Microindentation and nanoindentation of human fingernails at

varying relative humidity

Laura Farran
Materials Science Centre, School of Materials, University of Manchester, Manchester M1 7HS, United Kingdom
A. Roland Ennos
Faculty of Life Sciences, University of Manchester, Manchester, M1 3NJ, United Kingdom
Stephen J. Eichhorna)
Materials Science Centre, School of Materials, University of Manchester, Manchester M1 7HS, United Kingdom

(Received 22 July 2008; accepted 30 October 2008)

The indentation properties of human fingernails at varying humidity are reported. The
samples were indented using both microindentation, to obtain their Vickers hardness and
also nanoindented using a Berkovich indenter tip. The relative humidity (RH) of the
samples was controlled by using salt solutions with a sealed and enclosed environment
surrounding the testing equipment. It was shown that the Vickers hardness of the samples
is sensitive to RH, with recovery of the nail material more readily occurring for nails
tested at >55% RH. This recovery mechanism is discussed in terms of the structure of the
nails, and this approach is also suggested as a technique for following recovery
mechanisms in natural materials under varying humidity. The hardness obtained by
nanoindentation is similar to previously published data, but does not change with
humidity. The modulus of the nails is also insensitive to relative humidity, but in the
same range as the value derived from the microindentation tests.

I. INTRODUCTION than laterally, deflecting cracks laterally away from the

Fingernails are an anatomical feature of primates that
are used for a variety of functions including foraging and
gripping. The mechanics of nails, in particular finger-
nails, have not been studied to a large extent. However,
recent studies have investigated their anatomy and frac-
ture properties. The nail plate comprises three histologi-
cal layers of keratinous tissue that are deposited down by
the nail matrix at the base of the nail unit beneath the
skin.1,2 These layers are arranged in a sandwichlike
structure as shown in Fig. 1. The intermediate layer, in
which the keratin fibers are oriented laterally, makes up
approximately two-thirds of the thickness of the nail. It
is enclosed by the thinner dorsal layer, which makes up
approximately a quarter of the thickness of the nail, and
the even thinner ventral layer that together wrap around
the edge of the intermediate layer. In both of these outer
layers the keratin fibers show no preferred orientation.
The function of the three layers was enlightened by
Farren et al.,3 who carried out cutting tests with instru-
mented scissors. They showed that this design is admira-
bly suited to limit and control fracture. The lateral
orientation of the fibers in the thick intermediate layer
ensured that the work of fracture was greater proximally
a)
Address all correspondence to this author.
e-mail: s.j.eichhorn@manchester.ac.uk
DOI: 10.1557/JMR.2009.0105
nail bed and allowing self-trimming. The outer layers of
the sandwich meanwhile protected the intermediate layer
and prevented cracks forming at its edge. A disadvan-
tage was that the nail layers tend to peel apart at their
boundaries, particularly when wet.3
Little is currently known, however, about other me-
chanical properties of nails, or the effect of humidity on
them. The microindentation of nails was first reported by
Ramrakhiani4 who, using a spherical indenter, measured
their recovery times using an optical technique. Since
then only one report on the indentation, in this case
nanoindentation, of nails has been reported.5 They stud-
ied the mechanical properties of nails by nanoindenta-
tion to detect the onset of osteoporosis and related this to
disulfide content using Raman spectroscopy. In terms of
the indentation of keratinous materials, the first exam-
ples were studies on the bill of a European starling6 and
on Willow Ptarmigan feathers.7 It was subsequently
shown that bill keratin had a higher microhardness (by
the Vickers indentation) than both claw and feather ker-
atin.8 Much later studies of toucan beaks9,10 have shown
that nanoindentation measurements of the stiffness cor-
roborate with tensile testing data on the same material.
Nonavian keratin materials have also been reported in
the literature, namely wool11,12 and human hair.13
The physical properties of nails are commonly known
to change when they become wet, or conversely dry,

980 J. Mater. Res., Vol. 24, No. 3, Mar 2009 © 2009 Materials Research Society
 L. Farran et al.: Microindentation and nanoindentation of human fingernails at varying relative humidity
FIG. 1. Schematic of a human fingernail showing the layered structure.
(a) Dorsal (upper). (b) Intermediate (middle). (c) Ventral (underside).
although this has never been fully quantitatively ana-
lyzed. One study has shown that the number of flexions
that a nail can withstand before fracture, using a custom-
ized flexure device, increased with immersion time in
water.14 This flexibility was also prolonged by immers-
ing the nail samples in a phospholipid-water solution.14
This study presents data on the indentation properties of
nails, examines how the material relaxes back after an
indentation at different relative humidity (RH) levels, and
discusses the possible mechanisms that give rise to these
relaxations. Both micro and nanoindentation tests were
carried out at a range of humidity levels to achieve this.
II. MATERIALS AND METHODS
A. Materials selection
Thirty randomly selected fingernail clippings from
more than 50 healthy young adult human subjects were
taken using nail clippers and then first transferred to plas-
tic bags and subsequently stored in controlled relative
humidity environments after mounting on testing blocks.
B. Microindentation of nail material
Nail samples were sectioned into 3 mm squares and
secured to small Bakelite blocks using cyanocrylate ad-
hesive. These samples were then lightly ground using a
fine-grade grinding paper and finished off using 1 mm
grain size polishing discs to give a flat and smooth sur-
face. Care was taken to ensure that no more than 10% of
the material was ground or polished to prevent the dorsal
and ventral layers being removed by this grinding and
polishing process. Samples were imaged to check that
the dorsal layer was still intact after this process. Sam-
ples were prepared with either the dorsal (upper) or the
ventral (lower) surface uppermost, and stained with tolu-
idine blue to give improved contrast under the light
microscope. These mounted samples were then trans-
ferred to controlled humidity environments.
Saturated salt solutions were used to control humidity
and subsequently the moisture content of the nails,
as has been described for biological samples.15 Small
containers with a range of saturated salt and aqueous
J. Mater. Res., Vol. 24, No. 3, Mar 2009
FIG. 2. (a) Schematic of the pyramidal indenter used for the micro-
hardness experiments and (b) end-on view of the base of the micro-
hardness indenter. (c) Optical microscope view of a typical
microhardness indent on the dorsal surface of a fingernail specimen.
solutions along with a dessicator were used. These
were distilled water, potassium chloride, sodium nitrite,
calcium nitrate, magnesium chloride hexahydrate, and
silica gel, which gave rise to relative humidities of 100,
85, 65, 55, 33, and 0%. The RH values obtained were
tested using a portable hygrometer. Five nail samples
were prepared and tested at each hydration level.
An Instron Wilson Tukon 2100 instrument (Norwood,
MA) with a four-sided pyramidal indenter was used to
perform the indentation experiments. This instrument is
equipped with a standard Vickers microhardness indenter
with pyramidal geometry [see Fig. 2(a)]. A load of 1 N
(100 g) was used; lower loads were found to produce
indents that recovered too quickly to be measured accu-
rately. The indenter was allowed to remain in contact
with the surface for a period of 10 s, and then each
impression was measured immediately after indentation,
after 60 s and then again after 300 s. The sizes of the
indents were measured using an optical microscope
equipped with a 50
objective lens. To calculate the
hardness (H) the equation
L
H ¼ 2 sinð136 =2Þ ; ð1Þ
dH 2
was used, where dH is the diagonal length of the indent
[see Fig. 2(b)] and L is the load. To maintain the desired
relative humidity during testing, the indenter and
stage were enclosed with clear plastic sheeting and the
saturated salt solutions were placed below the stage.
A portable hygrometer was used to verify the relative
humidity during testing.
981
 L. Farran et al.: Microindentation and nanoindentation of human fingernails at varying relative humidity

C. Nanoindentation of nail material Vickers microhardness values decrease with increasing

Fingernail samples were sectioned and prepared using
the same methods used for the microhardness indentation
experiments. These samples were then attached to stain-
less steel blocks using superglue. A NanoIndenter XP
with a three-sided Berkovich indenter (Agilent, Berkshire,
UK) was used to perform instrumented indentation testing.
The indenter is of a different geometry than the Vickers
indenter used for microhardness indentation, and the equa-
tions used to calculate the mechanical properties take this
into account. To calculate the hardness (Hn) the equation
L The apparent Vickers microhardness number decreases
Hn ¼ ; ð2Þ
A with increasing relative humidity from 28.5 to 4.3
where L is the load applied and A is the projected contact
area at the point of loading was used. To determine
stiffness the approach of Oliver and Pharr was used,
where the gradient of the unloading curve is deter-
mined.16 A series of 25 indentations were made per
specimen to acquire average values. Humidity was con-
trolled by placing a container filled with the same salt
solutions used for microindentation beneath the apparatus
and was checked using a hygrometer.
III. RESULTS
A. Microindentation of nail material
An optical image of a typical indent obtained from
microindentation tests is shown in Fig. 2(c). This partic-
ular image was taken at 0% RH and was photographed
less than 5 min after indentation. The diagonal lines
on the image are artefacts from the polishing process.
The hardness of the dorsal (upper) and ventral (bottom)
layers of the nail are presented in Fig. 3(a). These data
show no significant differences between the two layers
at ambient relative humidity [F(2,9) = 1.86, P = 0.124].
Data obtained from Vickers microindentation at dif-
ferent RHs are shown in Fig. 3(b). These show that the
RH and increase with increasing time elapsed, postin-
dentation. Microhardness values ranged from 4 kg/
mm2 (0.04 GPa) to around 30 kg/mm2 (0.29 GPa). There
was a significant difference between mean Vickers
microhardness numbers at all relative humidities [at 0 s
F(5,24) = 285.81, P < 0.001, at 60 s F(5,24) = 551.96, P <
0.001, at 300 s F(5,24) = 1055.1, P < 0.001]. Tukey tests
showed that the values for microhardness at all humidity
levels were significantly different after 300 s. Differ-
ences between all humidity levels, except 55% and 65%
RH, were significant at both 0 s and 60 s postindentation.
kg/mm2 (0.28–0.04 GPa) if the impression is measured
immediately, from 29.0 to 9.6 kg/mm2 (0.28–0.09 GPa)
after 60 s postindentation, and from 29.4 to 14.6 kg/mm2
(0.29–0.14 GPa) after 300 s postindentation.
The results of repeated measured ANOVA with Bon-
ferroni post-hoc tests showed that there was a significant
difference in Vickers microhardness over time in sam-
ples tested at 55 to 100% RH. Time-elapsed postindenta-
tion did not have a significant effect on the
microhardness values obtained for samples at 0% and
33% RH.
B. Nanoindentation of nail material
A typical load–displacement trace for a finger-
nail sample undergoing nanoindentation is shown in
Fig. 4(a). Modulus and hardness values were calcu-
lated using the continual stiffness measurement (CSM)
approach where these data are determined as a func-
tion of depth; both modulus and hardness were found
not to vary considerably over the depth ranges inves-
tigated (200–1000 nm).
Average Young’s modulus for the fingernail samples,
as derived by nanoindentation, was found to be 4.62
GPa [0.41 GPa, standard deviation (SD*)]. The data

FIG. 3. (a) Vickers microhardness of fingernails as determined from the indentation of the dorsal and ventral layers of nail specimens at ambient
relative humidity. (b) Vickers microhardness values of fingernails at various humidity levels with measurements taken at 0, 60, and 300 s
postindentation. Groups denoted by the same character are not significantly different from each other with respect to the RH (P < 0.05) according
to Tukey’s post-hoc tests. Data labeled with an * are significantly different from one another with respect to time with RH groups according to
Bonferroni post-hoc tests. All errors in (a) and (b) are standard errors from the mean. The scales on the right were determined by multiplying the
Vickers hardness number by 0.009807.

982 J. Mater. Res., Vol. 24, No. 3, Mar 2009

 L. Farran et al.: Microindentation and nanoindentation of human fingernails at varying relative humidity

FIG. 4. (a) A typical load–displacement trace for nanoindentation performed on the dorsal layer of a human fingernail. (b) Modulus of finger-
nails, as derived by nanoindentation, as a function of relative humidity. (c) Hardness of fingernails, as derived by nanoindentation, as a function of
relative humidity. All errors in (b) and (c) are standard errors from the mean.

obtained over the range of relative humidities are shown
in Fig. 4(b). As can be seen, the hardness does not vary
significantly over the relative humidity range. On aver-
age the hardness of nails, determined by nanoindenta-
tion, is 0.18 GPa (0.05 GPa, SD) and did not vary
significantly over the humidity range [see Fig. 4(c)].
Values obtained at 65 and 85% RH appear lower than
for any other humidity, but due to the high variability
and the small sample sizes there was no significant dif-
ference in hardness at the different relative humidities
[F(5,45) = 1.95, P < 0.12].
IV. DISCUSSION
Vickers microhardness and nanoindentation tests
proved to be useful techniques for measuring the proper-
ties of fingernails. Under dry conditions the fingernail
material, which is primarily keratin, remains plastically
deformed. For this type of response it is easier to derive
a hardness value that can be compared with other mate-
rials. The dorsal (upper) and ventral (lower) layers were
found to have similar hardness values at ambient humid-
ity. The dorsal layer is likely to be more susceptible to
changes in the environment in vivo. For this reason all
subsequent indentation tests were performed on the up-
per or dorsal layer.
When the samples were placed in humidity-controlled
environments, then indentations into the material were
found to relax, particularly above 55% RH. Although it
is not possible to derive a comparative hardness value at
this point, the values obtained reflect the ability of the
material to recover the plastic deformation when mois-
ture is present. This recovery may be the result of plasti-
cization of the matrix material. Keratinous materials,
such as hair, have been postulated to comprise smaller
fibrillar units embedded in an amorphous rubbery matrix
of the same keratinous material.17 When moisture enters
the nail structure it is possible that the disulfide bridges

J. Mater. Res., Vol. 24, No. 3, Mar 2009 983

 L. Farran et al.: Microindentation and nanoindentation of human fingernails at varying relative humidity
between molecular chains will be disrupted, enabling
them to become more entangled. This entanglement
could be a source of the recovery mechanism via an
increase in entropy. This indentation approach is there-
fore useful for understanding the properties of natural
materials and their recovery mechanisms under a chang-
ing environment.
Taking the values of Vickers microhardness at ambi-
ent humidity after 60 s, the results are similar to, if
slightly higher than, values obtained by Bonser7 for
feather keratin of around 10 to 14 kg/mm2 (0.10 to 0.14
GPa), and close to the values obtained for bill keratin of
12 to 20 kg/mm2 (0.12 to 0.20 GPa).6 The value of the
load used for this study (100 g) was, however, much
higher than the load used by Bonser6,7 (5 g), which could
explain why our data are slightly higher in magnitude.
Ramrakhiani4 found a hardness value of 8 kg/mm2 (0.08
GPa) for nails tested using a load of 500 g, which is
slightly lower than our values.
Nanoindentation tests also gave comparable results to
previous tests and to the results of the microindentation
tests. The value obtained for hardness of 0.18 GPa is
similar to a value recently reported for healthy nails of
0.23 GPa.5 When the hardness of the nails at ambient
humidity, as derived by microindentation, is converted to
the same units a similar value of 0.20 GPa is also obtained.
The main contrast between the results of the microin-
dentation and nanoindentation tests is that the stiffness
and hardness measured by nanoindentation does not
change with relative humidity, whereas that measured
by microindentation falls, especially at the highest hu-
midity levels. Surface roughness may have played a role
in this since the indents are small (<1 mm), but the
results may be explicable in terms of the structure of the
nail. Since the indents are so small, the tests will only
affect the dorsal layer, and it is possible that the proper-
ties of this layer are unaffected by humidity. Certainly
such a property would be useful in a protective layer that
has to withstand scratching and chipping and might be
expected in a layer that is composed of flat pancakelike
cells with closely packed orthogonally arranged fibers.3
In contrast, the deformation due to microindentation is
likely to penetrate to the more open fibrous intermediate
layer, which is likely to give more in response to stress,
particularly at high relative humidity. Unfortunately, it
was not possible to test the indentation properties of the
naked intermediate layer because it proved impossible to
polish it to a flat surface, and this remains a topic for
future research.
984 J. Mater. Res., Vol. 24, No. 3, Mar 2009
ACKNOWLEDGMENTS
The authors wish to thank the EPSRC DTA (Doctoral
Training Account) and the University of Manchester for
funding a Ph.D. scholarship and Mr. Andrew Forrest for
assistance with the nanoindentation experiments.
REFERENCES
1. G. Achten: Histpathology of the nail, in The Nail, edited by
M. Pierre (Churchill Livingstone, 1981), p. 1.
2. R. Caputo, G. Gasparini, and D. Contini: A freeze-fracture
study of the human nail plate. Arch. Dermatol. Res. 272, 117
(1982).
3. L. Farren, S. Shayler, and A.R. Ennos: The fracture properties
and mechanical design of human fingernails. J. Exp. Biol. 207,
735 (2004).
4. M. Ramrakhiani: Indentation and hardness studies of human
nails. Indian J. Biochem. Biophys. 15, 341 (1978).
5. P. Moran, M.R. Towler, S. Chowdhury, J. Saunders,
M.J. German, N.S. Lawson, H.M. Pollock, I. Pillay, and
D. Lyons: Preliminary work on the development of a novel detec-
tion method for osteoporosis. J. Mater. Sci. Mater. Med. 18, 969
(2007).
6. R.H.C. Bonser and M.S. Witter: Indentation hardness of the bill
keratin of the European starling. Condor 95, 736 (1993).
7. R.H.C. Bonser: Melanin and the abrasion resistance of feathers.
Condor 97, 590 (1995).
8. R.H.C. Bonser: Comparative mechanics of bill, claw, and feather
keratin in the common starling Sturnus vulgaris. J. Avian Biol. 27,
175 (1996).
9. Y. Seki, M.S. Schneider, and M.A. Meyers: Structure and
mechanical behavior of a toucan beak. Acta Mater. 53, 5281
(2005).
10. Y. Seki, B. Kad, D. Benson, and M.A. Meyers: The toucan beak:
Structure and mechanical response. Mater. Sci. Eng., C 26, 1412
(2006).
11. A.N. Parbhu, W.G. Bryson, and R. Lal: Disulfide bonds in the
outer layer of keratin fibers confer higher mechanical rigidity:
Correlative nano-indentation and elasticity measurement with an
AFM. Biochem. 38, 11755 (1999).
12. M. Huson, J. Church, and G. Heintze: Spectroscopy, microscopy
and thermal analysis of the bi-modal melting of Merino wool.
Wool Technol. Sheep Breeding 50, 64 (2002).
13. G.H. Wei, B. Bhushan, and P.M. Torgerson: Nanomechanical
characterization of human hair using nanoindentation and SEM.
Ultramicroscopy 105, 248 (2005).
14. A.Y. Finlay, P. Frost, A.D. Keith, and W. Snipes: An assessment
of factors influencing flexibility of human fingernails. Br. J.
Dermatol. 103, 357 (1980).
15. P.W. Winston and D.H. Bates: Saturated solutions for the control
of humidity in biological-research. Ecol. 41, 232 (1960).
16. W.C. Oliver and G.M. Pharr: An improved technique for deter-
mining hardness and elastic-modulus using load and displace-
ment sensing indentation experiments. J. Mater. Res. 7, 1564
(1992).
17. J.W.S. Hearle: A critical review of the structural mechanics of
wool and hair fibers. Int. J. Biol. Macromol. 27, 123 (2000).