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ORIGINAL ARTICLE

Barrier Function and Water-Holding and Transport


Properties of Infant Stratum Corneum Are Different
from Adult and Continue to Develop through the First
Year of Life
Janeta Nikolovski1, Georgios N. Stamatas2, Nikiforos Kollias2 and Benjamin C. Wiegand1

Skin water barrier development begins in utero and is believed to be complete by week 34 of gestational age.
The goal of this investigation was to assess the dynamic transport and distribution of water of the stratum
corneum of infants and compare it to those of adults. The interaction of water with the stratum corneum was
assessed by measuring capacitance, transepidermal water loss (TEWL), rates of absorption–desorption as well as
Raman spectra as a function of depth (a total of 124 infants (3–12 months) and 104 adults (14–73 years)). The
results show that capacitance, TEWL, and absorption–desorption rates had larger values consistently for infant
stratum corneum throughout the first year of life and showed greater variation than those of adults. The Raman
spectra analyzed for water and for the components of natural moisturizing factor (NMF) showed the
distribution of water to be higher and have a steeper gradient in infants than in adults; the concentration of
NMF was significantly lower in infants. The results suggest that although the stratum corneum of infants may
appear intact shortly after birth (o1 month), the way it stores and transports water becomes adult-like only after
the first year of life.
Journal of Investigative Dermatology (2008) 128, 1728–1736; doi:10.1038/sj.jid.5701239; published online 17 January 2008

INTRODUCTION newborns (Harpin and Rutter, 1983; Evans and Rutter, 1986;
Human skin barrier development begins in utero during the Saijo and Tagami, 1991; Hoath, 1997; Kalia et al., 1998;
first trimester with stratification of the epidermis (Holbrook, Cartlidge, 2000; Yosipovitch et al., 2000; Kikuchi et al.,
1982; Cartlidge, 2000). The formation of vernix in the third 2006). Infants born prematurely (less than 34-week gesta-
trimester is thought to contribute to the final steps of barrier tional age) exhibit underdeveloped skin barrier function, and
maturation (Visscher et al., 2005). Epidermal cell maturation much research has focused on determining the effect of
occurs continually during this entire process, whereas the gestational age on barrier development (Rutter and Hull,
stratum corneum (SC) as well as the dermo-epidermal 1979; Wilson and Maibach, 1980; Lund et al., 1997; Kalia
undulations become visible at 34-week gestational age. It is et al., 1998). Given the dramatic transition from an aqueous
believed that at this point barrier maturation is near complete to a dry terrestrial environment at birth, studies have dealt
and the fetal epidermis begins to function as a barrier (Kalia with skin barrier adaptation within the first few days or the
et al., 1998; Segre, 2003; Mancini, 2004). It is the mature first month of life (Harpin and Rutter, 1983; Visscher et al.,
semipermeable SC that provides a protective epidermal 2000; Yosipovitch et al., 2000; Hernes et al., 2002; Hoeger
barrier, allowing for terrestrial life (Menon, 2002). and Enzmann, 2002). There is no clear consensus, however,
Much clinical evidence exists stressing the importance of about the state of the SC barrier in infants after the first month
the SC and its barrier function for infants and especially for of life (Chiou and Blume-Peytavi, 2004).
Reports have also varied on the point at which infants
1 acquire an adult-like SC. Based on transepidermal water loss
Advanced Technologies, Johnson & Johnson Consumer and Personal
Products Worldwide, Skillman, New Jersey, USA and 2Models and Methods, (TEWL) and percutaneous water absorption studies, SC
Johnson & Johnson Consumer and Personal Products Worldwide, Skillman, maturity has been reported to occur anywhere from 30 to
New Jersey, USA 37 weeks (Rutter and Hull, 1979; Harpin and Rutter, 1983;
Correspondence: Dr Benjamin C. Wiegand, Johnson & Johnson Consumer Kalia et al., 1998). Others believe that even for term infants
and Personal Products Worldwide, 199 Grandview Road, SB226, Skillman,
New Jersey 08558, USA. E-mail: bwiegan@cpcus.jnj.com
the SC water holding and water transport properties are in a
state of flux in contrast to adults (Holbrook, 2000; Visscher
Abbreviations: NMF, natural moisturizing factor; SC, stratum corneum; SD,
standard deviation; TEWL, transepidermal water loss et al., 2000). Some investigators have reported TEWL values
Received 19 April 2007; revised 5 September 2007; accepted 7 November of term infants to be at adult levels and concluded that term
2007; published online 17 January 2008 infants are born with a functionally mature SC (Rutter,

1728 Journal of Investigative Dermatology (2008), Volume 128 & 2008 The Society for Investigative Dermatology
J Nikolovski et al.
Infant Skin Barrier Maturation

2000b). SC hydration measured by skin conductance seems pulled from five independent studies as explained in the
to consistently show reduced levels upon birth and elevated Materials and Methods section. A post hoc analysis has
levels thereafter (Visscher et al., 2000; Yosipovitch et al., shown that in all cases where infant data are compared to
2000; Giusti et al., 2001; Hoeger and Enzmann, 2002). adult data, the calculated statistical power exceeds 0.95,
Despite reports of normal basal barrier function at birth, except where noted otherwise.
full-term infant skin is known to exhibit a greater tendency to
develop irritant/allergic contact dermatitis, as well as to be Static (steady-state) measurements
prone to higher percutaneous absorption, prompting some to Skin conductance is higher in infants. Skin hydration was
suggest that barrier function is not fully developed at birth evaluated using skin conductance measurements (NOVA
(Rutter, 2000a; Behne et al., 2003). Natural moisturizing DPM meter) on the upper ventral (inner) arms and lower
factors (NMFs) in the SC are known to be involved in barrier dorsal (outer) arms. Skin conductance values as a function of
function and serve as efficient humectants. NMF concentra- age for the upper ventral arm site are shown in Figure 1a.
tion has been shown to decline with age and to be impacted Infant skin displays higher conductance values than adult
by skin disease and environmental damage (Rawlings and skin. Similar results were found for the lower dorsal arm area
Harding, 2004). (data not shown). The data were grouped by age (3–6 months,
The goal of this study was to investigate the water storing 7–12 months, 13–48 months, and adults). Conductance
and water transport properties of the SC of infants in the first values of infant skin (3–12 months) were significantly higher
year of life with traditional tools, such as TEWL, as well as when compared to adult skin (Figure 1b and c). There were
skin capacitance, absorption–desorption, and Raman con- no significant differences between infants aged 3–6 months
focal spectroscopy, and to compare these to the properties of and those aged 7–12 months. Linear regression analysis of the
adult SC. In the rest of the document, we will use the entire data set showed age to be a significant factor
expression ‘‘water-handling properties’’ to signify the water contributing to hydration values and accounting for 44% of
transport and water storing properties of the SC. the variation.
A large intersubject variability was found in the skin con-
RESULTS ductance measurements of the infant population (Figure 1a).
The water-handling properties of infant and adult skin were The coefficient of variance for all of the infant groups
measured using both a static and a dynamic approach. Static was 2–5 times higher compared to that for the adult group
or steady-state measurements relate to the presence and for the lower dorsal site (Table 1). Similarly, the coefficient
distribution of water within the SC, whereas the dynamic of variance for the conductance measurements on the
measurements refer to the uptake and release of water from upper ventral arm site was higher for the 3- to 6- and 7- to
the SC. Note that the data presented are for a population 12-month-old groups than the older groups.

500
450
400
(NOVA DPM, a.u.)

350
Conductance

300
250
200
150
100
50

0 10 20 30 40 50 20 40 60 80
Age (months) Age (years)


300 300 ∗
(NOVA DPM, a.u.)

(NOVA DPM, a.u.)

250 250
Conductance

Conductance

200 200

150 150

100 100

50 50

0 0
3–6 7–12 13–48 Adult 3–6 7–12 13–48 Adult
months months months months months months

Figure 1. Skin conductance is higher in infants. (a) Age distribution of skin conductance measurements (a.u. ¼ arbitrary units) on the ventral side of the
upper arm. (b) Data taken from the upper ventral arm were grouped by age: 3–6 months (N ¼ 21), 7–12 months (N ¼ 46), 13–48 months (N ¼ 17), and
adults (N ¼ 71). Skin conductance values of the upper ventral arm decrease after the first 12 months of life (*Po0.0005, 3–12 months vs adult). (c) Data taken
from the lower dorsal arm were averaged similarly. Skin conductance values of the lower dorsal arm decrease after the first 12 months (*Po0.0005,
3–12 months vs adult). Data shown as mean±SD.

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J Nikolovski et al.
Infant Skin Barrier Maturation

SC water content is higher in infants. Skin hydration was found to have both a steeper water gradient in the SC (the
assessed by in vivo Raman confocal spectroscopy. The gradient was calculated as the slope of the linear portion of
confocal arrangement allows the acquisition of Raman the concentration profile, 4–14 mm) compared to adults and a
spectra as a function of depth into the skin with a lateral higher water content within the first 20 mm from the surface of
resolution of 1 mm (x and y directions) and a depth resolution the SC (Figure 2b and c). Note that in the case of Figure 2b,
of 5 mm (z direction). The spectra were then analyzed for a post hoc analysis showed that the statistical power for the
water content, and water concentration profiles were comparison between infant and adult data is 0.831. The
calculated as a function of depth into the skin. Although strength of the conclusion for a difference in the water
water profiles have been reported from adult subjects profiles, however, is supported by the data in Figure 2c,
(Caspers et al., 2001; Hellemans et al., 2005), this method where the statistical power is 0.988.
was applied for the first time on infant skin in this study.
Measurements on infant skin revealed differences in water Dynamic measurements
distribution in the SC as compared to adults (Figure 2a). The Endogenous water loss through the skin is higher in infants.
infant group showed a higher amount of water on the skin Measurements of TEWL on the upper ventral arm as a function
surface (0 mm depth), as well as within the SC and throughout of age are shown in Figure 3a. Similar results were obtained
the first 26 mm from the skin surface. No significant for the lower dorsal arm area (data not shown). Infant skin
differences in water concentration as a function of depth appears to lose water at higher rates than adult skin. The
were found between infants of less than 1 year of age average values of measurements of TEWL for all infant groups
compared to infants older than 1 year of age. Infants were and for both skin sites investigated were significantly higher
than for adults (Figure 3b and c). No significant difference was
found between the infant groups of 3–6 and 7–12 months for
Table 1. Coefficients of variance for the skin the upper ventral arm site, where a decreasing trend was seen
conductance measurements for the lower dorsal arm site vs age.
Coefficient of variance Upper ventral Lower dorsal N A large intersubject variability in TEWL measurements was
found among the infant subjects (Figure 3a). Calculation of
3–6 months 0.53 0.38 21
coefficients of variance for each age group revealed higher
7–12 months 0.56 0.50 46 values of variance in the younger group (3–6 months old)
13–48 months 0.19 0.55 30 compared to either the 7- to 12-month-old or the adult group
Adult 0.09 0.08 71
(Table 2). The coefficient of variance of all infant TEWL
measurements from the upper ventral arm site was higher
than that of adults.

80
Water concentration (mass %)

Infant

∗∗ ∗ ∗
70 Adult
∗∗
60 ∗

50 ∗

40 ∗
∗∗
30
20
10

0 10 20 30 40
Depth (μm)

3 Infant 1,200 Infant


∗ Adult Adult
2.5 1,000 ∗
(slope from 4 –14 mm)

AUC (0 –20 mm)


Water gradient

2 800

1.5 600

1 400

0.5 200

0 0

Figure 2. Water content of the infant SC is higher than that of the adult SC. (a) The average distribution of water on the lower ventral arms of infants
(aged 3–33 months, N ¼ 13) and adults (N ¼ 13) was measured within the top 40 mm of the skin by Raman confocal microspectroscopy. Infant SC contains more
water than adult SC (*Po0.05 up through the first 26 mm). Data shown as mean±SD. (b) The water distribution within infant SC demonstrates a steeper
slope (at depths of 4–14 mm). Data shown as mean±SD, *Po0.005. (c) Infant SC contains higher concentration of water within the top 20 mm than
adult SC, calculated by the area under the curve (AUC). Data shown as mean±SD, *Po0.0005.

1730 Journal of Investigative Dermatology (2008), Volume 128


J Nikolovski et al.
Infant Skin Barrier Maturation

60

50

TEWL (g m −2 hour −1)


40

30

20

10

0 2 4 6 8 10 12 14 10 20 30 40 50 60 70 80
Age (months) Age (years)

∗ ∗
45 45 ∗
40 40
TEWL (g m −2 hour −1)

TEWL (g m −2 hour −1)


35 35
30 30
25 25
20 20
15 15
10 10
5 5
0 0
3–6 7–12 Adults 3–6 7–12 Adults
months months months months

Figure 3. TEWL values are higher in infants. (a) TEWL from the upper ventral arm for each subject was measured and plotted against age. (b) Data taken
from the upper ventral arm were averaged for infants ranging in age from 3 to 6 months (N ¼ 19), 7 to 12 months (N ¼ 31), and adults (N ¼ 71). TEWL values from
the upper ventral arm are higher for the first 12 months of life compared to adult (*Po0.0005 for both age ranges). (c) Data taken from the lower dorsal
arm were averaged similarly. TEWL values from the lower dorsal arm are higher in the youngest subjects (*Po0.01, 3–6 vs 7–12 months) and higher among
all infants tested than that of adults (*Po0.0005, 3–6, 7–12 months vs adult). Data shown as mean±SD.

Table 2. Coefficients of variance for the TEWL The shape of the desorption curves implies exponential
measurements kinetics. The adult data can be fit with a single exponential,
Coefficient of variance Upper ventral Lower dorsal N
but for the infant data the initial rapid decline imposes a
better fit with a double exponential function. Infant skin
3–6 months 0.40 0.59 19 appears to absorb a greater amount of water than adult skin
7–12 months 0.43 0.33 31 and it also gives it up within the first 45 seconds after blotting;
Adult 0.37 0.38 71 thereafter, it behaves similarly to adult skin. The time
constant for the water desorption in adults was
TEWL, transepidermal water loss.
t ¼ 30 seconds. For the infants, the two time constants were
t1 ¼ 12 seconds and t2 ¼ 30 seconds corresponding to an
initial rapid decay and a secondary slower phase.
Water absorption into the SC was also investigated with
Exogenous water absorption and desorption were higher in in vivo Raman confocal microspectroscopy. Measurements of
infants. Water absorption and desorption were measured on the water concentration throughout the epidermis were taken
the lower arm of both infants and adults. The changes in skin before and after a 10-second application of water to the SC
hydration due to the addition of water and its subsequent (Figure 5). This maneuver resulted in significantly higher
evaporation/dissipation were monitored with measurements amounts of water in the top layers of the SC of infants aged
of skin conductance. Conductance measurements at each 3–12 months and had no effect on the water profiles
time point before and after the addition of a drop of water measured in adult skin (Figure 5a and b). The changes in
onto SC of the lower dorsal arm were taken and then water concentration following this maneuver in infant skin
averaged among infants (ages 3–12 months) and then adults. were found to be significant within the first 8 mm of the SC
The measured values of conductance were normalized to the (Po0.05 before vs after).
initial baseline readings (Figure 4a). The skin of infants was
found to absorb water at a faster rate compared to adults. NMF concentration is lower in infants
Similarly, the initial water desorption rate was higher for the Water in the SC may be taken up by corneocytes and/or by-
infant population (Figure 4b), and it approximated the rate of products of protein degradation called natural moisturizing
adults for times longer than 45 seconds. factors. In vivo Raman confocal microspectroscopy was used

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J Nikolovski et al.
Infant Skin Barrier Maturation

to measure NMF concentration profiles through the skin as a months) show significantly lower amounts of NMF in the first
function of depth following the method of Caspers et al. 12 mm of the SC surface than that from adults (Figure 6).
(2001) (Hellemans et al., 2005). Profiles from infants (3–12
DISCUSSION
500
The goal of this study was to investigate the water-handling
properties of the SC of infants in the first year of life with
traditional tools, such as TEWL, as well as skin capacitance,
Change in conductance (a.u.)

400
absorption–desorption, and Raman confocal spectroscopy,
300 Infant and to compare these to the properties of adult SC. This

Adult
information on water handling in the SC might be of value in
200
enhancing our understanding of the barrier function of the SC
and its maturation postpartum.
100
SC hydration has been reported to be low at birth in term
0 neonates and to increase with increasing post-natal age
0 30 60 90 120
(Visscher et al., 2000; Yosipovitch et al., 2000; Giusti et al.,
−100 2001; Hoeger and Enzmann, 2002; Behne et al., 2003). The
Time (seconds)
duration of this increase in SC hydration ranges from 2 weeks
20
after birth (Visscher et al., 2000) to 30–90 days after birth
(Hoeger and Enzmann, 2002). The results of this investigation
15 indicate that the SC of infants (3–12 months of age) is
Infant
significantly more hydrated than adult SC. This conclusion is
Rate of change of conductance

∗ Adult
10 supported by both measurements of skin conductance and
water distribution using Raman confocal microspectroscopy.
5 It should be noted that hydration measurements using
Desorption
(30–60 seconds) skin conductance might be confounded when comparing
0 skin sites of unequal thicknesses. The SC of infants has been
Absorption reported to be thinner than that of adults and this was also
(0– 30 seconds)
−5 confirmed in vivo in our experiments (data not shown).
∗ However, measurements of water content with increasing
−10 depth into the skin using Raman confocal microspectroscopy
confirm that the epidermis of an infant contains significantly
−15 more water on the surface, as well as throughout the first
Figure 4. Exogenous water absorption and desorption rates are higher in 26 mm, than adult epidermis. The greatest difference between
infants. The absorption and subsequent desorption of exogenously applied the infant and adult epidermal water concentration was seen
water was monitored by measuring skin conductance before and 10 seconds between 10 and 14 mm from the surface (Figure 2).
after water application to the skin of the lower dorsal arm. (a) Infant skin Infant SC exhibited a significantly steeper water gradient
(N ¼ 88) shows a higher change in conductance values from baseline than
and higher total water amount than adult SC. This may be
adult skin (N ¼ 97; *Po0.001 at 30 seconds). (b) Rates of change were
calculated and averaged from initial to maximum value (rate of absorption) attributed in part to a thinner SC in infants. These findings are
and from maximum to the value at 60 seconds (rate of desorption). Infant skin particularly noteworthy considering the biological signifi-
demonstrates a higher rate of water absorption (*Po0.002) and desorption cance and implication of this water gradient on SC
(*Po0.0001) than adult skin. Data shown as mean±SD. maturation and on cellular differentiation in infants. Skin

80 Infants 80 Adults
Water concentration (mass %)
Water concentration (mass %)

70 70
60 60
∗ Before Before
50 ∗∗ ∗ After 50 After

40 40
30 30
20 20
10 10

0 10 20 30 40 0 5 10 15 20 25 30 35 40
Depth (μm) Depth (μm)

Figure 5. Water absorption profiles. The absorption of exogenously applied water was monitored by in vivo Raman confocal microspectroscopy
10 seconds after water application to the skin of the lower ventral arm. Data were averaged before and after water application. (a) A significant amount of water
absorption was found within the SC of infants less than 12 months old (N ¼ 5; *Po0.05 before vs after for first 8 mm). (b) In contrast, no significant water
absorption was found in adult skin (N ¼ 7) after water application. Data shown as mean±SD.

1732 Journal of Investigative Dermatology (2008), Volume 128


J Nikolovski et al.
Infant Skin Barrier Maturation

2 60
∗ Infant
∗ Infant
1.8 ∗ Adult
Adult
∗ 50
NMF concentration (mass %)

1.6
∗ ∗

TEWL (g m −2 hour −1)


1.4
40
1.2
1 30
0.8
20
0.6
0.4
10
0.2
0
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 0 200 400 600
Depth (μm) Conductance (NOVA DPM, a.u.)

Figure 6. NMF concentration is lower in infants. The average distribution of Figure 7. A combination graph (TEWL vs conductance) demonstrates the
NMF on the lower ventral arms of infants (aged 3–12 months, N ¼ 8) and differences in water-handling properties of infant SC vs adult SC. TEWL and
adults (N ¼ 15) was measured within the top 28 mm of the skin. Infants have conductance values from the upper ventral arm for each subject were
less NMF in the SC and upper epidermis of their arms as compared to adults measured and plotted against each other. The graph demonstrates the high
(*Po0.05 up through the first 12 mm). Data shown as mean±SD. variability of the state of the infant SC (being in a ‘‘state of flux’’) compared to
the well-established water-handling control mechanisms of adult SC.

surface morphology, the desquamation process, and epider- Chilcott et al., 2002). These factors were controlled in this
mal expression of keratins and other proteins are influenced study by performing measurements in a temperature- and
by the amount and distribution of water in the SC (Rawlings humidity-controlled room, allowing an acclimation period to
et al., 1994; Pierard et al., 2000; Sato et al., 2000; Bouwstra stabilize subjects before measurements were performed, and
et al., 2003; Fluhr et al., 2004; Rawlings and Matts, 2005). refraining from taking data from visibly distressed or crying
Measurements of TEWL have been used extensively as an infants. Despite these efforts, a relatively high amount of
indicator of barrier function. Although it is generally variability and high TEWL values were seen within data sets
appreciated that gestational age has a profound effect on (Tables 1 and 2). We believe that the high TEWL values shown
TEWL, the values reported for infants vary (Harpin and Rutter, for some subjects may be indicative of a mildly stressed
1983; Saijo and Tagami, 1991; Lund et al., 1997; Kalia et al., subject rather than suggestive of a sustained high water flux.
1998; Yosipovitch et al., 2000; Visscher et al., 2002; Agren However, the data presented here highlight a very interesting
and Sedin, 2006). These inconsistencies may be attributed to and consistently apparent observation that infant skin does not
differences in environmental acclimation times, number and seem to exist at a constant, steady state regarding its water
age range of subjects, body site studied, or the type of holding properties. The TEWL data were further plotted vs the
instrument used to measure TEWL (that is, closed vs open conductance values measured at the corresponding sites
chamber techniques). Studies in which open chamber (Figure 7). This figure demonstrates both the intersubject
instruments were used to assess TEWL for term infant skin variability in infants as well as the tight clustering of adult data
have reported TEWL values that were lower than (lower left part of the figure corresponding to lower values of
10 g m2 hour1, which are lower than or equal to those TEWL and skin conductance).
reported from adults (Harpin and Rutter, 1983; Saijo and The conclusions drawn based on the measurements of
Tagami, 1991; Kalia et al., 1998). On the basis of those data, TEWL were confirmed by the water absorption–desorption
it was concluded that term infants are born with a studies. Water absorption and desorption studies have been
functionally mature SC. Higher TEWL values have also been performed to monitor the rate at which water is taken up by
reported from term infants (Lund et al., 1997; Yosipovitch the superficial layers of the SC and then released (Tagami
et al., 2000). TEWL measurements performed in this study et al., 1982; Segre, 2003; Fluhr et al., 2004; Elias, 2005).
show significant differences between the values obtained Infant skin was found to absorb significantly more water than
from infants less than 1 year of age and those obtained from adult skin, and the initial rate of desorption was higher for
adults. The average TEWL values reported in this study for infants than for adults; at later time points, the two responses
infants range from 15 to 30 g m2 hour1, consistent with were identical. Interestingly, water desorption was found to
some published reports (Lund et al., 1997; Yosipovitch et al., follow a different process in infant skin as demonstrated by
2000). These data may support the notion that infant skin two distinct rate constants. This implies that a different
barrier is not the same as adults and continues to develop mechanism of water desorption may exist in infant skin that is
during the first year of life. absent or present to a lesser degree later in life. Whereas in
Measurements of TEWL can often be confounded by adult skin water desorption seems to follow a single process,
factors such as environment (that is, ambient humidity), sweat possibly a relatively slower diffusion through the tissue, water
gland activity, and subject stress (Wilson and Maibach, 1982; desorption from infant skin seems to occur first via a distinct

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J Nikolovski et al.
Infant Skin Barrier Maturation

rapid process followed by the slower diffusive process similar et al., 2000). On the basis of the results of this study, the
to that seen in adults. These conclusions depend strongly on stabilization or adaptation/maturation period may extend
the conductance values corresponding to the maximum beyond the first year of life. In contrast to the relative steady-
amount of water absorbed. The difference between adult and state nature of adult skin, infant skin barrier should not
infant values at that time point is statistically significant at be thought of as deficient but may exist in a state of
confidence levels of Po0.001. It can be speculated that optimization, balancing growth, thermoregulation, water
water desorption through the superficial layers of the SC in barrier, and protective functions. This adaptive flexibility
infants is giving rise to this rapid process, indicating once can be attributed to the higher rate of skin growth in infants
more the different nature of the infant SC water holding and is evidenced further by the age-related decline in the
properties. ability to restore barrier during adulthood.
The phenomenon of water absorption was also investi- On the basis of these results, we conclude that the
gated with Raman confocal microspectroscopy. Spectra were properties that make infant skin unique continue to persist at
taken from specific skin sites before and after a 10-second least through the first 12 months of life. Infant barrier function
application of water onto the skin. A significant increase in and water-handling properties of SC continue to be in a state
water content in the first 8 mm of the SC was found in infants of flux until more stable regulation mechanisms are deve-
after application of water, whereas no change was measured loped later in life.
in adult skin during the same experiment. No differences
were seen in water absorption using this method with infants MATERIALS AND METHODS
older than 1 year of age (data not shown), although further Clinical protocol
experiments need to be performed with more subjects to Five independent studies were performed on healthy Caucasian
better elucidate the transition point in age. male and female infants and female adults without any history of
The water barrier function of skin is localized in the SC skin disorders. Only healthy infants were recruited. In terms of atopic
(Norlen, 2001; Bouwstra et al., 2003). Water has been shown dermatitis, this corresponds to ‘‘no eruptions,’’ Hanifin scale of 0
to be inhomogeneously distributed within adult hydrated SC, (Hanifin et al., 2001). The adult population was with Fitzpatrick skin
where it concentrates within corneocytes and lacunae. The types I–III and the infant population was fair complexioned and
data presented in this study suggest the possibility of distinctly equally distributed between males and females. The studies were
different SC barrier and water-handling properties in the skin performed with an independent Institutional Review Board approval
of infants less than 1-year old compared to adult skin. Natural and following the Declaration of Helsinki Principles. Subjects and/or
moisturizing compounds, found in high concentrations in the parents or legal guardians signed a written informed consent before
SC and within corneocytes, are hygroscopic and act as very the start of each study. The studies were performed in New Jersey,
efficient humectants (Rawlings and Harding, 2004). Distribu- and variations due to the time of year were found to be
tion profiles of NMF as a function of skin depth showed that comparatively small in magnitude and did not significantly
infant SC contains significantly smaller amounts of NMF than contribute to the results. A total of 124 infants (3 months to 4 years
adult SC. The lower amounts of NMF in infant skin could of age) and 104 adults (14–73 years of age) were studied.
impact the SC’s water-handling properties and is one likely For all studies, subjects were instructed to avoid use of skin care
mechanism contributing to the present results demonstrating products on their arms for at least 24 hours before the study.
faster water desorption from infant skin. However, less NMF Measurements on the arms were taken after subjects acclimated to
does not sufficiently explain the higher capacity for water an environmentally controlled room (20–25 1C and 40% relative
uptake seen in infant skin, suggesting a more hygroscopic humidity) for a minimum of 15 minutes (except for Raman studies
environment. Skin barrier function and water transport outlined below). Measurements were not taken from visibly
through the skin involve a complex interplay of multiple distressed or crying infants.
factors such as corneocyte maturity/hydrophilicity, lipid
amount and phase, density of appendages, surface micro- Instrumentation
relief, as well as diffusion path length, which could all play a Skin conductance was measured to evaluate the hydration state of
role in the differences between infant and adult skin the SC using the NOVA DPM (NOVA, Portsmouth, NH), which
properties. measures an electrical parameter directly relevant to hydration of the
Skin barrier function is undoubtedly influenced by the upper SC layers (Berardesca, 1997). This instrument will typically
environment and the dramatic changes that occur with the report a reading between 90 and 999 DPM units, according to the
shift from an aqueous and constant temperature environment manufacturer’s instruction. TEWL measurements were made using
in utero to the arid, cooler, and variable conditions of the the closed chamber technique with the Delfin VapoMeter (Delfin,
extra-uterine world. It has been suggested that interaction Kuopio, Finland). Measurements were standardized following the
with the external environment fosters barrier stabilization European Group on Efficacy Measurement of Cosmetics and Other
(Visscher et al., 2000). The adaptation period of adult Topical Products (EEMCO) guidelines (Rogiers, 2001). Measure-
skin barrier function to environmental changes has been ments were performed on the lower dorsal and upper ventral sites of
shown to be in the order of weeks for mice (Denda et al., randomly chosen arms of each subject, except where noted. We
1998; Elias, 2005) and weeks to months for humans (Chou chose the lower dorsal and upper ventral arm sites to examine
et al., 2005). It has been suggested that post-natal adaptation possible early effects of sun exposure (with the upper inner site being
occurs within the first month in newborn infants (Visscher more protected than the lower dorsal site). TEWL values were

1734 Journal of Investigative Dermatology (2008), Volume 128


J Nikolovski et al.
Infant Skin Barrier Maturation

compared from 65 infants (pooled from 2 independent studies) and determined using Student’s t-test, after confirmation of normal
71 adults. Baseline skin hydration values were compared from distribution using MINITAB’s normality test, and statistical signifi-
97 infants (pooled from 2 independent studies) and 71 adults. cance was assumed at the level of P ¼ 0.05. The data sets that were
Dynamic water-handling properties of the SC were measured not normally distributed (skin conductance and TEWL data) were first
using an absorption–desorption technique on the lower dorsal arm transformed to create normally distributed data so that we could
(Tagami et al., 1982). After a baseline reading (using the NOVA DPM perform Student’s t-test. The transformation used in this case was 1/x 2.
meter) was taken, a drop of water was applied to the skin for
10 seconds, followed by blotting with a paper towel. Serial CONFLICT OF INTEREST
conductance measurements were subsequently taken starting The authors state no conflict of interest.
30 seconds after initial water application up to 180 seconds at
15-second intervals. The rate of change was calculated for both the ACKNOWLEDGMENTS
We gratefully acknowledge Diana Friscia and Dr Laura McCulloch for their
absorption (uptake) and desorption (loss) of water from the skin. The
efforts on study design and execution, Dr Anthony Rawlings for insightful
rate of absorption for each subject was calculated from the initial discussions and editing of the manuscript, and River Diagnostics (Rotterdam,
reading to the maximum reading after 30 seconds. The rate of The Netherlands) for guidance with the in vivo Raman microspectroscopy
desorption was calculated similarly from the maximum reading at measurements.
30–60 seconds. Water desorption curves (conductance vs time) were
also fitted to exponential functions. Rates of water absorption and
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1736 Journal of Investigative Dermatology (2008), Volume 128