]. Soc. Cosmet.Chem.

, 25, 253-262 (May 1974)

Skin Moisturizers. II. The Effects of

Cosmetic
Ingredients
on Human
Stratum Corneum
MARTIN M. RIEGER, Ph.D., and DONALD E. DEEM, M.S.*

Synopsis-Thefour parametersdescribedin Part I of this study,i.e., elasticmodulus,re-

laxationfunction,waterabsorption,
andwater vaportransmission, havebeenusedto study
the effect of typical COSMETIC INGREDIENTS on human STRATUM CORNEUM.
The elasticmodulusand the stressrelaxationmodulusare usefulmeasures of the ability
of various cosmetic materials to alter the VISCOELASTIC BEHAVIOR of stratum
corneum.It has been demonstratedthat typical cosmeticHUMECTANTS increasethe
rate of transepidermal
waterlossin vitro,and an attemptis madeto explainthisphenom-
enon.

INTRODUCTION

A varietyof humectantsand of occlusive

lipidshasbeenusedfor many
yearsto improvehumanskinand to protectit againstdamage,eventhough
theprinciples
underlying
theseapproaches to skintreatmentwerenot clearly
understood.Blankshowedin 1952howimportantwateris to thewell-beingof
human skin and, in effect, establisheda rational basisfor these time honored
methodsfor skinconditioning( 1). In hisvery thoughtfulreview,Chudzikow-
skicomments
on the undesirability
of highlyocclusive
barriersbecause
they
havea tendencyto lead to edema;he a]sorejectsthe traditiona]treatmentof
dry skinwith humectants because theywill absorbmoisturenot onlyfromthe
air but alsofrom the skin (2). It is apparent,therefore,that the cosmetic
benefitsof eithertreatmentare debatableand that basicknowledgeon how
skinmoisturizerswork is lacking.
In thefirstpartof thisstudy,in vitromethods for determining the response
of humanstratumcorneumto moisturewere developed.It is the purposeof
this secondpart to describethe influenceof commoncosmeticmoisturizers
on the properties
of stratumcomeurn.

*Warner-Lambert ResearchInstitute, 170 Tabor Rd., Morris Plains, N.J. 07950.

253
9.54 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

ExrEmMENT•m

The techniquesare thosedescribedin Part I (3) with minormodifications

if required.Generally,stripsof stratumcorneum(with the attachedtabs)
were immersed for one hour into the test material or solution. Thereafter the
strip was floatedonto a stainlesssteelwire mesh,drained,and carefullyblot-
ted. The strip was then air-dried and placed into the appropriaterelative
hum'•ditychamberfor at least 24 hoursbefore testing.Sometimes,the test
substancewas appliedto the stratumeorneumwith a cottonswab.Any ex-
cesswascarefullyblottedoff beforethe stratumeorneumwasconditioned.
For the determinationof water sorptionby humectants,50% aqueoussolu-
tionsof the humeetantswere appliedto smallpreweighedpiecesof Gelman
Type A glassfilter paper.After conditioningat 0% RH, the weight of humee-
tant was determinedand the weight gain at a given RH was based on this
"dry"weight.The glassfilterpaperwasshownto absorblessthan 1%of water
at 90%RH, andthissorptionwasgenerallyneglected.

RESULTS AND DISCUSSION

MechanicalProperties
Many cosmeticmaterialsare reportedto softenskin and to act as emol-
lients.It was expected,therefore,that thesematerialswould exert somein-
fluenceon the mechanicalpropertiesof stratumcorneum.Of particularinter-
est is glycerolwhich is widely usedby cosmeticformulatotsas a humectant
andskinmoisturizer.It is generallyrecognized(2) that glycerolis hygroscop-
ic and will absorbwater until it reachesequilibriumwith the ambient RH.
Glycerolhasbeenemployedfor manyyearsto "heal"chappedskin,and this
effectis attributedto its abilityto holdwaterin contactwith the skin.
Values of the modulusAxE obtained at 22øC on strips of stratum cor-
neum immersedin mixturesof glyceroland water are summarizedin the
curveshownin Fig. 1. The experimentally obtainedvaluesof AxE are nor-
realizedby dividing them by AxE of the samestrip of stratumcomeurn
immersedin distilledwaterto yield the plottedvalueof E,. The resultsindi-
catethat glyceroldoesnotincrease the elasticityof stratumcorneumby itself
but actuallystiffensthe stratumcorneumat concentrations aboveabout30%,
possiblyby removingstratumcorneum-hydrating water.Theseresultsare in
accord with Blank'sfindingsthat water is the only plasticizerof horny
material (4).
It was considered
possiblethat the interactionof the stratumcorneum/
glycerol/watersystemunder investigation was temperaturesensitive.In
orderto studythisaspect,the effectof temperatureon the valueof AxE was
firstdeterminedby extendingstripsof stratumcorneum, takenfromthe same
specimen, underwatermaintained at varioustemperatures between2ø and
 EFFECTS OF COSMETIC INGREDIENTS 255

9O

• 7O

=' •0'
-g
ß= 40,

i,-,,,

E
o
30,

2O

•o •o Jo 40 •o •o lO •6 •o '•do

• •ate•

Figure 1. Effect of water concentration on normalized modulus values of stratum comeurn

(extendedin glycerol/watermixturesat 22 øC)

37øC. The valueswere divided by the value of AxE at 37øC under water,
and the resultsare includedin Fig. 2. It is apparentthat the extensibility
of
humanstratumcorneumunder the conditionsof this experiment(immersed
in water) showsvery little changebetween 37øC and about 5øC. Middleton
and Allen (5) alsoobservedonly a modestchangein extensibilityof guinea
pig foot pad stratumcorneumbetweenabout 20ø and 40øC under different
experimentalconditions(in air of RH between60 and 80%). Below about
5øC,stratumcorneumappearsto be stiffer,i.e., harder to stretch.
Figure 2 also includesthe resultsof stretchingin a solutionof 1:1 water:
glycerol(wt/wt) and a solutionof 6M potassiumiodide. The plotted data
pointswere obtainedby dividingthe AxE valuesby the AxE valuesfound
at 37øCin the respectivesolutions.Glycerolwas selectedas a typical humec-
tant, while potassiumiodide (6M) is known to break the structureof water
(6).In viewof the'similarity
of theresults,
it islikelythattheeffectof glycer-
ol is due to the breakingof the water structurein the stratum corneum;this
might reduce hydrationof the protein and make the stratum corneumstiffer
or more difficult to stretchat temperaturesbelow 18øC. The temperature
dependence
of stratumcorneumelasticityobservedhere is probablytoo low
256 JOURNAL OF THE SOCIETY OF COSMETIC CtlEMISTS

oo-'1- -t I I I '• I ool o-t-

+l+l+l+l+l+l+l+l+l+l+l+l+l+lffl+l+l+l
 EFFECTS OF COSMETIC INGREDIENTS 257

11.0

10.0

9,0

8,0

7,0

6.0
5,Q
4.,0
3.0

2.0

1,0

-10 -5 0 5 10 15 20 25 30 35

Temperature
tn øC
Figure 2. Effect of temperatureon normalizedmodulusvalues.Stratum corneumextended
under solution and normalized to the value for water at 37øC
-e-e-e-glycerol:water (1:1)

to exertanyinfluenceon chappingor relatedskindisorders,sincethe temper-

ature of the face and handsdoesnot reach 18øC until the air temperature
dropsto about -12øC [extrapolatedfrom data of Phelpsand Vold (7)].
However, the temperatureof the lower extremitiescan reach 18øC at an
ambienttemperatureashigh as 19øC(8).
The effectsof a variety of cosmeticingredientsand of a few chemicaltreat-
mentson the physicalpropertiesof stratumcorneumat 31% RH are summa-
rized in Table I. In examiningthesedata, it mustbe rememberedthat only
horizontalcomparisons are possiblesincethe "control"and "treated"samples
258 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

arefromthe samestratumcorneumspecimen.
A comparison
withinthe vari-
ous columnsmay lead to erroneous.conclusions.
A decreasein the elastic
modulusbelowthat of the control(recordedas +) is indicativeof an in-
creasein the elasticityof the stratumcorneum.In the caseof the stressrelaxa-
tion modulus,an increasein the value over that of the control(recorded
as +) describes
the skin'sability to relievea strainby viscoelastic
response.
The wide variations in the values for the controls are due to the fact that dif-
ferentspecimens of epidermis
wereused,but thisin noway reducesthe value
of the data.Eachvaluerecordedin Table I represents an averageof 4 to 6
determinations on the samespecimenof humanepidermis.Treatmentwith
eitheracid or baseincreasesthe elasticityof the stratumcorneumsubstantial-
ly but doesnot appreciably alter its ability to relievestress.Generally,the
elasticand the stressrelaxationmoduliresultsafter treatmentparallel each
other.Particularlynoticeableis the remarkable"plasticizing" effectof dilute
solutionsof sodiumpyrrolidonecarboxylateand of sodiumlactate.It is be-
lievedthat the influenceof cosmetic ingredients on the mechanicalperform-
ance of stratum corneumcorrelatesdirectly with their beneficialeffect in
skinpreparations.
MoistureAbsorption
The equilibriummoistureabsorptionof severalimportantcosmeticmois-
turizerson glassfilter clothwasstudiedafter equilibrationat severalrelative
humidities.The data in Table II show that sodiumpyrrolidonecarboxylate
andsodiumlactateare capableof holdingrelativelylargequantitiesof water
even at intermediate relative humidities, i.e. between 50 and 70% RH. It
provedimpossible
to conductsimilarexperimentation
with propyleneglycol
becauseof its unexpectedlyhigh volatility. Data for stratum corneumand
somehydrophobic materialsare includedmerelyasa matterof orientation.
In view of the large effectof glycerinon the mechanicalbehaviorof stratum
corneumasa functionof temperature(Fig. 2), it was considered conceivable
that there is a specificinteractionbetweenglycerin,stratumcorneum,and
water which may increasethe water-holdingpropertiesof stratumcorneum.
Similarly,the highhygroscopicity of sodiumpyrrolidonecarboxylate makesit
possiblethat it too mightimpart somespecialwater-holding
propertiesto
stratumcorneum.In orderto determinewhetherany suchinteractionsoccur,
the isothermalabsorptions of water by combinations of known weightsof
unextractedstratumcorneumand glycerolor sodiumpyrrolidonecarboxylate
were determined.The resultsshownin Table III clearly show that water
absorption by stratumcorneumis merelyadditiveto that by the humectant.
In other words,the moistureabsorptionby stratumcorneumtreated with a
humectantis the sumof the absorptions of the individualcomponents,with
no evidencefor any synergisticeffects.Theseresultsare in agreementwith
the data reportedby Fox et al. (9).
 EFFECTS OF COSMETIC INGBEDIENTS 9.59

Table II
Equilibrium Moisture Absorption
Water Absorbed at
Material 31% RH 52% RH 76% RH 90% RH
Stratum corneton 7 9 13 22
Glycerol 11 26 67 240
Sodium lactate 19 40 104 ...
Sodium pyrrolidone carboxylate 17 45 210 ...
Mineral oil 0.2 0.4 0.9 ...
Safflower oil 0.2 0.5 1.0 ...

Table III
Weight Gain (Water Content) of Stratum Corneum
and Stratmn Corneum Plus Hnmeetant

Amount of Water (rag)

after Equilibra•ion at
Sample Initial Weighta (mg) 35% RH 75% RH
Stratum corneum 3.816 0.226 0.530
Glycerolø 3.058 0.339 2.058
Stratum corneumq- glycerol
Experimental 6.874 0.b'•72 2.575
Calculated . . . 0.15615 2.588
Stratum corneum 3.784 0.183 0.515
Sodiumpyrrolidonecarboxylate
c 3.468 0.720 3.488
Stratum corneumq- sodium
pyrrolidonecarboxylate
Exp•Hmental 7.252 0.892 3.892
Calculated 0.903 4,003

•At equilibrium with air at 0% RH.

•U.S.P.
CApp.•iedfrom a 50% solution.

Water Vapor Transmission

The lossof moisturefrom the skin to the atmosphereis a continuouspro-
cess,and excessive lossat low humidity and low temperaturesis generally
associated with skin drynessand chapping.It seemedparticularlyimportant
to determinehow cosmetic ingredients
appliedto stratumcorneummight al-
ter this tissue'sability to "transpire"water to an essentiallydry atmosphere.
Sometypicalin vitro water vaportransmission ratesthroughstratumcorneum
are summarized in Table IV. This table also includes some common cosmetic
humectants,someocclusivelipid materials,and Lotion #78. This last prep-
aration,which is Formula78 describedby Barnett (10), was employedhere
becauseit wa• alsostudiedin vivo by Berubeet al. (11).
g60 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

Table IV
Vitro •Vater Vapor Transmissionthrough Stratum Corneum
Rate (mg cm-'• hr-•) Ratio
Material Untreated Treated (Treated/ Untreated)

25% glycerol 0.264 0.518 1.96

4% sodium lactate 0.294 0.372 1.27
4% sodiumpyrrolidone carboxylate 0.142 0.227 1.60
4% propylene glycol 0.223 0.267 1.20
Light tech. mineral oil 0.274 0.205 0.75
Safflower oil 0.309 0.281 0.91
Lotion 78 a 0.335 0.1'81 0.54

"Formula 78 from Barnett (10).

The resultsobtainedhere are only directionalin view of severalexperimen-

tal uncertainties:Experimentally,stratumcorneumwas first equilibratedin
the diffusioncell against0% humidityto yield an "untreated"rate. The mate-
rial of interestwasthen appliedto the exposedstratumcorneumsurfacewith
the aid of a cotton swab. An effort was made to remove excess material while
maintaininga continuous film, but there is no quantitationof the amountof
substanceactually remaining on the stratum corneum.The "treated"rate
recordedin Table IV is the rate resultingafter completeequilibrationof the
treated stratum corneumagainstthe dry environment.As expected,lipid
materials reduce water vapor transmission,whereas humectantscause a
marked increase. The results with Lotion #78 confirm the in vivo data of
Berubeet al. who foundthat heavyapplicationof this preparationwas re-
quiredto producethe occlusiveeffect(11). It is particularlynotedthat the
small amountof sodiumpyrrolidonecarboxylateremainingon the stratum
corneumafter applyinga 4% aqueoussolutionstill causesa very large in-
creasein the rate of water vaportransmission.
Theseresultsclearlyconfirm
the earlierdata presentedby Powersand Fox (12) that humectantsincrease
transepidermal moistureloss.
Traditionally,humectants, suchas glycerol,have beenusedfor "improving
drynessof skinand chapping."Publishedcontrolledclinicaldata attestingto
the utility of humectantsare missing,althoughthe authorshave accessto a
monadicclinicalin-usestudysuggesting stronglythat a preparationcontain-
ing a high concentration of glycerolwithout any occlusivepropertiesallevi-
ates the dryness/chapping syndrome(13). It is difficult to reconcilethe
evidentlybeneficialeffectsof glycerolwith its ability to increasetransepider-
mal waterloss.Only a verytentativehypothesis is offeredhere:Stratumcor-
neumis commonlythoughtof as a homogeneous layer of cornifiedepithelial
cells.The assumption is probablyincorrect,sincethe outer portionof the
stratumcorneummay have sufferedsomedamagedue to wear and tear and
the dissolution
of lipid- and water-soluble
constituents.
As a result,the outer
 EFFECTS OF COSMETIC INGREDIENTS 261

barrier to the evaporationof water from stratumcorneumis not the top of the
stratum corneumbut lies somewherebetween the topmostand innermost
cellularlayersof the stratumcorneum.Glycerolis known not to penetrate
stratumcorneumappreciablyand presumablyits applicationwill resultin the
formationof a layer at or near the top of this epidermalstructure.By virtue
of its hygroscopicity,glycerolwill unquestionably attract water from the low-
er layersof the stratumcorneumwheneverambienthumidity conditionspre-
clude absorptionfrom the air. This will result in continuousmigration of
water moleculesthroughall layersof stratumcorneumtowardsthe glycerol
layer which, in turn, readily loseswater to the atmosphere.The benefitsde-
rived from the applicationof glycerolthen residein its ability to move the
evaporativelayer of skinmoisturefrom somewhere in the centerof the strat-
um corneumto the very top of the stratumcorneum.

CONCLUSIONS

1. The stress relaxation modulus and the elastic modulus are sensitive mea-
suresof the ability of variouscosmetictreatmentsto affect the viscoelastic
behavior of stratum corneum.
2. The water-holdingcapacityof a systemcomprisinga humectantand
stratumcorneumis the sumof the componentparts.No synergisticeffect
could be demonstrated.
3. The applicationof typicalcosmetic humectantsto the exposed
sideof the
stratumcorneumincreases the rate of transepidermalwaterloss.
4. A modelto explainthe beneficialeffectsof a moisturizeron the surface
of the stratumcorneumis proposed.
( ReceivedNovember13, 1973)

REFERENCES

(1) Blank, I. H., Factors which influence the water content of the stratum comeurn,
J. Invest. Dermatol., 18, 433-40 (1952).
(2) Chudzikowski,R. J., Skin versusthe weather,Mir. Chem. AerosolNews, 44, No. d,
35-41 (1973).
(3) Rieger,M. M., and Deem, D. E., Skin Moisturizers.I. Methodsfor measuringwater
regain, mechanicalproperties,and transe½idermal moisturelossof stratumcorneum,
.l. Soc. Cosmet.Chem., 25, 239-52 (1974).
(4) Blank, I. H., Further observations on factorswhich influencethe water contentof the
stratumcorneum,J. Invest. Dermatol.,21, 259-71 (1953).
(5) Middleton, J. D., and Allen, B. M., The influence of temperature and hmnidity on
stratumcorneumand its relationto skin chapping,J. Soc.Cosmet.Chem., 24, 239-43
(1973).
(6) Hertz, H. G., and Klute, R., The slowingdown of proton exchangein aqueoussolu-
tions of structurebreaking ions, Z. Phys. Chem., Neue Folge, 69, 101-7 (1970).
(7) Phelps,E. B., and Void, A., Studieson ventilation,I. Skin temperatureas related to
atmospherictemperatureand humidity, Amer. J. Pub. Health, 24, 959-65 (1934).
(8) Sheart,C., et al., Investigationson the exchangesof energybetween the body and its
,environment,
Trans.Amer.Soc.Heat Vent. Eng.,43, 115-20, (1937).
262 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS

(9) Fox, C., et al., Modificationof the water-holdingcapacityof callusby pretreatment

with additives,I. Soc.Cosmet.Chem., 13, 263-79 (1962).
(10) Barnett,G., in Balsam,M.S., and Sagarin,E., Cosmetics:
Scien•ce
and Technolog•l,
2nd Ed., Wiley Interscience,New York, 1972, p. 76.
(11) Berube, G. R., et al., Measurementin vivo of transepidermalmoisture loss, I. Soc.
Cosme•.Chem. 22, 361-8 (1971).
(12) Powers,D. H., and Fox, C., A studyof the effectof cosmeticingredients,creamsand
lotionson the rate of moisturelossfrom the skin, Proc. Sci. Sect. Toilet GoodsAss.,
28, 21-6 (1957).
(13) Unpublishedresultsin the files of Warner-Lambert Co., Morris Plains,N.J.