CHAPTER

Skin Structure and Function

1
Golara Honari and Howard Maibach

INTRODUCTION
Skin as one of the largest organs in body has multiple key features
required to interact dynamically with the environment. Primary func-
tions include a barrier function against environmental hazards such as
ultraviolet (UV) radiation, chemical and physical insults, and microor-
ganisms. Skin also prevents dehydration, regulates temperature, and
has self-healing properties. Dynamic and complex arrangement of vari-
ety of cells, element of the extracellular matrix, vascular, appendageal,
and nervous structures each play a role. Knowledge of skin penetration
pathways is essential in assessment of chemical safety, drug delivery
systems, and formulation of cosmetic products.

These chapter overviews essential structural elements of skin, their

key functions relevant to dermatotoxicology, skin absorption path-
ways, and key devices and methods to measure skin properties.

STRUCTURE AND FUNCTION

Normal skin consists of three main layers: epidermis, dermis, and
hypodermis.
• Epidermis: Barrier function, innate immunity, UV protection.
• Dermis: The largest component of skin, dermis is an integrated sys-
tem of fibrous cellular and acellular matrix. Many cell types reside
in dermis including fibroblasts, macrophages, mast cells. Vascular,
lymphatic, and nervous networks are present in dermis.
• Hypodermis (subcutis): Provides mechanical and physiologic sup-
port and contains larger source of vessels and nerves.
There are regional variations in the skin thickness and presence of dif-
ferent appendages such as hair, sebaceous, and sweat glands, which can
Applied Dermatotoxicology. DOI: http://dx.doi.org/10.1016/B978-0-12-420130-9.00001-3
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2 Applied Dermatotoxicology

affect the functional properties. For example, hair baring skin is typically
thinner and more permeable than nonhair baring skin of palms and soles.

Epidermis
Epidermis is the outermost layer and is about 0.05 1 mm in thickness
depending on body part. Three main populations of cells reside in
the epidermis: keratinocytes, melanocytes, and Langerhans cells.
Keratinocytes are the predominant cells in the epidermis, which are
constantly generated in the basal lamina and go through maturation,
differentiation, and migration to the surface. As keratinocytes differen-
tiate, they form three layers above the basal layer known as stratum
spinosum (SP), stratum granulosum (SG), and stratum corneum (SC)
(Figure 1.1). Keratinocyte transit time from basal layer up to SC is
about 14 days1 and turn over time within SC is also around 14 days,2
certain inflammatory conditions can affect these turn over times.
SC is the outer layer of the epidermis and serves as the main func-
tional barrier. A theoretical model is “brick and mortar” like structure
where bricks represent terminally differentiated nonviable keratino-
cytes, also known as corneocytes embedded in intercellular lipid mem-
branes.3 As corneodesmosomes (protein bridges between corneocytes)
degrade, lacunar spaces are created within the SC referred to as

Stratum corneum

Stratum granulosum

Langerhans cell

Stratum spinosum

Melanocyte

Merkel cell

Basal cell layer

Figure 1.1 Schematic of epidermis—basal cell layer is the deepest layer of epidermis differentiating to spinous
cells then to granular cells and eventually terminally differentiate to SC.
 Skin Structure and Function 3

aqueous “pore” pathway. These spaces can extend and form continues
networks, creating a pathway for penetration across the SC.4

Major components of the SC lipid membranes are free fatty acids, cer-
amides, and esterols.5 Melanocytes are neural crest-derived, pigment syn-
thesizing dendritic cells that reside primarily in the basal layer. Merkel
cells are mechanosensory receptors also present in basal layer.
Langerhans cells are dendritic antigen-processing and antigen-presenting
cells in the epidermis.6 They form 2 8% of the total epidermal cell popu-
lation, mostly found in a suprabasal position. The dermal epidermal
junction (DEJ) is a basement membrane zone that forms the interface
between the epidermis and dermis. The major functions of the DEJ are to
attach the epidermis and dermis to each other and to provide resistance
against external shearing forces.

Dermis
The dermis is an integrated system of fibrous cellular and acellular matrix
that accommodates nervous and vascular structures as well as epidermally
derived appendages. Many cell types reside in the dermis including fibro-
blasts, macrophages, mast cells, and circulating immune cells. Dermis is
responsible for skin elasticity, pliability, and tensile strength and provides
protection against mechanical injury, retins water, and aids in thermal
regulation. Dermis also contains and supports receptors of sensory stimuli
and a key element in wound healing.7

Hypodermis
Hypodermis is primarily composed of adipose tissue, which insulates
the body, serves as a reserve energy supply. It cushions and protects
skin and supports nerves, vessels, and lymphatics located within the
septa, supplying the overlying region.

SKIN APPENDAGES
Skin appendages include nails, hair, sebaceous glands, eccrine (sweat)
glands, and apocrine glands. They have two distinct components:
superficial and deeper components in the dermis, which are down
growths of epidermis. Dermal component regulates differentiation of
the appendage. During embryonic development, dermal epidermal
interactions are critical for the induction and differentiation of these
structures.
4 Applied Dermatotoxicology

SKIN A ROUTE OF ENTRY

The SC is 3 20 µm in thickness, composed of 15 25 layers of corneo-
cytes. It provides an effective barrier against transcutaneous water loss
and entry of exogenous materials.
Extracellular lipids contribute to barrier function and the route
taken through the SC by all molecules. Arrangement of extracellular
lipids, their hydrophobicity, composition, and distribution of key com-
ponents (ceramides, cholesterol, and free fatty acids) provide more
barrier function.8 Skin absorption varies between different body parts,
and between individuals, these regional intra- and inter-individual
variations are partly related to variations in lipid composition and SC
thickness.8 A range of biological factors can influence the rate and
extent of percutaneous penetration including anatomical site, age,
appendageal density, SC morphology, and composition. Routs through
which chemicals can cross the SC (Figure 1.2) include9:

Mechanical- Transfollicular
delivery methods Intracellular Intercellular
Stratum corneum

Hair Follicle
Viable epidermis

Sebaceous
gland

Dermis

Figure 1.2 Schematic pathways of penetration into the skin with arrangement of corneocytes in a “brick and
mortar” model.9
 Skin Structure and Function 5

• Intercellular (or extracellular), in which chemicals pass exclusively

through the lipid matrix
• Intracellular or transcellular, in which chemicals pass through both
the lipid matrix and the corneocytes themselves.
• Through skin appendages
• Mechanical methods to remove SC such as stripping, ablation,
micro needles, etc.

In vivo measurements skin absorption to quantify and rate the

extent of absorption is of fundamental importance in the risk assess-
ment of compounds that are active via the dermal route of entry,
though the details are beyond the scope of this chapter.

MEASURING SKIN
Measuring physical characteristics of skin using biophysical instru-
ments provides key information about various skin parameters. Many
noninvasive techniques and equipment are available with increasing
applications within dermatotoxicology, allowing study of skin in real
time and providing objective, quantitative data.
Parameters measured with these techniques provide information
about a particular aspect of skin. Using multiple parameters measured
simultaneously, along with clinical assessment provides more compre-
hensive analysis. Histologic studies may be used to complement these
analyses. Few parameters and techniques are briefly introduced in this
chapter; additional texts are available for comprehensive study.10,11

Skin Surface pH
Skin pH is normally acidic, ranging in pH values of 4 6, while the PH
in body’s internal environment is near-neutral, ranging from 7 to 9.12
The term “acid mantle” refers to inherent acidic nature of the SC. Skin
pH affects barrier function and SC cohesion. Elevation of pH in nor-
mal skin creates a disturbed barrier.13 Measurement of skin surface pH
is used to assess the acidity of the skin’s surface which skin surface pH
can vary according to the time of day, skin site, and between indivi-
duals. There are several instruments available for measurement of skin
pH; basically any standard, portable pH meter with a planar electrode
should suffice.
6 Applied Dermatotoxicology

Sebum
Sebum is a light yellow viscous fluid, composed of triglycerides, free
fatty acids, squalene, wax and sterol esters, and free sterols. Sebum is
produced by sebaceous glands and contributes to moisture balance in
the SC. Sebum production is mainly influenced by androgens and
varies among individuals and races, but the average rate in adults is
approximately 1 mg/10 cm2 every 3 h.14 Sebum production less than
0.5 mg/10 cm2 every 3 h is associated with dry skin, and values of
1.5 4.0 mg/10 cm2 every 3 h associated with seborrhea.15 Sebum can
be measured using a gravimetric or a variety of photometric
techniques.

Methods used in the past for measurement of skin lipids included

swabbing skin by absorbing pads followed by soaking them in solvents
or by weighing the absorptive tapes (gravimetric analysis).16 Also pho-
tometric analyses such as “grease-spot” photometry or UV Visible
spectrophotometry are used. Photometric techniques can provide addi-
tional information, such as droplet size and distribution.8 Multiple
commercially available devices can provide quantitative measures of
sebum secretion.

Desquamation
Loss of superficial SC is a natural process called desquamation, which
can be affected in certain disease. Desquamation is used to assess
structure and biological dynamics of SC and to investigate effect of
drugs and topical products on skin. Techniques for desquamation mea-
surement usually involve stripping surface layers for analysis or visual
assessment through skin imaging.

Squamometry involves striping of corneocytes form surface of SC

using adhesive disks followed by staining with toluidine blue-basic
fuchsin ethanol-based solution and reading the colorimetric variable.17
Image analysis systems are also used to asses desquamation.18 20
These techniques are rapid, reliable, and sensitive.

Thickness
Epidermal and SC thickness can be measured in vivo, using noninva-
sive methods such as ultrasound-based devices, confocal microscopy
and spectroscopic techniques.21 25 These measurements have investiga-
tive and clinical applications.
 Skin Structure and Function 7

The Measurement of Transepidermal Water Loss

Transepidermal water loss (TEWL) is the amount of water that pas-
sively evaporates through skin to the external environment due to
water vapor pressure gradient on both sides of the skin barrier and is
used to characterize skin barrier function. The average TEWL in
human is about 300 400 mL/day; however, it can be affected by envi-
ronmental and intrinsic factors. In high humidity, the amount of water
loss will decrease due to the drop in the water vapor pressure gradient.
TEWL varies in different anatomic sites and is inversely related to the
corneocyte size. Skin sites with smaller corneocytes have higher TEWL
values.26 28 Multiple instruments are commercially available to mea-
sure TEWL, providing valuable data with applications in clinical set-
tings, toxicology, and product development. TEWL is a sensitive
indicator of skin irritation and is widely used in objective analysis of
irritancy potential or protective properties of topical products. The
accuracy of TEWL measurements can be influenced by environmental
factors such as humidity, temperature, ventilation, and intrinsic fac-
tors. It is essential that these measurements be conducted under stan-
dard conditions.29

Hydration Measurement
Water content of SC in normal conditions is estimated around 5 10%
in the outer layer and about 30% near viable epidermis.30 Water con-
tents of skin influence its physical properties, viscoelastic characteris-
tics, and functional properties such as the drug penetration and barrier
function. In disease states such as atopic dermatitis, SC cannot stay
properly hydrated. Hydration status of SC is considered a measure of
toxicity. Hydration measurements are extensively used in assessment of
topical product safety and efficacy.

Various methods have been introduced to indirectly measure hydra-

tion. One method uses electrical properties of skin like capacitance,
impedance, resistance, and conductance, to calculate hydration
levels.31 Another method measures transient thermal transfer (TTT),
by using a probe to transfer a constant generated thermal pulse to the
epidermis, while simultaneously obtains high precision measurements
of skin temperature. Water content of SC is calculated based on
changes detected in the temperature.32 The third technique, nuclear
magnetic resonance (NMR) spectroscopy, directly measures the total
water content of the epidermis and the outer dermis, using magnetic
8 Applied Dermatotoxicology

fields.33 NMR provides direct measurements and is considered a refer-

ence technique for skin hydration studies. However, all the available
techniques can complement each other.34

Measurement of Vascular Perfusion

Skin microcirculation and perfusion can be affected by a number of
exogenous and endogenous factors. Changes in cutaneous vascular
perfusion may reflect a physiologic response such as thermoregulation,
or a pathologic response such as inflammation caused by exposure to
chemical irritants and concomitant release of inflammatory mediators.
Also exposure to topical vasoactive drugs can affect skin circulation.
Laser Doppler instruments measure the Doppler shift induced by the
laser light that’s being scattered by moving red blood cells. A signal is
quantified based on average red blood cell concentration and velocity.
Since this measure is not an exact measure, it is referred to as flux,
which has a linear relationship with the actual flow.35 37 Two distinct
laser Doppler tools are used: laser Doppler flowmetry (LDF), which
has a small probe touching the skin, measuring blood flow over a
small volume (1 mm3 or smaller). The second method is laser Doppler
imaging (LDI), in which the laser beam is emitted at a certain distance
above the skin surface and reflected by a computer-driven mirror to
scan an area of skin. DLI provides two-dimensional images mapping
the perfusion but is a slow method. Quantifying fast changes in blood
flow is easier with DLF compared to DLI.35
Laser speckle contrast imaging (LSCI) is a more recent, noncon-
tact imaging technique that provides information on skin perfusion
over large areas (up to 100 cm2) with a high frequency (up to 100
images/s). These images are obtained based on the generation of a
high contrast grainy pattern when laser hits a matt surface. This pat-
tern is called speckled pattern, which fluctuates when objects move.
Images created by LSCI reflect fast changes in skin blood flux over
wide skin areas.35,38

These techniques along with other methods briefly mentioned in

this chapter are among many tools to assess skin parameters and to
provide objective measures for investigators and clinicians. Many bio-
engineering methods are subject to operational guidelines and have
limitations. Data obtained from various methods can complement
each other.
 Skin Structure and Function 9

REFERENCES
1. Weinstein G, Van Scott E. Autoradiographic analysis of turnover times of normal and psori-
atic epidermis. J Invest Dermatol. 1965;45(4):257 262.
2. Bergstresser P, Taylor J. Epidermal “turnover time”—a new examination. Br J Dermatol.
1977;96:503 509.
3. Michaels AS, Chandrasekaran SK, Shaw JE. Drug permeation through human skin. Theory
and in vitro experimental measurements. AICHE J. 1975;21(5):985 996.
4. Menon GK, Elias PM. Morphologic basis for a pore-pathway in mammalian stratum cor-
neum. Skin Pharmacol. 1997;10(5 6):235 246.
5. Lee D, Ashcraft JN, Verploegen E, Pashkovski E, Weitz DA. Permeability of model stratum
corneum lipid membrane measured using quartz crystal microbalance. Langmuir. 2009;
25(10):5762 5766.
6. Mutyambizi K, Berger CL, Edelson RL. The balance between immunity and tolerance: the
role of Langerhans cells. Cell Mol Life Sci. 2009;66(5):831 840.
7. Chu DH. Development and structure of skin. Fitzpatrick’s Dermatology in General Medicine.
8th ed., USA: McGraw-Hill; 2012.
8. Elias PM. The epidermal permeability barrier: from the early days at Harvard to emerging
concepts. J Invest Dermatol. 2004;122(2):xxxvi xxxix.
9. Prausnitz MR, Mitragotri S, Langer R. Current status and future potential of transdermal
drug delivery. Nat Rev Drug Discov. 2004;3(2):115 124.
10. Fluhr WJ, Elsner P, Berardesca E, Maibach HI. Bioengineering of the skin: water and the
stratum corneum. Dermatology: Clinical & Basic Science. 2nd ed. USA: CRC Press; 2004.
11. Agache P, Humbert P. Measuring the Skin. Heidelberg: Springer; 2004.
12. Ali SM, Yosipovitch G. Skin pH: from basic science to basic skin care. Acta Derm Venereol.
2013;93(3):261 267.
13. Hachem JP, Crumrine D, Fluhr J, Brown BE, Feingold KR, Elias PM. pH directly regulates
epidermal permeability barrier homeostasis, and stratum corneum integrity/cohesion. J Invest
Dermatol. 2003;121(2):345 353.
14. Plewig G, Kligman A. Acne and Rosacea. Berlin: Springer; 2000.
15. Bolognia JL, Jorizzo JL, Schaffer JV. Dermatology. 3rd ed. China: Elsevier; 2012.
16. Pande SY, Misri R. Sebumeter. Indian J Dermatol Venereol Leprol. 2005;71(6):444 446.
17. Pierard-Franchimont C, Henry F, Pierard GE. The SACD method and the XLRS squamo-
metry tests revisited. Int J Cosmet Sci. 2000;22(6):437 446.
18. Black D, Boyer J, Lagarde JM. Image analysis of skin scaling using D-squame samplers:
comparison with clinical scoring and use for assessing moisturizer efficacy. Int J Cosmet Sci.
2006;28(1):35 44.
19. Wilhelm KP, Kaspar K, Schumann F, Articus K. Development and validation of a semiau-
tomatic image analysis system for measuring skin desquamation with D-squames. Skin Res
Technol. 2002;8(2):98 105.
20. Xhauflaire-Uhoda E, Loussouarn G, Haubrechts C, Leger DS, Pierard GE. Skin capacitance
imaging and corneosurfametry. A comparative assessment of the impact of surfactants on
stratum corneum. Contact Dermatitis. 2006;54(5):249 253.
21. El Gammal S, El Gammal C, Kaspar K, et al. Sonography of the skin at 100 MHz enables
in vivo visualization of stratum corneum and viable epidermis in palmar skin and psoriatic
plaques. J Invest Dermatol. 1999;113(5):821 829.
10 Applied Dermatotoxicology

22. Kaspar K, Vogt M, Ermert H, Altmeyer P, el Gammal S. 100 MHz sonography in the visu-
alization of the palmar stratum corneum after application of various creams and ointments.
Ultraschall Med. 1999;20(3):110 114.
23. Corcuff P, Bertrand C, Leveque JL. Morphometry of human epidermis in vivo by real-time
confocal microscopy. Arch Dermatol Res. 1993;285(8):475 481.
24. Caspers PJ, Lucassen GW, Carter EA, Bruining HA, Puppels GJ. In vivo confocal Raman
microspectroscopy of the skin: noninvasive determination of molecular concentration pro-
files. J Invest Dermatol. 2001;116(3):434 442.
25. Egawa M, Hirao T, Takahashi M. In vivo estimation of stratum corneum thickness from
water concentration profiles obtained with Raman spectroscopy. Acta Derm Venereol.
2007;87(1):4 8.
26. Rougier A, Lotte C, Corcuff P, Maibach HI. Relationship between skin permeability and
corneocyte size according to anatomic site, age and sex in man. J Soc Cosmet Chem.
1988;39:15 26.
27. Machado M, Salgado TM, Hadgraft J, Lane ME. The relationship between transepidermal
water loss and skin permeability. Int J Pharm. 2010;384(1 2):73 77.
28. Hadgraft J, Lane ME. Transepidermal water loss and skin site: a hypothesis. Int J Pharm.
2009;373(1 2):1 3.
29. Sotoodian B, Maibach HI. Noninvasive test methods for epidermal barrier function. Clin
Dermatol. 2012;30(3):301 310.
30. Blank IH, Moloney III J, Emslie AG, Simon I, Apt C. The diffusion of water across the stra-
tum corneum as a function of its water content. J Invest Dermatol. 1984;82(2):188 194.
31. Berardesca E, Borroni G. Instrumental evaluation of cutaneous hydration. Clin Dermatol.
1995;13(4):323 327.
32. Berardesca E, Fideli D, Borroni G, Rabbiosi G, Maibach H. In vivo hydration and water-
retention capacity of stratum corneum in clinically uninvolved skin in atopic and psoriatic
patients. Acta Derm Venereol. 1990;70(5):400 404.
33. Foreman MI. A proton magnetic resonance study of water in human stratum corneum.
Biochim Biophys Acta. 1976;437(2):599 603.
34. Girard P, Beraud A, Sirvent A. Study of three complementary techniques for measuring cuta-
neous hydration in vivo in human subjects: NMR spectroscopy, transient thermal transfer
and corneometry—application to xerotic skin and cosmetics. Skin Res Technol. 2000;
6(4):205 213.
35. Roustit M, Cracowski JL. Assessment of endothelial and neurovascular function in human
skin microcirculation. Trends Pharmacol Sci. 2013;34(7):373 384.
36. Stern MD. In vivo evaluation of microcirculation by coherent light scattering. Nature.
1975;254(5495):56 58.
37. Ahn H, Johansson K, Lundgren O, Nilsson GE. In vivo evaluation of signal processors for
laser Doppler tissue flowmeters. Med Biol Eng Comput. 1987;25(2):207 211.
38. Briers JD. Laser Doppler, speckle and related techniques for blood perfusion mapping and
imaging. Physiol Meas. 2001;22(4):R35 R66.