International Journal of Cosmetic Science, 2005, 27, 17–34
Ultraviolet radiation and skin aging: roles of reactive
oxygen species, inflammation and protease
activation, and strategies for prevention of
inflammation-induced matrix degradation – a review
S. Pillai, C. Oresajo and J. Hayward
Engelhard Corporation, Long Island, New York, NY, U.S.A.
Received 3 June 2004, Accepted 28 September 2004
Keywords: antipotease, elastase, matrix metalloprotease, neutrophils, serine protease
Synopsis
Inflammation and the resulting accumulation of
reactive oxygen species (ROS) play an important
role in the intrinsic and photoaging of human skin
in vivo. Environmental insults such as ultraviolet
(UV) rays from sun, cigarette smoke exposure and
pollutants, and the natural process of aging con-
tribute to the generation of free radicals and ROS
that stimulate the inflammatory process in the
skin. UV irradiation initiates and activates a com-
plex cascade of biochemical reactions in human
skin. In short, UV causes depletion of cellular anti-
oxidants and antioxidant enzymes (SOD, catalase),
initiates DNA damage leading to the formation of
thymidine dimmers, activates the neuroendocrine
system leading to immunosuppression and release
of neuroendocrine mediators, and causes increased
synthesis and release of pro-inflammatory media-
tors from a variety of skin cells. The pro-inflamma-
tory mediators increase the permeability of
capillaries leading to infiltration and activation of
neutrophils and other phagocytic cells into the
skin. The net result of all these effects is inflamma-
tion and free radical generation (both reactive oxy-
gen and nitrogen species). Furthermore, elastsases
and other proteases (cathepsin G) released from
neutrophils cause further inflammation, and acti-
Correspondence: Dr Sreekumar Pillai PhD, Engelhard Cor-
poration, 50 Health Sciences Drive, Stony Brook, NY
11790, U.S.A. Tel.: +1 631 380 2352; fax: +1 631 380
2515; e-mail: kumar.pillai@engelhard.com
ª 2005 Society of Cosmetic Scientists and the Société Française de Cosmétologie
vation of matrix metalloproteases. The inflamma-
tion further activates the transcription of various
matrixes degrading metalloproteases, leading to
abnormal matrix degradation and accumulation of
non-functional matrix components. In addition,
the inflammation and ROS cause oxidative damage
to cellular proteins, lipids and carbohydrates,
which accumulates in the dermal and epidermal
compartments, contributing to the aetiology of
photoaging. Strategies to prevent photodamage
caused by this cascade of reactions initiated by UV
include: prevention of UV penetration into skin by
physical and chemical sunscreens, prevention/
reduction of inflammation using anti-inflammatory
compounds (e.g. cyclooxygenase inhibitors, inhibi-
tors of cytokine generation); scavenging and
quenching of ROS by antioxidants; inhibition of
neutrophil elastase activity to prevent extracellular
matrix damage and activation of matrix metallo-
proteases (MMPs), and inhibition of MMP expres-
sion (e.g. by retinoids) and activity (e.g. by natural
and synthetic inhibitors).
Résumé
L’inflammation et l’accumulation de dérivés réac-
tifs de l’oxygène qui en résulte jouent un rôle
important sur le vieillissement normal (chronologi-
que) et pathologique (photovieillissement) de la
peau in vivo. Les attaques de l’environnement, tels
que les UV, l’exposition a la fumée de cigarettes et
17
Ultraviolet radiation and skin aging
a des polluants, et le procédé naturel du vieillisse-
ment, contribuent à la création de radicaux-libres
et de dérivés réactifs de l’oxygène qui stimulent
l’inflammation cutanée. Les radiations UV sont à
l’origine d’une cascade complexe de réactions bio-
chimiques sur la peau humaine. A court terme, les
UV dégradent les antioxydants cellulaires et les
enzymes antioxydants (SOD, catalase); endomma-
gent l’ADN provoquant la formation de dimères de
la thymidine; activent le système neuroendocrinien
engendrant une immunodépression et la sécrétion
de médiateurs neuroendocriniens; et enfin provo-
quent une synthèse accrue et la sécrétion de
médiateurs pro-inflammatoires par diverses cellules
cutanées. Les médiateurs pro-inflammatoires aug-
mentent la perméabilité des capillaires engendrant
l’activation et l’infiltration de neutrophiles et
autres cellules phagocytes dans la peau. Le résultat
de toutes ces réactions est l’inflammation et la
génération de radicaux-libres (à la fois oxygénés et
azotés). Puis, les élastases et autres protéases
(cathepsine G) issues des neutrophiles accroissent
l’inflammation et activent les métalloprotéases
matricielles (MPM). En effet, l’inflammation ainsi
créée active la transcription de diverses MPM
néfastes à la matrice extracellulaire, générant sa
dégradation anormale et l’accumulation de dérivés
non fonctionnels en son milieu. De plus, l’inflam-
mation et les dérivés réactifs de l’oxygène oxydent
et dégradent les protéines, lipides et sucres cellulai-
res qui s’accumulent dans le derme et l’épiderme,
contribuant à l‘étiologie du photovieillissement.
Les stratégies pour prévenir de la photo-dégrada-
tion générée par cette cascade de réactions causées
par les UV sont les suivantes: prévention de la
pénétration des UV dans la peau par des écrans
solaires physiques ou chimiques; prévention et
réduction de l’inflammation en utilisant des agents
anti-inflammatoires (tels les inhibiteurs de la cyclo-
oxygenase et les inhibiteurs de la production de la
cytokine); en complexant et bloquant les dérivés
réactifs de l’oxygène; inhibition de l’activité du
neutrophile élastase pour prévenir la dégradation
de la matrice extracellulaire et l’activation des
MPM; et inhibition de l’expression des MPM (ex:
par des rétinoı̈des) et de leur activité (ex: par des
inhibiteurs naturels et synthétiques).
Introduction
Inflammation and the resulting accumulation of
reactive oxygen species (ROS) play an important
18
S. Pillai, C. Oresajo and J. Hayward
role in the intrinsic and photoaging of human skin
in vivo. Environmental insults such as ultraviolet
(UV) rays from sun, cigarette smoke exposure and
pollutants, as well as the natural process of aging
contribute to the generation of ROS that stimulate
the inflammatory process in the skin. One of the
primary events in ROS-mediated inflammation is
the activation of transcription factors that regulate
the proteolytic degradation of the skin extracellu-
lar matrix (ECM). Photoaged skin shows increased
proteolytic activation and abnormal ECM turnover
leading to increased degradation of collagen and
elastic fibres in the dermis, resulting in a loss of
skin’s ability to resist stretching.
An interlinked network of enzymes that convert
ROS to harmless water and molecular oxygen
regulates skin’s antioxidant defence system. The
skin of aged and photoaged individuals has
reduced levels of these natural enzymatic and non-
enzymatic antioxidant defence mechanisms,
increased neutrophil infiltration into skin and
increased inflammation. The anti-aging technology
used today in skin care products relies heavily on
the use of antioxidants for prevention of UV- and
ROS-mediated inflammation. New basic under-
standing on the mechanisms of neutrophil activa-
tion and emerging technologies on inhibition of
UV- and ROS-induced proteases have significantly
enhanced our ability to prevent UV-induced skin
aging.
Excellent reviews have been written on the free
radical theory of aging and the role of oxidative
stress and mitochondrial involvement in aging
process [1–3]. The purpose of this review is to
focus on the role of inflammation, especially that
caused by UV irradiation including neutrophil acti-
vation and proteolytic activation in skin aging and
methods to prevent skin aging by interfering with
these mechanisms.
Effects of UV on ROS generation,
inflammation, neutrophil activation and
matrix metalloprotease-induced ECM
degradation
UV, photoaging and skin changes
Skin changes with photoaging
Premature aging of the skin (photoaging) is a
well-documented consequence of exposure to
Ultraviolet A (UVA) and Ultraviolet B (UVB). Pho-
toaged skin is biochemically characterized by the
ª 2005 International Journal of Cosmetic Science, 27, 17–34
Ultraviolet radiation and skin aging
presence of abnormal elastic fibres in the dermis
and by a dramatic decrease of distinct collagen
types. The elastic fibres demonstrate a dystrophic,
amorphic structure as a consequence of UV
directly increasing tropoelastin gene expression
[4]. Photoaged skin is coarsely wrinkled and asso-
ciated with skin thickening (both epidermal and
dermal compartments), dyspigmentation and
telangiectasia [5–7]. Biochemically, the photoaged
skin shows increased proteolytic activation, and
abnormal ECM turnover leading to increased deg-
radation of collagen and elastic fibres in the der-
mis, resulting in a loss of skin’s ability to resist
stretching. Environmental insults such as UV rays,
cigarette smoke exposure and pollutants, and the
natural process of aging contribute to the genera-
tion of ROS that stimulate the inflammatory pro-
cess in the skin. The skin of aged and photoaged
individuals has reduced levels of natural enzymatic
and non-enzymatic antioxidant defence mecha-
nisms [8] increased neutrophil infiltration into skin
and increased inflammation [9].
Cellular mechanisms activated by UV irradiation
The acute effects of UV irradiation on human skin
are well characterized. The immediate reaction to
UV is mediated by several mechanisms including
direct effects on keratinocytes to release pro-
inflammatory cytokines such as interleukin-1
(IL-1) and tumour necrosis factor-a (TNF-a) [10];
direct effects of photons on DNA to cause DNA
breakage [11]; depletion of cellular antioxidants
and generation of ROS including hydrogen perox-
ide, hydroxyl radical, singlet oxygen and peroxyl
radicals [5, 8]; and generation of prostaglandins
and other inflammatory mediators such as hista-
mine and leucotrines by mast cells in skin [12].
UV-induced inflammation and the resulting accu-
mulation of ROS play an important role in the
intrinsic and photoaging of human skin in vivo
[13]. At the cellular level, UV radiation triggers
cytokine production [14], induces surface expres-
sion of adhesion molecules [15] and affects cellular
mitosis, apoptosis and necrosis [16]. These media-
tors also increase the skin’s sensitivity to irritants.
The free radical-induced peroxidation of membrane
lipids contributes to increased phospholipase A2
activity leading to more production of prostaglan-
dins [17]. Morphologically acute UV irradiation is
followed by epidermal keratinocytes damage,
depletion of Langerhans cells, dermal edema,
endothelial swelling, mast cell degranulation and
ª 2005 International Journal of Cosmetic Science, 27, 17–34
S. Pillai, C. Oresajo and J. Hayward
cellular infiltration with neutrophils and mono-
cytes within dermis and epidermis. Biochemical
changes include release of histamine and accumu-
lation of cyclooxygenase- and lipooxygenase-
derived products of arachidonic acid, kinins and
cytokines derived from keratinocytes and fibro-
blasts. These responses are more pronounced in
young subjects [18]. Aged skin demonstrated
decreased epidermal turnover, diminished inflam-
matory response to UV and impaired immune
function. In addition to the differences in immune
and inflammatory responses of young versus aged
skin, the response of barrier function and barrier
recovery is also impaired in older skin. In studies
with murine epidermis, it was demonstrated that
the skin becomes less sensitive to UVB-induced
barrier alterations with age and this decreased
sensitivity correlated with decreased DNA synthe-
sis following UV irradiation [19].
Acute and chronic UV exposure is associated
with protein oxidation in human skin in vivo, both
in the living layers as well as in the stratum cor-
neum [20, 21]. Human dermal fibroblasts play a
central role in connective tissue breakdown in
photoaging. UV irradiation induces UV-responsive
genes of fibroblasts, among them matrix metallo-
proteases, which degrade macromolecules of the
ECM [22]. UVB modulates several signal transduc-
tion pathway(s) leading to generation of ROS that
induces the transcription of metalloproteases
[13, 23]. Recent data also suggest the potential
involvement of reactive nitrogen species (RNS) in
addition to ROS in UVB-mediated expression of
interstitial collagenase [matrix metalloprotease-1
(MMP-1)] and stromelysin-1 (MMP-3) [23]. One of
the early responses of keratinocytes to UV irradi-
ation is release of pro-inflammatory cytokines in a
similar fashion to response to injury.
Release of inflammatory mediators in response to
UV
Cytokines released in response to UV
UV, inflammation, tape stripping and even
mechanical stimulation of skin can cause release
of IL-1 from stratum corneum to initiate the
inflammatory reactions within the living layers of
skin [10, 24]. In addition, UV irradiation stimu-
lated TNF-a synthesis and release from keratino-
cytes into blood stream, suggesting epidermal cell
derived cytokines may mediate systemic inflamma-
tory reactions [25]. These two ‘primary cytokines’,
19
Ultraviolet radiation and skin aging
can in turn induce the synthesis and release of
other pro-inflammatory cytokines in response to
injury or UV radiation [14, 26, 27]. UV irradiation
dramatically induced the production and secretion
of IL-1, IL-3 and IL-6 that are normally low in un-
irradiated cells. Additionally, UV irradiation of
human skin in vivo also induced the expression of
IL-1, IL-10 and IL-7 [28]. IL-10 and IL-12 are
regulated differentially by UVB irradiation [29].
UVB induced both IL-10 and IL-12, however, UVA
appears to induce only IL-10, not IL-12 in kera-
tinocytes. UV irradiation also induced the IL-10
and IL-12 expression in lymph nodes [30] suggest-
ing the systemic role of UV in inflammatory
responses. In addition to ILs, TNF-a and granulo-
cyte–macrophage colony-stimulating factor (GM–
CSF) levels are also increased in response to UV
[31]. All these factors, in addition to mediating
inflammation, also cause immunosuppression.
Other growth factor agonists/antagonists secreted
by keratinocytes in response to UV include: IL-1
receptor antagonist [26]; alpha-melanocyte-stimu-
lating hormone (a-MSH) [32] vascular endothelial
growth factor (VEGF) [33]; nitric oxide (NO) [34,
35]; basic fibroblast growth factor (b-FGF); nerve
growth factor (NGF), endothelin-1 and the proopi-
omelanocortin (POMC)-derived peptides – MSH,
ACTF, beta-LPH and beta-endorphin [36].
Influence on other cell types in skin
Leukocyte/keratinocyte cell adhesion is mediated
by intercellular adhesion molecule-1 (ICAM-1) on
the keratinocyte cell surface. ICAM-1 serves as the
counter-receptor for lymphocyte function associ-
ated antigen-1 for adhesion to keratinocytes [15].
IL-1a released in response to UV irradiation stimu-
lates the ICAM-1 expression of keratinocytes,
thereby promoting the leukocyte adhesion to skin
and generation or ROS by the leukocytes. a-MSH
produced by keratinocytes can in turn act as an
immunosuppresent or as a stimulant of melano-
cytes to produce more melanin for protection of
skin from further UV damage [32]. The immuno-
suppressive capacity of a-MSH is mediated by its
effect on melanocyte and macrophage functions.
Another mediator of immunity and skin pigmenta-
tion, nitric oxide is also produced in keratinocytes
in response to UV exposure. UV stimulates the
expression of inducible nitric oxide synthase
(iNOS), the enzyme that produces NO in response
to UV or inflammation. The NO produced by kera-
tinocytes appears to play a role in UV-induced skin
20
S. Pillai, C. Oresajo and J. Hayward
reactions such as sunburn reaction and photo-
induced immune alterations [35]. Other cytokines
such as b-FGF, NGF, endothelin-1 and POMC-
derived peptides also stimulate melanocyte growth,
melanogenesis and skin tanning [36]. Cytokines
generated by the keratinocytes in the acne lesions
and comedonal part of acne may also influence
sebum production and inflammation of acne
lesions [37]. UV irradiation induced the production
of IL-1a, IL-1 receptor antagonist (IL-1 ra), Il-6
and IL-10 in acne patients. UV-induced IL-1 ra
and IL-10 may have a role as anti-inflammatory
regulators in acne. The levels of these two cyto-
kines were elevated in stationary acne patients,
suggesting that UV irradiation may benefit the sta-
tionary acne patients.
ROS, inflammation and protease activation
ROS generation and inflammation
Generation of ROS is associated with life under
aerobic conditions. ROS are generated under phy-
siological and pathological conditions. Superoxide,
hydrogen peroxide and hydroxy radicals are pri-
marily produced by phagocytic cells and polynu-
clear lymphocytes in response to inflammation
[38]. Phagocytic cells such as macrophages and
lymphocytes produce large quantities of ROS by
expressing high amounts of NADPH oxidase
enzyme [39]. Involvement of NADPH oxidase-
induced ROS in host defence response of these cells
is well established. Skin is uniquely vulnerable to
the damage caused by ROS, being rich in unsatur-
ated fatty acids and exposed to high oxygen ten-
sion and UV light, both of which promote ROS
generation. Additionally, the respiratory burst of
infiltrating polymorphonuclear leukocytes and
macrophages in inflamed skin produces high local
levels of superoxide and can release catalytic ion
from ferritin. The iron in combination with ROS
can cause additional damage to skin [38]. ROS,
especially the hydroxyl radical, are capable of
damaging biological macromolecules such as
DNA, carbohydrates, lipids and proteins. Among
the most susceptible targets are polyunsaturated
fatty acids, where ROS initiates the process of lipid
peroxidation. Damage to the lipids in the cellular
membrane can cause cellular damage, leakage of
cellular components and ultimately cell death.
Lipid aldehydes produced during peroxidation can
also react with amino acid side chains and nucleic
acids to cause further damage.
ª 2005 International Journal of Cosmetic Science, 27, 17–34
Ultraviolet radiation and skin aging
ROS generated at the injury site play an import-
ant role in modulating the inflammatory response
under acute and chronic injury conditions. One of
the primary events in ROS-mediated inflammation
is the activation of transcription factors. Nuclear
factor kappa B (NFjB) and activator protein-1
(AP-1) are transcription factors that are regulated
by cellular redox levels, and involved in regulation
of gene expression [40]. These factors are co-ordi-
nately responsible for a huge range of extracellular
signalling molecules responsible for inflammation,
tissue remodelling, oncogenesis and apoptosis, pro-
cesses that orchestrate many of the degenerative
processes associated with aging [41]. UV exposure
also induces the expression of inducible cyclooxyge-
nase (COX-2) and lipooxygenase (LOX) in human
skin, leading to increased production of pro-inflam-
matory mediators such as prostaglandins and
thromboxanes [35].
ROS and activation of neutrophils
Phagocytic cells such as neutrophils and monocytes
infiltrate into skin from capillaries in response to
UV-induced production of cytokines such as IL-1 or
TNF-a [42]. Large amounts of IL-1 are stored in
stratum corneum, the top layer of human epidermis
[43]. In addition to keratinocytes, the phagocytic
cells themselves secrete cytokines (TNF-a, IL-1,
IL-8, etc.) that further enhance recruitment of
inflammatory cells [44]. The phagocytic cells recrui-
ted and activated by these inflammatory cytokines
generate ROS and NO as part of their defence mech-
anisms. Both compounds can combine to form per-
oxynitrite, a reactive species capable of inducing
lipid peroxidation and damage [45]. In addition,
these cells generate a variety of other molecular
oxygen species (O2), ÆOH and H2O2) that assist in the
killing of microorganisms. They also release oxida-
tion products of membrane fatty acids (e.g. arachi-
donate), which are converted to thromboxanes and
prostaglandin that are additional mediators of
inflammation [46]. Phagocytic cells induce the ker-
atinocytes to synthesize and secrete elafin, an inhib-
itor of human neutrophil elastase, which eventually
limits the damage caused by the inflammatory neu-
trophils to skin [44]. Elafin (also known as skin-
derived anti-leukoprotease/trappin-2) is highly
induced in inflamed skin such as in psoriasis, after
wounding and in skin tumours [47]. In addition to
being an inhibitor of elastase, elafin is also incorpor-
ated in the cornified envelope in psoriatic skin
lesions [48].
ª 2005 International Journal of Cosmetic Science, 27, 17–34
S. Pillai, C. Oresajo and J. Hayward
Chronic Protease-Antiprotease
Inflammation Balance
Degrades
Anti-
Neutrophils Elastase proteases
+
IL-8
Inhibits
Inactivates TIMP
Activation of macrophages,
keratinocytes
Activates
Degradation
Release of IL-1, TNF-a,
prostaglandins and MMPs of ECM
leukotrines
Figure 1 Role of neutrophil elastase in inflammation
and matrix metalloprotease activation.
Inflammation and neutrophil protease activation
Damage to connective tissues is a major complica-
tion of the inflammatory response. Inflammatory
tissue injury contributes to the pathological chan-
ges in various tissues and organs including skin in
photodamage. [49]. Proteases released by leuko-
cytes play a major role in tissue injury associated
with inflammation (Fig. 1). Polymorphonuclear
leukocyte (PMN) secretes lysosomal enzymes from
open phagocytic vacuoles regulated by cytoplasmic
microtubules. Some of the major enzymes secreted
by PMN in response to inflammation are neutro-
phil elastase, interstitial collagenase (MMP-1), and
cathepsin G [46]. The serine protease, neutrophil
elastase is activated in response to inflammatory
tissue injury. Serine proteases have substrate spe-
cificities and substrate binding characteristics that
differ entirely from those of the MMPs, as discussed
in detail in Direct inhibition of MMPs section [49].
Polymorphonuclear serine proteases
Serine proteases have been classified by their sub-
strate specificity into three types: trypsin-like,
chymotrypsin-like and elastase-like. Serine prote-
ase elastase prefers substrates with small aliphatic
chains. The mode of action of serine proteases
involves the amino acid serine, which has a
hydroxyl group that acts as a nucleophile for
hydrolytic cleavage [49]. In contrast, a metal ion
(usually zinc), coordinates and activates the target
protein amide carbonyl for hydrolysis by MMPs.
Therefore, serine protease elastase differs funda-
mentally from MMP elastase. Several matrix com-
ponents such as collagens, gelatins, elastins,
fibronectin, laminin and glycosaminoglycans are
excellent substrates for both serine proteases and
metalloproteases.
21
Ultraviolet radiation and skin aging
Neutrophil (serine) elastase, which is released
during neutrophil infiltration of the epidermis, has
been shown to induce hyperproliferation in
keratinocytes in vitro and in vivo [50]. Neutrophil
infiltration has been demonstrated in hyperproli-
ferative skin conditions such as psoriasis, especially
in pustular psoriasis [51, 52]. The secreted ela-
stase may be one mechanism by which hyperpro-
liferative skin conditions persist in psoriasis [51].
Hyperproliferation of keratinocytes can be induced
by neutrophil elastase both in vitro and in vivo,
and topical application of elastase inhibitors can
prevent elastase-induced keratinocyte hyperprolif-
eration [50]. The mechanism by which elastase
induces keratinocyte proliferation is believed to be
mediated by TGF-a stimulation and EGF receptor
activation [53]. Inhibition of UV-induced neutro-
phil elastase prevented UV-mediated skin tumour
formation in hairless mice [54]. Another elastase
activity, fibroblast elastase, also has been shown to
be involved in skin wrinkle formation. Inhibition
of this enzyme has been demonstrated to prevent
UV-induced wrinkle formation in skin [55].
Protease–antiprotease balance
Metalloproteinases and serine proteases can work
in combination to directly attack most of the ele-
ments of the ECM including interstitial stroma and
basement membranes. In addition, these enzymes
activate one another through cleavage of the pro-
enzyme forms and can also cause degradation of
protease inhibitors that maintains the appropriate
protease–antiprotease balance in normal skin
(Fig. 1). For example, cathepsin G (a serine prote-
ase) can activate MMP-8; human leukocyte ela-
stase (a serine protease) can inactivate the major
endogenous tissue inhibitors of matrix metallopro-
teases (TIMPs); and MMP-8 and MMP-9 can inacti-
vate a-1-protease inhibitor, the major endogenous
inhibitor of human leukocyte elastase [56, 57].
Thus, by contributing to protease activation and
inactivating endogenous inhibitors, the two classes
of proteases can skew the protease–antiprotease
balance toward pathological tissue degradation.
While the protease–antiprotease balance is strictly
regulated under normal conditions, a breakdown of
the control mechanism occurs in various disease
conditions characterized by excess serine protease
activity. However, other elastase inhibitors, secre-
tory leukocyte protease inhibitor and elafin are
resistant to degradation by MMP-8 [58]. In addi-
tion, the secretory leukocyte protease inhibitor can
22
S. Pillai, C. Oresajo and J. Hayward
also suppress the expression of MMP [59]. The
complex interaction between proteases and their
inhibitors can regulate the ECM breakdown in neu-
trophil-mediated inflammatory skin diseases.
Role of UV in MMP activation
MMPs and their endogenous inhibitors, TIMPs
MMPs are a group of zinc-dependent endopeptidas-
es capable of degrading ECM components and are
involved in the turnover and remodelling of the
dermis [60]. In normal skin, the MMPs are
expressed in very low levels and are kept in their
inactive form bound to endogenous inhibitors.
Inflammation, UV irradiation and normal process
of aging can activate MMPs leading to increased
matrix degradation.
The MMP family of genes consists of at least
two-dozen individual enzymes in human tissues.
Each is structurally distinct and has the ability to
degrade a particular subset of matrix proteins
[60]. MMP expression is organ-dependent, and dif-
ferent MMPs are expressed in different develop-
mental stages and in different diseases. For
example, MMPs are involved in tumour progres-
sion and inflammatory disorders. MMPs can be
broadly classified into four major classes depending
on substrate specificity and structural and expres-
sion characteristics: collagenases, which break
down collagens (MMP-1 and MMP-8); gelatinases,
(which degrade denatured collagens – MMP2,
MMP 9); stromelysins, which have broad spectrum
specificity, and membrane bound MMPs, which are
located mainly on tumour cells. In addition to col-
lagens, MMPs also degrade other ECM components
gelatine, laminin, fibronectin and elastin. Most
skin cell types (fibroblasts, keratinocytes, melano-
cytes) synthesize MMPs. The activities of MMPs
within the skin are regulated by endogenous
inhibitors, small molecular weight proteins called
tissue inhibitor of metalloproteases (TIMPs). TIMPs
bind MMPs in a 1 : 1 ratio. Skin cells produce sev-
eral classes of TIMPs, and the activity of the MMPs
is determined by the balance of MMPs to TIMPs
within the cells. In addition, the activity and the
production of MMPs are regulated by various
cytokines that are either generated within skin or
enter skin from the blood. The collagen breakdown
that occurs during inflammation, wound healing,
tumour invasion and chronological and photoag-
ing is regulated via cytokine modulation of MMP
expression. Prevention or blocking these conditions
ª 2005 International Journal of Cosmetic Science, 27, 17–34
Ultraviolet radiation and skin aging
using anti-inflammatory drugs, antioxidants, sun-
screens, etc. is an indirect way to prevent MMP
activation and reduce matrix breakdown.
MMPs in skin
Human skin expresses three distinct collagenases,
MMP-1, -8 and -13. Using UV-exposed organ cul-
tures of human skin, it was demonstrated that
although several MMPs including MMP-1 and -13
are stimulated by UVB, only MMP-1 inhibition
using specific antibodies prevented UV-enhanced
collagen breakdown in this model [61]. MMP-8 is
also stimulated by UVB in human skin in vivo
[62]. UVB, even at suberythemal doses, induced
expression of MMP-1, -3 and -9 in normal human
epidermis [63]. In addition, UVA has been shown
to induce the expression of MMP-1 in dermal fibro-
blasts in vivo and MMP-1, -2 and -3 in cultured
fibroblasts.
MMP-1 initiates cleavage of fibrillar collagen
types I and III in the dermis, which is then further
degraded by MMP-2 and -9 [60]. In addition, sim-
ultaneous expression of MMP-2, -3 and -9 could
result in degradation of non-collagenous compo-
nents of dermal ECM, including basement mem-
brane glycoproteins and proteoglycans. Solar
irradiation may exacerbate chronological aging by
further increasing the elevated levels of MMPs in
aged skin. Furthermore, MMPs in aged skin may
already be in a more active state because of the
decreased levels of TIMPs [64].
Molecular mechanisms of MMP activation
UV and ROS can activate the protein kinase cas-
cades to stimulate MMP gene expression [63, 65].
ROS also directly stimulate MMP zymogen activa-
tion by inducing the cleavage of the cysteine resi-
due association with the Zn active site. It is
suspected that in addition to the ROS, RNS also
are capable of enhancing MMP activities [23].
Selected transcription factors such as NFjB, AP-
1 and AP-2 are activated by UV irradiation [40].
These molecular mechanisms operate in an inter-
dependent manner. For example, NFjB activation
can in turn lead to the expression of a variety of
genes involved in immunological and inflamma-
tory responses, including genes that encode cyto-
kines, matrix degrading metalloproteases, adhesion
molecules and regulators of cell growth, differenti-
ation and cell death. MMPs are one of the
UV-responsive genes that degrade macromolecules
of the ECM. UV induces MMP transcription by
ª 2005 International Journal of Cosmetic Science, 27, 17–34
S. Pillai, C. Oresajo and J. Hayward
five-fold, however TIMP expression is either not
altered or only slightly elevated by UV [66, 67].
Figure 2 below summarizes the roles of UV and
ROS and the possible mechanisms of regulation of
MMP in skin.
MMP activity is regulated at multiple steps from
gene transcription to enzyme activation. The fol-
lowing sections detail some of the critical steps in
the regulation of MMPs.
Transcriptional activation
The regulatory sites in the MMP genes (5¢ flanking
region) contain an AP-1 regulatory element. Sev-
eral growth factors and cytokines stimulate the
expression of AP-1 transcription factors (Jun and
Fos) that form dimers, bind to the AP-1 binding site
of MMP genes and activate their gene expression
[68]. In addition to the AP-1 signalling system, the
TGF-b signalling system and the NFjB activated sys-
tem also play roles in MMP induction in the skin.
The mechanisms by which the cytokines and
growth factors regulate Jun and Fos involve three
distinct classes of protein kinases: mitogen-activated
protein kinases (MAPKs), extracellular stimulus-
regulated kinases (ERKs) stress-activated protein
kinase/Jun N-terminal kinases (SAPK/JNKs). NFjB
is especially important in UV- and inflammation-
induced activation of MMPs because of the role of
ROS in activating NFjB. Generation of ROS via
activation of NADPH oxidase also is necessary for
stimulation of MMP expression through the MAPK-
signalling pathways.
Zymogen activation
Most MMPs are synthesized and secreted as latent
precursors or zymogens. These have to be proteo-
lytically activated in the extracellular space. The
UV
Kinase Cascades
(MAPK,ERK, SAPK/JNK)
NFkB activation
MMP gene expression
C fos C jun ProMMPs
5’ AP-1 3’
Zymogen activation
Collagenases
Gelatinases
MMP activity stromelysin
Photoaging Imperfect repair Collagen breakdown
Figure 2 Matrix metalloprotease regulation by UV irra-
diation in skin.
23
Ultraviolet radiation and skin aging
latent MMPs are kept inactive by a ‘cysteine
switch’ formed by the interaction of a conserved
cysteine residue in the propeptide with the Zn in
the active site of the enzyme [69]. Disruption of
the cys-Zn bond and the cleavage of the prodo-
main lead to a conformational change and activa-
tion of the pro-MMPs. Examples of proteolytic
activation are the activation of collagenase
(MMP-1) by plasmin, trypsin, and stromelysin, and
activation of collagenase 3 (MMP-13) by plasmin,
stromelysin and gelatinase (MMP-2). The activa-
tion cascade between MMPs and other proteases
form a complex network in regulating tissue pro-
teolysis.
Regulation of proteolytic activity of MMPs
The third step in regulation of MMP activity in
skin is their endogenous inhibition. In the natural
state, MMPs are bound to non-specific inhibitors
such as a-2 macroglobulin and a-1 antiprotease
or to specific inhibitors such as the TIMPs [70].
Four different TIMPs have been identified. TIMP-1
and -2 are secreted in soluble form; TIMP-3 and
-4 are bound to the ECM. TIMPs bind MMPs in a
1 : 1 stoichiometry. There is a certain degree of
specificity for TIMP towards different MMPs. For
example TIMP-1 potentially inhibits most MMPs
except MMP-2 and MMP-14. TIMP-2 inhibits most
MMPs except MMP-9 [71]. The balance between
TIMPs and MMPs is critical in determining the
degradative potential of cells in normal and patho-
logical conditions. This balance is altered in dis-
eased states such as cancer development, tumour
invasion, angiogenesis and arthritis and in normal
conditions such as foetal development and chrono-
logical aging.
Neuroendocrine system and UV-induced skin
inflammatory responses
UV irradiation also causes alterations in the skin
immune system. There is increasing experimental
evidence that UV-induced skin inflammation is
influenced by the sensory nervous system and
the neuroendocrine system in the skin [72]. The
resulting complex network of cytokines, chemo-
kines, neuropeptides, neuropeptide-degrading
enzymes, neurohormones, and other inflamma-
tory mediators are involved in the infiltration of
neutrophils into skin and cutaneous inflamma-
tion. Neuropeptides such as substance P (SP) and
calcitonin gene-related peptide (CGRP) are
24
S. Pillai, C. Oresajo and J. Hayward
released from sensory nerves innervating the skin
upon UV exposure. In addition, a variety of cells
in the skin produce increased neuroendocrine
hormones such as proopiomelanocortin (POMC)
peptides and their receptors as well as neurotro-
phins after UV exposure. Neuropeptides and neu-
rohormones are capable of directly or indirectly
mediating UV-induced cutaneous inflammation by
the induction of vasodilatation, plasma extravasa-
tion, and augmentation of UV-induced cytokine,
chemokine, or cellular adhesion molecule expres-
sion required for activation and trafficking of
inflammatory cells into the inflamed tissue. Neu-
ropeptides and neurotrophins may also play a
role in the repair of cutaneous UV injury.
Protective role of UV
The ability of UV to induce pigmentation in vivo
and in vitro is well documented, but the intracel-
lular signals that trigger this response are poorly
understood. In addition to the roles of keratino-
cyte-produced NO and a-MSH (described in Influ-
ence on other cell types in skin section) in
protecting skin from further damage by UV, UV
radiation also stimulates melanocytes directly to
produce more melanins. A variety of UV-induced
cytokines produced by keratinocytes such as
basic fibroblast growth factor (b-FGF), nerve
growth factor (NGF), endothelin-1 and the
POMC-derived peptides MSH, ACTF, beta-LPH
and beta-endorphin stimulate melanocyte mitosis,
increase melanogenesis, enhance melanocyte
dendricity and prevent apoptotic cell death fol-
lowing UV injury [36]. A novel mechanism by
which UV damage to cellular DNA induces skin
melanogenesis and skin tanning was elucidated
by pioneering work from Dr Gilchrest’s laborat-
ory [73]. At the molecular level, UV radiation
causes cellular DNA damage [36] and activates
cellular-signalling pathways [7]. Increasing DNA
repair after irradiation enhanced UV-induced
melanogenesis. This response comes from the
small DNA fragment thymine dinucleotides
(pTpT), selectively excised during the repair of
UV-induced DNA photoproduct. pTpT stimulated
melanogenesis in mammalian pigment cells and
intact skin, mimicking the effects of UV irradi-
ation on skin [11]. The mechanism by which
pTpT activates melanogenesis appear to be medi-
ated by activation of p53 gene transcription,
leading to nuclear accumulation of this protein
ª 2005 International Journal of Cosmetic Science, 27, 17–34
Ultraviolet radiation and skin aging
and increasing the specific binding of this tran-
scription factor to its DNA consensus sequence
[74]. Thymidine oligonucleotides also induced
protein kinase C beta levels suggesting that this
DNA excised fragment induces tanning by upreg-
ulating the levels of p53 gene mediated at least
in part by stimulating protein kinase cascade
[75].
Strategies to prevent photoageing and
inflammation-induced matrix
degradation
UV-induced inflammation and the resultant photo-
aging can be prevented by various strategies out-
lined below.
Prevention of UV penetration to skin
Ultraviolet A radiation of wavelength 320–
400 nm penetrates the epidermis resulting in
damage to the dermis. This causes skin to age pre-
maturely, with effects that include roughening,
blotchiness, sagging and wrinkles. UVA also con-
tributes to the development of skin cancer but it is
only during the last decade that the damaging
effects of UVA have been fully appreciated. UVA
absorbers are chemicals that absorb radiation in
the 320–360 nm region such as benzophenones,
anthranilates and dibenzoyl methanes. Microfine
zinc oxide is a physical blocker and is particularly
effective in this region (Table I).
Ultraviolet B is radiation between 290 and
320 nm and causes sunburn and skin cancer. UVB
radiation inhibits DNA, RNA and protein synthesis
and induces erythemal responses. The principal
UVB absorbers are para-aminobenzoic acid deriva-
tives, salicylates, cinnamates and camphor deriva-
tives plus microfine titanium dioxide (Table I).
Ultraviolet C radiation from 100 to 290 nm is
filtered out by the ozone layer and does not reach
the earth’s surface, hence the current concern
about ozone depletion. The only sunscreen that is
Table I Commonly used sunscreens in cosmetics and OTC products and their UV absorption range
UV spectrum (nm) Sunscreens approved for topical use
UVA (320–400) Benzophenones (benzophenone 3), anthranilates, dibenzoyl derivatives (avobenzone or parsol 1789)
UVB (290–320) PABA derivatives, salicylates (octyl salicylate, homosalate), cinnamates (octylmethxycinnamate)
UVC (100–290) Physical UV blockers such as microfine titanium dioxide, zinc oxide
ª 2005 International Journal of Cosmetic Science, 27, 17–34
S. Pillai, C. Oresajo and J. Hayward
effective in this region would be microfine titanium
dioxide or zinc oxide of sufficiently small particle
size (Table I).
Evidence that a full-UV spectrum sunscreen can
protect human skin against the biological changes
occurring in photoaging was presented by work
performed by L’Oreal [76]. Daily use of a cream
containing a photostable combination of UVB and
UVA absorbers providing a continuous absorption
through the entire UV spectrum protected skin of
volunteers from damage from solar simulated radi-
ation (SSR) damage. Daily exposure to SSR for
6 weeks caused skin erythema, hyperpigmenta-
tion, increased skin thickness because of elastolysis
and decrease in collagen, increase in MMP-2
expression and increased deposition of lysozyme
and a-1-antitrypsin on elastic fibres. Daily use of
the sunscreens prevented all these biological chan-
ges [76]. Prevention of UV penetration using other
physical sunscreens should also prevent activation
of MMPs in the epidermis and dermis by UVA and
UVB.
Prevention of inflammation using antioxidants
or anti-inflammatories
Antioxidants
Topical antioxidants can prevent generation of
ROS and subsequent inflammatory reactions. ROS
are generated either by the normal process of
metabolism whereby excess electrons generated in
the mitochondrial respiratory chain are donated to
molecular oxygen to generate superoxide anions,
or by UV-induced non-enzymatic conversion of
molecular oxygen into superoxide anion radical.
In addition, under certain circumstances UV and
visible lights are capable of causing excitation of
molecular oxygen in the skin to form highly react-
ive singlet species.
An interlinked network of enzymes that convert
ROS to harmless water and molecular oxygen
regulates skin’s antioxidant defence system. How-
ever, it is clear from recent studies that the levels
25
Ultraviolet radiation and skin aging
of these major antioxidant enzymes are decreased
with age and in photoaging [8]. In addition, the
levels of endogenous antioxidants such as tocophe-
rols, ascorbic acid and glutathione, the activities of
enzymes that generate these antioxidants such as
glutathione reductase and glutathione peroxidase,
and the activities of other antioxidant enzymes
that directly inactivate ROS such as superoxide
dismutase and catalase are all reduced in intrinsic
aging and in photoaging in human skin [8, 20,
21, 77, 78]. Sun exposure variation because of
seasonal changes (summer versus winter) also
affects the levels of antioxidant enzyme activities
in the stratum corneum. Levels of catalase in stra-
tum corneum are lesser in summer compared with
winter, although SOD did not follow this pattern
[79]. An acute exposure to UVA or UVB also
reduced catalase activity without affecting the SOD
activity [79]. The recovery of the catalase enzyme
after UV exposure occurred in 3–4 weeks, follow-
ing an age-dependent fashion. Recovery was faster
in younger individuals than in older individuals
[79]. In normal human skin, there is an antioxid-
ant gradient in the stratum corneum with lower
concentrations in the outer layers and higher con-
centrations in the deeper layers. This gradient is
disrupted with UV and ozone exposure, suggesting
that oxidative stress disrupts the antioxidant distri-
bution of skin [78].
As the levels of endogenous anti-oxidants such
as reduced glutathione (GSH) decrease with age, it
was suggested that addition of exogenous antioxi-
dants might reduce inflammatory responses to UV
radiation and the aging process itself. Topical
application of several biologically relevant antioxi-
dants such as vitamin E, ascorbic acid and carote-
noids have been shown to provide photoprotection
and reduce photodamage in human skin [80]. In
addition, a variety of naturally occurring antioxid-
ant compounds from plant origin have been dem-
onstrated to be beneficial by quenching free
radical generation and preventing photodamage in
skin [80, 81]. Using in vitro models it was demon-
strated that the dietary antioxidant quercetin,
reverses age-related GSH depletion and oxidative
stress [82]. In fact, several polyphenolic antioxi-
dants of plant origin have been shown to be bene-
ficial in preventing photodamage and skin
inflammation. These include polyphenols from
green tea [9, 83, 84], grape seed [85], pomegran-
ate [86] and a variety of other plant extracts
[87, 88]. An excellent recent review summarizes
26
S. Pillai, C. Oresajo and J. Hayward
the effects of natural phenolic compounds and
their role in the prevention of UV-induced skin
damage [89]. A list of the natural polyphenolic
antioxidants is shown in the Table II. All these
studies suggest that both dietary and topical anti-
oxidants could play a significant role in the reduc-
tion of inflammatory responses.
Anti-inflammatories
Anti-inflammatory actives (both steroidal and non-
steroidal) have been demonstrated to prevent skin
inflammation caused by UV irradiation [90]. These
agents generally act by suppressing the generation
of inflammatory agents such as prostaglandins,
leukotrines or inflammatory cytokines. Among the
most commonly used are topical retinoids, which
have been shown to block the UV-induced activa-
tion of inflammation mediated by transcription fac-
tors [91]. The recent introduction of several
analogues with increased potency and reduced
irritation, retinoid therapy has become a common
treatment for reversing the signs of photoaging
[92–94]. Other therapies include use of vitamin D
[95], corticosteroids and newly developed non-ste-
roidal immunosuppressive and anti-inflammatory
agents such as tacrolimus and pimecrolimus
[96, 97]. Although these agents are currently
administered mostly for inflammatory skin dis-
eases, there is potential for their use as an anti-
inflammatory in acute UV damage.
Inhibition of gene activation of MMPs
Among the most used therapies for prevention and
reversal of photodamage are topical retinoids
[98–100]. Retinoids are a group of precursors of
retinoic acid that can be converted to retinoic acid
within the skin under normal physiological condi-
tions. These include retinol, retinal, retinyl esters
with various fatty acids and other glycosidic pre-
cursors of retinoic acid Topical treatment with all-
trans retinoic acid prevents UV induction of MMPs
by multiple mechanisms. One mechanism is by
prevention of UV-induced accumulation of c-Jun
protein, which is required for MMP expression. In
addition all-trans retinoic acid may also stimulate
the breakdown of jun protein through ubiquitin–
proteasome-mediated pathways [101]. Topical
all-trans retinoic acid prevented UVB induction of
MMP-1, -3 and -9. Retinoic acid has no effect on
erythema suggesting that the effect on gene expres-
sion of MMP is specific. Topical glucocorticoids
ª 2005 International Journal of Cosmetic Science, 27, 17–34
 Table II Natural polyphenolic antioxidants and anti-inflammatory compounds

Compounds (and their class) Natural sources Biological activities, skin benefits
Ultraviolet radiation and skin aging

Caffeic acid ferulic acid Basil, thyme, taqragon, asafetida, Protects membranes from lipid peroxidation, protects skin from UV-induced erythema,
(cinnamic acid derivative) chia, apple, peanuts photoprotective agent
Quercetin (flavonoid) Evening primerose leaves, onion, tea A natural flavonoid, powerful antioxidant, MMP inhibitor, Protects skin antioxidant systems,
anti-inflammatory, protects UV-induced lipid peroxidation
Apigenin (flavonoid) Apples, beans, onions, broccoli, etc. Prevents UV-induced carcinogenesis, inhibits ODC activity, antioxidant
Genistein (isoflavone) Soybean, Lima bean Tumor protector, phytoestrogen, protects from UV-induced hydrogen peroxide generation,
reduce inflammation
Resveratrol (phytoalexin) Grape skin, grape seed, peanuts Protects from UV-induced tumor initiation, LOX, ODC and COX inhibitor,

ª 2005 International Journal of Cosmetic Science, 27, 17–34

Nordihydroguaiaretic acid
Carnosic acid, ursolic acid,
rosemarinic acid (triterpinoids)
Silymarin (flavonolignan)
Epicatechin, epicatechin-3-gallate,
epigallocatechin and epigallocatechin gallate
Procyanidins, proanthocyanidins,
gallotannins and ellagotannins (tannins)
Pycnogenol
Chaparral leaves
Sage, rosemary, periwinkle, thyme
Milkthistle
Tea leaves (Camellia sinesis) in
general,
green tea in particular
Red grape skin, eyebright
bark of pine tree (pinus maritime)
protects from UV-induced lipid peroxidation, phytoestrogen,
Powerful antioxidant, MMP inhibitor, LOX and COX inhibitor
Chemoprotective, elastase inhibitor
Inhibition of UV-induced erythema, COX inhibition, ODC induction,
prevents UV-induced infiltration of leukocytes, inhibits NFjB activation by UVB
Protects UV-induced carcinogenesis, UV protection, blocks tumor invasion,
prevents UV-induced edema, lipid peroxidation, general protection from UV-induced skin aging
Produced in plants to protect plant against UV radiation. Reduces hydrogen peroxide generation,
decrease tumor incidence, inhibit induction of ODC, and epidermal DNA synthesis
Prevent UV-induced tumor induction, anti-inflammatory

COX, cyclooxygenase; LOX, lipooxygenase; MMP, matrix metalloprotease; NFjB, nuclear factor kappa B; ODC, ornithine decarboxylase; UV, ultraviolet.
S. Pillai, C. Oresajo and J. Hayward

27
Ultraviolet radiation and skin aging S. Pillai, C. Oresajo and J. Hayward

may also control MMP gene activation mediated
by its anti-inflammatory actions [102]. As dis-
cussed in the UV, photoaging and skin changes
section, prevention of UV penetration to skin using
sunscreens can also prevent MMP induction in
response to UV [76].
Direct inhibition of MMPs
Many natural or synthetic inhibitors can directly
block MMPs (Table III). Natural products inhibit
MMPs non-specifically. Synthetic compounds are
designed for specific interaction with the enzymes,
mostly by chelating the Zn atom from the active
site. Inhibition of MMPs using this strategy would
inhibit all the MMPs, without any specificity. This
strategy carries a risk in that activities of some
MMPs are required for cleavage and removal of
damaged ECM components. For example, collage-
nase-1 (MMP-1) is activated by UV and is respon-
sible for dermal matrix damage. However,
gelatinases (MMP-2 and -9) not only are not
induced by UV, their activity is needed to ‘clear
away’ the damage produced by MMP-1. Therefore,
a more preferred strategy is to selectively inhibit
certain MMPs (e.g. MMP-1) while sparing other
MMPs (MMP-2 and -9). Currently such selective
inhibitors are not available; however, such a strat-
egy has been patented [103]. Recently, a biotech-
nologically derived human identical TIMP-2 has
been introduced as a cosmetic ingredient [104].
Natural non-specific inhibitors include: flavo-
noids [105, 106] avocado and soybean unsaponi-
fyables [107]; green tea polyphenols [9, 108];
plant terpenoids such as quercetin [109, 110];
polymethoxy flavonoid, nobiletin, from citrus
depressa [111]; fucose and fucose-rich polysaccha-
rides [112]; and some plant extracts such as spa-
tholobi caulis [91] euonymus alatus [113] and
Rhizoma notopterygii [114].
Several synthetic inhibitors of MMPs have been
synthesized. These include hydroxamic acid deriva-
tives (Batimastat, Marimastat, AG 3340), butanoic
acid analogues (BAY 12-9566) and others [115].
One of the most potent inhibitor so far described is
Ilomastat (aka Galardin, GM-6001), a hydroxamic
acid derivative of a pseudo-dipeptide. Ilomastat
inhibits MMPs in the nanomolar range by chelat-
ing the Zn metal ion from the enzyme active site
[116, 117]. Other drugs in development are Neov-
astat an active from shark cartilage (AE-941; from
Aeterna Laboratories, Quebec, Canada) and a vari-
ety of synthetic inhibitors from British Biotech
Pharmaceuticals Ltd, Oxford, UK [118].
Inhibitors of leucocyte proteases
Considerable energy has been devoted to develop-
ing inhibitors of serine proteinase elastase (human
leukocyte elastase or HLE) and Cathepsin G. Inhi-
bition of neutrophil elastase could in addition to
blocking ECM degradation, also prevent the prote-
ase–antiprotease imbalance observed in inflamma-
tion and photoaging.
In vitro inhibition of human leukocyte elastase
by non-peptidic natural products of plant origin
has been described [119, 120]. Pentacyclic triterp-
enoid metabolites of plant origin are inhibitors of

Table III Natural and synthetic matrix metalloprotease (MMP) inhibitors

Inhibitor (class) Mechanism/Specificity

Neovastat (shark cartilage fraction) General inhibitor of MMP, possibly by chelation of Zn

Galardin or Ilomastat
(hydroxamate derivative)
Batimastat, Marimastat, AG 3340
(hydroxamate derivatives)
BAY 12-9566 (butanoic acid analogues
Green tea polyphenols
Several plant extracts
Quercetin
Nordihydroguaiaretic acid
Nobilin (citrus polyphenol)
Highly specific high affinity inhibitor of MMPs, chelates Zn from active site.
General MMP inhibitor, Zn chelation
Zn chelation
Mode of action may be related to reducing inflammation; also directly
inhibits MMP by protein interaction
Inhibition maybe by interfering with transcription regulation by interfering with reactive
oxygen species- or inflammation-mediated MMP gene induction
Possibly by interfering with inflammation-induced MMP gene transcription activation
Same as above
Same as above

28 ª 2005 International Journal of Cosmetic Science, 27, 17–34

Ultraviolet radiation and skin aging S. Pillai, C. Oresajo and J. Hayward

Table IV Leukocyte elastase inhibitors

Compound Sources Mechanism of inhibition

Ursolic acid and other plant terpenoid derivatives Apple, rosemary, basil, Holy basil Reversible inhibition
Fatty acids, bile acids Fish oils, vegetable oils Non-specific inhibition
Pyrene trisulfonic acid Synthetic Non-specific inhibition
N-Phenylphosphoonyl-leucyl-tryptophan Synthetic Specific inhibitor of skin fibroblast elastase

hydrolysis of both synthetic peptide substrates and
elastin by HLE. Ursolic acid, the most potent of
these compounds, has an inhibition constant of
4–6 lm for hydrolysis of peptide substrates [119].
Other non-specific inhibitors of HLE are fatty
acids, bile acids, and pyrene trisulfonic acid
(Table IV) [121–124]. These compounds are all
reversible inhibitors, and form no covalent interac-
tions with the enzyme. Because they bind at the
extended substrate-binding site, they are competit-
ive inhibitors of enzymatic hydrolysis of larger
oligopeptide substrates and proteins such as elas-
tin, but are non-competitive inhibitors of hydroly-
sis of the smallest synthetic substrates, which bind
only in the immediate vicinity of the catalytic triad
within the active site.
Specific inhibition of skin fibroblast elastase
activity has been demonstrated by a group at Kao
Corporation to block UV-induced wrinkle forma-
tion [55]. Topical application of N-phenetylphos-
phonyl-leucyl-tryptophane, an inhibitor of skin
fibroblast elastase appears to inhibit wrinkle for-
mation by UV in rats. Six weeks of UVB irradiation
significantly enhanced elastase activity in rat skin,
whereas topical N-phenetylphosphonyl-leucyl-
tryptophane prevented this increase. These find-
ings suggest that skin fibroblast elastase plays an
equally important role as HLE in mediating UV
effects on skin. Strategies to block both the HLE
and fibroblast elastase would therefore provide
enhanced protection.
Summary
UV irradiation initiates and activates a complex
cascade of biochemical reactions in human skin
(Fig. 3). In short, UV causes depletion of cellular
antioxidants and antioxidant enzymes (SOD, cat-
alase), initiates DNA damage leading to the forma-
tion of thymidine dimmers, activates the
neuroendocrine system leading to immunosuppres-
sion and release of neuroendocrine mediators and
causes increased synthesis and release of pro-
inflammatory mediators from a variety of skin
cells. The pro-inflammatory mediators increase the
permeability of capillaries leading to infiltration
and activation of neutrophils and other phagocytic
cells into the skin. The net result of all these effects
is inflammation and free radical generation (both
reactive oxygen and nitrogen species).

Protective Neuroendocrine Release of

Inflammatory mediators ROS generation
role release

DNA damage
UV IL-1 alpha and TNF-a release
from keratinocytes
Activation of skin
Neuroendocrine system Synthesis and release of prostaglandins
Thymidine And release of neuropeptides, leucotrines, secondary cytokines
Dimer formation histamines (IL-6, IL-8) from skin cells Depletion of
cellular antioxidants

Melanocyte Inflammation Activation of neutrophils

Macrophages
activation
Cellular protein, lipid
Activation of transcription and CHO oxidation
Free Radical Generation
Factors (AP-1, AP-1, NFkB) Accumulation of ROS, NOS
Skin tanning/
darkening
MMP transcription Release of Neutrophil
activation elastase
Cell death,
Protection from lipid peroxidation
further UV damage Abnormal matrix Disruption of
Figure 3 A summary of the differ- degradation Protease-antiprotease
balance
ent mechanisms initiated by UV that
Accumulation of non-
Accumulation of
lead to reactive oxygen species gen- Functional matrix components
abnormal proteins,
cellular debris
eration, inflammation and photoag-
ing in skin. Photoaging

ª 2005 International Journal of Cosmetic Science, 27, 17–34 29

Ultraviolet radiation and skin aging
Elastases and other proteases (cathepsin G)
released from neutrophils cause further inflamma-
tion and activation of matrix metalloproteases.
The inflammation further activates the transcrip-
tion of various matrix degrading metalloproteases,
leading to abnormal matrix degradation and accu-
mulation of non-functional matrix components.
Furthermore, the inflammation and ROS cause
oxidative damage to cellular proteins, lipids and
carbohydrates, which accumulates in the dermal
and epidermal compartments, contributing to the
aetiology of photoaging.
Strategies to prevent photodamage caused by
this cascade of reactions initiated by UV include:
prevention of UV penetration into skin by physical
and chemical sunscreens, prevention/reduction of
inflammation using anti-inflammatory compounds
(e.g. cyclooxygenase inhibitors, inhibitors of cyto-
kine generation); scavenging and quenching of
ROS by antioxidants; inhibition of neutrophil ela-
stase activity to prevent ECM damage and activa-
tion of MMPs, and inhibition of MMP expression
(e.g. by retinoids) and activity (e.g. by natural and
synthetic inhibitors).
References
1. Melov, S. Mitochondrial oxidative stress: physiolog-
ic consequences and potential for a role in aging.
Ann. NY Acad. Sci. 908 219–225 (2000).
2. Ashok, B.T. and Ali, R. The aging paradox: free
radical theory of aging. Exp. Gerontol. 34, 293–303
(1999).
3. Wickens, A.P. Ageing and the free radical theory.
Respir. Physiol. 128, 379–391 (2001).
4. Bernstein, E.F., Brown, D.B., Urback, F. et al. UV
radiation activates the human elastin promoter in
transgenic mice: a novel in vivo and in vitro model
of cutaneous photoaging. J. Investig. Dermatol. 105,
269–273 (1995).
5. Lavker, R.M. Structural alterations in exposed and
unexposed aged skin. J. Invest. Dermatol. 73, 559–
566 (1979).
6. Lavker, R.M. and Kligman, A.M. Chronic helio-
dermatitis: a morphological evaluation of chronic
actinic dermal damage with emphasis on the role
of mast cells. J. Invest. Dermatol. 90, 325–330
(1988).
7. Gilchrest, B.A. Skin aging and photoaging: an over-
view. J. Am. Acad. Dermatol. 21, 610–613 (1989).
8. Rhie, G., Shin, M., Seo, J.Y. et al. Aging and photo-
aging dependent changes of enzymic and non-enzy-
mic antioxidants in the epidermis and dermis of
30
S. Pillai, C. Oresajo and J. Hayward
human skin in vivo. J. Invest. Dermatol. 117, 1212–
1217 (2001).
9. Katiyar, S.K. and Mukhtar, H. Green tea polyphenol
(-)-epigallocatechin-3-gallate treatment to mouse
skin prevents UVB-induced infiltration of leukocytes,
depletion of antigen-presenting cells, and oxidative
stress. J. Leukoc. Biol. 69, 719–726 (2001).
10. Wood, L.C., Elias, P.M., Calhoun, C. et al. Barrier
disruption stimulates interleukin-1 alpha expres-
sion and release from a pre-formed pool in mu-
rine epidermis. J. Invest. Dermatol. 106, 397–403
(1996).
11. Gilchrest, B.A. and Eller, M.S. DNA photodamage
stimulates melanogenesis and other photoprotec-
tive responses. J. Investig. Dermatol. Symp. Proc. 4,
35–40 (1999).
12. Gonzalez, S. and Pathak, M.A. Inhibition of UV
induced formation of ROS, lipid peroxidation, ery-
thema and skin photosensitization by polypodium
leucotomos. Photodermatol. Photoimmunol. Pho-
tomed. 12, 45–56 (1996).
13. Kawaguchi, Y., Tanaka, H., Okada, T. et al. The
effects of UV A and reactive oxygen species on the
mRNA expression of type IV collagenase and its tis-
sue inhibitor in cultured human dermal fibroblasts.
Arch. Dermatol. Res. 288, 39–44 (1996).
14. Takashima, A. and Bergstresser, P.R. Impact of UV
radiation on the epidermal cytokine network. Phot-
ochem. Photobiol. 63, 397–400 (1996).
15. Krutmann, J. and Grewe, M. Involvement of cytok-
ines, DNS damage and reactive oxygen intermedi-
ates in UV induced modulation of intercellular
adhesion molecule-1 expression. J. Invest. Dermatol.
105 (Suppl. 1), 67S–70S (1995).
16. Bielenberg, D.R., Bucana, C.D., Sanchez, R. et al.
Molecular regulation of UVB induced cutaneous
angiogenesis. J. Invest. Dermatol. 111, 864–872
(1998).
17. Hruza, L.L. and Pentland, A.P. Mechanisms of UV-
induced inflammation. J. Invest. Dermatol. 100,
35S–41S (1993).
18. Soter, N.A. Acute effects of UV on the skin. Semin.
Dermatol. 9, 11–15 (1990).
19. Haratake, A., Uchida, Y., Mimura, K., Elias, P.M.
and Holleran, W.M. Intrinsically aged epidermis
displays diminished UVB-induced alterations in bar-
rier function associated with decreased prolifer-
ation. J. Invest. Dermatol. 108, 319–323 (1997).
20. Thiele, J.J. Oxidative targets in the stratum cor-
neum, a new basis for antioxidative strategies.
Skin Pharmacol. Appl. Skin Physiol. 1, 87–91
(2001).
21. Sander, C.S., Chang, H., Salzmann, S. et al. Photo-
aging is associated with protein oxidation in
human skin in vivo. J. Invest. Dermatol. 118, 618–
625 (2002).
ª 2005 International Journal of Cosmetic Science, 27, 17–34
Ultraviolet radiation and skin aging
22. Kang, S., Fisher, G.J. and Voorhees, J.J. Photoaging:
pathogenesis, prevention, and treatment. Clin. Geri-
atr. Med. 17, 643–659 (2001).
23. Brenneisen, P., Sies, H. and Scharffetter-Kochanek,
K. UV-B irradiation and matrix metalloproteinases:
from induction via signaling to initial events. Ann.
NY Acad. Sci. 973, 31–43 (2002).
24. Kupper, T.S. and Groves, R.W. The interleukin-1
axis and cutaneous inflammation. J. Investig. Der-
matol. 105, 62S–66S (1995).
25. Kock, A., Schwarz, T., Kirnbauer, R. et al. Human
keratinocytes are a source for tumor necrosis factor
alpha: evidence for synthesis and release upon sti-
mulation with endotoxin or UV light. J. Exp. Med.
172, 1609–1614 (1990).
26. Hirao, T., Aoki, H., Yoshida, T., Sato, Y. and Ka-
moda, H. Elevation of interleukin-1 receptor antag-
onist in the stratum corneum of sun-exposed and
UV B-irradiated human skin. J. Invest. Dermatol.
106, 1102–1107 (1996).
27. Shreedhar, V., Giese, T., Sung, V.W. and Ullrich,
S.E. A cytokine cascade including prostaglandin E2,
IL-4, and IL-10 is responsible for UV-induced sys-
temic immune suppression. J. Immunol. 160,
3783–3789 (1998).
28. Brink, N., Szamel, M., Young, A.R., Wittern, K.P.
and Bergemann, J. Comparative quantification of
IL-1 beta, IL-10, IL-10r, TNF alpha and IL-7
mRNA levels in UV-irradiated human skin in vivo.
Inflamm. Res. 49, 290–296 (2000).
29. Kondo, S., The roles of keratinocyte-derived cytok-
ines in the epidermis and their possible responses to
UVA-irradiation. J. Investg. Dermatol. Symp. Proc. 4,
177–183 (1999).
30. Katiyar, S.K. Skin photoprotection by green tea:
antioxidant and immunomodulatory effects. Curr.
Drug Targets Immune Endocr. Metabol. Disord. 3,
234–242 (2003).
31. Schwarz, T. and Luger, T.A. Effect of UV irradiation
on epidermal cell cytokine production. J. Photochem.
Photobiol. B 4, 1–13 (1989).
32. Luger, T.A., Scholzen, T. and Grabbe, S. The role of
alpha melanocyte stimulating hormone in cutane-
ous biology. J. Investig. Dermatol. Symp. Proc. 2,
87–93 (1997).
33. Mildner, M., Weninger, W., Trautinger, F., Ban, J.
and Tschachler, E. UVA and UVB radiation differ-
entially regulate vascular endothelial growth factor
expression in keratinocyte-derived cell lines and in
human keratinocytes. Photochem. Photobiol. 70,
674–679 (1999).
34. Lee, S.C., Lee, J.W., Jung, J.E. et al. Protective role
of nitric oxide-mediated inflammatory response
against lipid peroxidation in UVB-irradiated skin.
Br. J. Dermatol. 142, 653–659 (2000).
ª 2005 International Journal of Cosmetic Science, 27, 17–34
S. Pillai, C. Oresajo and J. Hayward
35. Seo, S.J., Choi, H.G., Chung, H.J. and Hong, C.K.
Time course of expression of mRNA of inducible
nitric oxide synthase and generation of nitric oxide
by UV B in keratinocytes cell lines. Br. J. Dermatol.
147, 655–662 (2002).
36. Gilchrest, B.A., Park, H.Y., Eller, M.S. and Yarr,
M. Mechanisms of UV induced pigmentation. Photo-
chem. Photobiol. 63, 1–10 (1996).
37. Suh, D.H., Kwon, T.E. and Youn, J.I. Changes of
comedonal cytokines and sebum secretion after UV
irradiation in acne patients. Eur. J. Dermatol. 12,
139–144 (2002).
38. Trenum, C.W., Blake, D.R. and Morris, C.J. Skin
inflammation: reactive oxygen species and the role
of iron. J. Invest. Dermatol. 99, 675–682 (1992).
39. Chlopicki, S., Olszanecki, R., Janiszewski, M. et al.
Antioxid. Redox Signal. 6, 691–698 (2004).
40. Abeyama, K., Eng, W., Jester, J.V. et al. A role for
NF-kappa B dependent gene transactivation in sun-
burn. J. Clin. Invest. 105, 1751–1759 (2000).
41. Rittie, L. and Fisher, G.J. UV-light-induced signal
cascades and skin aging. Ageing Res Rev. 1, 705–
720 (2002).
42. Krishnadasan, B., Naidu, B.V., Byrne, K. et al. The
role of proinflammatory cytokines in lung ischemia-
reperfusion injury. J. Thorac. Cardiovasc. Surg. 125,
261–272 (2003).
43. Hales, J.M. and Camp, R.D. Potent T cell stimulato-
ry material with antigenic properties in stratum
corneum of normal human skin. J. Invest. Dermatol.
110, 725–729 (1998).
44. Tanaka, N., Fujioka, A., Tajima, S., Ishibashi,
A. and Hirose, S. Elafin is induced in epidermis in
skin disorders with dermal neutrophilic infiltration:
interleukin-1 beta and tumor necrosis factor-alpha
stimulate its secretion in vitro. Br. J. Dermatol. 143,
728–732 (2000).
45. Drew, B. and Leeuwenburgh, C. Aging and the role
of reactive nitrogen species. Ann. NY Acad. Sci.
959, 66–81 (2002).
46. Weissmann, G., Smolen, J.E. and Hoffstein, S. Poly-
morphonuclear leukocytes as secretory organs of
inflammation. J. Investig. Dermatol. 71, 95–99
(1978).
47. Pol, A., Pfundt, R., Zeeuwen, P., Molhuizen, H. and
Schalkwijk, J. Transcriptional regulation of the ela-
fin gene in human keratinocytes. J. Invest. Derma-
tol. 120, 301–307 (2003).
48. Nakane, H., Ishida-Yamamoto, A., Takashashi, H.
and Ilzuka, H. Elafin, a secretory protein, is cross-
linked into the cornified cell envelopes from the
inside of psoriatic keratinocytes. J. Invest. Dermatol.
119, 50–55 (2002).
49. Simon, S.R., Roemer, E.J., Golub, L.M. and Rama-
murthy, N.S. Serine proteinase inhibitory activity
31
Ultraviolet radiation and skin aging
by hydrophobic tetracycline. US Patent no.
5773430 (1998).
50. Rogalski, C., Ulf, M.-H., Proksch, E. and Wiedow,
O. Human leukocyte elastase induces keratinocyte
proliferation in vitro and in vivo. J. Invest. Dermatol.
118, 49–54 (2002).
51. Piskin, G., Tursen, U., Bos, J.D. and Teunissen, M.B.
IL-4 expression by neutrophils in psoriasis lesional
skin upon high-dose UVB exposure. Dermatology
207, 51–53 (2003).
52. Schaerli, P., Britschgi, M., Keller, M. et al. Charac-
terization of human T cells that regulate neutrophi-
lic skin inflammation. J. Immunol. 173, 2151–
2158 (2004).
53. Meyer-Hoffert, U., Wingertszahn, J. and Wiedow, O.
Human leukocyte elastase induces keratinocyte pro-
liferation by epidermal growth factor receptor activa-
tion. J. Investig. Dermatol. 123, 338–345 (2004).
54. Starcher, B., O’Neal, P., Granstein, R.D. and Beis-
sert, S. Inhibition of neutrophil elastase suppresses
the development of skin tumors in hairless mice.
J. Invest. Dermatol. 107, 159–163 (1996).
55. Tsukahara, K., Takema, Y., Moriwaki, S. et al.
Selective inhibition of skin fibroblast elastase elicits
a concentration-dependent prevention of UV B-
induced wrinkle formation. J. Invest. Dermatol. 117,
671–677 (2001).
56. Mallya, S.A., Hall, J.E., Lee, H.M. et al. Interaction of
matrix metalloproteinases with serine protease
inhibitors. Ann. NY Acad. Sci. 732, 303–314 (1994).
57. Rinehart, A.R., Mallya, S., Simon, S.R. et al.
Human alpha-proteinase inhibitor binds to extracel-
lular matrix in vitro. Am. J. Respir. Cell Mol. Biol. 9,
666–679 (1993).
58. Henry, M.T., McMahon, K., Costello, C., Fitz-
gerald, M.X. and O’Connor, C.M. Secretory leuko-
cyte proteinase inhibitor and elafin are resistant
to degradation by MMP-8. Exp. Lung Res. 28,
85–97 (2002).
59. Zhang, Y., DeWitt, D.L., McNeely, T.B., Wahl, S.M.
and Wahl, L.M. Secretory leukocyte protease inhib-
itor suppresses the production of monocyte PGH
synthase-2, PGE2 and MMPases. J. Clin. Invest. 99,
894–900 (1997).
60. Kahari, V.M. and Saarialho-Kere, U. Matrix metal-
loproteinases in skin. Exp. Dermatol. 6, 199–213
(1997).
61. Brennan, M., Bhatti, H., Nerusu, K.C. et al. Matrix
metalloproteinase-1 is the major collagenolytic
enzyme responsible for collagen damage in UV-irra-
diated human skin. Photochem. Photobiol. 78,
43–48 (2003).
62. Fisher, G.J., Choi, H.C., Bata-Csorgo, Z. et al. UV
irradiation increases matrix metalloproteinase-8
protein in human skin in vivo. J. Invest. Dermatol.
117, 219–226 (2001).
32
S. Pillai, C. Oresajo and J. Hayward
63. Fisher, G.J., Datta, S.C., Talwar, H.S. et al. Molecu-
lar basis of sun-induced premature skin ageing and
retinoic antagonism. Nature 379, 335–339 (1996).
64. Ashcroft, G.S., Herrick, S.E., Tarnuzzer, R.W. et al.
Human ageing impairs injury-induced in vivo
expression of tissue inhibitor of matrix metallopro-
teinases (TIMP)-1 and -2 proteins and mRNA.
J. Pathol. 183, 169–176 (1997).
65. Scharffetter, K., Wiaschek, M., Hogg, A. et al. UVA
irradiation induces collagenase in human dermal fi-
broblasts in vitro and in vivo. Arch. Dermatol. Res.
283, 506–511 (1991).
66. Herrmann, G., Wlasehek, M., Lange, T.S. et al. UVA
irradiation stimulates the synthesis of various mat-
rix metalloproteinases in cultured human fibro-
blasts. Exp. Dermatol. 2, 92–97 (1993).
67. Petersen, M.J., Hansen, C. and Craig, S. Ultraviolet
A irradiation stimulates collagenase production in
cultured human fibroblasts. J. Invest. Dermatol. 99,
440–444 (1992).
68. Mauviel, A. Cytokine regulation of metalloprotein-
ase gene expression. J. Cell Biochem. 53, 288–295
(1993).
69. Van Wart, H.E. and Birkedal-Hansen, H. The cys-
teine switch: a principle of regulation of metallopro-
teinase activity with potential applicability to the
entire matrix metalloproteinase gene family. Proc.
Natl Acad. Sci. USA 87, 5578–5582 (1990).
70. Birkedal-Hansen, H. Moore, W.G.I., Bodden, M.K.
et al. Matrix metalloproteinases: a review. Crit. Rev.
Oral Biol. Med. 4, 197–250 (1993).
71. Watson, S.A. and Tierney, G. Matrix metalloprote-
inase inhibitors: a review. BioDrugs 9, 325–335
(1998).
72. Scholzen, T.E., Brzoska, T., Kalden, D.H. et al.
Effect of UV light on the release of neuropeptides
and neuroendocrine hormones in the skin: media-
tors of photodermatitis and cutaneous inflamma-
tion. J. Investig. Dermatol. Symp. Proc. 4, 55–60
(1999).
73. Eller, M.S., Ostrom, K. and Gilchrest, B.A. DNA
damage enhances melanogenesis. Proc. Natl Acad.
Sci. USA 93, 1087–1092 (1996).
74. Eller, M.S., Maeda, T., Magnoni, C., Atwal, D. and
Gilchrest, B.A. Enhancement of DNA repair in
human skin cells by thymidine dinucleotides: evi-
dence for a p53-mediated mammalian SOS
response. Proc. Natl Acad. Sci. USA 94, 12627–
12632 (1997).
75. Wu, C., Marks, S., Prk, H.Y., Howard, C. and Gilch-
rest, B.A. T-oligos induce mitf, tyrosinase and pkc-
beta in cultured human melanocytes. Pigment Cell
Res. 17, 452 (2004).
76. Seite, S., Colige, A., Piquemal-Vivenot, P. et al. A
full-UV spectrum absorbing daily use cream pro-
tects human skin against biological changes occur-
ª 2005 International Journal of Cosmetic Science, 27, 17–34
Ultraviolet radiation and skin aging
ring in photoaging. Photodermatol. Photoimmunol.
Photomed. 16, 147–155 (2000).
77. Thiele, J.J. Photoaging is associated with protein
oxidation in human skin in vivo. J. Invest. Dermatol.
118, 618–625 (2002).
78. Weber, S.U., Thiele, J.J., Cross, C.E. and Packer, L.
Vitamin C, uric acid, and glutathione gradients in
murine stratum corneum and their susceptibility to
ozone exposure. J. Invest. Dermatol. 113, 1128–
1132 (1999).
79. Hellemans, L., Corstjens, H., Neven, A., Declercq, L.
and Maes, D. Antioxidant enzyme sactivity in
human stratum corneum shows seasonal variation
with an age-dependent recovery. J. Invest. Dermatol.
120, 434–439 (2003).
80. Dreher, F. and Maibach, H. Protective effects of top-
ical antioxidants in humans. Curr. Probl. Dermatol.
29, 157–164 (2001).
81. Podda, M. and Grundmann-Kollmann, M. Low
molecular weight antioxidants and their role in skin
ageing. Clin. Exp. Dermatol. 26, 578–582 (2001).
82. Skaper, S.D., Fabris, M., Ferrari, V., Dalle Carbon-
are, M. and Leon, A. Quercetin protects cutaneous
tissue-associated cell types including sensory neu-
rons from oxidative stress induced by glutathione
depletion: cooperative effects of ascorbic acid. Free
Radic. Biol. Med. 22, 669–678 (1997).
83. Katiyar, S.K., Afaq, F., Perez, A. and Mukhtar,
H. polyphenol (-)-epigallocatechin-3-gallate treat-
ment of human skin inhibits UV radiation-induced
oxidative stress. Carcinogenesis 22, 287–294
(2001).
84. Vayalil, P.K., Elmets, C.A. and Katiyar, S.K. Treat-
ment of green tea polyphenols in hydrophilic cream
prevents UVB-induced oxidation of lipids and
proteins, depletion of antioxidant enzymes and
phosphorylation of MAPK proteins in SKH-1 hair-
less mouse skin. Carcinogenesis 24, 927–936
(2003).
85. Zhao, J., Wang, J., Chen, Y. and Agarwal, R. Anti-
tumor-promoting activity of a polyphenolic fraction
isolated from grape seeds in the mouse skin
two-stage initiation-promotion protocol and identifi-
cation of procyanidin B5-3¢-gallate as the most
effective antioxidant constituent. Carcinogenesis 20,
1737–1745 (1999).
86. Afaq, E., Saleem, M., Brans, R. and Mukhtar, H.
Novel agents and targets for skin cancer chemopre-
vention: studies with pomegranate fruit extract.
J. Investig. Dermatol. 121 (2003) (abstract no.
140).
87. Phillipson, J.D. 50 years of medicinal plant research
– every progress in methodology is a progress in
science. Planta Med. 69, 491–495 (2003).
88. Russo, A., Izzo, A.A., Cardile, V., Borrelli, F. and
Vanella, A. Indian medicinal plants as antiradicals
ª 2005 International Journal of Cosmetic Science, 27, 17–34
S. Pillai, C. Oresajo and J. Hayward
and DNA cleavage protectors. Phytomedicine 8,
125–132 (2001).
89. Svobodova, A., Psotova, J. and Walterova, D. Nat-
ural phenolics in the prevention of UV-induced skin
damage: a review. Biomed. Pap. 147, 137–145
(2003).
90. Bissett, D.L., Chatterjee, R. and Hannon, D.P. Pho-
toprotective effect of topical anti-inflammatory
agents against UV radiation-induced chronic skin
damage in the hairless mouse. Photodermatol.
Photoimmunol. Photomed. 7, 153–158 (1990).
91. Kang, I.C., Kim, S.A., Song, G.Y. et al. Effects of the
ethyl acetate fraction of Spatholobi caulis on tumor
cell aggression and migration. Phytother. Res. 17,
163–167 (2003).
92. Thacher, S.M., Vasudevan, J., Tsang, K.Y., Nagpal,
S. and Chandraratna, R.A. New dermatological
agents for the treatment of psoriasis. J. Med. Chem.
44, 281–297 (2001).
93. Verschoore, M., Bouclier, M., Czernielewski, J. and
Hensby, C. Topical retinoids, their uses in dermatol-
ogy. Dermatol. Clin. 11, 107–115 (1993).
94. Guenther, L.C. Topical tazarotene therapy for psori-
asis, acne and photoaging. Skin Therapy Lett. 7,
1–4 (2002).
95. Griffiths, C.E. Retinoids and vitamin D analogues:
action on nuclear transcription. Hosp. Med. 59,
12–16 (1998).
96. Gupta, A.K. and Chow, M. Pimecrolimus: a review. J.
Eur. Acad. Dermatol. Venereol. 17, 493–503 (2003).
97. Niedner, R. Topical corticosteroids versus topical
inhibitors of calcineurin. Hautarzt 54, 338–341
(2001).
98. Voorhees, J.J. and Fisher, G.J. Method of inhibiting
photoaging of skin. US Patent no. 5837224
(1998).
99. Fisher, G.J., Voorhees, J.J. and Kang, S. Composi-
tions and methods for inhibiting photoaging of
skin. US Patent no. 6365630 (2002).
100. Varani, J., Fisher, G.J., Voorhees, J.J. and Kang, S.
Methods and compositions for preventing and treat-
ing chronological aging in human skin. US Patent
application no. 20010053347 (2001).
101. Fisher, G.J., Talwar, H.S., Lin, J. and Voorhees, J.J.
Molecular mechanisms of photoaging in human
skin in vivo and their prevention by all-trans reti-
noic acid. Photochem. Photobiol. 69, 154–157
(1999).
102. Mauviel, A., Halcin, C., Vasiloudes, P., Prks, W.C.,
Kurkinen, M. and Uitto, J. Unco-ordinate regulation
of collagenase, stromelysin and TIMP genes by
PGE2: selective enhancement of collagenase gene
expression in human dermal fibroblasts in culture.
J. Cell Biochem. 54, 465–472 (1994).
103. Varani, J., Fisher, G.J. and Voorhees, J.J. Method for
protecting and restoring skin using selective MMP
33
Ultraviolet radiation and skin aging
inhibitors. US Patent application no. 20020119107
(2002).
104. Pentapharm news release. IFSCC news release from
Phenta pharm at http://www.pentapharm.com on
PEPHA-TIMP, a product to inhibit MMP and stimu-
late collagen synthesis.
105. Hantke, B., Lahmann, C., Venzke, K. et al. Influence
of flavonoids and vitamins on the MMP- and TIMP-
expression of human dermal fibroblasts after UVA
irradiation. Photochem. Photobiol. Sci. 1, 826–833
(2002).
106. Stab, F. and Tschesche, H. Influence of flavonoids
and vitamins on the MMP- and TIMP-expression of
human dermal fibroblasts after UVA irradiation.
Photochem. Photobiol. Sci. 1, 826–833 (2002).
107. Kut-Lasserre, C., Miller, C.C., Ejeil, A.L. et al. Effect
of avocado and soybean unsaponifiables on gela-
tinase A (MMP-2), stromelysin 1 (MMP-3), and tis-
sue inhibitors of matrix metalloproteinase (TIMP- 1
and TIMP-2) secretion by human fibroblasts in cul-
ture. J. Periodontol. 72, 1685–1694 (2001).
108. Demeule, M., Brossard, M., Page, M., Gingras,
D. and Beliveau, R. Matrix metalloproteinase inhi-
bition by green tea catechins. Biochim. Biophys.
Acta 1478, 51–60 (2000).
109. Morrow, D.M., Fitzsimmons, P.E., Chopra, M. and
McGlynn, H. Dietary supplementation with the
anti-tumour promoter quercetin: its effects on mat-
rix metalloproteinase gene regulation. Mutat. Res.
1, 480–481 (2001).
110. Song, L., Xu, M., Lopes-Virella, M.F. and Huang,
Y. Quercetin inhibits matrix metalloproteinase-1
expression in human vascular endothelial cells
through extracellular signal-regulated kinase. Arch.
Biochem. Biophys. 391, 72–78 (2001).
111. Sato, T., Koike, L., Miyata, Y. et al. Inhibnition of
AP-1binding activity and PIP-3-kinase pathway by
nobiletin, a polymethoxy flavonoid, results in aug-
mentation of TIMP-1 production and suppression of
production of MMP-1 and -9 in human fibrosarco-
ma HT-1080 cells. Cancer Res. 62, 1025–1029
(2002).
112. Isnard, N., Peterszegi, G., Robert, A.M. and Robert,
L. Regulation of elastase type endoprptidase activ-
ity, MMP-2 and MMP-9 expression and activation
in human dermal fibroblasts by fucose and fucuse-
34
S. Pillai, C. Oresajo and J. Hayward
rich polysaccharide. Biomed. Pharmacother. 56,
258–264 (2002).
113. Cha, B.Y., Bpark, C.J., Lee, D.G. et al. Inhibitory
effect of methanol extract of Euonymus alatus on
MMP-9. J. Ethanopharmacol. 85, 1637 (2003).
114. Sun, Y. and Xu, Q. Aqueous extract from Rhizoma
notopterygii reduces conteact sensitivity by inhibiting
lymphocyte migrationvia down-regulating MMP
activity. Pharmacol. Res. 46, 333–337 (2002).
115. Rothenberg, M.L., Nelson, A.R. and Hande, K.R.
New drugs on the horizon: matrix metalloprotein-
ase inhibitors. Oncologist 3, 271–274 (1998).
116. Holleran, W.M., Galardy, R.E., Gao, W.N. et al.
Matrix metalloproteinase inhibitors reduce phorbol
ester-induced cutaneous inflammation and hyperpl-
asia. Arch. Dermatol. Res. 289, 138–144 (1997).
117. Yamamoto, M., Tsujishita, H., Hori, N. et al. Inhibi-
tion of membrane-type 1 matrix metalloproteinase
by hydroxamate inhibitors: an examination of the
subsite pocket. J. Med. Chem. 41, 1209–1217
(1998).
118. Brown, P.D. Ongoing trials with matrix metallopro-
teinase inhibitors. Exp. Opin. Investig. Drugs 9,
2167–2177 (2000).
119. Ying, Q.L., Rinehart, A.R., Simon, S.R. and Cheron-
is, J.C. Inhibition of human leucocyte elastase by
ursolic acid: evidence for a binding site for pentacy-
clic triterpenes. Biochem. J. 277, 521–526 (1991).
120. Sartor, L., Pezzato, E. and Garbisa, S. (-)Epigallo-
catechin-3-gallate inhibits leukocyte elastase:
potential of the phyto-factor in hindering inflamma-
tion, emphysema, and invasion. J. Leukoc. Biol. 71,
73–79 (2002).
121. Simon, S., Vered, M., Rinehart, A., Cheronis, J. and
Janoff, A. Inhibition of human neutrophil elastase
by polyguanylic acid and other synthetic polynucle-
otides. Adv. Exp. Med. Biol. 240, 65–74 (1988).
122. Tyagi, S. and Simon, S.R. Parinaric acids as probes
of binding domains in neutrophil elastase. J. Biol.
Chem. 266, 15185–15191 (1991).
123. Tyagi, S. and Simon, S.R. Inhibitors directed to
binding domains in neutrophil elastase. Biochemis-
try 20, 9970–9977 (1990).
124. Tyagi, S. and Simon, S.R. Interaction of neutrophil
elastase with hydrophobic polyanoinic chelators.
Biochem. Cell Biol. 69, 624–629 (1991b).
ª 2005 International Journal of Cosmetic Science, 27, 17–34