figure 2. Formation photodegradation of products the hair fibercross in section. Absorbance scans were

made 330nmacross fiber at hair cross sections (top) after before and (bottom) exposure. UV

intosoluble, low-molecular-weight peptides, which capable diffusing lower are of into cuticle cell layers out of the fiber)while in a highlyswollen (or stateduringthe
humidificationcycleat 95% RH.

PHOTODEGRADATION

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109

Figure 3. FESEMmicrographs hair fibersafterexposure UV radiation.(a) control,0 h; (b) 100 h; (c) of to 300 h; and (d) 700 h UV exposure.
FUSION OF THE CUTICULAR SHEATH

We werealsointerested exploringthe changes the physical in in natureof the cuticular sheath,which had shownsuchthinning and fusionof the surface cuticle cell during long-termUV exposure. Cross-sectional viewsof the freeze-fractured untreated hair fiber showedthe individual layersof the cuticular sheathto be easily identifiable (Figure 5al,a2). However,after 300 hoursof UV exposure, fusionof all cuticularlayersinto a monolithic,brittle unit had occurred preferentially the sideof the hair fiber oriented on towardsthe damagingraysof the light source (Figure 5b). In that regionof the fiber, individualcuticlecellswereno longeridentifiable.Also,fibersexposed 300 hoursof to UV light exhibit deep radial cracks, which are parallel to eachother and absolutely perpendicular the fiber axis(Figure6), fracturing to easilyunderminimal amounts of stress during bendingor extension.

More drasticresults were observed hair fibersexposed an evenlongertime (700 in for h) to UV light undersimilar conditions. Upon extension hair fibersexposed 700 of for hoursto UV irradiation/humidification cycling,mostfibersfailedinstantaneously the at start of extension. These fibers had becomeso brittle that they were incapableof extension due to lossof all original elasticproperties.Upon failure, thesehair fibers displayed unusualfracturepattern.Figure 7 shows an this fracturephenomenon, which occurs the fiberssnapapart. Fusionof the completecuticulaand possiblythe outer as

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JOURNAL OF COSMETIC SCIENCE

0.3

0.1

0.0

100

200

300

400

500

600

700

UV
irradiation.

Irradiation

(h)

Figure 4. Progressive thinningandfusion the surface of cuticlecellsasa function exposure of time to UV

layerof the cortexinto one rigid unit causes brittle failurecircumferentially disand places stress extension those the of to regions the hair fiber that arestill untouched of by thisprogressive fusion. Multiple,successive fractures develop individual at sites along
the corticalcell boundaries, then changedirection and travel radially across individual

cortical cells,and this patterncontinues until it tapers towards coreof the fiber, off the therebycreatingwhat we call the "cathedral spire"fracturepattern(Figure7a). The opposite, corresponding of the "cathedral site spire"fracture shows hollowopening a (Figure7b), surrounded a firmly fused by wall. Higher magnification shows that this wall consists a firmly fusedcuticularsheath of andpossibly alsothe outerlayerof cortical cells(Figure7c,d). Somecuticularregions, preferentially thoseon the sideof the fiber orientedtowardsthe damaginglight source, havebecome indistinguishable, rigid, and very brittle. The "cathedral spire"fracture patternclearlyshows primarylevels photodegradation of in the fiberperiphery (cuticular sheath) a drop-off lesser and to levels degradation of in
the fiber interior.

PHOTOCHEMICAL

VERSUS

CHEMICAL

OXIDATION

Differences between chemical photochemical and oxidation hairproteins melanins of and havebeenwidely discussed the literature.Robbins(9) reports in that both chemical and photochemical oxidation attackboth the hair pigments and proteins, and within the proteins, primarilythe aminoacidcystine. to 25% of the disulfide Up bonds human in hairaredegraded "normal" by bleaching, 45% of the disulfide and bonds maybebroken

PHOTODEGRADATION

OF HUMAN

HAIR

111

- .

TRI

Figure 5. Representative cross-sectional of the cuticular views sheath an untreated of (al,a2) and a 300-h UV-exposed hair fiber. (b)

duringsevere bleaching. However, chemical oxidation the hairpigments of occurs faster than the degradation the proteins. of
UV irradiation the wavelength in regionbetween 254 and400 nm hasbeenshown (9) to degrade hair proteinand pigment.Althoughboth hair proteins and pigmentsabsorb light in the UV/visible region,the longerwavelengths havebeenfound to be less effectivein causing photodamage. with chemicaloxidation,photodegradation As of cystine the mostextensive is phenomenon because the reactivity the disulfide of of bond.
Examination cross of sections hair fibersexposed to 700 hoursUV-irradiation/ of up humidification cyclingshowed, quite surprisingly, the physical that appearance the of melaningranules not changed had much,if at all. Figure8a,b shows melaningranules in cross sections untreated of hair fibers.Thesegranules of various are sizes, spherical or ellipticalin shape, and appear consist smallergranularentities.The granules to of are housed smallcavities in and appear be connected the cell walls by someintercelto to
lular material.

Evenafterlong-term UV-irradiation/humidification cycling,the physical natureof the melaningranules appears intact and undegraded (Figure 9a,b). In unpublished work
carried out at TRI, we have shownthat hair fibers retained their dark brown color and

that only a few had beenfadedslightly to a lighter brown color.Sincethe melanin granules retainedtheir physical bulk and appearance, melaninpigment was prothe

112

JOURNAL OF COSMETIC SCIENCE

30509

50

kV

x4.0k"'ff25'&

..

Figure6. a-d: Typicallongitudinal views a long-term of UV-exposed fiber,displaying hair severe radial
cracking, typicalradiation-induced a damage phenomenon.

tectedfromdegradation, which,in turn, prevents loss hair color.The assumption a of

wasmadethat photochemical oxidation the melaninchromophor of wouldhaveto occur
in situ if color loss was to occur at all.

This is quite in contrast the swellingand dissolution the melaningranules to of by

chemical oxidationwith hydrogen peroxide. After 1 hour of oxidationwith 6% alkaline

hydrogen peroxide, moderate disintegration the melaningranules occurred of had (Figure 10b,c).The subgranular structures, whichmakeup the melanin granules, appear
more pronounced. There are variousstages degradation, of from mile to severe, due to

partialsolubilization themelanoprotein. of Solubilization appears occur to preferentially from the centerof the granules (Figure10c).

Afterthehairfibers were exposed fourhours oxidation for to with 6% alkaline hydrogen peroxide, mostof the granular cavities wereempty(Figure10d).The assumption was madethat the majorityof the cavities wereempty because granules the weredissolved away theperoxide. by Progressive solubilization themelanoproteins prerequisite of isthe for solubilization and/or dispersion the melanin of pigments. With the melanoprotein dissolved diffused and into the bulk of the hair fiber,the spread-out melanin pigment becomes unrestricted the targetof bleaching loss colorof the humanhair.Wolfram or in and Hall (10) haveobserved similarbehavior with melaningranules isolated from hair
and exposed peroxidebleaching. to

PHOTODEGRADATION

OF HUMAN

HAIR

113

Figure 7. a-d: The "cathedral spire"fracture patternseen uponextension hair fibersexposed 700 h of to of UV irradiation/humidification cycling.

Using microspectrophotometry, have shown that under the selectedconditionsof we progressive exposure UV radiation/humidification to cyclingin the QUV, no or insignificantloss color of occurs aftershort exposure timesandthat onlymoderate lightening of the coloris observed after700 hours UV exposure. melaningranules of The remain
intact as observedin the FESEM. Chemical oxidation, on the other hand, results in

increased lighteningof the hair colorwith increased exposure time, while solubilization
of the melanin granulesoccurs.

Sincemicrospectrophotometrya non-destructive is technique, change hair color the in is measured transmittedlight at the very same(previously in marked)locations the on verysame hair fibersunderidenticalinstrumental settings before and after the various exposure times to UV radiation.Increases transmission in intensityare directlyproportionalto lossin hair color.The same technique usedon hair fibersexposed chemical is to oxidation.Figure 11 compares decrease hair colorcaused chemical the in by oxidation with 6% alkalinehydrogen peroxide versus progressive irradiation. UV

MANIFESTATION

OF ADVANCED

UV DEGRADATION

OF HAIR PROTEINS (CUTICULA

AND MELANIN

GRANULES) DURING SUBSEQUENT HYDROGEN PEROXIDE TREATMENT

Hair fibersexposed long-term(700 h) UV irradiation/humidification to cycling,and

114

JOURNAL OF COSMETIC SCIENCE

Figure 8. a,b: Melanin granules cross in sections untreated of hair fibers.

subsequently subjected treatmentwith alkaline hydrogenperoxide,revealsignifito cantlymoredamage the hair fiber than hadoriginallybeensuspected. to Short-term alkalineperoxide bleaching long-termUV-irradiatedhair fibersresults of in instantaneous disintegration dissolution elements the cuticular and of of cells,fusingthe hair fibersfirmly together. After drying,these fiberscanno longerbe freelyseparated, andforcefully pulling them apartresults their tearingandfibrillation(Figure12a,b). in

PHOTODEGRADATION

OF HUMAN

HAIR

!!5

"6"K';i . :" ' 2.'

,B.
;..

Figure9. a,b:Afterlong-term UV-irradiation/humidification thephysical cycling, nature themelanin of

granulesappears intact.

The cross-sectional of these view fibersconfirms observations duringthe the made longitudinal study. cross The sections partially show disintegrated, dissolved, fused and surface cuticle (Figure cells 12c,d). also We observed some themelanin that of granules appear havebeensolubilized to aftermerely15 minutes peroxide of treatment. Such solubilizationthemelanin of granules untreated occurs in hair usually several after hours (4 h) of bleaching. Long-term exposure without UV has, doubt, severely degraded the

116

JOURNAL OF COSMETIC SCIENCE

Figure 10. Intact(a) andpartially(b,c)andcompletely solubilized (d) melaningranules before after 1 and h and 4 h of oxidationwith 6% alkalinehydrogen peroxide, respectively.

hair fibers (cuticle cells and melanin granules)and preconditioned them further to accelerated disintegration.

After oneand two hoursof alkalineperoxide bleaching long-termUV-irradiatedhair of fibers, the cuticulahas completelydisintegratedand dissolved into an unrecognizable, thin, film-like layer of cuticularmembranes enveloping outer cellsof the cortex the (Figure 13a,b). The disintegration, dissolution and fusionof the cuticle cellsare so advanced that the individual cuticle cells are no longer identifiable.The original topographyof the hair fiber no longerexists.Insteadof surface cuticle cellswith their characteristic scale-like structure, bumpy surface a covered a thin film is displayed. by This thin film may consist cuticularremnants, cell membranes, of the and the CMC, which existsbetweencuticulaand cortex.The corresponding cross-sectional views of thesefibersdemonstrate progressive the damageinflictedupon the hair fiber by longterm UV exposure, (Figure 13c,d). The low magnification cross section(Figure 13c) showsthe fusedcuticula and merely a cortex containingempty cavities(Figure 13d) where oncethe melanin granuleswere housed.Besides severe oxidativedamageto the cuticula, long-term U irradiation has also photochemically degradedthe melanin granules, and haspreconditioned them for accelerated dissolution during subsequent short-termtreatmentwith alkaline hydrogenperoxide.
These observations clearly show that long-term UV exposure causes severechemical

PHOTODEGRADATION

OF HUMAN

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117

Transmission Intensity (%FS)

60 50 40 30 20 10 0

Peroxide Treatment

(h)

Transmission Intensity (%FS)

60

40

20

100

300

500

700

QUV Exposure ( hours )

Figure 11. Comparison loss haircolor to chemical of in due oxidation with 6% alkaline hydrogen peroxide (top) and UV irradiation (bottom).

degradation onlyof the cuticula melanin not and granules, also the hairfiberin but of
general.

Optical microscopy. unusual The results demonstrated the SEM studyproduced in several questions aboutthe drastic changes the physical in natureof the fiber'stopography. Therefore, long-term UV-irradiated hair fibers wereobserved the opticalmicroscope in duringalkaline peroxide treatments, whichprovided some answers questions to about

118

JOURNAL OF COSMETIC SCIENCE

006020

5. 0 RV

xG00

50. O,mtl.

doGo 1

kV x

B.

Figure 12. Longitudinal (a,b) and cross-sectional viewsof long-termUV-irradiatedhair fibersafter (c,d) subsequent 15-rnintreatment with 6% alkalinehydrogen peroxide.

this extremecuticulardisintegration during chemicaloxidationof UV-exposedfibers. Long-term UV irradiationcauses extensive oxidativedegradation the proteinsof the of cuticulaas manifested an Allw6rden-like reactionduring subsequent in chemicaloxi-

dationof UV-exposed fibers(Figure14a-f). After only seconds alkalineperoxide hair of treatment(Figure 14b,c), small bubblesdevelopalong the hair surface. Thesesmall bubblesor half domesresemble famousAllw6rden sacs the formedby the Allw6rden reaction.Allw6rdenfirst detected thesesacs the surface wool fibersduring treatat of
ment with chlorinewater. He thusalsodiscovered epicuticlesurrounding cuticle the the cell. The epicuticle is a semipermeable, partly proteinaceous membrane,-25 A in thickness, whichallowssmallmolecules (water)to diffuse into the cell, rapidlyswelling degraded, hydrophilic, low-molecular-weight peptidefractions. The highly hydrophilic molecules are not capableof diffusing out and remain trapped inside the cell. In Allw6rden'sreaction,the hydrophilicmolecules continueto attract water from the outsidesolution,and thus thesesacs formedby the resultingosmoticpressure. are

In our studyconcerning long-termUV exposure humanhair, the bubbles of resembling Allw6rdensacs formedby diffusion peroxide, are of ammonium hydroxide, and water through the epicuticleinto the cuticle cell, where the alreadyphotolyticallydegraded proteins nowfurtherdegraded the peroxide swollen water.The hydrophilic are by and by proteinfractions still too bulky to diffuse are out throughthe semipermeable epicuticle. They remaintrappedwithin the cuticlecell, attractingwater and forming the sac-like

PHOTODEGRADATION

OF HUMAN

HAIR

1 I9

J.

B0011

5.0

kV

Figure 13. Longitudinal(a,b) and cross-sectional viewsof long-termUV-irradiated hair fibersand (c,d) subsequent treatment 2-h with 6% alkalinehydrogen peroxide.

structures (Figure 14b,c). Eventually,thesesac-likestructures (swollencuticle cells) burst and their contents drain (Figure 14d), leavingbehindmembranes shellsof or cuticlecells(Figure14e),whichmay detach from the hair fiber anddrift away(Figure 14f), or may remain attachedand form a thin film-like layer enveloping the outer corticalcells, as has beenshownin the SEM study.
EFFECTS OF RELATIVE HUMIDITY AND SPECTRAL ENERGY DISTRIBUTION ON

PHOTOCHEMICAL

OXIDATION

Moistureplaysan importantrole in the photobleaching human hair (3,11,12). Phoof tochemical oxidation(alsotermed"weathering" "photo-bleaching") or damages hair at anygiven RH and increases durationof exposure. wet mechanical with The properties of photo-bleached fibersdecrease hair with increasing extentof damage (11). Reduction in wet mechanical properties foundto be the mostsevere was whenhair is weathered at high or low RH. Thesefiber properties leastaffected are when hair is exposed a RH at
of 30%.

We havealso investigated effectsof both the relativehumidity and the spectral the energy distribution photochemical on oxidation hair fibers.We compared results of the obtained various at relativehumiditiesin two differentfadingunits, namely,the QUV

accelerated weathering tester andthe AtlasWeather-Ometer .

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Figure 14. Longitudinal views long-term of UV-irradiated hairfibers andsubsequent exposure various for timesto 6% alkalinehydrogen peroxide: no peroxide (a) post-treatment, 5 s, (c) 20 s, (d) 15 rain, (e) 15 (b) rain (focused domeof fiber), and (f) 60 rain of peroxide on post-treatment.

UV exposure the QUV. Fibersexposed continuous in to UV light at constant10% humidity in the QUV do not showthinning and fusionof the cuticlecells(Figure 15a), aswasobserved our earlierstudiesin which hair had beenexposed UV irradiation/ in to humidification(at 95% RH) cyclingin the QUV. However, uponsubsequent immersionin water, collapse fusionof the surface and cuticlecell occurs (Figure 15b), with a severitysimilar to that observed hair fibers exposed irradiation/humidification in to cyclingin the QUV. This indicates that photodegradation occurred a low conhad at centration water in the fiber structure, of but that the degraded proteinfractions (unlike in the swollenfiber), werenot ableto diffusefrom the cuticlecell due to lack of adequate amountsof water to promoteswelling. This thinning and fusionphenomenon can be easilyexplained.At very low relative

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Figure 15. Longitudinal views hairfibers of exposed (a) 300 h of continuous irradiation constant to UV at 10% RH, 50C,and (b) subsequent immersion lukewarmdeionized 1-h in water.

humidities,below25%, watermolecules principallyboundto hydrophilic are sitesof the keratinfiberby hydrogen bonds, the wateris not mobile(13). As the humidity and increases, additionalwateris sorbed, occupying remainingadsorption the sitesassociated with the protein.At highRH, whenall the adsorption of the keratinareoccupied, sites water molecules no longerbound and are more mobile within the fiber. This are facilitates diffusion degraded, of highlysoluble, low-molecular-weight peptidefractions

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from the cuticlecell. This accelerates degradation because now the radiationcanattack new proteinsratherthan the alreadydegraded proteins.Similar phenomena havebeen observed Ratnapandian al, (11). by et

Exposure theso/atspectrum the "AW." Fibersexposed continuous to in to irradiationat constant20%, 50%, and 70% relative humidity in the "AW" show the extent of photodegradation be less to than in hair fibersexposed the QUV (to eithercontinuous in or cyclingconditions), evenafter subsequent immersionin water. This wassomewhat unexpected, especially since hair fiberswereexposed UV irradiation the 250-400 to in nm rangeaswell asto visiblelight at 400-800 nm wavelengths. While fibersexposed to continuous UV light at constant10% RH in the QUV show extremecollapse and fusionof the cuticulaafter subsequent immersionin water, fibersexposed the "AW" in displaymoremoderate cuticularcollapse fusion.Micrographs and depictlargeporesin the scale faces (Figure16b) andsmallpores openings the fused or at scale edges (Figure 16b,c),mostlikely areas wheremoisture and solubilized materials escaped during immersion water.We concluded the increased in that damage fibersexposed the QUV to in mustbe dueto radiation emittedat }tmof 340 nm in the UV-A range, since is that it wavelengthat which the radiation energy in the QUV is approximatelythree times greaterthan that of the "AW."
There is no SEM evidenceof melanin granule degradation,even in the presence of moisturefor either wavelengthrange,(QUV or "AW"). After long-term exposure to irradiation in both fading units, the melanin granulesappearphysicallyintact, even thoughin someinstances melaningranules the may havebecome slightly"grainier" at moderate levelsof degradation. long as the melaningranules intact, lossof hair As are
color does not occur.

Loss hair co/or. of Using microspectrophotometry transmittedwhite light, we have in measured in hair colorasa functionof exposure loss time to radiationin both the QUV and "AW." Sincethe transmission intensity was measured the samelocationon the at same hair fibers beforeand after the variousexposuretimes, the true color losswas monitored, example, heavilyor weaklypigmented for for hair fibers.Figure17 shows the increases transmission in intensity(TIt-TIo), indicativeof lossin colorasa functionof exposure time to radiation in both the QUV and "AW." The followingsummarizes effects boththe RH andthe spectral the of natureof radiation on the photolysis hair pigment: of (a) At low RH conditions (10%, 20%, and 50% RH), independentthespectral of range, there is a lack of lossin hair color in both the UV range of the QUV and the UV/visible range of the "AW." In other words, at low RH, neither UV nor UV/visible radiationappearto cause significantphotodegradation melanin in of
brown hair.

(b) Moderate lossof hair coloris observed whenlong-termexposure radiation the to in UV/visible rangeis combined with a higher relativehumidity (70% RH). (c) Moderatelossof hair coloris alsoobserved upon long-term exposure alternating to three-hourcycles humidificationat 95% RH and UV radiation,during which of the relative humidity is decreased within 30 minutes from 95% to a low of 10% RH, then remainingat the 10% RH for the durationof the radiationcycle. From this microspectrophotometric study it can be concluded that the high relative hmidity is theprimarycontributing factorto accelerated in hair color, loss sincelow RH

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Figure 16. Longitudinal viewsof hair fibersexposed (a) 300 h of continuous radiationat constant to UV 20% RH and 50C in the "AW," and (b,c) subsequent immersionin lukewarm deionizedwater. 1-h

conditions not resultin significantlossin hair color,evenafter long-term exposure. do Increased exposure time and spectral rangeappearto be secondary factors. Also, there appears be an initial periodof greatresistance fading,since to to colorloss does occur not during shortexposure times under any of the conditions investigated. However,it is clearlyshown that the combination both the high RH conditions the light in the of and visible rangehasthe most damagingeffectson hair color.

According Wolfram and Albrecht(2), partial solubilization the melanoproteins to of is a prerequisite the decolorization disintegration for by and dispersion the melanin of pigmentswithin the hair fiber.

CONCLUSIONS

Long-termexposure UV irradiationin the QUV resultsin photo-oxidative to degradation of the hair proteins, especially cystine, of which occurs its highestconcentration at in the cuticulardomains wherethe degradation most severe. is UV irradiation(QUV) results aminoacid degradation indicatedby shiftsin the UV absorbance in as spectrum to higherwavelengths development an absorbance and of shoulder. Cycling UV irradiationat 10% RH with humidificationat 95% RH (QUV) leadsto extensive "thinning"of the surface cuticlecell and "fusion"of its scaleedge to the

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20
t6
12

20
16

Figure 17. Loss hair colordue to progressive n light exposure the QUV and "AW," shownasincrease in in transmission intensity(%) of hair fibersdue to light exposure.

underlyingcuticlelayer.The characteristic differentiation the surface of cuticlecell of untreated hair gradually disappears with increasingexposure. Besidesthinning and fusionof the surface cuticlecell, therealsooccurs fusionof the complete cuticularsheath into a rigid, brittle unit. While intercellularcohesion within the cuticularsheathis high, possibly due to free radical-initiated crosslinking the CMC, the cuticle cells of themselves brittle. A newlyobserved are fracture patternof long-termUV-exposed fibers suggests fusionof the regions attackedby UV light into one rigid and brittle mass, incapable extension of due to lossof all originalelasticproperties. While chemical oxidation results partial(1 h H202) andthencomplete in solubilization (4 h H102) of themelanin granules, photochemical oxidation produces entirelydifferent results. Even after long-term exposureto severeconditions of UV irradiation/
humidification cycling, the melanin granulesappearphysicallyintact with little change

in their physical nature.Loss colordoes occuraslong asthe melaningranules of not are

intact.

Long-termUV irradiation/humidification cyclinghasseverely damaged hair proteins the and preconditioned them for accelerated disintegration during subsequent treatments with alkaline hydrogenperoxide.Suchcontactrestfitsin rapid disintegrationand dissolutionof elements the cuticlecells, restiltingin formationof Allw/Srden of sacs via osmosis. UV irradiationhas alsoseverely damagedthe melanoproteins and preconditioned them for accelerated disintegration and solubilization the alkalinehydrogen by peroxide, indicated SEMmicrographs theemptycavities as by of where once melanin the granules were housed.

While photochemical oxidationof hair occurs all humidity conditions, appears at it to be morepronounced rapid at high relativehumidity and/orhumidificationcycling. and

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ContinuousUV irradiation at constant10% RH (QUV), without alternating cyclesof humidification, does not show the thinning and fusion of the surfacecuticle cells. However,photochemical degradation occurred, subsequent has as immersionin water causes cuticle thinning and fusionof a magnitudesimilar to that experienced fibers by exposed UV irradiation/humidification to cycling(QUV).

Low and moderate RH conditions during exposure unfilteredsolarlight in the "AW" to alsofail to produce cuticlethinning and fusion,and hair fibersstill displaythe characteristic cuticle cell differentiation. Subsequent immersionin water showsonly moderatecuticlethinning and fusion.This suggests that the solarspectrum the "AW" of causes lesssevere levelsof photo-oxidative degradation the proteinsthan the UV of irradiation in the QUV.

High relative humidity is the primary contributingfactor to accelerated in hair loss color.The combinationof both the high RH conditions and the unfilteredsolarlight havethe most damagingeffects hair color. on

REFERENCES

(1) E. Hoting, M. Zimmermann, and S. Hilterhaus-Bong,Photochemical alterationsin human hair. I. Artificial irradiation and investigations hair proteins,J.Soc. of Cosmet. Chem., 85-99 (1995). 46, (2) L.J. Wolfram and L. Albrecht,Chemicaland photobleaching brownand red hair,J. Soc. of Cosmet.
Chem.,82, 179-191 (1987).

(3) C. Dubief, Experiments with hair photodegradation, Cosmet. Toilerr.,107, 95 (1992). (4) J. B. Speakman P. R. McMahon, and The actionof light on woolandrelated fibers, Z.J. Sci.Tech., N.
20 (1939).

C. Robbins and M. Bahl,J. Soc. Cosmet. Chem., 379-390 (1984). 35, S. B. Ruetsch, Kamath, and H.-D. Weigmann,unpublished Y. observations. C. R. Robbins,J. Soc. Cosmet. Chem., 22, 339 (1971). W. E. Savigeand J. A. Maclaren,in The Chemistry Organic of Salfar Compoands, Kharash,C.Y. N. Meyers, Eds.(Pergamon Press, New York, 1966), Vol. 2, pp. 367-402. (9) C. R. Robbins, Chemical Physical and Behavior HamanHair, 2nd ed. (Springer-Verlag, of New York, 1994), pp. 103-104, 108. (10) L.J. Wolfram and K. Hall,J. Soc. Costa. Chem., 247 (1975). 26, (11) S. Ratnapandian, B. Warner,andK. Kamath,Photodegradation humanhair,J. Soc. S. of Cosmet. Chem.,
49, 309-320 (1998).

(5) (6) (7) (8)

(12) F. Leroy,A. Deftandre,and J. C. Garson,Photoaging human hair, 7th International of Hair Science Symposiam (Bad-Neuenahr,1990). (13) C. R. Robbins,Chemical Physical and Behavior HamanHair, 3rd ed. (Springer-Verlag, of New York, 1994), p. 78.