Photochemistry and Photobiology, 20**, **: **
Invited Review
Damaging Effects of Ultraviolet Radiation on the Cornea
Naomi C. Delic1,2, J. Guy Lyons1,2,3, Nick Di Girolamo4 and Gary M. Halliday*1
1
Discipline of Dermatology, Bosch Institute, University of Sydney, Camperdown, NSW, Australia
2
Immune Imaging Program, Centenary Institute for Cancer Medicine and Cell Biology, Camperdown, NSW, Australia
3
Sydney Head and Neck Cancer Institute, Cancer Services, Royal Prince Alfred Hospital, Camperdown, NSW, Australia
4
Department of Pathology, School of Medical Sciences, University of New South Wales, Randwick, NSW, Australia
Received 12 September 2016, accepted 18 October 2016, DOI: 10.1111/php.12686
ABSTRACT
The cornea sits at the anterior aspect of the eye and, like the
skin, is highly exposed to ultraviolet radiation (UVR). The
cornea blocks a signicant proportion of UVB from reaching
the posterior structures of the eye. However, UVA can pene-
trate the full thickness of the cornea, even reaching the ante-
rior portion of the lens. Epidemiological data indicate that
UVR is a contributing factor for a multitude of diseases of
the cornea including pterygium, photokeratitis, climatic dro-
plet keratopathy and ocular surface squamous neoplasia
(OSSN), although the pathogenic mechanisms of each require
further elucidation. UVR is a well-known genotoxic agent,
and its effects have been well characterized in organs such as
the skin. However, we are only beginning to identify its
effects on the cornea, such as the UVR signature C ? T and
CC ? TT transversions identied by sequencing and
increased proliferative and shedding rates in response to
UVR exposure. Alarmingly, a single low-dose exposure of
UVR to the cornea is sufcient to elicit genetic, molecular
and cellular changes, supporting the consideration of using
protective measures, such as wearing sunglasses when out-
doors. The aim of this review was to describe the adverse
effects of UVR on the cornea.
THE CORNEA
The human eye is a roughly spherical organ that measures
approximately 2.5 cm in diameter and contains two internal
cavities: the anterior and posterior chambers. The cornea is
located at the most anterior aspect of the eye, overlying the iris,
pupil and anterior chamber, and its health is essential for exqui-
site vision (Fig. 1a). Structurally, the mammalian cornea consists
of ve layers (Fig. 1b): beginning at the most anterior position
(a) a multilayered corneal epithelium, (b) a basement membrane-
like structure known as Bowmans layer, (c) the corneal stroma,
maintained by resident mesenchymal cells known as keratocytes,
(d) a second basement membrane, known as Descemets
*Corresponding author email: gary.halliday@sydney.edu.au (Gary M. Halliday)
This article is part of the Special Issue honoring Dr. Hasan Mukhtars 70th birth-
day and his outstanding contributions to various aspects of photobiology research,
including photocarcinogenesis and chemoprevention.
2016 The American Society of Photobiology
1
membrane, and (e) a monolayer of specialized corneal endothe-
lial cells. Located at the periphery of the cornea is a narrow tran-
sitional band of tissue approximately 1 mm wide, known as the
limbus, which separates the cornea from the conjunctiva. The
corneal epithelium is perpetually self-renewing, whereby the pro-
geny of stem cells residing in the limbus replaces terminally dif-
ferentiated cells that are constantly shed from the corneal surface
into the tear pool (14). These limbal epithelial stem cells
(LESCs) are unipotent as they give rise to corneal, but not con-
junctival epithelia. In vivo studies combined with mathematical
modeling have described a striking spoke-like pattern that forms
as corneal epithelia migrate from the limbus, moving cen-
tripetally toward the apex of the cornea (Fig. 1c,d) (1,5). Mathe-
matical modeling of this process has demonstrated that the
development of this pattern requires no external cues; rather, it is
purely population pressure-driven (5). Lineage tracing experi-
ments using a mosaic mouse model have recently shown that a
single precursor cell is responsible for a unique spoke that forms,
conrming the role of stem cells in the maintenance of the cor-
neal epithelium (1). The mosaic model (Confetti) used a tamox-
ifen-inducible system so that any one or two of four uorescent
proteins (mCFP, nucGFP, cytoYFP and cyto-tDimer2) could be
expressed in K14-expressing cells, including the limbal epithelial
stem cells, giving each of these cells a distinct color. These lim-
bal stem cells give rise to progeny (stem cell and/or corneal
epithelial cell) that inherit the same uorescent protein therefore
making this system ideal for lineage tracing.
EXPOSURE OF THE EYE TO ULTRAVIOLET
RADIATION (UVR)
The sun is the main source of UVR emitting a wide spectrum of
electromagnetic radiation. However, only the ultraviolet (UV),
visible and infrared bands reach the Earths surface to make up
what is known as terrestrial sunlight, with UVR constituting
approximately 5% (6) (Fig. 2a). UVR can be further divided into
three spectral regions; UVC (100290 nm), UVB (290320 nm)
and UVA (320400 nm). UVC is absorbed by ozone in the earths
atmosphere(7), and thus, the UVR component of terrestrial light
consists of approximately 4% UVB and 96% UVA with irradi-
ances differing depending on cloud cover (Fig. 2b) (8).
The eyes and skin are the organs most highly exposed to
UVR and its potentially damaging effects. When discussing
2 Naomi C. Delic et al.
(a) (b) UV Penetrance
C B A
Pupil Sclera
Retina
Conjunctiva
Lens
Iris
Cornea
(c) (d)
Limbus
Cornea
Figure 1. Schematic of the human eye (a) the limbus is a narrow area, located at the periphery of the cornea, where the stem cells reside; (b) cellular
structure of the cornea, depicting the ve distinct layers and the depth of penetration of each ultraviolet (UV) wavelength; (c) colored arrows depict the
direction that the epithelial progeny move from the limbus into the epithelium; (d) A K14CreERT2-Confetti murine eye, 50-week post-tamoxifen injec-
tion (1,5) with uorescently labeled corneal epithelial cells, derived from stem cells in the limbus. The bright halo in the center is autouorescence of
the crystalline lens. Scale bar is 200 lm.
exposure to solar radiation, it must be acknowledged that there
are two components: direct and diffuse. Direct UVR exposure
involves looking directly at the sun, something that humans natu-
rally avoid. However, diffuse (or albedo) UVR exposure is mul-
tidirectional due to photoscattering and reectance by gas or
aerosol molecules in the atmosphere. Due to the directional inci-
dence of diffuse UVR, most ocular UVR exposure is due to this
component, which poses a more difcult task in minimizing its
exposure. In Toowoomba, Australia (27.56 S, 151.95E), the
proportion of diffuse UVB varies from 23% in spring to 59% in
winter, whereas diffuse UVA varies from 17% in spring to 31%
in winter (9). This change in proportion of diffuse UVR can be
affected not only by the season, but also by the time of day. For
example, Parisi et al. (9) noted that cloud cover increases in the
afternoon, and therefore, diffuse UVR readings are often higher
during this time for both UV wavelengths. In addition to the
atmosphere and clouds, the physical environment (such as snow,
buildings, ground coverings) contributes to the scattering effects
observed in diffuse UV. Furthermore, eyelid opening must be
considered when estimating the dose of UVR reaching the ocular
surface. Sliney (10) performed extensive measurements which
were used to design an algorithm that took into account the
Corneal epithelium
Bowman s Layer
Corneal stroma
Descemet s Membrane
Endothelium
reectivity of surfaces in the immediate environment and the
extent of eyelid opening, when estimating the dose of UVR that
was received by the ocular surface. The use of a mannequin
dosimetry system allowed for the evaluation that sunglasses
could attenuate UVR irradiance to the corneal surface by 80%,
compared to the absence of sunglasses (10). Most impressively,
UVB exposure was reduced to 1% when wrap-around style sun-
glasses were employed (10), thus conrming the importance of
choosing the appropriate eyewear when in a sunny environment.
In recent years, UVA exposure has also been developed for
clinical treatments. UVA in conjunction with riboavin has been
approved for cross-linking stromal collagen bers as a therapy for
keratoconus, a degenerative eye disease that is often characterized
by coning of the cornea due to thinning of stromal collagen (11).
In general, shorter wavelengths of UVR and visible light pen-
etrate tissues less than longer wavelengths. Thus, UVB is primar-
ily absorbed by the corneal epithelium, whereas UVA can
penetrate the entire thickness of the cornea (12) (Fig. 1b). The
age of the eye is important to its UVR penetrance. In humans, a
cornea from a 24-year-old blocks over 90% of UVB (290
315 nm) and 45% of UVA (315400 nm) (12). However, cor-
neas from the elderly (80+ years) are more efcient at blocking
 Photochemistry and Photobiology 3

(a)
Radio Microwave Infrared Visible Ultraviolet X-Ray Gamma

Wavelength 103 10-8

10-2 10-5 10-6 10-10 10-12
(meters)

UVC UVB UVA

100-290nm 290-320nm 320-400nm

Frequency 104 108 1012 1015 1018 1020

(Hz)

(b)

Figure 2. (a) Schematic demonstrating the wide spectrum of electromagnetic radiation (EM), with ultraviolet radiation (UVR) measured 100400 nm;
(b) an example of solar irradiance measured on a sunny day (red line) compared to an overcast day (black line). Spectrums were measure on the roof of
The Centenary Institute, Camperdown, Australia, using an Optronics OL-754 spectroradiometer. The boundaries of the UVC, UVB, UVA and visible
(VIS) regions are shown.

UVA (60%) but less efcient at ltering shorter wavelengths
(80%) (12). Furthermore, corneas from younger individuals
(<24 years) transmit even more UVA, therefore rendering them
and the ocular medium behind this tissue more vulnerable to
UVR damage (12).
MEDICAL CONDITIONS OF THE CORNEA
ASSOCIATED WITH UVR EXPOSURE
Photokeratitis
UVR has long been known to cause damage to the cornea, the
acute reaction to UVR-induced burns being referred to as pho-
tokeratitis. The most common accounts of photokeratitis are
snowblindness and welders ash. Snowblindness results
from excessive exposure to naturally occurring UVB in an envi-
ronment of high reectivity; such conditions are encountered
when skiing, at the beach or at high altitude. Conversely,
welders ash results from exposure to articial UVB (and some-
times UVC) sources such as those from a welders arc.
Photokeratitis remains asymptomatic until several hours post-
UVR exposure with the earliest symptoms presenting as a gritty
ocular sensation, followed by photophobia and tearing. These ini-
tial symptoms are attributed to the loss and damage of epithelial
cells in the supercial layers of the cornea. Corneal edema
results in haze and vision impairment. Further exposure to UVR
results in epithelial exfoliation, which leads to agonizing pain. In
1916, Verhoeff and Bell calculated that a threshold dose of
approximately 500 mJ cm 2 of UVR from an articial source
was needed to cause pathological changes in the cornea and that
the main wavelengths responsible within the terrestrial sunlight
band were in the UVB part of the spectrum (13). Ren et al. (14)
showed that suprathreshold doses of UVB did in fact disrupt the
normally ordered process of epithelial desquamation from the
corneal surface, by elevating it to such a rate that the subepithe-
lial nerve endings underlying the cornea are exposed, thus giving
rise to the characteristic pain associated with photokeratitis.
Climatic droplet keratopathy (CDK)
An association of CDK with chronic exposure to UVR has been
conrmed by epidemiological studies (15,16). This condition
occurs when soluble plasma proteins photochemically react with
UVR and deposit beneath the corneal epithelium, within the
4 Naomi C. Delic et al.
(a)
(c)
corneal epithelium
pterygium epithelium
BV
BV
BV
Bowman s Layer
stroma
x1000
Figure 3. (a) Clinical image of a patient with a pterygium; (b) a low power histological micrograph demonstrating the encroaching edge of the ptery-
gium (hatched box); (c) magnied micrograph of the hatched box from b, BVblood vessels; (d) demonstrates stromal elastosis, a feature often seen in
tissue chronically exposed to ultraviolet radiation (UVR).
Bowmans layer and the supercial stroma, causing opacication
of the cornea and visual impairment (17,18). Recent advances in
confocal laser scanning microscopy for studying CDK described
further abnormalities in addition to those already dened.
Patients suffering from moderate-to-advanced CDK display an
increased reectivity of the supercial corneal epithelium as well
as alterations in the density and morphology of the sub-basal
nerve plexus (19). Tear uid from patients with CDK has been
analyzed and found to contain a myriad of inammatory cytoki-
nes including interleukin (IL)-1b, IL-5, IL-6, IL-7, IL-8, mono-
cyte chemoattractant protein-1 (MCP-1), macrophage
inammatory protein-1 beta (MIP-1b) and tumor necrosis factor-
alpha (TNF-a) (18). The matrix metalloproteinase (MMP) proen-
zymes, proMMP-2 and proMMP-9, are found in higher quantity
in the tear uid of CDK patients, while a higher level of active
MMP-9 is found in the corneal epithelial basement membrane,
compared to healthy subjects (18). The signicant increase in
inammatory cytokines and MMPs in corneas of patients suffer-
ing from CDK indicates that its pathology is attributable to a sig-
nicant chronic inammatory response and disruption of the
extracellular matrix (ECM) (18).
Pterygium
Pterygia (Fig. 3a) are common benign lesions of the ocular sur-
face (20), characterized by local displacement of normal epithe-
lium by abnormal epithelial cells. Histologically (Fig. 3b,c), a
demarcation zone exists that distinguishes the normal corneal
epithelium from the advancing edge of the pterygium head, which
often displays regions of elastotic connective tissue (21). Pterygia
(b)
normal corneal epithelium
x200
(d)
pterygium epithelium
stroma
elastosis
x1000
commonly arise from the nasal limbus and form characteristic
wing-shaped lesions that can grow bilaterally and, if left
untreated, restrict eye movement and vision (22). The Coroneo
Effect has aided in explaining the predominance of nasally
located pterygia (and cortical cataracts), by showing that the ante-
rior chamber is capable of focusing light that enters the eye tem-
porally and is internally reected onto the opposing nasal side of
the corneolimbal region. Epidemiological studies have indicated
that UVR plays a major role in the pathogenesis of pterygia
(23,24). This association is supported by the increase in preva-
lence in equatorial regions and among individuals that work out-
doors in an environment with high reectance compared to
indoor workers (regardless of latitude). This risk was increased
20-fold when the surface was sealed concrete and several hun-
dred-fold when working on sand (23). Further evidence of the
role of UVR in pterygia is seen by the formation of elastic tissue
in the pterygium stroma, similar to the solar elastosis that is often
seen in the dermis after prolonged UVR exposure (Fig. 3d) (25).
Ocular surface squamous neoplasms (OSSN)
Ocular surface squamous neoplasms (OSSN) are the most com-
mon form of nonmelanocytic lesions in the conjunctiva and cor-
nea and refer to a disease spectrum comprising mildly dysplastic
to invasive lesions that originate from squamous epithelium (26
28). Patients with these lesions present with a pathological mass
that can be raised and gelatinous, irritation, red eye and leuko-
plakia, originating from the limbus before involving the cornea
and conjunctiva (27,29). Peak incidence of OSSN occurs at a lat-
itude of 16 south (30,31). Two predominant and distinct main

groups of patients are affected. The rst comprises of older
males, who live in temperate climates and are not associated with
human immunodeciency virus (HIV) or human papillomavirus
(HPV) infection (31). The second group includes younger indi-
viduals (both males and females) that reside in the tropics, but
are associated with HIV or HPV infection (31). The association
with HIV suggests that immunosuppression plays a role in pre-
disposing these patients to UV-induced OSSNs, whereas the nat-
ure of the association with, and identities of specic subtypes of
HPV involved, are still unknown (31). OSSN and UVR are also
linked via patients suffering from xeroderma pigmentosa, a rare
autosomal-recessive genetic disorder in which the DNA damage
repair machinery is faulty (32). These patients often develop
OSSNs as well as skin cancer at a young age (26,32).
UVR-INDUCED GENETIC CHANGES
It is well established that solar radiation is the main culprit in the
initiation and progression of several types of skin cancers (squa-
mous cell carcinomas, basal cell carcinomas, melanomas) (23).
Specically, UVR has been proven to be a highly genotoxic
agent (33) by inducing DNA damage in the form of two well-
described photoproducts: cyclobutane pyrimidine dimers (CPDs)
and pyrimidine (6-4) pyrimidone photoproducts (6-4 PPs). UVB
photons can be directly absorbed by DNA bases, largely by the
pyrimidine constituents. This energetic absorption induces the
formation of covalent bonds between two adjacent pyrimidine
bases, with CPDs being the most prevalent photolesion. Because
of their inefcient removal and abundance, CPDs are considered
promutagenic. Misincorporation of adenine instead of guanine
opposite a cytosine in CPDs results in the UV signature C ? T
and CC ? TT transversions (12). As the cornea is at the air
interface and absorbs approximately 90% of the UVB that irradi-
ates the surface of the eye, it is expected that it will sustain
DNA damage. Studies exploring DNA damage in the mam-
malian cornea have observed the presence of photolesions.
Experimental models using human and rabbit eyes described the
rate of CPD formation versus CPD distribution in the cornea,
according to different UV wavelengths (12,34). UVA penetrates
as far as the corneal endothelium and supercial layers of the
crystalline lens; however, CPDs are induced at a much lower rate
by UVA than by UVB (12,34). Conversely, although little UVB
penetrates beyond the anterior third of the corneal stroma, this
region has a signicantly higher rate of CPD formation (12,34).
Although physiologically irrelevant, UVC-induced CPDs are
highly concentrated in the supercial layers of the epithelium
and are virtually undetectable in the remaining 90% of the
stroma (12). Those studies demonstrated a shield-like role the
cornea plays in protecting the posterior structures of the eye from
UVR-induced damage. In addition to CPDs, UVB-induced 6-4
PPs were more abundant in the epithelial layers of the rat cornea.
However, when broadband UVR (280380 nm) doses were used,
equating to well over two minimal erythemal doses (i.e. twice
the dose needed to cause sunburn to exposed skin), evidence of
6-4 PPs was detected throughout all ocular structures, including
the corneal stroma, iris, lens and retina (35). Notably, transmit-
tance of UVR through various ocular structures is highly
dependent on age.
Much remains to be learnt about genetic changes that occur in
the corneal epithelium as a result of overexposure to UV. Recent
advances in understanding the effects of mutagenic changes to
Photochemistry and Photobiology 5
specic genes in the cornea have come from Moloney et al. (36)
and Hassan et al. (37). In these investigations, BRM was found
to be mutated or have reduced expression in human non-
melanoma skin carcinomas (NMSC) (36,38). This gene encodes
one of two ATPase subunits in the SWI/SNF complex involved
in unraveling DNA for factors requiring access to it to mediate
their functions, such as repair and transcription. In a photocar-
cinogenesis study, using a BRM knockout mouse, it was shown
that the absence of this gene resulted in the increased incidence
of both eye and skin tumors, implying that it functions as a
tumor suppressor (39). Both carcinomas and sarcomas of the eye
were induced by chronic UVR exposure in Brm- and/or Tp53-
decient mice. Subsequent studies (37) examined corneas of
mice chronically exposed to UVR (i.e. beyond 25 weeks). Cor-
neas of Brm knockout mice displayed increased thickening,
hyperplasia and decreased atrophy when compared to Brm wild-
type mice, the damage further exacerbated when Brm knockout
mice lost a p53 allele. It was also noticed that irradiated Brm
knockout mice demonstrated increased proliferation and regener-
ation of the corneal epithelium, which was conrmed by Ki-67
staining (37). Brm knockouts showed an increased number of
Ki-67+ corneal epithelial cells compared to Brm wild types after
only 2 weeks of irradiation, which was maintained after
25 weeks of irradiation (37). Of signicance is that proliferation
in the Brm-decient corneas displayed disorganization, with pro-
liferative cells observed in the suprabasal layer of the epithelium,
rather than being conned to the basal layer, which is typical in
the Brm wild-type corneas with or without irradiation (37).
Taken together, the corneal thickening and cellular proliferation
data rmly suggest that Brm plays a signicant role in prevent-
ing hyperproliferation in response to acute or chronic UVR expo-
sure, thus conrming its tumor suppressor status in the cornea.
As with NMSCs, TP53 mutations are observed in UVR-
induced corneal sarcomas. Studies in the opossum found that
these mutations were the hallmark UVR signature C ? T con-
version located in codons equivalent to those found in mutational
hot spots of human UVR-induced SCCs (40). Similar studies
were conducted in human conjunctival SCCs, where a high
prevalence of CC ? TT transversions, a signature mutation of
UVR, was found (41). The location of these mutations at sites of
DNA photoproduct formation suggested that the TP53 mutations
in these OSSN were a result of UVR directly acting on DNA
(40,41).
Although no TP53 mutations were identied, Galor et al. (42)
were rst to perform whole-exome sequencing on OSSN. Of the
seven samples included in their analysis, they identied muta-
tions in Titin (TTN), Neuron Navigator 2 (NAV2), FAT atypical
cadherin 2 (FAT2), hepatocyte growth factor (HGF), dynein
axonemal heavy-chain 8 (DNAH8) and CREB binding protein
(CREBBP)(42). The functional signicance of these mutant genes
remains to be elucidated.
UVR-INDUCED MOLECULAR CHANGES
Multiple molecular factors that have altered activity in response
to UV have been identied in the cornea, including nuclear fac-
tor-jB (NF-jB), cytokines, MMPs and PAX6. NF-jB has been
identied as an initiator of cell death in the cornea and is also
known to be a key driver of cytokine production in response to
injury. Likewise, cytokines can upregulate MMP production
(43,44). Although the current literature describes these
6 Naomi C. Delic et al.
independently of each other, it is likely that these factors work in
concert and in a cascade-type arrangement to exhibit these identi-
ed molecular changes. The relationship of PAX6 to the NF-jB/
cytokine/MMP pathway in the cornea remains to be explored.
NF-jB
Although cell death does not appear to be the primary route for
removal of UVR-exposed corneal epithelia (see the section on
cellular changes in response to UV irradiation), NF-jB may play
a role in initiating cell death. NF-jB is a transcription factor that
remains inactive via ubiquitous expression of its inhibitor IjB or
contiguous latency peptide sequence (45). Targeted phosphoryla-
tion and subsequent degradation of IjB or latency peptide allow
activation of NF-jB followed by its translocation to the nucleus
where its dimers are free to bind to its target DNA elements.
This activates transcription of genes involved with responses to
inammation or cell growth. Using the SV40 immortalized cor-
neal epithelial cell line T-HCEC, Lee et al. (46) showed that cell
death followed nuclear translocation of NF-jB after UVR irradi-
ation. Furthermore, cell death could be blocked by treating cell
lines with the potent NF-jB inhibitors sulfasalazine and SN-50
prior to UVR exposure. Immunocytochemical analysis conrmed
that NF-jB was inhibited from translocating to the nucleus after
irradiation, and downstream cell death was blocked. Elec-
trophoretic mobility shift assays (EMSA) in conjunction with
immunocytochemical staining showed that NF-jB translocated to
the nucleus prior to cell death being observed, and its binding
activity remained increased for 2 h postirradiation, suggesting that
NF-jB indeed plays an early role in the cellular death cascade.
PAX6
PAX6 plays a major role in structural development and ocular
cell differentiation in the eye of vertebrates and has therefore
been coined a master oculogenic control gene (47,48). Its
expression is essential for development, maintenance and repair
of the eye. PAX6 is transcriptionally regulated by CTCF, a zinc
nger phosphoprotein that binds to the CCCTC motif, located
upstream of the PAX6 P0 promoter. Because overexpression of
the CTCF gene in corneal epithelial cells resulted in a downregu-
lation of PAX6 activity, Wu et al. (49) explored PAX6 expres-
sion regulation by CTCF in cultured rabbit and human corneal
epithelial cells that were stimulated with UV. Within 8 hours of
exposure to UV, PAX6 expression increased while CTCF
decreased, both detectable by Western blot analysis. The upregu-
lation of PAX6 by decreased activity of CTCF was conrmed by
EMSA that showed that the binding activity of CTCF on PAX6
P0 upstream enhancers was signicantly decreased after UVR
exposure. Finally, they showed that CTCF had a direct effect on
PAX6 using site-directed mutagenesis on the 80-bp PAX6 P0
enhancer sequence to genetically alter the ve binding motifs,
resulting in decreased binding activity of CTCF.
Matrix metalloproteinases
MMPs are a family of proteolytic enzymes that are important for
tissue remodeling and wound healing. They are synthesized as
inactive zymogens and become activated once proteolytically
cleaved at their propeptide domain on the N-terminus. Once
active, MMPs are capable of either degrading or processing
extracellular matrix components and other bioactive proteins
such as cytokines (50). In normal circumstances, MMPs are
expressed at low levels and their activity is tightly regulated by
their natural inhibitors (TIMPs); therefore, an overexpression and
activation of MMPs are associated with excessive degradation of
the ECM, which can be potentially devastating for the structure
and clarity of the cornea (51). The gelatinases, MMP-2 and
MMP-9, and two of the collagenases, MMP-1 and MMP-8, have
been studied in the cornea in depth (5256). MMP-2 and MMP-
9 were originally identied as enzymes involved in the degrada-
tion by epithelial cells of type IV collagen, a principal
component of basal membranes (57,58).
MMP-1 has been demonstrated to be one of the most abun-
dant MMPs present in pterygia. Di Girolamo et al. (52) showed
that UVB could signicantly increase MMP-1 production in both
pterygium epithelial cell lines and ex vivo pterygia explants. It is
regulated via the UVR-inducible ERK1/2 pathway. MMP-2,
MMP-9, MMP-7, MMP-8 and to a lesser degree MMP-14 have all
been shown to be upregulated by UVB exposure in vivo
(51,59,60), but their expression remains unaltered when corneas
are exposed to UVA (51). MMP-14 is active in multiple roles such
as the cleavage of ECM, activation of other MMPs and activating
signaling pathways for cell migration during wound healing in the
cornea (6163), but interestingly, its expression is only mildly
increased after UVB exposure in rabbit corneal epithelium (51).
Cytokines
UV-induced changes in the cornea can be heavily attributed to
the upregulation of pro-inammatory cytokines including IL-1,
IL-6, IL-8 and TNF-a in the corneal epithelium and stroma (64
67). Specically, it was found that IL-6 and IL-8 were expressed
in the supercial epithelium, with some IL-8 expression in the
vascular endothelium of pterygium tissue that was exposed to
low-dose UVB (40 mJ cm 2) (66). In cultured pterygium epithe-
lial cells that were irradiated with UVB, expression of IL-6 and
IL-8 increased in a time- and dose-dependent manner (66). Nor-
mal limbal broblasts and corneal epithelial cells upregulate
pro-inammatory and macrophage recruiting cytokines, TNFa,
interferon gamma (IFNc) and MCP-1 after low-dose
(20 mJ cm 2) UVB (64). Conversely, Notara et al. (64) also
showed that UVB treatment of limbal broblasts downregulates
their production of soluble factors usually involved in mediating
lymphatic and blood endothelial activity, most likely a regulatory
mechanism to avoid neovascularization of the limbus and cornea.
CELLULAR CHANGES IN RESPONSE TO UV
IRRADIATION
Corneas of mice that are chronically exposed to low doses
(equivalent to two minimal erythemal doses) of UVR display
abnormalities from as early as 25 weeks with epithelial hyper-
plasia, dysplasia, atrophy of the corneal epithelial center, signi-
cant hypocellularity of the stroma, brosis, vascularization and
inammation (37,68). These changes eventually culminate in the
formation of OSSNs.
The acute histological changes to corneas exposed to UVR
occur in a dose-dependent manner (37,68,69). Pitts and col-
leagues (69) showed that UVR enhanced the premature shed-
ding/sloughing of the outermost epithelial cell layers and
increased cell death 24 hours after radiant exposure. Gradually

increasing the radiant exposure caused loss of corneal epithelia;
keratocytes became fragmented and the endothelium became dis-
organized, vacuolated and detached from the stroma (69).
Through the use of ex vivo organ cultivated corneas, Ren et al.
(14) showed that a single dose of 1.05J cm 2 of UVB increased
the shedding rate of epithelial cells from the ocular surface. They
found that, after irradiation, there was a delay of approximately
3 h before shedding was increased and maintained for the ensu-
ing 3 h (14).
Using computer modeling, Lobo and colleagues (5)
recently demonstrated that the well-documented spoke pattern that
corneal epithelial cell clones form as they move from the limbus
(1,70) requires no external physiological cues. Rather, this pattern
can be simply explained by population pressure from the periph-
ery. However, the spoke formation does not rule out the role of
UV-regulated factors, such as signaling molecules, neurons and
biophysical cues in response to inammation and injury (5).
Using the Confetti mosaic mouse model, it was shown that a
single dose of 150 mJ cm 2 UVB increases the spoke growth
rate from the control value of approximately 10 lm day 1 to
approximately 50 lm day 1 after UVR exposure (1,5). Mathe-
matical modeling (5), together with in vivo experiments
(5,68,71,72), has shown that, although apoptosis peaks at around
24-hour post-UV exposure, it is insufcient to account for the
signicant inux of cells in response to UVR exposure without
causing a substantial thickening of the corneal epithelium. On
the contrary, the corneal epithelium thins following UVR expo-
sure (14).
A hypothesis to explain this lack of apoptotic cell death in
response to UVB comes from Leerar et al. (73). Following
UVB, human corneal epithelial cells lose potassium ions (K+)
that would normally activate caspases, triggering the apoptotic
cascade (7375). However, tears are naturally high in K+, and
therefore, it was argued that they would reduce the loss of intra-
cellular K+, preventing apoptosis (73,74,76). Additionally, Ren
and Wilson (14) observed that cells which were shed in response
to UVR were smaller than a typically differentiated cell, suggest-
ing that the epithelial wing cells were failing to fully terminally
differentiate before leaving the epithelium.
Thus, although the cornea is anatomically simple, the mecha-
nisms which it uses to maintain its integrity are complex and
remain relatively unexplained. While apoptosis and increased cell
shedding/terminal differentiation appear to be means of stabiliz-
ing the structure after a UV insult, the current evidence indicates
these mechanisms may not be wholly responsible, and other reg-
ulatory mechanisms await discovery.
IMMUNOGENICITY AND THE CORNEA
The eye is considered an immune privileged site. It was origi-
nally hypothesized that the anterior chamber of the eye was lack-
ing in lymphatic vessels, protecting this organ from an
overwhelming immune response if an antigen was detected. This
was proposed to explain the results of studies performed on
transplanted cells from genetically different hosts, which sur-
vived signicantly longer in the anterior chamber than in other
sites of the body (77,78). However, this hypothesis was chal-
lenged when Kaplan and Streilein (79) found the presence of
hemagglutinating antibodies in serum from rats, 4 days after
being immunized intracamerally with allogeneic lymphoid cells,
concluding that antigens present in the anterior chamber can
Photochemistry and Photobiology 7
indeed elicit a systemic immune response (7880). Today, it is
well established that the cornea contains resident antigen-present-
ing cells of dendritic and macrophage lineages that seem to be
entrenched in the limbal transition zone, perhaps playing a key
role as rst-line defense cells able to sample antigen at a sight
prone to infection, inammation and tumorigenesis (81).
Although much is now known about the immune system in
the eye, little is known about the immune response following
UVR irradiation. Immunosuppression by UVR has been well
studied in the skin, with these effects being attributed partially to
cis-urocanic acid (UCA) which is naturally abundant in the skin
as the trans isomer (82). UV irradiation causes its isomerization
from trans to cis. Treatment with a topical form of cis-UCA on
a corneal allograft in BALB/c mice suppressed the immune sys-
tem therefore prolonging the graft survival time compared to
untreated groups (83). This presence of cis-UCA inhibits the
expression of the cytokines, IL-6 and IL-8, and general cytotoxi-
city associated with inammation (84). In human corneal epithe-
lial cells, cis-UCA is able to inhibit UVB-induced apoptosis and
latent proliferation by blocking the binding of the transcription
factors, c-Jun and c-Fos, but not NF-jB (84). It has also been
proposed that UVR-induced upregulation of JunB is able to
antagonize the pro-apoptotic activity of c-Jun (85). Additionally,
in cultured spleen cells, cis-UCA increases the production of IL-
10, an important factor in immunosuppression because it inhibits
antigen presentation and production of T helper 1 (TH1) cytoki-
nes by epidermal antigen-presenting cells (83).
Transplantation is often the only solution for treating patients
with limbal stem cell deciency, which is frequently caused by
chemical and thermal burns. Rama and colleagues showed that
grafts generated from the patients own limbal stem cell popula-
tion were a potential solution, although there was still a 10%
failure rate and cited inammation as one of the contributing
complications (86). Unlike the skin, an inux of immune cells
and chronic inammation in the cornea can potentially cause
irreparable damage, and so, understanding the immunogenicity
of the cornea is imperative for positive advancements in thera-
peutic applications.
CONCLUSIONS AND FUTURE NEEDS
It is clear from the available data that the level of UVR required
to provoke a response in the cornea is low and, when provoked
chronically, will result in clinical disease. UVR-protective eye-
wear should be worn whenever outdoors, with the need being
greater on days with cloud cover and in surroundings with highly
reective surfaces, due to increased reectance and scatter.
There is currently a need for deeper understanding of the
genetic, cellular and immunological changes that occur in the
cornea after UV exposure, compared to the greater amount of lit-
erature on the skin. Galor and colleagues (42) have pioneered
the area of somatic genetic changes of the cornea by identifying
mutational changes in human patients with OSSNs. Although a
very small number of samples were analyzed, they demonstrate
the potential for identifying mutations and correlating them with
efcient therapies. The mechanism for the nonapoptotic cell loss
that results in thinning of UVR-exposed corneas remains to be
elucidated. Identifying the molecular pathways involved might
shed insights into novel means of treating degenerative condi-
tions involving the corneal epithelium. Studies identifying the
semi-immunosuppressive state of the eye and its correlation with
8 Naomi C. Delic et al.
OSSN demonstrate the complexity of the immune system. This
requires considerably more analysis. There are many mouse
models available in which immune cells are uorescently tagged
and these would assist in teasing out mechanisms of the ocular
immune response.
Mathematical models might prove to be useful in improving
therapies such as UVA cross-linking and limbal stem cell trans-
plants to optimize locations of treatment and avoiding more dam-
age due to the treatment (87). Lobo et al. (5) demonstrated that
their model was efcient in replicating the biology of the epithe-
lial cells under homeostatic and experimental conditions. Mathe-
matical simulations could potentially be programmed to
incorporate a vast number of experimental parameters to identify
appropriate targets. This could be used to help design experi-
ments, reducing costs associated with large-scale experimental
testing and optimizing therapeutic strategies. Therefore, not only
would mathematical modeling be benecial for developing treat-
ment strategies, but also for planning research programs.
AcknowledgementsN.C. Delic was supported by Australian
Postgraduate Award and Sydney Catalyst Top-Up Scholarship from
Cancer Institute NSW. This work was supported by Human Frontiers
Science Program Grant RGP0041. The authors thank Professor Minas
Coroneo (Department of Ophthalmology, Prince of Wales Hospital,
Randwick, Sydney, Australia) for providing the clinical image of a
pterygium.
REFERENCES
1. Di Girolamo, N., S. Bobba, V. Raviraj, N. C. Delic, I. Slapetova, P.
R. Nicovich, G. M. Halliday, D. Wakeeld, R. Whan and J. G.
Lyons (2015) Tracing the fate of limbal epithelial progenitor cells in
the murine cornea. Stem Cells 33, 157169.
2. Dua, H. S. and A. Azuara-Blanco (2000) Limbal stem cells of the
corneal epithelium. Surv. Ophthalmol. 44, 415425.
3. Zhao, J., V. Mo and T. Nagasaki (2009) Distribution of label-retain-
ing cells in the limbal epithelium of a mouse eye. J. Histochem.
Cytochem. 57, 177185.
4. Thoft, R. A. and J. Friend (1983) The X, Y, Z hypothesis of cor-
neal epithelial maintenance. Invest. Ophthalmol. Vis. Sci. 24, 1442
1443.
5. Lobo, E. P., N. C. Delic, A. Richardson, V. Raviraj, G. M. Halliday,
N. Di Girolamo, M. R. Myerscough and J. G. Lyons (2016) Self-
organized centripetal movement of corneal epithelium in the absence
of external cues. Nat. Commun. 7, 12388.
6. Diffey, B. L. (2002a) Sources and measurement of ultraviolet radia-
tion. Methods 28, 413.
7. Parisi, A. V. and D. Turnbull (2005) Diffuse solar ultraviolet radia-
tion. Int. Ophthalmol. Clin. 45, 1927.
8. Diffey, B. L. (2002b) Human exposure to solar ultraviolet radiation.
J. Cosmet. Dermatol. 1, 124130.
9. Parisi, A. V., A. Green and M. G. Kimlin (2001) Diffuse solar UV
radiation and implications for preventing human eye damage. Pho-
tochem. Photobiol. 73, 135139.
10. Sliney, D. H. (1995) UV radiation ocular exposure dosimetry. J.
Photochem. Photobiol., B 31, 6977.
11. Mastropasqua, L. (2015) Collagen cross-linking: when and how? A
review of the state of the art of the technique and new perspectives.
Eye Vis. (Lond) 2, 19.
12. Mallet, J. D. and P. J. Rochette (2013) Wavelength-dependent ultra-
violet induction of cyclobutane pyrimidine dimers in the human cor-
nea. Photochem. Photobiol. Sci. 12, 13101318.
13. Verhoeff, F. H., L. Bell and C. B. Walker (1916) The pathological
effects of radiant energy on the eye. Proc. Am. Acad. Arts Sci. 51,
627818.
14. Ren, H. and G. Wilson (1994) The effect of ultraviolet-B irradiation
on the cell shedding rate of the corneal epithelium. Acta Ophthalmol.
(Copenh) 72, 447452.
15. Johnson, G. J. (1981) Aetiology of spheroidal degeneration of the
cornea in Labrador. Br. J. Ophthalmol. 65, 270283.
16. Taylor, H. R., S. K. West, F. S. Rosenthal, B. Munoz, H. S. New-
land and E. A. Emmett (1989) Corneal changes associated with
chronic UV irradiation. Arch. Ophthalmol. 107, 14811484.
17. Cullen, A. P. (2002) Photokeratitis and other phototoxic effects on
the cornea and conjunctiva. Int. J. Toxicol. 21, 455464.
18. Holopainen, J. M., A. Robciuc, T. A. Cafaro, M. F. Suarez, Y. T.
Konttinen, H. M. Alkatan, K. F. Tabbara, T. Tervahartiala, T. Sorsa,
J. A. Urrets-Zavalia and H. M. Serra (2012) Pro-inammatory
cytokines and gelatinases in climatic droplet keratopathy. Invest.
Ophthalmol. Vis. Sci. 53, 35273535.
19. Serra, H. M., J. M. Holopainen, R. Beuerman, K. Kaarniranta, M. F.
Suarez and J. A. Urrets-Zavalia (2015) Climatic droplet keratopathy:
an old disease in new clothes. Acta Ophthalmol. 93, 496504.
20. Grossniklaus, H. E., W. R. Green, M. Luckenbach and C. C. Chan
(1987) Conjunctival lesions in adults. A clinical and histopathologic
review. Cornea 6, 78116.
21. Chui, J., M. T. Coroneo, L. T. Tat, R. Crouch, D. Wakeeld and N.
Di Girolamo (2011) Ophthalmic pterygium: a stem cell disorder with
premalignant features. Am. J. Pathol. 178, 817827.
22. Young, R. W. (1994) The family of sunlight-related eye diseases.
Optom. Vis. Sci. 71, 125144.
23. Mackenzie, F. D., L. W. Hirst, D. Battistutta and A. Green (1992)
Risk analysis in the development of pterygia. Ophthalmology 99,
10561061.
24. Yam, J. C. and A. K. Kwok (2014) Ultraviolet light and ocular dis-
eases. Int. Ophthalmol. 34, 383400.
25. Austin, P., F. A. Jakobiec and T. Iwamoto (1983) Elastodysplasia
and elastodystrophy as the pathologic bases of ocular pterygia and
pinguecula. Ophthalmology 90, 96109.
26. Kheir, W. J., M. T. Tetzlaff, M. L. Pfeiffer, K. Mulay, O. Ozgur, G.
Morrell and B. Esmaeli (2016) Epithelial, non-melanocytic and mela-
nocytic proliferations of the ocular surface. Semin. Diagn. Pathol.
33, 122132.
27. Lee, G. A. and L. W. Hirst (1995) Ocular surface squamous neo-
plasia. Surv. Ophthalmol. 39, 429450.
28. Shields, C. L., H. Demirci, E. Karatza and J. A. Shields (2004) Clin-
ical survey of 1643 melanocytic and nonmelanocytic conjunctival
tumors. Ophthalmology 111, 17471754.
29. Tunc, M., D. H. Char, B. Crawford and T. Miller (1999) Intraepithe-
lial and invasive squamous cell carcinoma of the conjunctiva: analy-
sis of 60 cases. Br. J. Ophthalmol. 83, 98103.
30. Gichuhi, S., S. Ohnuma, M. S. Sagoo and M. J. Burton (2014)
Pathophysiology of ocular surface squamous neoplasia. Exp. Eye
Res. 129, 172182.
31. Gichuhi, S., M. S. Sagoo, H. A. Weiss and M. J. Burton (2013) Epi-
demiology of ocular surface squamous neoplasia in Africa. Trop.
Med. Int. Health 18, 14241443.
32. Gupta, N., R. Sachdev and R. Tandon (2011) Ocular surface squa-
mous neoplasia in xeroderma pigmentosum: clinical spectrum and
outcome. Graefes Arch. Clin. Exp. Ophthalmol. 249, 12171221.
33. Lucas, R., T. McMichael, W. Smith and B. Armstrong (2006)
Solar ultraviolet radiation: global burden of disease from solar
ultraviolet radiation. In Environmental Burden of Disease Series no.
13, World Health Organization. (Edited by A. Pruss-Ust un, H.
Zeeb, C. Mathers and M. Repacholi). pp. 1250. Geneva,
Switzerland.
34. Mallet, J. D. and P. J. Rochette (2011) Ultraviolet light-induced
cyclobutane pyrimidine dimers in rabbit eyes. Photochem. Photobiol.
87, 13631368.
35. Estil, S., W. M. Olsen, H. S. Huitfeldt and E. Haaskjold (1997)
UVB-induced formation of (6-4) photoproducts in the rat corneal
epithelium. Acta Ophthalmol. Scand. 75, 120123.
36. Moloney, F. J., J. G. Lyons, V. L. Bock, X. X. Huang, M. J. Bugeja
and G. M. Halliday (2009) Hotspot mutation of Brahma in non-mel-
anoma skin cancer. J. Invest. Dermatol. 129, 10121015.
37. Hassan, N. M., N. Painter, C. R. Howlett, A. W. Farrell, N. Di Giro-
lamo, J. G. Lyons and G. M. Halliday (2014) Brm inhibits the pro-
liferative response of keratinocytes and corneal epithelial cells to
ultraviolet radiation-induced damage. PLoS ONE 9, e107931.
38. Bock, V. L., J. G. Lyons, X. X. Huang, A. M. Jones, L. A. McDon-
ald, R. A. Scolyer, F. J. Moloney, R. S. Barnetson and G. M. Halli-
day (2011) BRM and BRG1 subunits of the SWI/SNF chromatin

remodelling complex are downregulated upon progression of benign
skin lesions into invasive tumours. Br. J. Dermatol. 164, 12211227.
39. Halliday, G. M., Y. Zhou, P. W. Sou, X. X. Huang, S. Rana, M. J.
Bugeja, N. Painter, R. A. Scolyer, C. Muchardt, N. Di Girolamo and
J. G. Lyons (2012) The absence of Brm exacerbates photocarcino-
genesis. Exp. Dermatol. 21, 599604.
40. Kusewitt, D. F., T. E. Sherburn, K. B. Miska, G. B. Tafoya, J. M.
Gale and R. D. Miller (1999) The p53 tumor suppressor gene of the
marsupial Monodelphis domestica: cloning of exons 4-11 and muta-
tions in exons 5-8 in ultraviolet radiation-induced corneal sarcomas.
Carcinogenesis 20, 963968.
41. Ateenyi-Agaba, C., M. Dai, F. Le Calvez, E. Katongole-Mbidde, A.
Smet, M. Tommasino, S. Franceschi, P. Hainaut and E. Weiderpass
(2004) TP53 mutations in squamous-cell carcinomas of the conjunc-
tiva: evidence for UV-induced mutagenesis. Mutagenesis 19, 399
401.
42. Galor, A., C. L. Karp, D. Sant, M. Joag, N. Shalabi, C. B. Gustafson
and G. Wang (2016) Whole exome proling of ocular surface squa-
mous neoplasia. Ophthalmology 123, 216217, e211.
43. Ito, A., Y. Itoh, Y. Sasaguri, M. Morimatsu and Y. Mori (1992)
Effects of interleukin-6 on the metabolism of connective tissue com-
ponents in rheumatoid synovial broblasts. Arthritis Rheum. 35,
11971201.
44. Luca, M., S. Huang, J. E. Gershenwald, R. K. Singh, R. Reich and
M. Bar-Eli (1997) Expression of interleukin-8 by human melanoma
cells up-regulates MMP-2 activity and increases tumor growth and
metastasis. Am. J. Pathol. 151, 11051113.
45. Won, M., H. S. Byun, K. A. Park and G. M. Hur (2016) Post-trans-
lational control of NF-kappaB signaling by ubiquitination. Arch.
Pharm. Res. 39, 10751084.
46. Lee, D. H., J. K. Kim and C. K. Joo (2005) Translocation of nuclear
factor-kappaB on corneal epithelial cells induced by ultraviolet B
irradiation. Ophthalmic Res. 37, 8388.
47. Gehring, W. J. (1996) The master control gene for morphogenesis
and evolution of the eye. Genes Cells 1, 1115.
48. Gehring, W. J. and K. Ikeo (1999) Pax 6: mastering eye morphogen-
esis and eye evolution. Trends Genet. 15, 371377.
49. Wu, D., T. Li, Z. Lu, W. Dai, M. Xu and L. Lu (2006) Effect of
CTCF-binding motif on regulation of PAX6 transcription. Invest.
Ophthalmol. Vis. Sci. 47, 24222429.
50. Rodriguez, D., C. J. Morrison and C. M. Overall (2010) Matrix met-
alloproteinases: what do they not do? New substrates and biological
roles identied by murine models and proteomics. Biochim. Biophys.
Acta 1803, 3954.
51. Ardan, T. and J. Cejkova (2012) Immunohistochemical expression of
matrix metalloproteinases in the rabbit corneal epithelium upon UVA
and UVB irradiation. Acta Histochem. 114, 540546.
52. Di Girolamo, N., M. T. Coroneo and D. Wakeeld (2003) UVB-eli-
cited induction of MMP-1 expression in human ocular surface
epithelial cells is mediated through the ERK1/2 MAPK-dependent
pathway. Invest. Ophthalmol. Vis. Sci. 44, 47054714.
53. Ma, D. H., J. K. Chen, W. S. Kim, Y. X. Hao, H. C. Wu, R. J. Tsai,
D. G. Hwang and F. Zhang (2001) Expression of matrix metallopro-
teinases 2 and 9 and tissue inhibitors of metalloproteinase 1 and 2 in
inammation-induced corneal neovascularization. Ophthalmic Res.
33, 353362.
54. Smith, V. A., H. B. Hoh and D. L. Easty (1999) Role of ocular
matrix metalloproteinases in peripheral ulcerative keratitis. Br. J.
Ophthalmol. 83, 13761383.
55. Xue, M. L., D. Wakeeld, M. D. Willcox, A. R. Lloyd, N. Di Giro-
lamo, N. Cole and A. Thakur (2003) Regulation of MMPs and
TIMPs by IL-1beta during corneal ulceration and infection. Invest.
Ophthalmol. Vis. Sci. 44, 20202025.
56. Yang, Y. N., D. Bauer, S. Wasmuth, K. P. Steuhl and A. Heiligen-
haus (2003) Matrix metalloproteinases (MMP-2 and 9) and tissue
inhibitors of matrix metalloproteinases (TIMP-1 and 2) during the
course of experimental necrotizing herpetic keratitis. Exp. Eye Res.
77, 227237.
57. Lyons, J. G., B. Birkedal-Hansen, W. G. Moore, R. L. OGrady and
H. Birkedal-Hansen (1991) Characteristics of a 95-kDa matrix metal-
loproteinase produced by mammary carcinoma cells. Biochemistry
30, 14491456.
58. Salo, T., J. G. Lyons, F. Rahemtulla, H. Birkedal-Hansen and H.
Larjava (1991) Transforming growth factor-beta 1 up-regulates type
Photochemistry and Photobiology 9
IV collagenase expression in cultured human keratinocytes. J. Biol.
Chem. 266, 1143611441.
59. Cejkova, J., T. Ardan, C. Cejka and J. Luyckx (2011) Favorable
effects of trehalose on the development of UVB-mediated antioxi-
dant/pro-oxidant imbalance in the corneal epithelium, proinamma-
tory cytokine and matrix metalloproteinase induction, and heat shock
protein 70 expression. Graefes Arch. Clin. Exp. Ophthalmol. 249,
11851194.
60. Chandler, H. L., D. F. Kusewitt and C. M. Colitz (2008) Modulation
of matrix metalloproteinases by ultraviolet radiation in the canine
cornea. Vet. Ophthalmol. 11, 135144.
61. Chakraborti, S., M. Mandal, S. Das, A. Mandal and T. Chakraborti
(2003) Regulation of matrix metalloproteinases: an overview. Mol.
Cell. Biochem. 253, 269285.
62. Joo, C. K. and Y. Seomun (2008) Matrix metalloproteinase (MMP)
and TGF beta 1-stimulated cell migration in skin and cornea wound
healing. Cell Adh. Migr. 2, 252253.
63. Smine, A. and J. J. Plantner (1997) Membrane type-1 matrix metal-
loproteinase in human ocular tissues. Curr. Eye Res. 16, 925929.
64. Notara, M., N. Refaian, G. Braun, P. Steven, F. Bock and C. Cur-
siefen (2015) Short-term UVB-irradiation leads to putative limbal
stem cell damage and niche cell-mediated upregulation of macro-
phage recruiting cytokines. Stem Cell Res. 15, 643654.
65. Corsini, E., N. Sangha and S. R. Feldman (1997) Epidermal strati-
cation reduces the effects of UVB (but not UVA) on keratinocyte
cytokine production and cytotoxicity. Photodermatol. Photoimmunol.
Photomed. 13, 147152.
66. Di Girolamo, N., R. K. Kumar, M. T. Coroneo and D. Wakeeld
(2002) UVB-mediated induction of interleukin-6 and -8 in pterygia
and cultured human pterygium epithelial cells. Invest. Ophthalmol.
Vis. Sci. 43, 34303437.
67. Kennedy, M., K. H. Kim, B. Harten, J. Brown, S. Planck, C.
Meshul, H. Edelhauser, J. T. Rosenbaum, C. A. Armstrong and J. C.
Ansel (1997) Ultraviolet irradiation induces the production of multi-
ple cytokines by human corneal cells. Invest. Ophthalmol. Vis. Sci.
38, 24832491.
68. Newkirk, K. M., H. L. Chandler, A. E. Parent, D. C. Young, C. M.
Colitz, D. A. Wilkie and D. F. Kusewitt (2007) Ultraviolet radiation-
induced corneal degeneration in 129 mice. Toxicol. Pathol. 35, 819
826.
69. Pitts, D. G., J. P. Bergmanson and L. W. Chu (1987) Ultrastructural
analysis of corneal exposure to UV radiation. Acta Ophthalmol.
(Copenh) 65, 263273.
70. Mort, R. L., T. Ramaesh, D. A. Kleinjan, S. D. Morley and J. D.
West (2009) Mosaic analysis of stem cell function and wound heal-
ing in the mouse corneal epithelium. BMC Dev. Biol. 9, 4.
71. Alov, I. A. (1959) The mechanism of the diurnal periodicity of mito-
sis. Bull. Exp. Biol. Med. 48, 14181421.
72. Ren, H. and G. Wilson (1996) Apoptosis in the corneal epithelium.
Invest. Ophthalmol. Vis. Sci. 37, 10171025.
73. Leerar, J. R., C. D. Glupker, M. P. Schotanus and J. L. Ubels
(2016) The effect of K+ on caspase activity of corneal epithelial
cells exposed to UVB. Exp. Eye Res. 151, 2325.
74. Glupker, C. D., P. M. Boersma, M. P. Schotanus, L. D. Haarsma
and J. L. Ubels (2016) Apoptosis of corneal epithelial cells caused
by ultraviolet B-induced loss of K(+) is inhibited by Ba(2.). Ocul.
Surf. 14, 401409.
75. Schotanus, M. P., L. R. Koetje, R. E. Van Dyken and J. L. Ubels
(2011) Stratied corneal limbal epithelial cells are protected from
UVB-induced apoptosis by elevated extracellular K(+). Exp. Eye
Res. 93, 735740.
76. Ubels, J. L., C. D. Glupker, M. P. Schotanus and L. D. Haarsma
(2016) Involvement of the extrinsic and intrinsic pathways in ultravi-
olet B-induced apoptosis of corneal epithelial cells. Exp. Eye Res.
145, 2635.
77. Medawar, P. B. (1948) Immunity to homologous grafted skin; the fate
of skin homografts transplanted to the brain, to subcutaneous tissue,
and to the anterior chamber of the eye. Br. J. Exp. Pathol. 29, 5869.
78. Perez, V. L., A. M. Saeed, Y. Tan, M. Urbieta and F. Cruz-Guilloty
(2013) The eye: a window to the soul of the immune system.
J. Autoimmun. 45, 714.
79. Kaplan, H. J. and J. W. Streilein (1977) Immune response to immu-
nization via the anterior chamber of the eye. I. F. lymphocyte-
induced immune deviation. J. Immunol. 118, 809814.
10 Naomi C. Delic et al.

80. Stein-Streilein, J. and J. W. Streilein (2002) Anterior chamber associ- Nick Di Girolamo is
ated immune deviation (ACAID): regulation, biological relevance, Director of the Ocular
and implications for therapy. Int. Rev. Immunol. 21, 123152. Diseases Research and
81. Wilkinson, A., N. Kawaguchi, C. Geczy and N. Di Girolamo (2016) Head of the Mechanisms
S100A8 and S100A9 proteins are expressed by human corneal stro- of Disease and Transla-
mal dendritic cells. Br. J. Ophthalmol. 100, 13041308. tional Research at the
82. Noonan, F. P. and E. C. De Fabo (1992) Immunosuppression by School of Medical Sci-
ultraviolet B radiation: initiation by urocanic acid. Immunol. Today ences, University of New
13, 250254. South Wales, Australia.
83. Krulova, M., L. Kuffova, A. Zajicova, M. Filipec and V. Holan His research interests
(1999) IL-10 is an effector molecule mediating urocanic acid-induced include diseases that arise
immunosuppression. Transplant. Proc. 31, 12181219. on the ocular surface, pre-
84. Jauhonen, H. M., A. Kauppinen, T. Paimela, J. K. Laihia, L. Leino, dominantly the cornea.
A. Salminen and K. Kaarniranta (2011) Cis-urocanic acid inhibits His team recently devel-
SAPK/JNK signaling pathway in UV-B exposed human corneal oped a novel technique to
epithelial cells in vitro. Mol. Vis. 17, 23112317. deliver stem cells to cor-
85. Wickert, H., K. Zaar, A. Grauer, M. John, M. Zimmermann and F. nea using a therapeutic
Gillardon (1999) Differential induction of proto-oncogene expression contact lens. His team
and cell death in ocular tissues following ultraviolet irradiation of also developed a trans-
the rat eye. Br. J. Ophthalmol. 83, 225230. genic mouse model to
86. Rama, P., S. Matuska, G. Paganoni, A. Spinelli, M. De Luca and G. study the activity of cor-
Pellegrini (2010) Limbal stem-cell therapy and long-term corneal neal stem cells from
regeneration. N. Engl. J. Med. 363, 147155. development to aging.
87. Nejad, T. M., C. Foster and D. Gongal (2014) Finite element mod-
elling of cornea mechanics: a review. Arq. Bras. Oftalmol. 77, 6065.

AUTHOR BIOGRAPHIES Gary M. Halliday is a

Professor of Dermatology
at the University of Syd-
ney. He obtained his PhD
Naomi C. Delic is a PhD from Monash University
student at the University in 1981 and his Doctor of
of Sydney. She received Science from the Univer-
her Bachelor of Science sity of Sydney in 2001.
with rst class honors His research is directed
from the University of toward understanding the
New South Wales in role of sunlight in skin
2006. Her research inter- carcinogenesis, particu-
ests include cancer biol- larly ultraviolet radiation
ogy of the skin; however, suppression of immunity,
her current research is induction of gene muta-
focused on understanding tions, skin cancer cell
the response of the cornea biology and development
to ultraviolet radiation of effective prevention.
exposure, using intravital His research group is part
lineage tracing techniques. of the Sydney Cancer
Centre, the Bosch Insti-
tute, the department of
Dermatology of Royal
Prince Alfred Hospital
and the Central Clinical
School of the University
J. Guy Lyons received of Sydney.
his PhD from the Univer-
sity of Sydney in 1988.
He is a cancer biologist
with a long-term interest
in epithelial carcinogene-
sis. Squamous cell carci-
nomas of the skin, oral
mucosa and ocular surface
are of particular interest.
He is currently residing at
the Centenary Institute,
where he is using intravi-
tal lineage tracing and
genetic manipulation tech-
niques to study clonal
evolution during develop-
ment and carcinogenesis.