International Journal of Cosmetic Science, 2011, 33, 171–182
Grey hair: clinical investigation into changes in hair fibres with loss
of pigmentation in a photoprotected population
P. D. Kaplan*, T. Polefka , G. Groveà, S. Daly , L. Jumbelic , D. Harper*, M. Nori*, T. Evans*, R. Ramaprasad* and R. Bianchini
*TRI/Princeton, 601 Prospect Ave, PO Box 625, Princeton, NJ 08542,  Johnson and Johnson, 199 Grandview Rd, Skillman, NJ 08588 and
à
Cyberderm, Inc., 700 Parkway Dr, Broomall, PA 19008, U.S.A.
Received 23 May 2010, Accepted 21 June 2010
Keywords: ageing hair, grey hair, material properties of hair, pigmented/unpigmented hair
Synopsis
Loss of pigmentation in hair fibres is one of the most obvious phe-
notypic changes with ageing and has been a topic of increasing
interest in the study of follicle biology. The onset of greying brings
cosmetic complaints that grey fibres are wild or difficult to manage.
Of course, these perceptions may be the consequence of visual
obviousness rather than underlying physical or chemical differ-
ences. Although several studies have compared pigmented and
unpigmented fibres, few have tried to control genetic and ethnic
difference as well as extrinsic factors such as photoexposure and
chemical treatment. We have recruited subjects with salt-and-
pepper hair from a population of Old Order Mennonites who, for
cultural reasons, are not only prohibited from chemically treating
their hair but also limit their exposure to sunlight. Hair samples
were examined for elemental composition, surface energy, Young’s
modulus, break stress, bending modulus, shear modulus and water
sorption/desorption isotherm. The parameters were evaluated sta-
tistically for global differences, individual differences and typical
individual differences. Consistent with previous published literature,
few global differences were found between pigmented and unpig-
mented hair across the population. We do find that many indivi-
dual subjects had differences between pigmented and unpigmented
fibres. These differences tend to be more pronounced in bulk than
in surface properties. The small differences in mechanical properties
and moisture uptake and loss lend support to the perception by
consumers that grey hair is wilder, drier and less manageable.
Résumé
La perte de pigmentation des fibres capillaires est un des change-
ments de phénotypes les plus évidents avec le vieillissement et reste
un sujet d’intérêt croissant dans l’étude de la biologie du follicule.
Le début du grisonnement s’accompagne de désagréments cosmé-
tiques avec des fibres grises plus difficiles à coiffer. Bien sûr, ces
Correspondence: Peter Kaplan, TRI/Princeton, 601 Prospect Ave, Prin-
ceton, NJ 08540, U.S.A. Tel.: +1 609 430 4839; e-mail: pkaplan@tri-
princeton.org
Current address:
R. Bianchini, Merck & Co., One Merck Drive, Whitehouse Station, NJ
08889, U.S.A.
T. Polefka, Life Science Solutions, 79 Ellison Road, Somerset, NJ
08873, U.S.A.
ª 2011 TRI/Princeton. Journal compilation
ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
doi: 10.1111/j.1468-2494.2010.00614.
 
perceptions peuvent être la conséquence de l’évidence visuelle
plutôt que de réelles différences physiques ou chimiques. Bien que
plusieurs études aient comparé des fibres pigmentées et non pig-
mentées, peu ont essayé d’évaluer le rôle de la différence génétique
ou ethnique aussi bien que des facteurs extrinsèques comme la
photo exposition ou le traitement chimique. Nous avons recruté
des sujets avec des cheveux poivre et sel d’une population du Vieil
Ordre Mennonites qui, pour des raisons culturelles, s’interdisent les
traitements chimiques capillaires, mais également limitent leur
exposition à la lumière du soleil. Sur les échantillons de cheveux,
ont été examinés: la composition élémentaire, la tension de surface,
le module d’Young, la capacité à se casser, le module de courbure,
le module de cisaillement et l’isotherme d’absorption/désorption de
l’eau. Le traitement statistique des paramètres évalués a permis de
mesurer des différences globales, des différences inter individuelles
et des différences intra individuelles. En accord avec la littérature
déjà publiée, peu de différences globales ont été trouvées entre des
cheveux pigmentés et non pigmentés dans cette population. Nous
constatons surtout que pour beaucoup chez le même sujet, des
différences entre des fibres pigmentées et non pigmentées sont
présentes et ont tendance à être plus prononcées que celles mises
en évidence sur les propriétés de surface. Les petites différences
observées pour les propriétés mécaniques et l’assimilation ou la
perte d’eau semblent être le support de la perception par les
consommateurs de cheveux grisonnants de cheveux plus secs et
moins faciles à coiffer.
Introduction
According to Yaar and Gilchrest [1], ‘aging is a basic biological
process characteristic of all living organisms…and widely acknowl-
edged to be the consequences of both a genetic program and cumu-
lative environmental wear and tear.’ One of the most obvious signs
of human ageing is the loss of hair pigmentation, greying [2]. Hair
greying, also known as canities, is a progressive loss of pigment
from growing hair shafts caused by one or more impairments in
melanocyte–keratinocyte functions including (i) reduction in tyrosi-
nase activity in the melanocyte, (ii) suboptimal melanocyte–kerati-
nocyte interactions, (iii) senile loss of melanocytes [3] and (iv)
defective migration of melanocyte cells from their reservoir in the
outer root sheath to their functional location near the dermal
papilla [4]. Although the mechanism is poorly understood [5],
onset is guided by inheritance and begins in the mid-30s for
Caucasians, late 30s for Asians, and mid-40s for Africans [6].
171
Clinical study of gray hair fibers P. D. Kaplan et al.

The hair follicle provides an interesting opportunity to study an Table I Demographic data
age-associated functional loss, because at some point in almost
every individual’s life, they will exhibit both high- and low-melanin Subject Age Sex
phenotypes, colloquially known as, salt-and-pepper hair. Indeed,
several investigators studied the material differences between pig-
mented and unpigmented hair fibres drawn from the same indivi- 1 59 M
2 56 M
dual [7–10]. Mathur [11] found that pigmented hair was
3 62 F
characterized by a larger cross-sectional diameter and greater dry 4 61 F
tensile modulus and break stress than unpigmented hair. Exposure 5 59 F
to UV radiation increased these differences. Hollenfelder, et al. [10] 7 53 F
observed that unpigmented hair fibres were coarser and more 8 60 F
‘wavy,’ and also exhibited greater weathering and more reactivity 10 54 F
11 62 M
than their pigmented counterparts. Wei [9] showed that unpigmen-
12 65 F
ted fibres exposed to UV radiation degraded faster and to a greater 13 58 F
extent than pigmented fibres. Gao and Bedell [8] reported findings
confirming those of Wei. While studying hair thickness, medulla-
tion and growth rate, Van Neste [7] observed that the growth rate clarifying shampoo lacking quaternary conditioning ingredients.
of unpigmented hair was approximately 10% greater than pigmen- They were instructed not to apply any other hair products nor
ted hair. Moreover, unpigmented hair was significantly thicker and tamper with their hair in any way during the 1-week condition-
characterized by a more prominent medulla. ing period.
In the current report, we analyse the material (mechanical and
chemical) properties of the hair fibres drawn from a group
Hair samples
(N = 11) of ageing (53–65 years of age) religious Mennonites who
for cultural reasons protect their hair from sunlight (eight women) A female technician clipped enough hair, as close to the scalp as
and avoid chemical treatments of any kind. Unlike earlier studies, possible, to include approximately 500 strands of each pigmenta-
we conduct a more detailed statistical analysis to determine global tion type and placed them in a plastic bag. No attempt was made
differences (i.e. differences that may describe all unpigmented fibres to separate the hairs into pigmented or unpigmented fibres at this
across the population), individual differences (i.e. difference within time, although extreme care was taken to ensure that the clippings
a particular individual), and typical individual differences (i.e. dif- were always oriented in the same direction. The strands were kept
ferences between pigmented and unpigmented fibres in most indivi- in the orientation by gluing (Loctite Super Glue Gel, Henkel, Wes-
duals). Consistent with previous published literature, few global tlake, OH, U.S.A.) the group(s) of hair near the midpoint of a
differences were found between pigmented and unpigmented hair pinched hair bundle. All subsequent measurements avoided the
across the population. However, more differences were observed glued portions of the hair. Swatches were placed into plastic bags
within individuals. after folding a strip of waxed paper over the glued portion. Strands
(at least 10 cm long) were manually sorted into pigmented and
unpigmented bundles prior to single fibre testing. Some tests were
Materials and methods
not possible on the hair of some subjects because of the length of
the hair and the location of the glue. A description of which tests
Panelists
were performed on hair from which subjects is found in Table II.
All volunteers were recruited from within a strict Mennonite com-
munity located in Lancaster, Pennsylvania. These volunteers were
Statistical analysis and presentation
chosen because (i) they have similar ethnic background (i.e. Swed-
ish), (ii) they are culturally prohibited from dyeing or otherwise As a complex biomaterial, hair fibres are quite variable in shape,
treating their hair, and (iii) in almost all cases, the women protect diameter and other characteristics. However, previous published
their hair from UV exposure (i.e. do not exhibit their hair outside reports [7–10] suggested that mechanical differences between
the home). The panel consisted of healthy Caucasian women
(n = 8) and men (n = 3) between the age of 45–65 years, having
salt-and-pepper hair (approximately, 50% of hairs not pigmented) Table II Analyses – performed by subject
long enough to obtain samples 2–4 inches in length, most samples
were considerably longer. Although these subjects have always Subject Surface XPS Friction Tensile Bending Torsion DVS
refrained from chemical treatments, they were specifically inter-
viewed to ensure that they had not chemically treated their hair in
1 X X X X – X –
the past 6 months. Panelists with curls tighter than 5 cm were 2 X X – X – – –
excluded from participating in this study. Demographic data for 3 X X X X X X X
these panelists are listed in Table I. 4 X X X X X X –
All volunteers signed a consent form after being informed of 5 X X X X X X X
their obligations and risks that they might encounter as a partici- 7 X X X X X X X
8 X X X X X X X
pant in this study. Upon being selected into the study, the pane-
10 X X X X X X –
lists were reminded of the general nature and purpose of the 11 – X X X – – –
study. Each candidate was given Neutrogena Anti-Residue 12 X X X X X X X
Shampoo (Los Angeles, CA, U.S.A.) to use for the 1 week prior to 13 X X X X X X X
hair sample collection. This formula was chosen because it is a

ª 2011 TRI/Princeton. Journal compilation

ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
172 International Journal of Cosmetic Science, 33, 171–182
Clinical study of gray hair fibers P. D. Kaplan et al.

pigmented and unpigmented hair fibres were relatively modest. The Table III Surface energy – cos(h) of pigmented and unpigmented hair by
fact that both pigmentation states can be collected from the same subject (*P < 0.05)
individual is the key to designing a sensitively balanced study. For
each parameter discussed in this report, three types of differences Pigmented Unpigmented
were examined: (i) global difference, which supports statements of
the form: pigmented hair has a higher value of a particular attri-
Subject Std Std P
bute than unpigmented hair (e.g. trivially, the statement, ‘pigmen- # Dcos(h) (2 sided)
cos(h) dev CV cos(h) dev CV
ted fibres are darker than unpigmented fibres’ refers to a global
difference), (ii) typical individual differences, which support state-
ments of the form: on most people, the pigmented hair is higher in 1 )0.254 0.068 )27 )0.136 0.047 )35 0.118 0.0004*
this attribute than unpigmented hair (e.g. we will see that unpig- 3 )0.192 0.106 )55 )0.085 0.06 )70 0.107 0.02*
4 )0.114 0.025 )22 )0.129 0.034 )27 )0.015 0.287
mented hair typically has a higher bending rigidity), and (iii) indivi-
5 )0.18 0.055 )30 )0.252 0.044 )18 )0.072 0.005*
dual difference, which support statements of the form: several 7 )0.244 0.037 )15 )0.171 0.071 )41 0.073 0.009*
subjects had a difference in an attribute between their pigmented 8 )0.203 0.085 )42 )0.197 0.044 )22 0.006 0.84
and unpigmented hair, (e.g. we will see that five of nine subjects 10 )0.234 0.039 )17 )0.206 0.011 )5 0.028 0.0485*
have pigmented hair that is more hydrophobic than their pigmen- 12 )0.252 0.053 )21 )0.266 0.03 )11 )0.014 0.477
ted hair). 13 )0.206 0.018 )9 )0.176 0.012 )7 0.03 0.0004*

Restated simply, it is common to look at global differences, finding

for example that grey hairs are not thicker than fully pigmented
hairs when all fibres are pooled. In this study, we also look at indi-
vidual differences to see if these hairs tend to be larger on an indi-
vidual head. Differences that are strongly observable in some (a)
people, but not on average in the population we call individual
differences. Individual differences do not describe the group, but do
describe phenomena that may be real and important to those with
the difference. When such differences are consistently found, we
say that we have found a typical individual difference.
The visualization of these three types of difference is not well
communicated using bar charts. Bar charts either represent the
average value of each measure or the individual value for each
subject, as illustrated in Fig. 1 (contact angle for each subject). This
bar chart, which compliments the same information shown in
Table III, does not make it easy to judge either overall averages
(global differences), patterns across subjects (typical individual differ-
ences) or the significance of particular long and short bars for single
subjects (individual differences). The presentation we have chosen to
use in this report is shown in Fig. 2. Global differences are deter-
mined by t-test (unpaired, two-sided) and discussed in the context
of the histogram showing the distribution of all measurements
(Fig. 2a). In this paper, all measurements of the pigmented hair
appear shaded in the histogram. A quick visual inspection of the
distribution of contact angles Fig. 2a suggests no obvious global
difference between the two hair types. To judge individual differences,
(b)
Contact angle
Subject ID
1 3 4 5 7 8 10 12 13
0

–0.05

–0.1

–0.15

–0.2

–0.25
Pigmented
–0.3 Unpigmented
Figure 2 Surface energy for pigmented (shaded bars) vs. unpigmented (open
Figure 1 Mean contact angle for pigmented and unpigmented hair fibres bars) hair (N = 50 fibres/subject-pigmentation state). (a) Distribution histo-
for individual subjects (N = 10 fibres/subject-pigmentation state). gram. (b) Within subject analysis.

ª 2011 TRI/Princeton. Journal compilation

ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
International Journal of Cosmetic Science, 33, 171–182 173
Clinical study of gray hair fibers P. D. Kaplan et al.

Table IV Elemental analysis of pigmented and unpigmented hair XPS/ESCA (a)

Hair type Subject C N O Na Si S Cl Ca

Unpigmented 1 79.9 4.9 11.6 0.3 1.2 1.7 0.1 0.3

2 79.7 4.7 12.3 0.6 0.9 1.4 0.1 0.3
3 79.1 5.4 12.4 0.3 0.6 1.9 0.1 0.3
4 78.8 5.2 12.8 0.1 0.8 1.6 0.0 0.6
5 80.0 4.6 12.2 0.8 1.1 1.2 0.1 0.1
7 81.8 3.9 11.3 0.4 0.9 1.2 0.1 0.6
8 78.1 6.0 12.4 0.6 0.6 2.0 0.1 0.2
10 85.3 4.0 8.5 0.6 0.3 1.1 0.1 0.2
11 78.6 5.2 12.9 0.5 0.7 1.5 0.2 0.3
12 79.9 5.5 11.5 0.3 0.5 1.8 0.1 0.5
13 79.5 6.2 11.2 0.2 0.3 2.2 0.1 0.4
Pigmented 1 81.3 4.7 11.0 0.3 0.9 1.5 0.0 0.2
2 79.9 4.9 12.3 0.4 0.6 1.4 0.0 0.5
3 83.8 3.5 10.3 0.2 0.6 1.2 0.0 0.5
4 80.1 5.1 11.6 0.6 0.7 1.6 0.1 0.2
5 78.7 5.1 12.9 0.6 1.3 1.3 0.0 0.1
7 79.0 5.4 12.4 0.2 0.9 1.7 0.0 0.4
8 80.9 4.8 10.8 1.0 0.5 1.7 0.1 0.2
10 80.6 5.3 11.3 0.7 0.4 1.5 0.1 0.2
11 78.5 5.5 12.9 0.3 0.7 1.5 0.1 0.5
12 81.6 4.6 11.2 0.1 0.3 1.4 0.0 0.7
13 79.8 5.8 11.3 0.2 0.4 2.0 0.0 0.4
(b)

the tabular data where P-values are presented show clearly which
subjects have a difference in the measured quantity. To illustrate
typical individual differences, we use the type of plot found in Fig. 2b,
which shows the individual differences plotted against the indivi-
dual means. Median lines are drawn on this figure (solid) and con-
fidence intervals are also indicated (dashed). For measures whose
confidence interval includes zero, the data do not support the state-
ment that there is a typical individual difference. But the final con-
clusion is calculated by means of paired t-test. Calculations are
performed using jmp version 7.0.1 from SAS (Cary, NC, U.S.A.).

Surface energy
The surface energy between hair and water, csl, was measured by
the Wilhelmy technique [12, 13]. The surface energy is generally
characterized by the cosine of the contact angle through Young’s
equation csa ) csl = cla cosh, where the energy between solid and
air is denoted csa and between liquid and air cla. Each fibre was
mounted between brass ferrules, and its elliptical area measured by
a laser micrometer from which we deduce the fibre’s perimeter p.
The fibre was then cut in half and one half, suspended from a
microbalance, was hung above a small cup of de-ionized water
(TRI Scan, TRI/Princeton, Princeton, NJ, U.S.A.). The cup, sitting
on a motorized table, rises until the fibre enters the water. The
microbalance reports the force of insertion F. This force divided by
the fibre perimeter gives the advancing or receding contact angle
through the equation F ¼ pcla cosðhÞ.
XPS/ESCA
X-ray Photoelectron Spectroscopy or Electron Spectroscopy for Che-
mical Analysis (XPS/ESCA) was performed by Evans Analytical
Group (East Windsor, NJ, U.S.A.). Briefly, the fibres were aligned in
the plane formed by the incident X-ray and detected electrons
paths. In addition, the hair samples were elevated above the platen
so that only the hair was in focus.
Figure 3 Single fibre friction analysis of pigmented (shaded) and unpigmen-
ted (open) hair fibres (N = 10 fibres/subject-pigmentation state). (a) Distribu-
tion histogram. (b) Within subject analysis.
X-ray Photoelectron Spectroscopy data was quantified using rela-
tive sensitivity factors and a model that assumed a homogeneous
layer. For analysis it was assumed that the volume probed was the
product of the analysis area (spot size or aperture size) and the
analysis depth. Photoelectrons are generated within the X-ray
penetration depth (typically many microns), but only the photo-
electrons within the top three photoelectron-escape-depths are
detected. Escape-depths are on the order of 15–35 Å, which leads
to an analysis depth of 50–100 Å. Typically, 95% of the signal
originates from within this depth. Note that because of the cylindri-
cal nature of hair, a larger surface area was probed than would be
from a flat sample. Analysis was performed using a PHI 5701 LSci
with a monochromated Al Ka X-ray source at 1486.6 eV, an
acceptance angle of ±7, and a take off angle of 65. The analysis

ª 2011 TRI/Princeton. Journal compilation

ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
174 International Journal of Cosmetic Science, 33, 171–182
Clinical study of gray hair fibers P. D. Kaplan et al.

Table V Friction analysis of pigmented vs. non-pigmented hair by subject Bending

The measurement of bending rigidity [14] was accomplished by
Hair type Difference lowering single fibres against a hook suspended from a microba-
lance (Cahn D-200). The fibres were oriented to bend about their
Subject ID Pigmented Unpigmented D P (1 sided) minor axis, in order to determine both the intrinsic modulus and
the extrinsic rigidity. In brief, 30 mm of each fibre was crimped
between two brass ferrules. Diameter measurements were made
1 0.17 0.16 0.01 0.31
using a laser micrometer (Mitutoyo LSM-6100) while orienting the
3 0.15 0.12 0.02 0.12
4 0.15 0.15 0.00 0.49 brass ferrules. The laser records a profile as a function of rotation
5 0.12 0.14 )0.02 0.06 angle, which was then used to identify the angle that the major
7 0.13 0.07 0.06 0.12 axes made with the ferrule. The fibres were then oriented so that
8 0.13 0.15 )0.02 0.28 they would bend about their minor axis. Each fibre was bent at
10 0.17 0.11 0.06 0.15 L = 5 mm from the ferrule at a rate of 1 lm min)1. The force dis-
11 0.11 0.12 )0.01 0.24
tance curve, F vs. D, is analysed to determine the fibre rigidity
12 0.08 0.08 )0.01 0.38
13 0.08 0.09 0.01 0.30 R ¼ EI ¼ FL3 3D where E is the bending modulus,  I the area-
moment of inertia about the minor axis I ¼ pb3 a 4 and a and b
are the semi-major and semi-minor radii. Ten fibres were measured
for each subject/pigmentation combination.
area is 2.0 · 0.8 mm and a charge correction used (C1s
284.8 eV). Torsion
Reference data values from published literature were used to
Following the methods of reference [15], each fibre (L = 30 mm)
assign chemical state for each element. However, chemical-state
was loaded in a torsional pendulum. After measurement of the
assignments must be considered tentative in cases where the
semi-major a and semi-minor b axes of the fibre, each fibre was
available reference data is limited or in cases where similar bind-
mounted in the instrument where it supported a weight (M =
ing energies are observed for a number of different chemical
5 g) with moment of inertia L = 8.851e)8 kg-m2. The weight
states.
was twisted through one revolution (shear strain of 12 per
Non-linear least squares (NLLS) curve fitting was applied to the
mm) and released. The instrument measures the period of oscil-
S2p high-resolution spectra to assist in possible chemical state
lation T and velocity of the pendulum at zero strain. The Shear
assignment. Results of the NLLS fits are shown on the individual
Modulus (G)
 of an elliptical fibre is given by
spectra. Curve fit routines are mathematical devices, and for a
G ¼ 16 p L M T 2 ða3 b þ ab3 Þ.
given peak a number of mathematically valid results may exist.

Dynamic vapour sorption

Friction
The water vapour sorption/desorption characteristic of pigmented
Low load friction was measured using the TRI/Scan microba-
and unpigmented hair was measured gravimetrically using a Model
lance. In this measurement, a 500 -mg weight hangs from a single
DVS-1 Dynamic Vapor Sorption instrument manufactured by Surface
hair fibre that was pushed slightly, forward by a steel wire so that
Measurement Systems Ltd. (Alperton, Middlesex, U.K.). Briefly, the
it makes an angle of 0.85 with the vertical. The wire moves down
hair (approximately 50 mg) sample was placed into the instrument
the fibre at 1 mm min)1. The forces recorded by the microbalance
set to 95% RH for several hours before being exposed to a dry stream
show that the fibre exhibits stick/slip motion. We identify the force
of nitrogen for 15 h. The relative humidity was then increased from
associated with each peak in the force–distance curve as a stick
0% to 90% RH in 10% increments followed by a single final step
event and average these events to determine the static coefficient of
increase to 95% RH. Once equilibrium at 95% RH was attained, the
friction.
humidity was decreased to 90% and then to 0% in increments of
10%. The humidity controller is set to proceed to the next step when
Tensile and cross-sectional area either equilibrium of the specimen weight was detected (<0.0015%
change in mass per min) or 360 min was exceeded.
Fibres, compression mounted in brass ferrules lined with an elas-
The equilibrium weight data at each target humidity during
tomeric plastic, were measured in sequence for dimensions, for
sorption phase (humidity increasing) and desorption phase (humid-
dry elastic (Young’s) modulus and then for wet tensile properties
ity decreasing) were converted to ‘Per cent change in mass’ (S, D)
including Young’s modulus, plateau yield and modulus, break
relative to the initial dry sample mass. For each target humidity
extension and stress. The dimensions were measured by Mitutoyo
level h between 10% and 90%, the hysteresis H was determined by
LSM-6100 laser micrometer (Aurora, IL, U.S.A.) that records
subtracting, H = D ) S. The hysteresis between the two curves is a
major and minor axis. Measurements are made at three slices
measure of the capacity for water binding.
along the fibre, an average cross-sectional area is calculated and
that averaged area is used in subsequent analysis. The dry modu-
lus was determined by performing a 3% extension at 65% RH.
Results
After allowing at least a day for the fibres to recover, they were
soaked in water and their wet tensile properties were measured.
Surface energy
These measurements were all performed and analysed using a
robotic tensile analyzer, Diastron model MTT-675 (Andover, Surface energies are listed in Table III, and the paired differences
Hampshire, U.K.). are plotted against the mean in Fig. 2. Ten pigmented and ten

ª 2011 TRI/Princeton. Journal compilation

ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
International Journal of Cosmetic Science, 33, 171–182 175
Clinical study of gray hair fibers P. D. Kaplan et al.

unpigmented fibres from each subject were measured. We see that XPS
all surfaces are hydrophobic [cos(h) < 0] and that there is no glo-
Table IV provides the atomic concentrations of the elements
bal difference between pigmented and unpigmented fibres.
detected by XPS. Values given were normalized to 100% using the
Although six of nine subjects showed individual differences (Table -
elements detected. No differences were detected.
III) whereby the pigmented hair in five of these subjects appeared
This negative result should be evaluated in terms of the sensitiv-
more hydrophobic, the magnitude of the differences was small
ity of detection. In particular, this technique does not detect H or
and as can be seen in Fig. 2b, not sufficiently consistent to con-
He. Detection limits are approximately 0.05–1.0 atomic per cent.
clude that there is a typical individual difference. It is possible that
The surfaces of all 22 (11 pigmented and 11 unpigmented) hair
residual sebum contributes to these surface energies and we have
samples contained similar concentrations of carbon, oxygen, nitro-
no evidence to suggest that it contributes differently for the pig-
gen, sulphur, and low to trace concentrations of silicon, sodium,
mented and unpigmented fibres.
calcium, and probably chlorine. The chemical states observed

Table VI Summary of tensile of pigmented vs. non-pigmented hair by subject (*P < 0.05)

(a) Cross-sectional area

Sub Area (pig) Area (unpig) D CSA Dry mod Dry mod D Dry Ratio Ratio D
ID ·1e)9 m2 ·1e)9 m2 ·1e)9 m2 P (GPa) (pig) (GPa), unpig mod P pig unpig Ratio P

1 4.45 4.31 )0.14 0.170 3.463 3.389 )0.074* 0.001 2.37 2.28 )0.083 0.05
3 2.76 2.82 0.06 0.400 3.719 4.305 0.586* 0.020 2.41 2.53 0.122 0.06
4 1.63 1.87 0.23* 0.001 3.245 3.611 0.367 0.094 2.36 2.49 0.130 0.30
5 3.97 4.15 1.80 0.370 3.194 3.281 0.087* 0.014 5.73 2.18 )3.547 0.31
7 4.54 4.95 0.40* 0.043 4.078 3.583 )0.496* 0.000 2.30 3.18 0.877* 0.000
8 4.97 5.55 0.57* 0.003 3.428 3.303 )0.125 0.060 2.13 2.19 0.058 0.20
10 3.84 3.87 0.03 0.420 3.949 3.918 )0.031 0.190 28.75 2.39 )26.366 0.12
12 2.81 3.00 0.20 0.090 3.573 4.131 0.558* 0.000 2.49 2.49 0.006 0.41
13 2.37 2.80 0.43* 0.000 3.939 4.027 0.088* 0.000 2.37 2.30 )0.076 0.24

b) Elastic modulus (wet, dry and ratio)

Dry mod Dry mod Wet mod Wet Mod

Sub (GPa) (GPa), D Dry (GPa), (GPa), D Wet Ratio Ratio
ID (pig) unpig mod P pig unpig mod P pig unpig D Ratio P

1 3.463 3.389 )0.074* 0.00 1.471 1.485 0.014 0.43 2.37 2.28 )0.083 0.05
3 3.719 4.305 0.586* 0.02 1.553 1.661 0.108* 0.02 2.41 2.53 0.122 0.06
4 3.245 3.611 0.367 0.09 1.379 1.463 0.084 0.22 2.36 2.49 0.130 0.30
5 3.194 3.281 0.087* 0.01 1.428 1.516 0.089 0.33 5.73 2.18 )3.547 0.31
7 4.078 3.583 )0.496* 0.00 1.814 1.791 )0.023 0.48 2.30 3.18 0.877* 0.000
8 3.428 3.303 )0.125 0.06 1.661 1.520 )0.141 0.02 2.13 2.19 0.058 0.20
10 3.949 3.918 )0.031 0.19 13.376 1.638 )11.738* 0.04 28.75 2.39 )26.366 0.12
12 3.573 4.131 0.558* 0.00 1.444 1.665 0.221* 0.00 2.49 2.49 0.006 0.41
13 3.939 4.027 0.088* 0.00 1.665 1.772 0.107* 0.00 2.37 2.30 )0.076 0.24

c) Break Stress

Sub ID Break Stress (MPa) pig Break Stress (MPa) unpig D break stress P

1 178.8 178.9 0.1 0.290

3 169.4 165.4 )4.0* 0.020
4 150.0 164.1 14.1 0.330
5 122.0 140.9 18.9 0.080
7 159.9 135.2 )24.7 0.390
8 161.4 176.8 15.4 0.055
10 174.1 174.0 )0.1* 0.020
12 148.1 56.5 )91.6* 0.000
13 183.2 186.7 3.4* 0.000

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176 International Journal of Cosmetic Science, 33, 171–182
Clinical study of gray hair fibers
(a)
(b)
Figure 4 Cross-sectional area of pigmented (shaded bars) and unpigmented
(open bars) fibres (N = 50 fibres/subject-pigmentation state). (a) Distribution
histogram. (b) Within subject analysis.
include: C–(C,H), C–(O,N), C=O, O–C=O, R–S, R–SOx, Si–(C,H),
Na+, Ca2+, and Cl).
Friction
In terms of friction, no obvious global differences between pigmented
and unpigmented hair are seen (Fig. 3a). Table V shows that one
subject (#5) had a significant individual but as can be seen in
Fig. 3b, which estimates typical individual differences, there is no
support for a general statement that unpigmented hair is rougher
or smoother than pigmented hair.
Tensile and cross-sectional area
The tensile measurements are composed of several parameters
including fibre cross-sectional area, wet and dry elastic modulus
ª 2011 TRI/Princeton. Journal compilation
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International Journal of Cosmetic Science, 33, 171–182
P. D. Kaplan et al.
(a)
(b)
Figure 5 Dry elastic modulus (GPa) for pigmented (shaded bars) and unpig-
mented (open bars) fibres (N = 50 fibres/subject-pigmentation state). (a)
Distribution histogram. (b) Within subject analysis.
(Young’s) and break stress. The subject specific results are found in
Table VI. The population-wide distributions of these quantities are
quite broad as illustrated in Figs 4–7.
We not only observed a global difference in cross-sectional area
(Fig. 4a, 2-sided t-test P = 0.01), but unpigmented hair for four
subjects was individually significantly larger than their pigmented
counterparts (Table VI). For this study group, these differences in
cross-sectional area are also typical (two-sided t-test, P = 0.03).
These results support the consumer-perceived notion that unpig-
mented hair is coarser than pigmented hair.
Despite differences in cross-sectional area, the intrinsic
mechanical properties of pigmented and unpigmented hair do
not show global differences (Figs 5–8a), although individual differ-
ences in six of nine subjects are observed for dry modulus, four
of nine for wet modulus, one for ratio of moduli, and four of
nine for break stress (Table VI). These differences (Figs 5–8b),
177
Clinical study of gray hair fibers
(a)
(b)
Figure 6 Wet elastic modulus (GPa) for pigmented (shaded bars) and unpig-
mented (open bars) fibres (N = 50 fibres/subject-pigmentation state). (a) Dis-
tribution histogram. (b) Within subject analysis.
however, were judged not to be typical (P-value ‡0.05). Further-
more, as these are all intrinsic parameters, and we have already
documented that unpigmented fibres tend to be coarser (higher
cross-sectional area), these do not contradict the anecdotal obser-
vation that grey hair is stiffer.
Bending
Although only three of eight subjects showed individual differences
in bending modulus (Table VII), the population-wide global differ-
ence was significant (two-sided P-value £0.001) and typical (two-
sided P-value = 0.015) (Fig. 9a,b), indicating that unpigmented
hair tends to exhibit approximately 15% higher bending modulus.
This counter intuitive result, a global difference in the face of little
individual difference, most often happens when the difference is
real, but just below the sensitivity of the individual test. Paired sta-
tistics, in this case, bring out the effect.
ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
178
P. D. Kaplan et al.
(a)
(b)
Figure 7 Ratio of dry to wet elastic modulus for pigmented (shaded bars)
and unpigmented (open bars) fibres (N = 50 fibres/subject-pigmentation
state). (a) Distribution histogram. (b) Within subject analysis.
Torsion
The distribution of shear moduli for pigmented and unpigmented
fibres (Table VIII) are plotted in Fig. 10a. Two individual subjects
(#1 and #8) exhibit significant individual differences in shear modu-
lus, but in opposite directions. Overall, the paired analysis
(Fig. 10b) shows no typical individual difference in torsion between
the two hair types (P-value ‡0.05). Note that the moduli are nor-
malized to the dimensions of each fibre’s elliptical cross section. As
for tensile measurements, the fact that unpigmented fibres are, on
average, a little bit thicker means that they will exhibit greater
torsional rigidity even with the same intrinsic torsional modulus.
Dynamic vapour sorption
Table IX and Fig. 11 shows the adsorption/desorption curves for
pigmented (solid dots) and unpigmented hair (hollow dots). The
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Clinical study of gray hair fibers P. D. Kaplan et al.

(a) (a)

(b)
(b)

Figure 9 Bending modulus (GPa) for pigmented (shaded bars) and unpig-
Figure 8 Break stress modulus (MPa) for pigmented (shaded bars) and mented (open bars) fibres (N = 10 fibres/subject/pigmentation combination).
unpigmented (open bars) fibres (N = 50 fibres/subject-pigmentation state). (a) Distribution histogram. (b) Within subject analysis.
(a) Distribution histogram. (b) Within subject analysis.

Table VII Bending modulus (GPa) by subject (*P < 0.05) Table VIII Shear modulus (GPa) analysis by subject (*P < 0.05)

Hair type Difference Subject ID Pigmented Unpigmented D P (1-sided)

Subject# Pigmented Unpigmented D P (1 sided) 1 0.92 0.99 0.07* 0.002

3 1.00 1.06 0.06* 0.004
4 1.03 1.00 )0.02 0.24
3 0.99 1.35 0.36* 0.02 5 0.77 0.81 0.03* 0.05
4 1.31 1.39 0.08 0.34 7 1.05 1.07 0.02 0.32
5 0.81 0.98 0.17 0.07 8 0.94 0.89 )0.06 0.05
7 1.34 1.19 )0.15 0.09 10 1.00 0.98 )0.02 0.15
8 1.01 1.23 0.22* 0.05 12 1.02 1.04 0.02 0.22
10 1.07 0.96 )0.11 0.20 13 0.99 1.00 0.00 0.48
12 0.86 1.06 0.21 0.004
13 0.99 1.56 0.57* 0.02

hysteresis between sorption and desorption is a well-known phe- the data in this chart represents a single measurement on several
nomenon. Hysteresis is the difference in the capacity of a sample to hundred fibres (10–30 mg), we tested only for global differences.
adsorb and retain water at the same chemical potential. Because At the very highest humidity (80–90% RH), we find that grey hairs

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International Journal of Cosmetic Science, 33, 171–182 179
Clinical study of gray hair fibers P. D. Kaplan et al.

(a) Adsorption isotherms for panelist # 3

25

20

Amount adsorbed (%)

15

10

0
0 10 20 30 40 50 60 70 80 90 100
Relative humidity (%)

Figure 11 Typical water adsorption/desorption isotherm for pigmented

(closed circles) and pigmented (open cicles) fibres taken from subject #3.

(b) Intrigued by this difference, we also examined the raw DVS

data to obtain kinetic information and observe the rates of water
loss following a humidity step. As we only had sufficient hair from
five subjects, we report these dynamics as suggestive rather than
conclusive and refrain from plotting them. Quite consistently,
changes in water content happen faster in unpigmented hair. This
could potentially be a consequence of the hygroscopic nature of
melanin [16]. It could also be a consequence of the grey hair’s
more prominent medullation, which could create shorter paths for
the diffusion of water from the centre of the fibre to the edges
[17]. Kinetic differences in water transport could also be signifi-
cant for the phenomenon of unruly grey hair. Simplistically, as
fibres take up water, they have a tendency to return to their nat-
ural curl. If one fibre dries faster than its neighbours, there will
be a force for it to pop out of the array because of a different
momentary curliness.

Figure 10 Torsional modulus (GPa) for pigmented (shaded bars) and unpig-
mented (open bars) fibres (N = 14 fibres/subject-pigmentation state). (a) Dis-
tribution histogram. (b) Within subject analysis.
consistently release more water on initial drying. Interestingly, this
small difference could possibly be related to the interaction of water
with melanin or because of the small differences in structure
caused by the absence of melanin in the unpigmented hair.
Discussion
Hair is a complex and highly variable biomaterial that, at some
point in the ageing process, looses pigmentation [1, 2, 5, 6, 18].
The variability caused by individual intrinsic difference (e.g.
genetic/ethnic) is typically greater than that associated with pig-
mentation. To control genetic and ethnic differences and reduce
the impact of two key extrinsic factors; namely, photoexposure
[8–11] and chemical treatment [7], we obtained pigmented and
unpigmented hair from a group of Old Order Mennonites who are

Table IX DVS data by subject, RH and pigmentation, summary statistics in last row (D = hystersis pigmented ) unpigmented) (*P < 0.05)

RH 10 20 30 40 50 60 70 80 90

Subject P U P U P U P U P U P U P U P U P U

3 1.38 0.33 1.59 0.64 1.63 1.79 1.57 1.68 1.53 1.57 1.51 1.36 1.37 1.23 1.11 0.93 0.67 0.39
5 1.38 1.02 1.63 1.25 1.8 1.41 1.9 1.36 1.9 1.57 1.79 1.51 1.54 1.27 1.45 1.16 0.66 0.63
7 1.57 1.56 1.87 1.92 1.93 2.05 1.95 2.04 1.69 2.11 1.76 2.02 1.41 1.79 1.72 1.5 1.19 0.77
8 1.21 1.44 1.57 1.59 1.5 1.75 1.55 1.84 1.57 1.68 1.21 1.78 1.63 1.58 1.56 1.26 1.4 1.12
12 1.08 1.17 1.33 1.41 1.54 1.54 1.6 1.52 1.72 1.5 1.73 1.44 1.62 1.28 1.39 1.07 0.64 0.45
13 1.49 1.38 1.79 1.6 1.89 1.66 1.95 1.63 1.84 1.49 1.85 1.59 1.68 1.47 1.39 1.18 0.99 1.01
D/P )0.20 0.33 )0.23 0.21 )0.02 0.88 )0.08 0.58 )0.06 0.67 )0.03 0.87 )0.11 0.36 )0.25* <0.001 )0.20* 0.03

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180 International Journal of Cosmetic Science, 33, 171–182
Clinical study of gray hair fibers
culturally prohibited from dyeing or treating their hair, and in
the case of women, are required to cover their hair whenever
outside the home. Moreover, and in contrast to previously pub-
lished studies [7–11], we analysed the differences in chemical and
physical properties of pigmented and unpigmented fibres in terms
of three kinds of difference: global difference (illustrated in a histo-
gram); individual differences (tabulated data analysed via a two-
sided t-test); and typical individual difference (in which our sample
population is tested by plotting the difference vs. the mean for
each subject and the population is analysed through a paired t-
test).
Global differences
By definition, Global Differences reflects an attribute difference found
to be true across the whole subject population. Apart from colour
differences, the only measurable Global Differences between pigmen-
ted and unpigmented hair was that the unpigmented hair was
slightly coarser (Table VI, Fig. 5) and stiffer (e.g. bending measure-
ment, Table VII, Fig. 9). These results are consistent with the
results reported by Hollfelder, et al. [10] but contrary to those of
Gao and Bedell [8]. These two differences are synergistic in creating
more rigid grey fibres. In light of the fact that unpigmented hair is
typically more highly medullated, one would expect grey hair to be
softer and weaker in tensile measurements. The fact that, experi-
mentally, the reverse is the case suggests that there is an underly-
ing cause to be discovered.
Individual differences
Many people complain that their grey hairs behave differently from
their naturally pigmented hair [2]. Although some of this is attri-
butable to the fact that unpigmented hair is more visible against
the pigmented hair background, we find that real differences exist
for some individuals. The detailed data for these comparisons are
summarized in Tables III–IX.
In particular, we found that it was not unusual to observe clear
differences in mechanical properties. For example, significant differ-
ences were observed in six of nine subjects with respect to dry
modulus, four of nine for wet modulus, and four of nine for break
stress (Table VI, Figs 5–8).
Interestingly, our analysis differs from that of Gao and Bedell
who used pooled data and mean values for four subjects ([8]cf.
Table I). In their analysis, no difference in break stress was
observed in a test for global differences. By comparing the results
within individual, and not pooling the data from the entire popula-
tion, we have been able to identify differences for individual sub-
jects.
Typical individual differences
Typical individual differences represent differences that most people
can expect to find between their own pigmented and unpigmented
fibres. We have seen, in a sample of subjects in their 50s and early
60s, that unpigmented fibres generally have higher cross-sectional
area. We also report a clear and typical difference in bending mod-
ulus that is not matched by typical differences in linear elastic
moduli (Young’s modulus) measured wet or dry. We also find no
typical differences in twisting (shear modulus). Similar intrinsic
moduli, coupled with larger diameter fibres will, of course, result in
more rigid unpigmented fibres. From a measurement point of view,
all three differences were directionally similar. It may be that
ª 2011 TRI/Princeton. Journal compilation
ª 2011 Society of Cosmetic Scientists and the Société Française de Cosmétologie
International Journal of Cosmetic Science, 33, 171–182
P. D. Kaplan et al.
because of each measure’s different sensitivity, we were found less
uncertainty for bending than for twisting or stretching. Given the
relatively small number of subjects, we are content to conclude
that there is strong evidence of individual difference and an overall
trend that is not inconsistent with the observation that grey fibres
are sometimes stiffer.
A typical difference in contact angles was detected. This result is
directionally the same as previously reported, but it is small and
not likely to be even cosmetically significant [11].
In terms of water absorption and retention, the hysteresis
between 80% and 90% RH is consistent across the five measured
subjects, with the unpigmented hair retaining statistically signifi-
cantly less water than the pigmented hair, and apparently releasing
it faster. Because melanin is hygroscopic and binds water [19], it is
reasonable to speculate that the absence of melanin is one possible
reason why unpigmented fibres hold less water. The role of the
medulla in the kinetics of water transport could provide another
explanation [17]. If this difference in absorption kinetics had an
effect on the surface energy measurement, it would be to make the
perimeter slightly larger for the unpigmented hairs, which would
have resulted in measuring a more hydrophobic unpigmented fibre.
The fact that this was not observed suggests that any kinetic differ-
ences are either small, fast or only, as observed, between 80% and
90% RH.
Conclusion
The age-associated phenomenon of depigmentation (greying) has
been studied in 11 subjects who avoid both sunlight and chemical
treatment of their hair and scalp. Data was analysed for global,
individual and typical differences. This study design provided
greater sensitivity than previous reports by Mathur [11], Hollfelder,
et al. [10], or Wei [9].
Greying occurs when melanin is no longer produced or trans-
ferred to the growing hair shaft either because of a sometimes-
reversible loss of communication of melanocytic processes or the
death of the melanocyte [3, 18, 20]. Greying is one of many
physiological signs of ageing in the follicle [21, 22]. It has been
reported that unpigmented hair is subject to greater damage on
exposure to UV light, because of the obvious absence of melanin
and the critical role it plays in preventing and/or removing free
radicals [10, 11]. The volunteers in this study generally cover
their head and always wore their hair in a tight wrap, protecting
it from the sun, so the differences reported here should be
a result differences in physiology rather than resilience to UV
exposure.
We found evidence for some systematic differences between pig-
mented and unpigmented fibres on individual subjects. These sub-
jects, overall, displayed signs that their grey hairs are coarser [10]
and stiffer than their pigmented hair. We observed possible kinetic
differences in interactions with water. And we found no chemical
or surface chemical differences between these fibres.
In short, it is not unusual to have salt-and-pepper hair where
the salty hair is coarser, stiffer and dry faster than the peppery
hair.
Acknowledgements
The authors greatly acknowledge the technical assistance of Mary
Jo Grove and Trish Alfano (cyberDerm, Inc), as well as J. Qi and K.
Yang (TRI/Princeton). Funding for this research was provided by
Johnson & Johnson Consumer Companies.
181
Clinical study of gray hair fibers
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