1996 - v goffin - antidandruffshampoosandthestratumcorneum[retrieved-2014!09!20]

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Journal of Dermatolog ical Treatment (1996) 7. 21 5 - 2 18

@ 1996 Journal

of

Dermatological Treatment.Al l rights resewed 0954 6634

Antidandruff shampoos and the stratum corneum

V Goffin, C Piirard-Franchimont G E Piirard

Belgian SSTC Research Center

5596,

Department

of

Dermatopathology. Uiriversiry

of

Liige. Liige. Belgium

The clinical efficacy of antidandruff shampoos containing

antifungal agents is correlated with their anti-Mdusseziu

activity. Other ingredients

of

the shampoos also affect the

efficacy and tolerability of the treatments. This study was

conducted to quantify the effect of eight proprietary

antidandruff shampoos on the human stratum corneum.

Corneosurfametry and squamometry were used to ra te the

interaction between the shampoos and the stratum

corneurn. Significant differences in mildness were dis-

closed between some test shampoos and SLES-7E0

selected as a mild reference surfactant. The activity

against corneocyte clumping also showed significant

differences among the test products. I t is concluded t hat

the formulation of shampoos for dandruff should not be

overlooked.

( J

Dermatol Treat 1 996) 7: 21

5 -

2 18)

Keywords: Antidandruff sham poos- Stratum corneum

-

Corneosurfametry- Squamometry

Rrceivrd 13th November 1995; accepted 31sr Jul

19%

Materials and methods

Test products

Eight proprietary antidandruff shampoos were used.

They contained one of the following active agents: eco-

nazole nitrate 1

YO,

Pevaryl, Cilag, Berchem, Belgium),

ketoconazole (2%, Nizoral, Janssen Pharmaceutica,

Beerse, Belgium), piroctone olamine (0.5%, Dercos,

Laboratoire D'Anglas, Paris France;

1

Yo

Anatel, La-

boratoire Roche Posay, La Roche Posay, France; 1%

Cystelle, Laboratoire Bailleul, Paris, France), selenium

sulphide (2.5%, Selsun, Abbott, Ottignies, Belgium) and

zinc pyrithione 1

YO,

ead and Shoulders, Procter and

Gamble, Strombeek Bever, Belgium; 1YO, inkan, Le-

derle Cyanamid. Louvain-la-Neuve, Belgium). Sodium

laureth sulfate EO 7: 1 (SLES-7EO) as a 2% aqueous

solution served as the reference product in the squamo-

metric and corneosurfametric tests because it exhibits

only mild ir ri tancy in V ~ V O . ' ~ . ' ~

Introduction

Modem shampoos designed for treating dandruff and

seborrhoeic dermatitis were developed following the

demon stration that the active ingredients are antifungal

agents which inhibit the growth of Malassezia ovalis

(formerly known as Pityrosportrni ov ale) . -3The ultimate

goal of these shampoo s is to remove scales and to alter

the corneocyte binding and multiplication of this re-

sident lipophilic yeast. Removing the scales is important

both for the immediate cosmetic effect and for blocking

the cyclic process during which the yeast accumulates,

releases proinflammatory mediators and activates an

immune r e~ po ns e, ~ hich in turn disturbs the pro-

liferation and maturation of the epidermis. The resulting

thick and porous horny layer is the niche in which the

yeast continues to flourish. The effect of shampoos on

intercorneocyte cohesion is largely unknown.

Several investigational methods have been developed

to rate desquamation and the potency of squamolytic

agents6-' Among them, squamometric and corneo-

surfametric bioassays give insight into both the cohe-

siveness and structural changes occurring in

co rne~cy tes . ' .~hese methods are particularly suited for

evaluating the interaction of surfactants with the hum an

stratum corneum.I0-l3

The aim of this study was to rate the effects of anti-

dandruff shampoos on the

stratum corneum. Non-

invasive squamometric and corneosurfametric tests were

conducted to evaluate the effects of eight proprietary

shampoos on the stratum corneum.

Correspondence: Prof G Pierird, DepJrrment of Dermatopathology.

CHU Sari Ti lman, 8-4000, iege, Belgium, Tel. 32-41-662408; Fax.

+ 32-41 -662976

Squamometric in vivo bioassay

A grou p of 12 volunteers, aged 23-47 years, entered a

double-blind within-subject comparison after signing an

informed consent. The iterative short-term patch pro-

tocol consisted of occlusive patches applied three times

for 30 min

at

2-h intervals to the volar forearms.'.' Each

time, a fixed amount (0.15 ml) of water, or 2% solutions

of the shampoos or SLES-7E0 was deposited into

19-mm Hill Top chambers. Two hours after the final

application of the test products, clear adhesive-coated

discs (D-Squame, Cuderm Corporation, Dallas, TX)

were placed under controlled pressure

(1

10 g/cmz) for

5 s

onto each test site. After careful removal, they were

stained for 1 min with

a

solution of basic fuchsin-

toluidine blue (PMS, polychrome multiple stain, De-

lasco, Council Bluff, Iowa). The discs were evaluated

colorimetrically as previously described for the squa-

mometric method. - Da ta are expressed as colour

differentials, AE*ab, between the material collected

from each test site and from the water-treated site. This

variable is referred to as the differential squamometry

index (D-SQMI). In addition, the same samples were

examined under the microscope for rating both the in-

tensity of the corneocyte PMS staining and the pattern

of intercorneocyte cohesion (Table I, Figure 1).

Corneosurfametric

ex

vivo bioassay

Th e 12 volunteers, who part icipated in the in vivo

squamometry study also participated in this corneo-

surfametric bioassay study. The method entailed col-

lection of cyanoacrylate skin surface strippings (CSSS)

from healthy individuals and a 2-h contact time

with

a


2/4

216 V

Go ffn et al

Table

I

Interval scales for microscopic grading o f D-Squame

adhesive-coated discs

Score Preilorninunt pat tern

Staining intensity

0 No

staining

1

Discrete between corneocyta

2

Spotted

on

corneocytes

3

4

Strong

in

most corneocytes

0

1

2

3

4

Sinnleltorn corneocvtes

Strong in single and/or clustered corneocyta

intercorneocyte loosening

Large and uniform sheet of corneocytes

Large clusters. few single corneocyta

Small clusters, some single corneocytes

Disrupted clusters, many single corneocytes

2% solution of the test

product^.'^-^'

After staining the

samples with PMS for 3 min, rinsing and air drying,

reflectance colorimetry (Chroma Meter, Minolta, Osa-

ka, Jap an) was used to derive values of L* nd Chroma

C .

The colour differential, AE*ab, was calculated

be

tween each sample and a reference CSSS sprayed with

water only. This value

is

referred to as the comeo-

surfametry index (CSMI) which increases with the irri-

tation potential o f the skin-cleansing produ ct being

Statistical analysis

The distribution of each numerical variable CSMI, -

SQMI) was characterized by the mean (M), SD and

coefficient of variatio n

V=

o2 SD/M). Differences be

tween the test and reference products were tested with

the paired Student's t-test. The Wilcoxon statistical

processing of ordinal scales signed ranks test was used

for the microscopic grading of the D-Squames. Re-

gression analysis was applied to determine the re-

lationship between variables. The linear, logarithmic,

exponential and power models were tested with calcu-

lation of the coefficient of correlation. All results were

considered to be significant at the 5 critical level

P< .05).

Results

The results of the squamometry and the comeosurfa-

metry are presented in Table 11.

A

significant linear

a

b

Figure

1

Microscopic aspect of squamometry samples after PMS

staining

see

Table

I).

(a) Example

of

score

1

in both staining

intensity

and

intercorneocyte

loosening.

b) Example of

score

3

in

staining intensity and score I in intercorneocyte loosening. (c)

Example of scon 3 in both staining intensity and intercorneocyte

loosening.

Table Corneosurfam etry and squamom etry. Colorimetric evaluation of

the

corneosurfametry index CSMI) nd of the differential

squamometry index (D-SQMI). Microscopic evaluation

of

the corneocyte dye binding and of

the

pattern

of

intercorneocyte cohesion

on D-

Squames

Brand name Antifungal agent CSMI

Wab)

D-SQMI A F a b ) D-ve binding Corneocyte loosening

Pevaryl Econazole nitrate

1

32.8f .5'

15.3 k5 .3 2.1 f0 .6

Nizoral

Ketoconazole 2

25.5f 6.3

12.6f4.7. 1.8f 0.6 1.7f0.5'

Dercos Piroctone olamine 0.5 28.4k7.7

14.9k8.5 1.7f0.9 1.6f0.3'

Anatel Piroctone olamine

1YO 24.6k8.3

9.2f

5.9

1.3f0.8 0.5f0 .3 .

Cystelle Piroctone olamine

1

37.8

f 4.2'

16.8f8.4 2.4f

.5.

0 .9 f 0 .8

Selsun Selenium sulphide

2.5

48.3f15.8'

l 7 . 9 k

8.3

2.6f0 .5* 1 .8k1.2

Head and Shoulders Zinc pyrithione

1%

27.7f9 .9 13.6f5 .4

1.7

f 0 . 7 1 . 1 f l . 3

Zinkan

Zinc pyrithione

1

24.5f .2

17. 45 7.1 2.0k0.9 1.3k1.5

Reference product SLES-7EO

23.1k6.8 16.3f .4 1.8f0.3 1.3f0 .4

Shampoo Corneosurfametry Squamome t r y

0 .9 f 0 .6

*Significantly different

(p


3/4

An t i d a n d r u f f Fharnpoos

and

Ihe

ctraturn

corneum

217

correlation

( r = 0 . 5 4 . ~ 0 . 0 5 )

as found between C SM I

and D-SQMI values of all the shampoos combined. The

interindividual coefficients of variation were lower for

CSMI than for D-SQMI, ranging from

25

to

38%

and

from 3 5 to 65%, respectively. CSMI was linearly cor-

related

r =

0.78.

Pc0.01)

ith the rating of dye binding,

but was unrelated

( r = O . 2 3 ,

NS) to the rating of cor-

neocyte loosening. A power relationship ( r

= 0.62,

P < .05) was found between D-SQMI and the intensity

of corneocyte loosening.

Some shampoos showed significant differences from

the reference produc t

SLES-7E0

(Table 11). Th e mildest

shampoos showing prominent anticlumping activity

(Anatel, Dercos, Nizoral) were identified by b oth low D-

SQMI values and high corneocyte loosening in the

squamometric test. Two shampoos (Cystelle, Selsun)

were significantly more irritan t than

SLES-7E0

because

they showed increased CSMI values and increased dye

binding. The other shampoos (Head and Shoulders,

Pevaryl, Zinkan) formed an intermediate category

of

products which globally resembled the reference

sur-

factant SLES-7E0.

Discussion

The pivotal role of M . ovalis in the development of

dandruff is generally acknowledged. It has been shown

that the activity of antidandruff shampoos of the anti-

fungal class is primarily linked to their activity against

Mafusse~ia.'

n the global clinical evaluation, the other

ingredients of sham poos m ay interfere with the efficacy

an d tolerability of the treatment. One aspect is related to

the ability to remove a nd disrupt clumps of corneocytes

without inducing irritation of the scalp. Surfactants

present in shampoos are interfacially active substances

which interact with the stratum corneum. The mechan-

ism(s) whereby surfactants elicit undesirable effects is

not fully understood.16 Many techniques for the esti-

mation of the potential irr itancy of surfactants have

been described over the past

30

years. Traditional eva-

luations usually entail assessing the degree of erythem a,

dryness and scaling. -19 M os t of the methods categorize

products into a few broad categories. However, they

lack the sensitivity to distinguish products within a

References

1. Shuster S, The aetiology of dandruff and the mode of

action of therapeutic agents. Br J D e r m a f o l ( l 9 8 4 ) 111:

2 . Marks

R, Pearse AD, Walker AP, The effects of a

shampoo containing zinc pyrithione on the control of

dandruff. Br J Dermatol (1985)

112:

41 5

-

2.

3.

Van Cutsem J, Van Gerven

F,

Fransen J et al, The

in

vitro antifungal activity of ketoconazole, zinc pyr-

ithione, and selenium sulfide against Pityrosporum and

their efficacy as a shampoo in the treatment of

experimental pityrosporosis in guinea pigs. J Am Acad

Dermatol (1990) 22: 9 9 3 - 8 .

4 . Kikuchi T, Hori I, Sakamoto T et al. Demonstration of

neutrophil chemotactic anaphylatoxin

in

human dan-

druff.

A r c h

Dernratol Res (1989)281: 2 8 2 - 6 .

5

Pierard-Franchimont

C ,

Arrese JE, PiCrard GE, Im-

munohistochemical aspects of the l ink between Mulus-

sezia ovalis and seborrheic dermatitis. J Eur

Acad

Dermatol Vener 1995 4: 14- 19.

235 -42 .

group. The addition of instrumental measurements to

the clinical assessments further increases the power of

tests. J.M.ZI

In the present study, the interactions between

shampoos and stratum corneum were compared with

the effect of the mild surfactant

SLES-7E0,

using two

simple and quick bioassays. In the corneosurfametric

and squamometric tests , measures of CSMI and D-

SQMI, and grades of corneocyte dye binding give an

insight into the potential irritation of the products.8.

The highest dye binding to corneocytes in the squa-

mometric test was found for products identified as

potentially irritant in the corneosurfametric bioassay.

A significant linear relationship was found between

CSMI and D-SQMI of the shampoos . The present

findings apparently challenge an earlier assumption

that the two variables are not correlated. The corre-

lation exists for mild products and may be lost for

harsh products. When the tests are performed at room

temperature, CSMI and D-SQMI appear to be more

sensitive for discriminating between irritant and mild

products, respectively. However, the corneosurfametric

bioassay performed at 40C instead

of

20C also pro-

vides good discrimination between mild personal-care

products.

The colorimetric assessment

of

CSMI and D-SQMI

were designed to discriminate small differences in irri-

tation potential, but they lack the ability to separate

products based upon the effect on corneocyte cohesive-

ness. Microscopic observations of the squamometry

samples, with

or

without morphom etric evaluations, are

needed to explore the corneocyte loosening. Moderately

low anticlumping activity was found for most of

the

mildest shampoos as rated by D-SQMI. However, de-

spite the overall power correlation between D-SQMI

an d . the rating of corneocytes loosening, two

of

the

mildest sha mpo os (Dercos, Nizoral) exhibited strong

activity against corneocyte clumping.

In conclusion, the present study shows differences

amo ng antidandruff s ham poo s in their interaction with

the stratum corneum. N o single test method correlated

precisely with the variety of events which occur during

the normal use of a sha mpo o. The combination of

bioassays and in vivo testing gives the best insight into

the biological action mechanisms.

6 . Marks

R ,

Nickolls

S,

Dru gs which influence the stra tum

corneum and techniques for their evaluation.

Clin

Exp

D e r m a t o l ( l 9 8 1 )

6 :

4 1 9 - 2 7 .

7 . Takahashi M, Black D, Hughes

B

et al, Exploration of a

quantitative dansyl chloride technique

for

measurement

of the rate of desquamation. Clin Exp D e r m a t o l ( l 9 8 7 )

8 . Pitrard G E, EEMCO guidance to the assessment of dry

skin (xerosis) and ichthyosis: evaluation

by

stratum

corneum strippings.

Skin

Res Techno1 (19 96) 2:

3

-

1.

9 . Piirard GE, Piirard-Fran chimon t C, Saint Leger D et al,

Squamometry: the assessment of xerosis by colorimetry

of D-Squa me adhesive discs.

J SOC

Cosmet Chem (199 2)

10.

PiCrard GE, Goffin

V,

Piirard-Franchimont C, Corneo-

surfametry: a predictive assessment of the interaction of

personal care cleansing products

with h u m a n

stratum

corneum. Dermorology

I

994)

189:

152 - 6 .

12: 2 4 6 - 9 .

41: 297- 305.


4/4

218 V G o l l i n et i l l

1 1 . Pikrard GE, Goffin

V,

Piirard-Franchimont C, Squa-

mometry and corneosurfametry in rating interactions of

cleansing prod ucts with stratum corneum. Soc

Cosmet

Cheni

(1994) 45: 269

-

77.

12.

Goffin V, Paye M , PiCrard GE, Comparison of in vitro

predictive tests for irritation induced by anionic

surfactants. Contact

Dermatitis (1995) 33: 38 -41 .

13.

PiCrard GE, Goffin

V,

Hermanns-LB

T

et al, Surfactant

induced dermatitis.

A

comparison of corneosurfametry

with predictive testing on human a nd reconstructed skin.

Am Acad

Dermatol(1995) 33: 462-9 .

14.

Rhein LD , Simion FA , Surfactant interactions with skin.

Surf Sci Ser (1991) 32: 33-49.

15. Paye M, Simion

A,

PiCrard GE, Dansyl chloride

labelling of stratum corneum: its rapid extraction fro m

skin can predict skin irritation due to surfactants and

cleansing products. Contact Dermatitis (1994) 3 0 9 1- .

16.

Froebe CL, Simion FA, Rhein LD et al, Stratum

corneum lipid removal by surfactants: relation to in

vivo irritation. Dermatologica

(1990)

181:

277- 83.

17. Frosch PJ, Kligman AM , The so ap chamber test. Am

Acad Dermatol( l979) 1: 35 41.

18. Lukacovic MD . D unlap FE , Michaels S E et a l , Forearm

wash test to evaluate the clinical mildness of cleansing

products.

SO Cosmef Chem (1988) 3 9 355

-

66.

19. Strube DD. Koontz

SW,

Muraha ta RI e t al, The flex

wash test: a method for evaluating the mildness of

personal washing products.

Soc

Cosmet

Chem

(1989)

20. Simion FA, Rhe in LD, Grove G L e t al, Sequential order

of skin responses to surfactants during a soap chamb er

test. Contact Dermatitis (1991) 27: 174-81.

21. Paye M, Morrison BM. Wilhelm KP, Skin irritancy

classification of body cleansing products. C omp ariso n of

two test methodologies. Skin Res Techno1

(1995)

1:

30-

5.

22.

Goffin V, Letawe C, PiCrard GE, Temperature-depen-

dent effect of skin cleansing produc ts on hu man str atum

corneum. Toxicol Cutan Ocular Toxicol (1996) 15:

40: 279

-

306.

125- 30.

1996 - v goffin - antidandruffshampoosandthestratumcorneum[retrieved-2014!09!20]

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