Recent Patents on Inflammation & Allergy Drug Discovery 2009, 3, 221-231 221

Corneal Neovascularization: Molecular Events and Therapeutic Options

Yadollah Shakiba*,1,2, Kamran Mansouri2, Delnia Arshadi2 and Nima Rezaei3

1
Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran, 2Medical
Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran, 3Growth and Development
Research Center, Pediatrics Center of Excellent, Children's Medical Center, Tehran University of Medical Sciences,
Tehran, Iran

Received: April 2, 2009; Accepted: June 3, 2009; Revised: August 20, 2009
Abstract: Corneal neovascularization (NV) is a significant complication of numerous infectious and non-infectious ocular
surface disorders. Presence of newly formed blood vessels in the cornea can compromise clarity and therefore vision.
Various growth factors and proteinases seem to be involved in the corneal NV. During corneal injury, angiogenic factors
are released from corneal epithelial and stromal cells as well as infiltrating immune cells like macrophages. In fact, the
balance between angiogenic and anti-angiogenic factors is shifted towards angiogenic molecules in the corneal NV.
Numerous investigations support this idea that vascular endothelial growth factor (VEGF) plays a pivotal role in corneal
NV by inducing endothelial cells proliferation, migration, and tubulogenesis. There is a growing body of evidence that
corneal NV can be reduced by using anti-VEGF agents. This article reviews the most known molecular events in corneal
NV and also some of the recent patents relevant to the field. Understanding the role of growth factors, proteinases and
inflammatory cytokines in corneal NV can help the investigators to design therapeutic options for controlling this
debilitating condition.
Keywords: Cornea, neovascularization, VEGF, MMP, inflammation.

INTRODUCTION occurs when the balance between angiogenic and anti-

Cornea
The cornea is the transparent avascular tissue of the eye,
which its transparency is essential for optimal clarity and
vision. The cornea is a dynamic structure that interacts with
its surrounding tissues and plasma through tear fluid,
aqueous humor and circulating immune cells. These inter-
actions play an important role in the maintaining of its
normal structure and functions. Moreover, the cornea is an
immune-privileged site; immune privilege is a dynamic
phenomenon in which the immune response to antigens is
absent in order to protect this highly organized structure of
the eye. The lack of blood and lymphatic vessels in a normal
cornea prevents entering innate and adaptive immune system
cells and behaves as a mechanical barrier. The corneal
avascularity is considered as corneal angiogenic privilege
[1]. Recent investigations indicate a close relationship in the
molecular mechanisms maintaining corneal angiogenic and
immune privilege. The cornea is supplied with oxygen and
nutrients by pericorneal plexus (limbal plexus), tear and
anterior chamber aqueous humor. Limbal plexus of the
cornea arises from the ciliary arteries, which are branches of
the ophthalmic artery that divide and end in the pericorneal
plexus in the limbus area [2].
In physiological conditions, the corneal avascularity
requires low levels of angiogenic factors and high levels of
anti-angiogenic factors. Corneal neovascularization (NV)
*Address correspondence to this author at the Department of Immunology,
School of Medicine, Tehran University of Medical Sciences, Tehran, Postal
Zip Code: 1417613151, Iran; Tel/Fax: +98-21-66419536;
E-mail: yshakiba@razi.tums.ac.ir; y_shakiba@yahoo.com
angiogenic factors is tilted towards angiogenic molecules.
This phenomenon occurs in a variety of pathological con-
ditions, such as hypoxia, chemical burns, ischemia, inflam-
mation, infection and trauma [3]. Presence of new blood
vessels in previously avascular cornea disturbs light passing
and influences visual acuity and clarity negatively. Newly
formed vessels are not completely functional. These vessels
have no basement membrane and pericytes; therefore, they
are very fragile and susceptible to hemorrhage. The leakage
of plasma components leads to the corneal edema. In
addition, these compounds can be deposited in the cornea
and impair its transparency permanently [4]. To date, many
studies have been conducted for clarifying mechanisms
underlying corneal NV and finding new therapeutic options
for controlling the formation of new blood vessels and
maintaining corneal avascularity. In this article, we review
the recent findings on the molecular mechanisms of corneal
NV, focusing on the pivotal role of inflammation and
therapeutic potential of anti-angiogenic drugs.
EPIDEMIOLOGY AND ETIOLOGY OF CORNEAL
NEOVASCULARIZATION
Corneal NV is a main cause of loss of vision and
blindness in a wide range of corneal diseases worldwide. It is
estimated that 4% of the US population has corneal NV and
every year 1.4 million patients in the US may develop
corneal NV [5]. Although, the etiology of corneal NV is not
completely identified, numerous pathological conditions
such as inflammatory, infectious, degenerative, and
traumatic disorders may induce NV. Among these different
etiologies, infectious disease of the cornea seems to be the

1872-213X/09 $100.00+.00 © 2009 Bentham Science Publishers Ltd.

222 Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3
most important cause of corneal NV leading to vision loss
[6]. A typical example is trachoma, a chronic infection of the
conjunctiva caused by the Chlamydia trachomatis, which is
the leading cause of preventable blindness in the world.
Untreated active trachoma is accompanied by corneal NV as
pannus (fibrovascular tissue) formation. The pannus may
extend to central cornea and cause severe disturbance in
corneal transparency. It has been estimated that about 5.9
million people are blind or have severe vision-loss due to
trachoma infection and 10 million are at high risk worldwide
[6,7]. Corneal NV is also a common manifestation of herpes
simplex keratitis (HSK), which is the most common
infectious cause of corneal blindness in the Western
countries [8]. Infection of corneal stroma by herpes simplex
virus (HSV) is usually associated with NV. In fact, it has
been demonstrated that formation of NV is the essential step
for the pathogenesis of HSK and anti-angiogenic agents can
be used for controlling herpes simplex induced corneal
angiogenesis [9]. Corneal NV is also a common manifesta-
tion of severe infections of the cornea by other micro-
organisms such as staphylococcus, streptococcus and pseu-
domonas [8]. Although, all infectious keratitis may induce
corneal NV, Acanthamoeba keratitis has been reported to
lack corneal NV even in relatively severe and long-standing
cases [10].
Corneal NV can affect the prognosis of corneal trans-
plantation. It has been reported that 20% of corneal
specimens obtained during corneal transplantation show
histopathological evidences of NV [11]. In fact, corneal NV
can significantly worsen the prognosis of penetrating
keratoplasty (PK). Previous studies showed that 41% of eyes
in low risk patients developed corneal NV 6 to 9 months
after PK. Active blepharitis and stimulation of corneal
stroma by sutures are considered as risk factors for evolution
of NV after PK [11,12]. Today, there is a growing body of
interests to prevent NV after PK using anti-angiogenic
agents [13].
Contact lenses have become one of the most common
etiologies of corneal NV in developed countries during past
two decades. The prevalence of soft hydrogel lenses induced
corneal NV is very higher than hard gas-permeable lenses
[14]. Although, the precise mechanism of contact lens
induced NV is not clear, there are numerous evidences that
support the role of hypoxia in its pathology. As noted above,
the cornea supplies oxygen from surface layer by diffusion;
thus contact lens, especially soft lenses, can induce hypoxia
by reducing corneal oxygen uptake.
Contact lens induced hypoxia causes significant changes
to the corneal cells such as decreased metabolic rate, stromal
edema, and decreased endothelial cell density and function.
Moreover, long term hypoxia upregulates angiogenic factors
that induce corneal NV [15]. In overall, corneal NV caused
by contact lens wear is generally mild and does not affect
vision in the aggressive manner that infections do. However,
it is a sign of corneal distress, which needs medical attention
[8].
Shakiba et al.
ANGIOGENIC MOLECULES INVOLVED IN
CORNEAL NEOVASCULARIZATION
Vascular Endothelial Growth Factor
Vascular endothelial growth factor (VEGF), also called
VEGF-A, belongs to a family of closely related growth
factors that are characterized by the presence of eight
conserved cysteine residues. Other members of this family
include VEGF-B, VEGF-C, VEGF-D, VEGF-E and placen-
tal growth factor. All of the VEGFs are expressed in
mammalian cells, except VEGF-E that is encoded by the orf
virus; a parapoxvirus that affects sheep, goats and sometimes
humans [16, 17]. VEGF is a homodimeric glycoprotein that
is encodeds by a gene resides on chromosome 6 [18]. The
human VEGF gene spans ~14 kb and contains eight exons
separated by seven introns [19, 20]. As a result of alternative
splicing, at least 4 types of monomeric VEGF containing
121, 165, 189, and 206 amino acid residues (VEGF121,
VEGF165, VEGF189, and VEGF206) have been detected.
Exons 1 to 5 are common in all VEGF isoforms, while exons
6 to 8 can be added to this backbone and form various types
of VEGF [21].
VEGF exerts its pivotal role in vasculogenesis (formation
of blood vessels from endothelial cells) and angiogenesis
(formation of new capillaries from pre-existing vessels)
through binding to specific receptors. The receptor binding
site of VEGF is located in exons 3 and 4; thereby all VEGF
isoforms can bind to their receptors [22]. VEGF is thought to
signals through at least two transmembrane high-affinity
receptor tyrosine kinases: VEGFR-1 (FLT-1; fms-like
tyrosine kinase) and VEGFR-2 (KDR; kinase domain
region). Binding of VEGF induces dimerization and
autophosphory-lation of the receptor, which leads to multiple
cellular out-comes, including: endothelial cell migration,
proliferation, survival, and enhanced vascular permeability
that occur during vasculogenesis and angiogenesis. In fact,
VEGF indu-ces vascular permeability 50,000 times more
than histamine [23-25].
There are several evidences that support a causal role for
VEGF in corneal NV [26-33]. It has been shown that VEGF
expression is significantly higher in vascularized corneas
than in normal corneas. Macrophages, infiltrating leuko-
cytes, corneal epithelial and stromal cells are the main
sources of VEGF production during corneal NV [34]. In fact,
it has been reported that application of recombinant VEGF
induces corneal NV in animal models. VEGF overexpression
in the cornea promotes several steps of angiogenesis,
including proteolytic activities, proliferation, migration, and
tube formation of endothelial cells [35]. The role of VEGF
and VEGFR-2 in the regulation of corneal NV and wound
healing was investigated in an animal model of alkali burn
[27]. Moreover the overexpression of VEGF and flt-1 has
been reported in miscellaneous morbid corneas that may play
an important role in the repair of corneal injuries [33].
The critical role of VEGF in the pathogenesis of NV
confirmed by studies that showed anti-VEGF agents can
inhibit corneal NV. It has been reported that VEGF
Corneal Neovascularization Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3 223
inhibition with mFlt(1-3)-immunoglobulin G diminishes
angiogenesis and the severity of lesions after HSV infection
[36]. Another study showed that stromal implantation of an
anti-VEGF blocking antibody can inhibit corneal NV in a rat
model [37]. In addition, it has been reported that VEGF
antagonists, like soluble form of the VEGF receptor and anti-
VEGF siRNA, inhibit herpes simplex induced corneal NV in
mice [20, 32]. Recently, Bevacizumab (Avastin), an anti-
VEGF monoclonal antibody, is considered as a safe and
effective therapeutic option for corneal NV. Avastin has
been investigated in both human and animal corneal NV and
the results are very promising. Many studies showed that
topical and subconjunctival application of Avastin inhibits
chemical and viral induced corneal NV [38-49]. In addition
to VEGF neutralizing agents, it has been reported that
tyrosine kinase inhibitors that blocks VEGF signaling
through its receptors can inhibit corneal NV and improve
corneal graft outcome in animal models [50,51]. To date,
there are numerous patents that have been focused on the
inhibitors of VEGF and its receptor for controlling ocular
NV conditions [52-54].
FIBROBLAST GROWTH FACTOR
Basic fibroblast growth factor (bFGF) is another
angiogenic factor that its role in induction of corneal NV has
been investigated in several studies. bFGF belongs to the
fibroblast growth factor family, with more than 20 members
[11, 55]. It has been localized in the corneal epithelial and
endothelial cells in low levels [56, 57]. Injured corneal
epithelial cells produce bFGF in response to external stimuli
[58]. To date, bFGF has been used extensively in corneal
angiogenesis models and is thought to be one of the major
factors involved in the induction of corneal angiogenesis [11,
59]. The angiogenic properties of bFGF are mediated by
tyrosine kinase receptors, denoted as FGFR-1, -2, -3, and -4.
Signal transduction through these receptors is essential for
development and angiogenesis, because FGFR-1 or FGFR-2
gene disruption leads to embryonic death [60-62]. Numerous
evidences showed that FGF can stimulate proliferation of the
corneal epithelial cells, stromal fibroblasts, and endothelial
cells. bFGF is expressed in the normal corneal epithelium,
but is not detectable in corneal epithelial cell basement
membrane (Bowman's layer). Injury to the corneal
epithelium results in the releasing of bFGF from epithelial
cells and subsequent binding to the Bowman's layer [63]. It
has been shown that FGF-2 is up-regulated in a corneal
stromal cell-vascular endothelial cell coculture [60]. bFGF
induced micropocket assay is one of the most reliable
methods for induction of NV in the cornea. Placement of low
levels of bFGF in the cornea in animal models induces new
blood vessels formation from limbal plexus. Anti-bFGF
agents can inhibit in vitro endothelial cell tube formation and
in vivo corneal NV, which can indicate the important role of
bFGF in angiogenesis [64-66].
MATRIX METALLOPROTEINASES
Matrix metalloproteinases (MMPs) are considered as
other key mediators of angiogenesis. They are a family of
zinc-dependent enzymes, essential for degradation of
extracellular matrix (ECM) and vascular basement mem-
brane during angiogenesis. Their substrates include most of
the extracellular matrix components such as collagen, fibro-
nectin, laminin and cell surface molecules. The MMP family
composed of at least 25 different proteinases that are divided
into five groups, including: the gelatinases, collagenases,
stromelysins, membrane-type MMPs, and others. Several
MMPs are believed to be important in angiogenesis, but
particular interest has been focused on the MMP-2 and
MMP-9 (gelatinase A and B, respectively), because they
preferentially degrade basement membrane components such
as type IV collagen [67,68]. In the cornea, both MMP-2 and
MMP-9 are produced by the endothelial cells and stromal
cells around the invading vessels.
MMP-2 is expressed in tissues where NV occurs,
including solid tumors, wound healing, and choroidal NV in
age-related macular degeneration [69]. There is a general
consensus that MMP-2 promotes angiogenesis by breaking
down either the vascular endothelial cell basal membrane or
the surrounding ECM [68]. The over expression of MMP-2
during corneal NV has been confirmed by numerous
investigations [70-73]. Kvanta et al. reported that MMP-2
expression increases during inflammation-associated corneal
NV in a rat model [74]. A recent study investigated corneal
NV in MMP-2-deficient mice compared with wild-type mice
and showed statistically significant delay of NV in MMP-2-
deficient mice [75]. Another study reported that MMP-2 is
required for optimal experimental choroidal NV in a mouse
model [76].
MMP-9 (gelatinase B) is one of the major degrading
enzymes in ocular surface inflammation. MMP-9 is
overexpressed in vascularized cornea and plays a role in
wound healing [77]. There is high level of MMP-9 in tear of
patients with recurrent corneal ulcers [78]. In addition, it has
been reported that in MMP-9-deficient mice, healing of
corneal wounds is accelerated as a result of their increased
keratinocyte proliferation rate [79]. Taken together, these
data indicate that both MMP-2 and MMP-9 have a role in the
disruption of the basement membrane, integrity of the
corneal epithelium and corneal NV; thus targeting them may
be a useful therapeutic option for inhibition of angiogenesis
in the cornea. In fact, promising studies showed that MMPs
inhibitors can inhibit corneal NV and help to maintain
corneal clarity [80-82].
In addition to these mediators, several hormones and
biologically active molecules may be involved in corneal
NV. Numerous studies showed that angiopoietin 1 and 2 [83-
85], insulin like growth factor (IGF) [86], placenta growth
factor (PGF) [87], platelet derived growth factor (PDGF)
[88], leptin [89] and hepatocyte growth factor (HGF) [90]
are contributed to the pathogenesis of corneal NV. Further
investigations are needed to clarify the exact role of these
molecules in corneal NV.
ANTI-ANGIOGENIC MOLECULES AND CORNEAL
NEOVASCULARIZATION
Thrombospondin
Thrombospondins (TSP) are glycoproteins encoding by a
family of five genes that behave as multifunctional proteins.
Among this family, TSP-1 and -2 show potent anti-angio-
224 Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3
genic properties. TSP-1 inhibits critical events in angio-
genesis, including endothelial cell proliferation, survival and
migration [91]. TSP-1 binds to CD36 on endothelial cell
surface and induces apoptosis [92-94]. Moreover it has been
reported that TSP-1 inhibits VEGF secretion and down
regulates MMP expression in endothelial cells [95, 96]. In
normal cornea, TSP-1 expresses in the Bowman’s layer,
endothelial cells, and Descemet’s membrane [97]. Interes-
tingly, it has been shown that corneas of TSP1,2-/- mice show
greater NV than normal mice in response to injuries.
Therefore TSP could be a major natural angiogenic inhibitor
in the cornea which plays an active role in preserving
angiogenic privilege [91]. In animal models of corneal NV,
TSP-1 derived peptide inhibits bFGF induced angiogenesis
[98]. Moreover, a recent study reported that topical appli-
cation of mesenchymal stem cells to injured corneas inhibits
NV through production of TSP-1 [99].
PIGMENT EPITHELIUM DERIVED FACTOR
Pigment epithelium derived factor (PEDF) is a 50 - kDa
natural anti-angiogenic and neurotrophic protein that was
firstly localized in fetal human retinal pigment epithelial
cells [100]. In the eye, PEDF can be detected in retinal cells,
iris, and the cornea [11]. PEDF is a member of serine
protease inhibitors family but in practice it does not inhibit
serine proteases [101]. In contrast to VEGF that is induced
by low oxygen levels; the expression of PEDF is suppressed
by hypoxia [102]. The anti-angiogenic properties of PEDF
have been confirmed by numerous studies [103-106].
Neutralization of PEDF in normal cornea by specific
antibodies induces NV, which shows its important role in
corneal angiogenic privilege [11, 102, 107]. It has been
reported that application of PEDF can inhibit bFGF induced
corneal NV in animal models [108]. Further studies are
needed to clarify the exact role of PEDF in corneal NV.
ANGIOSTATIN
Angiostatin is a 38 - kDa protein derived from enzymatic
digestion of plasminogen, which is a potent anti-angiogenic
factor. It was first found in urine of mice carrying Lewis
lung carcinomas [109]. Angiostatin inhibits endothelial cells
proliferation, migration and tubule formation [11, 110, 111].
Moreover, angiostatin induces vascular endothelial cell
apoptosis, and stops cell growth at the G2/M transition [112].
Plasminogen and its mRNA were found in all three layers of
the cornea. Interestingly, it has been reported that the corneal
cells have ability to digest plasminogen to angiostatin. In
fact, all of the essential MMPs for converting plasminogen to
angiostatin are found in the corneal cells [108]. It seems that
angiostatin has a role in maintaining corneal avascularity.
Angiostatin can be detected in the tear film of contact lens
bearing patients [113]. Depletion of intra-corneal angiostatin
molecules using specific antibodies increases the risk of NV
in animal models [114]. In addition, application of angio-
statin inhibits bFGF and angiogenin induced NV [110].
Altogether, these data indicate the importance of angiostatin
and its fragments in normal and injured cornea and bring it
to the mind as a potent therapeutic option for clinical uses
[115-119].
Shakiba et al.
ENDOSTATIN
Endostatin is a 20 - kDa fragment of collagen XVIII,
which shows anti-angiogenic properties. Endostatin was first
discovered in the Dr. Judah Folkman laboratory from
conditioned medium of hemangioendothelioma cells [120].
Anti-angiogenic mechanism of endostatin is mediated by
several mechanisms: 1) Endostatin blocks the activity of
gelatinases (MMP-2 and MMP-9) and MMP-13 [121]. 2)
Endostatin inhibits the binding of VEGF to its receptors as
well as its signal transduction. It blocks VEGF receptor
tyrosine kinase phosphorylation and downstream signaling
events like activation of ERK, p38 MAPK, and p125FAK
[122]. 3) Binding of endostatin to endothelial cells arrests
their cell cycle in G1 [123]. 4) Endostatin induces apoptosis
in endothelial cell by activation of caspase 3 [124]. 5)
Endostatin inhibits endothelial cell tubule formation [125].
The results of microarrays studies covering over 90% of the
human genome showed that endostatin regulated about 12%
of all genes in human endothelial cells [126]. In normal
cornea, collagen XVIII can be found naturally. It seems that
during corneal NV and wound healing, this type of collagen
can be digested by MMPs, leading to production of
Endostatin like fragments [11, 127,128]. Previous studies
confirmed the efficacy of endostatin application in control-
ling corneal NV. In animal models, endostatin inhibits bFGF
induced corneal NV [129-131]. Altogether, the results of
numerous studies showed that endostatin and its fragments
can be considered as potent and safe anti-angiogenic agents
in ocular NV disorders, including corneal NV [132-134].
THE ROLE OF INFLAMMATORY CYTOKINES IN
CORNEAL NV
As noted above, inflammatory and infectious diseases are
major etiologies of corneal NV. In fact, corneal NV is almost
always accompanied by inflammation. Previous investi-
gations showed that the corneal cells secret various cytokines
that are involved in ocular surface diseases. In in vitro
conditions, the corneal epithelial cells express IL-1 [135-
137]; the corneal epithelial, stromal and endothelial cells
produce IL-6 [137-139]; and the corneal epithelial and
stromal cells produce IL-8 [140]. It has been reported that
IL-1 is the main inflammatory cytokine produced by the
corneal epithelial cells. In the inflamed cornea, IL-1 is
associated with inflammatory and degradative functions.
This cytokine is chemotactic for fibroblasts and inflam-
matory cells and stimulates fibroblast proliferation and
collagenase expression. In addition to the corneal epithelial
cells, IL-1 is produced by infiltrating monocytes and
macrophages and it’s over expression (along with TNF-
) is
one of the early hallmarks of corneal inflammation [141].
Previous studies confirmed that the IL-1 receptor, which
both IL-1
and IL-1 bind, is expressed in the corneal
fibroblasts. Binding of IL-1 to the corneal fibroblast induces
the expression of various angiogenic factors [142]. It has
been demonstrated that IL-1 induces NF-B signaling in the
mouse cornea during NV. In the corneal alkali burns model,
NF-B is activated in the corneal epithelial and stromal cells
[143]. Inhibitor of NF-B (IB-
) is mainly phosphorylated
in the corneal stromal cells, suggesting that NF-B-activated
corneal stromal fibroblasts play an important role in corneal
inflammatory conditions, such as angiogenesis and wound
Corneal Neovascularization Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3 225
healing [144]. Recently, an interesting study showed that
topical administration of 2% IL-1 receptor antagonist (IL-
1Ra) inhibits inflammation induced corneal NV [141].
TNF-
is another important proinflammatory cytokine
involved in corneal NV. It is a pleiotropic proinflammatory
cytokine that mediates various immunoregulatory functions,
such as up-regulation of adhesion molecules, activation of
neutrophils, and activation of the NF-B signaling pathway
[145]. TNF-
is produced by various cells in the inflam-
matory site and induces angiogenic cytokines, including IL-
8, VEGF, and bFGF [146]. TNF-
exerts inflammatory
properties through its receptors, TNF-
R1 and TNF-
R2.
Binding of TNF-
to its receptors on the endothelial cells
increases the expression of many angiogenic molecules
[147]. Previous studies showed that TNF-
is a two-edged
sword in controlling angiogenesis. It has been demonstrated
that angiogenic properties of TNF-
is dose-dependent. In
low doses, TNF-
induces angiogenesis, whereas a high dose
inhibits it [148]. In the inflamed cornea, TNF-
is secreted
by the corneal resident cells [148,149]. In addition TNF-

significantly increases IL-8 expression in the corneal
endothelial and stromal cells. IL-8 is a potent chemo-
attractant factor that promotes angiogenesis in vitro as well
as in vivo [150]. It has been reported that intracorneal injec-
tion of TNF-
increases the number of Langerhans cells in
limbus and induces migration of them to the central cornea
[151]. More investigations are needed for clarifying the role
of TNF-
in corneal NV.
Transforming growth factor  (TGF-) is another main
cytokine that has an important role in corneal NV. TGF-
can act as pro- or anti-angiogenic factor depending on
context and concentration. In low doses, TGF- stimulates
endothelial cells proliferation, whereas excessively high
doses can inhibit it [152,153]. During corneal injuries, TGF-
 is produced by inflammatory cells and resident corneal
cells [154]. TGF- also stimulates angiogenesis by recruiting
inflammatory cells which, in turn, release other pro-
angiogenic cytokines [155]. It has been reported that three
isoforms of TGF- and their receptors are present in the
cornea [156-159]. In fact, TGF- is present on the edge of
wound healing in the cornea [160]. In animal models,
neutralization of TGF- inhibited corneal NV [156]. Further-
more, it seems that TGF- directly stimulates the production
of VEGF in various cells, including fibroblastic epithelial
cells [161]. Altogether, the exact role of TGF- in corneal
NV is not completely understood and there are controversial
reports regard its role in angiogenesis. Further investigations
are needed for understanding its role in normal and
vascularized cornea.
THERAPEUTIC OPTIONS
Surgical Treatments
To date, numerous surgical methods have been recom-
mended for controlling corneal NV. The first investigations
considered diathermy for destruction of newly formed blood
vessels in the cornea. In recent years, other techniques like
laser photocoagulation have been evaluated in the treatment
of corneal NV. The effect of laser coagulation is due to
energy absorption by hemoglobin in blood vessels. It seems
that this technique is effective in treatment of numerous
subsets of corneal NV, but it requires high amounts of
thermal laser energy to produce adequate destruction of
vessels, and there is a very high recurrence rate. Moreover,
this high energy can lead to some complications, including
increased inflammation and exacerbation of the NV
[162,163]. Laser photocoagulation is used by some investi-
gators for diminishing NV before and after penetrating
keratoplasty. Results from laser photocoagulation showed
that it is more useful for controlling low to moderate forms
of corneal NV [11].
Photodynamic therapy is another method that involves
the administration of a photosensitizing compound, which is
selectively absorbed by neovascular tissues. In this method,
the photosensitizer may be administered systemic or topical
on the neovascular cornea. Subsequent activation of this
compound by low energy laser light generates cytotoxic
mediators, which cause selective newly formed vessel
thrombosis and destruction [164]. This method can be
considered as a safe and effective procedure for controlling
corneal NV, because total used energy is 20 times less than
conventional thermal laser [165]. Moreover, thermal damage
to normal pericorneal tissues is less common. In recent
years, many investigations showed the efficacy of photo-
dynamic therapy in both human and animal models of
corneal NV [166,167].
CORTICOSTEROIDS
Corticosteroids inhibit inflammation and angiogenesis.
To date, corticosteroid therapy has been considered as the
standard anti-inflammatory and anti-angiogenic treatment in
the cornea. Previous studies supported this idea that anti-
angiogenic effect of corticosteroids is due to their anti-
inflammatory properties [168,169]. It has been clarified that
various signaling pathways are common in inflammation and
angiogenesis. In the injured cornea, infiltrating inflammatory
cells are the major sources of angiogenic molecules like
VEGF. In addition, these cells produce many cytokines like
IL-1 and TNF-
that are categorized as potent angiogenic
molecules [170]. The steroids exert their anti-inflammatory
effects, mainly by inhibiting the enzyme phospholipase and
thereby the arachidonic acid pathway. Inhibition of phos-
pholipase decreases production of prostaglandins from cell
membrane lipids. In addition to inflammatory properties,
some prostaglandins act as angiogenic factors. In fact,
steroids behave as anti-angiogenic agents by inhibiting
prostaglandin synthesis [171]. Systemically administered
steroids inhibit the transcription of several inflammatory and
angiogenic cytokines, including IL-1, IL-3, IL-6, and IL-8
[172]. Moreover, previous studies notified that reducing the
expression of VEGF may be another mechanism of steroids
anti-angiogenic effects [173].
The first investigated steroids had both anti-inflammation
and anti-angiogenic activities and some of them appeared to
require specific cofactors such as heparin or cyclodextran
[174]. Although, this type of steroids has anti-angiogenic
effects, they may cause significant ocular side effects like
development of posterior subcapsular cataracts and elevated
intra ocular pressure [175-177]. In recent years, new class of
steroids discovered, which has broad angiostatic and low
glucocorticoid activities. This class of compounds is consi-
226 Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3
dered as potential therapy for ocular neovascular disease,
since undesirable side effects, which are commonly asso-
ciated with glucocorticoids, would be avoided [178,179].
Anecortave acetate is a known member of this class of
steroids. This anti-angiogenic drug is derived from the
glucocorticoid cortisol acetate and has no glucocorticoid
activity. Anecortave acetate has broad based angiostatic
activity and inhibits critical steps of angiogenesis. It inhibits
the angiogenic proteolytic cascade at several points by
suppressing production of uPA and proMMPs from
endothelial cells and increasing the expression of PAI-1, an
endogenous inhibitor of uPA. Moreover anecortave acetate
decreases the expression of gelatinase A and B and inhibits
VEGF-induced bovine and human endothelial cell
proliferation and tube formation [180]. In animal models, it
has been reported that topical administration of anecortave
acetate suppresses LPS and FGF2 induced corneal NV. In
that study, it has been reported that application of 0.1% or
1% suspensions of anecortave acetate four times in day
almost totally inhibits corneal NV in rabbits [168]. After
confirming safety and efficacy by more investigations, ane-
cortave acetate can be considered for controlling corneal NV.
NONSTEROIDAL ANTI-INFLAMMATORY DRUGS
Nonsteroidal anti-inflammatory drugs (NSAIDs) are
widely used in ophthalmology for controlling pain and
inflammation in ocular surface disorders. NSAIDs inhibit the
activity of the cyclooxygenases (COX-1 and COX-2) that
convert arachidonic acid to prostaglandins (PGs). Previous
studies showed that COX-2 is an inducible isoform of
cyclooxygenases, which induces multiple events of
angiogenesis like proliferation and migration of endothelial
cells and inhibits apoptosis [181]. It has been clarified that
some of PGs, like PGE2, induces angiogenesis by increasing
VEGF and bFGF gene expression [182,183]. In fact, it has
been showed that inhibitors of COX-2 inhibit angiogenesis
in vitro and in vivo [184-186]. Both COX-1 and COX-2 are
found in the cornea and their role in the pathogenesis of NV
has been investigated [187].
Previous studies showed that NSAIDs can control
corneal NV. Indomethacin and ketoprofen, nonselective
COX-1 and COX-2 inhibitors, significantly suppress bFGF-
and VEGF-induced angiogenesis in mouse cornea [184].
Interestingly, this study notified that selective inhibitor of
COX-2 (celecoxib and rofecoxib) suppresses bFGF-induced
NV more effectively than VEGF induced NV. Thus it was
concluded that selective inhibitors of COX-2 should be used
in the early course of corneal NV and before the upregu-
lation of large amounts of VEGF. Once VEGF is present,
selective COX-2 inhibitors would be less active in inhibiting
VEGF-induced angiogenesis [184].
ANTI-VEGF AGENTS
Over the past decade, there were progressive attempts for
using anti VEGF agents for controlling corneal NV.
Numerous studies showed that Bevacizumab can inhibit
ocular NV in both human and animal models. Bevacizumab
is a recombinant humanized monoclonal antibody against the
VEGF molecule that binds and neutralizes it effectively
[188]. Today, Bevacizumab is commercially available and is
Shakiba et al.
approved by the US Food and Drug Administration (FDA)
for use in the treatment of metastatic colorectal cancer [189].
Previous studies confirmed that topical or subconjuctival
application of Bevacizumab controls new blood vessel
formation in the cornea [190-193]. It is reported that single
subconjunctival dose of Bevacizumab inhibits NV in alkali
burned rabbit corneas [194]. Moreover, several case reports
notified the therapeutic properties of Bevacizumab in
patients with corneal NV [49, 195]. Interestingly, the recent
studies showed the efficacy of Bevacizumab in inhibiting
herpetic corneal NV. A recent investigation reported that
subconjunctival application of Bevacizumab decreases
corneal NV dramatically 1 week after the injection in a
woman with herpetic stromal keratitis [196]. In addition to
Bevacizumab, recent evidences showed that inhibitors of
VEGF receptors can be efficient in controlling corneal NV.
In animal models, tyrosine kinase inhibitors decrease corneal
NV and prolong graft survival [51].
Taken together, these data indicate that Bevacizumab can
be a promising drug in treatment of cornea NV and graft
survival [197,198]. Anti-VEGF substances may increasingly
play a role in treatment of anterior segment NV in the future.
In addition to these well-known therapeutic strategies
described above, there are numerous agents that showed
inhibitory effects in corneal NV. It has been reported that
‘suleparoide [199], thalidomide [200], suramin [201],
genistein [202], somatostatin [203], octreotide [204],
cyclosporine [205], methotrexate [206], rapamycin [207] and
tacrolimus [208] can abolish corneal NV in animal models.
However, it seems that these medications exert their
therapeutic properties through reducing inflammatory and
angiogenic factors, mainly VEGF. Further investigations in
animal models and human cases are needed to place these
agents alongside corneal NV therapeutics.
CURRENT & FUTURE DEVELOPMENTS
During past two decades, paralle with our understanding
of molecular events underlying ocular NV, there was a
general interesting for using anti-VEGF angents as a potent
and safe theraputic option for controlling corneal
angiogenesis. It is expected that VEGF blocking agents or
tyrosin kinase inhibitors will play an imporant role in
increasing the graft survival and conserving vision after
keratoplasty in near future. Moreover, VEGF is known as a
main mediator involves in the pathogenesis of herpetic
keratitis and pterygium; therefore anti-VEGF drugs can
revolutionize their treatment strategies. Today, at least two
VEGF inhibitors are approved for treatment of angiogenesis-
related ophthalmic diseases; Lucentis (rhuFab V2;
Genentech, Inc.) and Macugen (pegaptanib sodium; Eyetech
Pharmaceuticals) and more are poised to enter into clinical
practice. Focus on determination of safe and effective
concentration, exposure time, and administration routes of
anti-angiogenic drugs can accelerate their entrance to
ophthalmology in near future.
ACKNOWLEDGEMENT
The authors would like to thank Dr. Amir Kiani and
Dr. Behrouz Nikbin for their valuable assistance.
Corneal Neovascularization Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3 227
CONFLICT OF INTEREST
No financial contribution to the work has been declared.
No patents are reported in any stage of legal litigations.
REFERENCES
[1] Niederkorn JY. Immune privilege in the anterior chamber of the
eye. Crit Rev Immunol 2002; 22(1): 13-46.
[2] Gipson IK, Inatomi T. Cellular origin of mucins of the ocular
surface tear film. Adv Exp Med Biol 1998; 438: 221-227.
[3] Hosseini H, Nejabat M. A potential therapeutic strategy for
inhibition of corneal neovascularization with new anti-VEGF
agents. Med Hypotheses 2007; 68(4): 799-801.
[4] Jain RK. Molecular regulation of vessel maturation. Nat Med 2003;
9(6): 685-693.
[5] Colby KA, Adamis AP. Prevalence of corneal neovascularization in
a general eye service population (Abstract). Invest Ophthalmol Vis
Sci 1996; 37: S593.
[6] Zhang SX, Ma JX. Ocular neovascularization: Implication of
endogenous angiogenic inhibitors and potential therapy. Prog Retin
Eye Res 2007; 26(1): 1-37.
[7] West SK. Trachoma: New assault on an ancient disease. Prog Retin
Eye Res 2004; 23(4): 381-401.
[8] Lee P, Wang CC, Adamis AP. Ocular neovascularization: An
epidemiologic review. Surv Ophthalmol 1998; 43(3): 245-269.
[9] Zheng M, Schwarz MA, Lee S, et al. Control of stromal keratitis by
inhibition of neovascularization. Am J Pathol 2001; 159(3): 1021-
1029.
[10] Kremer I, Cohen EJ, Eagle RC Jr, et al. Histopathologic evaluation
of stromal inflammation in Acanthamoeba keratitis. CLAO J 1994;
20(1): 45-48.
[11] Chang JH, Gabison EE, Kato T, et al. Corneal neovascularization.
Curr Opin Ophthalmol 2001; 12(4): 242-249.
[12] Dana MR, Schaumberg DA, Kowal VO, et al. Corneal
neovascularization after penetrating keratoplasty. Cornea 1995;
14(6): 604-609.
[13] Gerten G. Bevacizumab (avastin) and argon laser to treat
neovascularization in corneal transplant surgery. Cornea 2008;
27(10): 1195-1199.
[14] Rozenman Y, Donnenfeld ED, Cohen EJ, et al. Contact lens-related
deep stromal neovascularization. Am J Ophthalmol 1989; 107(1):
27-32.
[15] Liesegang TJ. Physiologic changes of the cornea with contact lens
wear. CLAO J 2002; 28(1): 12-27.
[16] Li X, Eriksson U. Novel VEGF family members: VEGF-B, VEGF-
C and VEGF-D. Int J Biochem Cell Biol 2001; 33(4): 421-426.
[17] Lyttle DJ, Fraser KM, Fleming SB, et al. Homologs of vascular
endothelial growth factor are encoded by the poxvirus orf virus. J
Virol 1994; 68(1): 84-92.
[18] Vincenti V, Cassano C, Rocchi M, et al. Assignment of the vascular
endothelial growth factor gene to human chromosome 6p21.3.
Circulation 1996; 93(8): 1493-1495.
[19] Houck KA, Leung DW, Rowland AM, et al. Dual regulation of
vascular endothelial growth factor bioavailability by genetic and
proteolytic mechanisms. J Biol Chem 1992; 267(36): 26031-26037.
[20] Tischer E, Mitchell R, Hartman T, et al. The human gene for
vascular endothelial growth factor. Multiple protein forms are
encoded through alternative exon splicing. J Biol Chem 1991;
266(18): 11947-11954.
[21] Robinson CJ, Stringer SE. The splice variants of vascular
endothelial growth factor (VEGF) and their receptors. J Cell Sci
2001; 114: 853-865.
[22] Ng YS, Krilleke D, Shima DT. VEGF function in vascular
pathogenesis. Exp Cell Res 2006; 312(5): 527-537.
[23] Senger DR, Connolly DT, Van de Water L, et al. Purification and
NH2-terminal amino acid sequence of guinea pig tumor-secreted
vascular permeability factor. Cancer Res 1990; 50(6): 1774-1778.
[24] Meadows KN, Bryant P, Pumiglia K. Vascular endothelial growth
factor induction of the angiogenic phenotype requires Ras
activation. J Biol Chem 2001; 276(52): 49289-49298.
[25] Takahashi T, Yamaguchi S, Chida K, et al. A single
autophosphorylation site on KDR/Flk-1 is essential for VEGF-A-
dependent activation of PLC-gamma and DNA synthesis in
vascular endothelial cells. EMBO J 2001; 20(11): 2768-2778.
[26] Singh N, Amin S, Richter E, et al. Flt- 1 intraceptors inhibit
hypoxia-induced VEGF expression in vitro and corneal
neovascularization in vivo. Invest Ophthalmol Vis Sci 2005; 46(5):
1647-1652.
[27] Gan L, Fagerholm P, Palmblad J. Vascular endothelial growth
factor VEGF and its receptor VEGFR-2 in the regulation of corneal
neovascularization and wound healing. Acta Ophthalmol Scand
2004; 82(5): 557-563.
[28] Joussen AM, Poulaki V, Mitsiades N, et al. VEGF-dependent
conjunctivalization of the corneal surface. Invest Ophthalmol Vis
Sci 2003; 44(1): 117-123.
[29] Zhou LH, Xing YQ, Chen CL, et al. Antisense vascular endothelial
growth factor suppressed corneal neovascularization in rats.
Zhonghua Yan Ke Za Zhi 2006; 42(5): 426-430.
[30] Murata M, Takanami T, Shimizu S, et al. Inhibition of ocular
angiogenesis by diced small interfering RNAs (siRNAs) specific to
vascular endothelial growth factor (VEGF). Curr Eye Res 2006;
31(2): 171-180.
[31] Lai CM, Spilsbury K, Brankov M, et al. Inhibition of corneal
neovascularization by recombinant adenovirus mediated antisense
VEGF RNA. Exp Eye Res 2002; 75(6): 625-634.
[32] Kim B, Tang Q, Biswas PS, et al. Inhibition of ocular angiogenesis
by siRNA targeting vascular endothelial growth factor pathway
genes: Therapeutic strategy for herpetic stromal keratitis. Am J
Pathol 2004; 165(6): 2177-2185.
[33] Chen J, Liu W, Liu Z, et al. Expression of vascular endothelial
growth factor and its receptors (FLT-1) in morbid human corneas
and investigation of its clinic importance. Yan Ke Xue Bao 2002;
18(4): 203-207.
[34] Seta F, Patil K, Bellner L, et al. Inhibition of VEGF expression and
corneal neovascularization by siRNA targeting cytochrome P450
4B1. Prostaglandins Other Lipid Mediat 2007; 84(3-4): 116-127.
[35] Phillips GD, Stone AM, Jones BD, et al. Vascular endothelial
growth factor (rhVEGF165) stimulates direct angiogenesis in the
rabbit cornea. In Vivo 1994; 8(6): 961-965.
[36] Zheng M, Deshpande S, Lee S, et al. Contribution of vascular
endothelial growth factor in the neovascularization process during
the pathogenesis of herpetic stromal keratitis. J Virol 2001; 75(20):
9828-9835.
[37] Amano S, Rohan R, Kuroki M, et al. Requirement for vascular
endothelial growth factor in wound- and inflammation related
corneal neovascularization. Invest Ophthalmol Vis Sci 1998; 39(1):
18-22.
[38] Uy HS, Chan PS, Ang RE. Topical bevacizumab and ocular surface
neovascularization in patients with stevens-johnson syndrome.
Cornea 2008; 27(1): 70-73.
[39] Mackenzie SE, Tucker WR, Poole TR. Bevacizumab (avastin) for
corneal neovascularization-corneal light shield soaked application.
Cornea 2009; 28(2): 246-247.
[40] Bahar I, Kaiserman I, McAllum P, et al. Subconjunctival
bevacizumab injection for corneal neovascularization in recurrent
pterygium. Curr Eye Res 2008; 33(1): 23-28.
[41] Gerten G. Bevacizumab (avastin) and argon laser to treat
neovascularization in corneal transplant surgery. Cornea 2008;
27(10): 1195-1199.
[42] Han YS, Lee JE, Jung JW, et al. Inhibitory effects of bevacizumab
on angiogenesis and corneal neovascularization. Graefes Arch Clin
Exp Ophthalmol 2009; 247(4): 541-548.
[43] Bock F, Onderka J, Rummelt C, et al. Safety profile of topical
VEGF-neutralization at the cornea. Invest Ophthalmol Vis Sci
2009; 50(5): 2095-2102.
[44] Hurmeric V, Mumcuoglu T, Erdurman C, et al. Effect of
subconjunctival bevacizumab (Avastin) on experimental corneal
neovascularization in guinea pigs. Cornea 2008; 27(3): 357-362.
[45] Chen WL, Lin CT, Lin NT, et al. Subconjunctival injection of
bevacizumab (avastin) on corneal neovascularization in different
rabbit models of corneal angiogenesis. Invest Ophthalmol Vis Sci
2009; 50(4): 1659-1665.
[46] Kim TI, Kim SW, Kim S, et al. Inhibition of experimental corneal
neovascularization by using subconjunctival injection of
bevacizumab (Avastin). Cornea 2008; 27(3): 349-352.
[47] Bahar I, Kaiserman I, McAllum P, et al. Subconjunctival
bevacizumab injection for corneal neovascularization. Cornea 2008;
27(2): 142-147.
[48] Doctor PP, Bhat PV, Foster CS. Subconjunctival bevacizumab for
corneal neovascularization. Cornea 2008; 27(9): 992-995.
228 Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3
[49] Kim SW, Ha BJ, Kim EK, et al. The effect of topical bevacizumab
on corneal neovascularization. Ophthalmology 2008; 115(6): e33-
38.
[50] Shakiba Y, Arshadi D. Inhibition of corneal neovascularization with
new tyrosine kinase inhibitors targeting vascular endothelial growth
factor receptors: Sunitinib malate and Sorafenib. Irn J Med
Hypotheses Ideas 2007; 1:1
[51] Hos D, Bock F, Dietrich T, et al. Inflammatory corneal (lymph)
angiogenesis is blocked by VEGFR-tyrosine kinase inhibitor ZK
261991, resulting in improved graft survival after corneal
transplantation. Invest Ophthalmol Vis Sci 2008; 49(5): 1836-1842.
[52] Kimihiko, F., Tatsuro, I., Tadahisa, K., Yukio, S.: EP0811711A1
(2006).
[53] Rosenblum, M.G., Campochiaro, P. A.: US2007025957A1 (2007).
[54] Adamis, A. P., Guyer, D. R., Modi, M. W.: US2006293270A1
(2006).
[55] Hayashi N, Nakayasu K, Okisaka S. Immunohistochemical
localization of acidic and basic fibroblast growth factor through
corneal neovascularization in vivo and in vitro. Nippon Ganka
Gakkai Zasshi 1996; 100: 587-591.
[56] Gan L, Fagerholm P, Palmblad J. Expression of basic fibroblast
growth factor in rabbit corneal alkali wounds in the presence and
absence of granulocytes. Acta Ophthalmol Scand 2005; 83(3): 374-
378.
[57] Saghizadeh M, Chwa M, Aoki A, et al. Altered expression of
growth factors and cytokines in keratoconus, bullous keratopathy
and diabetic human corneas. Exp Eye Res 2001; 73(2): 179-189.
[58] Adamis AP, Meklir B, Joyce NC. in situ Injury-induced release of
basic-fibroblast growth factor from corneal epithelial cells. Am J
Pathol 1991; 139: 961-967.
[59] Rogers MS, Birsner AE, D'Amato RJ. The mouse cornea
micropocket angiogenesis assay. Nat Protoc 2007; 2(10): 2545-
2550.
[60] Kojima T, Chang JH, Azar DT. Proangiogenic role of
ephrinB1/EphB1 in basic fibroblast growth factor-induced corneal
angiogenesis. Am J Pathol 2007; 170(2): 764-773.
[61] Xu X, Weinstein M, Li C, et al. Fibroblast growth factor receptor 2
(FGFR2)-mediated reciprocal regulation loop between FGF8 and
FGF10 is essential for limb induction. Development 1998; 125(4):
753-765.
[62] Deng CX, Wynshaw-Boris A, Shen MM, et al. Murine FGFR-1 is
required for early postimplantation growth and axial organization.
Genes Dev 1994; 8: 3045-3057.
[63] Soubrane G, Jerdan J, Karpouzas I, et al. Binding of basic fibroblast
growth factor to abnormal and neovascular rabbit cornea. Invest
Opthalmol Vis Sci 1990; 31: 323-333.
[64] Kurokawa M, Doctrow SR, Klagsbrun M. Neutralizing antibodies
inhibit the binding of basic fibroblast growth factor to its receptor
but not to heparin. J Biol Chem 1989; 264(13): 7686-7691.
[65] Xiang, J., Deng, N., Li, H.: CN1560082A (2004).
[66] Yoshitake, Y., Nishikawa, K., Osawa, M, Asano, M., Koda, A.,
Suzuki, H.: JP11049701A (1997)
[67] Visse R, Nagase H. Matrix metalloproteinases and tissue inhibitors
of metalloproteinases: Structure, function, and biochemistry. Circ
Res 2003; 928: 827-839.
[68] Pepper MS. Role of the matrix metalloproteinase and plasminogen
activator-plasmin systems in angiogenesis. Arterioscler Thromb
Vasc Biol 2001; 217: 1104-1117.
[69] Mignatti P, Rifkin DB. Plasminogen activators and matrix
metalloproteinases in angiogenesis. Enzyme Protein 1996; 49(1-3):
117-137.
[70] Kato T, Kure T, Chang JH, et al. Diminished corneal angiogenesis
in gelatinase A-deficient mice. FEBS Lett 2001; 5082: 187-190.
[71] Zhang H, Li C, Baciu PC. Expression of integrins and MMPs
during alkaline-burn-induced corneal angiogenesis. Invest
Ophthalmol Vis Sci 2002; 434: 955-962.
[72] Rigal-Sastourne JC, Tixier JM, Renard JP, et al. Corneal burns and
matrix metalloproteinases MMP-2 and -9: The effects of human
amniotic membrane transplantation. J Fr Ophtalmol 2002; 257:
685-693.
[73] Ma DH, Chen JK, Kim WS, et al. Expression of matrix
metalloproteinases 2 and 9 and tissue inhibitors of metallo-
proteinase 1 and 2 in inflammation-induced corneal neovas-
cularization. Ophthalmic Res 2001; 336: 353-362.
[74] Kvanta A, Sarman S, Fagerholm P, et al. Expression of matrix
metalloproteinase-2 (MMP-2) and vascular endothelial growth
Shakiba et al.
factor (VEGF) in inflammation-associated corneal neovas-
cularization. Exp Eye Res 2000; 704: 419-428.
[75] Samolov B, Steen B, Seregard S, et al. Delayed inflammation-
associated corneal neovascularization in MMP-2-deficient mice.
Exp Eye Res 2005; 802: 159-166.
[76] Berglin L, Sarman S, van der Ploeg I, et al. Reduced choroidal
neovascular membrane formation in matrix metalloproteinase
metalloproteinase-2-deficient mice. Invest Ophthalmol Vis Sci
2003; 441: 403-408.
[77] Ye HQ, Azar DT. Expression of gelatinases A and B, and TIMPs 1
and 2 during corneal wound healing. Invest Ophthalmol Vis Sci
1998; 39: 913-921.
[78] Sakimoto T, Shoji J, Yamada A, et al. Upregulation of matrix
metalloproteinase in tear fluid of patients with recurrent corneal
erosion. Jpn J Ophthalmol 2007; 51(5): 343-346.
[79] Swarnakar S, Ganguly K, Kundu P, et al. Curcumin regulates
expression and activity of matrix metalloproteinases 9 and 2 during
prevention and healing of indomethacin-induced gastric ulcer. J
Biol Chem 2005; 280(10): 9409-9415.
[80] Gonzales RP, D-Soares FS, Farias RF, et al. Demonstration of
inhibitory effect of oral shark cartilage on basic fibroblast growth
factor-induced angiogenesis in the rabbit cornea. Biol Pharm Bull
2001; 24: 151-154.
[81] Tamargo RJ, Bok RA, Brem H. Angiogenesis inhibition by
minocycline. Cancer Res 1991; 51(2): 672-675.
[82] Sucholeiki, I., Gege, C., Gallagher, B, Powers, T., Deng, H., Wu,
X., Steeneck, C., Kiely, A., Taveras, A.: US20080021024A1
(2008).
[83] Singh N, MacNamara E, Rashid S, et al. Systemic soluble Tie-2
expression inhibits and regresses corneal neovascularization.
Biochem Biophys Res Commun 2005; 332: 194-199.
[84] White RR, Shan S, Rusconi CP, et al. Inhibition of rat corneal
angiogenesis by a nuclease-resistant RNA aptamer specific for
angiopoietin-2. Proc Natl Acad Sci USA 2003; 100: 5028-5033.
[85] Oike Y, Ito Y, Maekawa H, et al. Angiopoietin-related growth
factor (AGF) promotes angiogenesis. Blood 2004; 103(10): 3760-
3765.
[86] Grant MB, Mames RN, Fitzgerald C, et al. Insulin-like growth
factor I acts as an angiogenic agent in rabbit cornea and retina:
Comparative studies with basic fibroblast growth factor.
Diabetologia 1993; 36: 282-291.
[87] Cao Y, Linden P, Shima D, Browne F, Folkman J. in vivo
Angiogenic activity and hypoxia induction of heterodimers of
placenta growth factor/vascular endothelial growth factor. J Clin
Invest 1996; 98(11): 2507-2511.
[88] Cohen RA, Gebhardt BM, Bazan NG. A platelet-activating factor
antagonist reduces corneal allograft inflammation and
neovascularization. Curr Eye Res 1994; 13(2): 139-144.
[89] Park HY, Kwon HM, Lim HJ, et al. Potential role of leptin in
angiogenesis: Leptin induces endothelial cell proliferation and
expression of matrix metalloproteinases in vivo and in vitro. Exp
Mol Med 2001; 33(2): 95-102.
[90] Xin X, Yang S, Ingle G, et al. Hepatocyte growth factor enhances
vascular endothelial growth factor-induced angiogenesis in vitro
and in vivo. Am J Pathol 2001; 158(3): 1111-1120.
[91] Cursiefen C, Masli S, Ng TF, et al. Roles of thrombospondin-1 and
-2 in regulating corneal and iris angiogenesis. Invest Ophthalmol
Vis Sci 2004; 45: 1117-1124.
[92] Nor JE, Mitra RS, Sutorik M, et al. Thrombospondin-1 induces
endothelial cell apoptosis and inhibits angiogenesis by activating
the caspase death pathway. J Vasc Res 2000; 37: 209-218.
[93] Jimenez B, Volpert OV, Crawford SE, et al. Signals leading to
apoptosis-dependent inhibition of neovascularization by
thrombospondin-1. Nat Med 2000; 6: 41-48.
[94] Vande Woude, G. F., Zhang, Y.W.: US20070020234A1 (2007).
[95] Zak S, Treven J, Nash N, et al. Lack of thrombospondin-1 increases
angiogenesis in a model of chronic inflammatory bowel disease. Int
J Colorectal Dis 2008; 23(3): 297-304.
[96] Zhou L, Isenberg JS, Cao Z, et al. Type I collagen is a molecular
target for inhibition of angiogenesis by endogenous
thrombospondin-1. Oncogene 2006; 25: 536-545.
[97] Hiscott P, Seitz B, Schlotzer-Schrehardt U, et al. Immunolo-
calisation of thrombospondin 1 in human, bovine and rabbit cornea.
Cell Tissue Res 1997; 289(2): 307-310.
Corneal Neovascularization Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3 229
[98] Kanda S, Shono T, Tomasini-Johansson B, et al. Role of
thrombospondin-1-derived peptide, 4N1K, in FGF-2-induced
angiogenesis. Exp Cell Res 1999; 252(2): 262-272.
[99] Oh JY, Kim MK, Shin MS, et al. The anti-inflammatory and anti-
angiogenic role of mesenchymal stem cells in corneal wound
healing following chemical injury. Stem Cells 2008; 26(4): 1047-
1055.
[100] Tombran-Tink J, Johnson LV. Neuronal differentiation of
retinoblastoma cells induced by medium conditioned by human
RPE cells. Invest Ophthalmol Vis Sci 1989; 30: 1700-1707.
[101] Becerra SP, Sagasti A, Spinella P, et al. Pigment epithelium-
derived factor behaves like a noninhibitory serpin. Neurotrophic
activity does not require the serpin reactive loop. J Biol Chem 1995;
270(43): 25992-25999.
[102] Dawson DW, Volpert OV, Gillis P, et al. Pigment epithelium-
derived factor: A potent inhibitor of angiogenesis. Science 1999;
285(5425): 245-248.
[103] Notari L, Miller A, Martinez A, et al. Pigment epithelium-derived
factor is a substrate for matrix metalloproteinase type 2 and type 9:
Implications for downregulation in hypoxia. Invest Ophthalmol Vis
Sci 2005; 46(8): 2736-2747.
[104] Abe R, Fujita Y, Yamagishi S, et al. Pigment epithelium-derived
factor prevents melanoma growth via angiogenesis inhibition. Curr
Pharm Des 2008; 14(36): 3802-3809.
[105] Iizuka H, Awata T, Osaki M, et al. Promoter polymorphisms of the
pigment epithelium-derived factor gene are associated with diabetic
retinopathy. Biochem Biophys Res Commun 2007; 361(2): 421-
426.
[106] Bouck N. PEDF: Anti-angiogenic guardian of ocular function.
Trends Mol Med 2002; 8(7): 330-334.
[107] Meyer C, Notari L, Becerra SP. Mapping the type I collagen-
binding site on pigment epithelium-derived factor. Implications for
its antiangiogenic activity. J Biol Chem 2002; 277: 45400-45407.
[108] Azar DT. Corneal angiogenic privilege: Angiogenic and
antiangiogenic factors in corneal avascularity, vasculogenesis, and
wound healing (an American Ophthalmological Society thesis).
Trans Am Ophthalmol Soc 2006; 104: 264-302.
[109] O’Reilly MS, Holmgren L, Shing Y, et al. Angiostatin: A novel
angiogenesis inhibitor that mediates the suppression of metastases
by a Lewis lung carcinoma. Cell 1994; 79: 315-328.
[110] Cao R, Wu HL, Veitonmaki N, et al. Suppression of angiogenesis
and tumor growth by the inhibitor K1-5 generated by plasmin-
mediated proteolysis. Proc Natl Acad Sci USA 1999, 96: 5728-
5733.
[111] Shin SH, Kim JC, Chang SI, et al. Recombinant kringle 1-3 of
plasminogen inhibits rabbit corneal angiogenesis induced by
angiogenin. Cornea 2000; 19: 212-217.
[112] Griscelli F, Li H, Bennaceur-Griscelli A, et al. Angiostatin gene
transfer: Inhibition of tumor growth in vivo by blockage of
endothelial cell proliferation associated with a mitosis arrest. Proc
Natl Acad Sci USA 1998; 95: 6367-6372.
[113] Sack RA, Beaton AR, Sathe S. Diurnal variations in angiostatin in
human tear fluid: A possible role in prevention of corneal
neovascularization. Curr Eye Res 1999; 18: 186-193.
[114] Gabison E, Chang JH, Hernandez-Quintela E, et al. Anti-
angiogenic role of angiostatin during corneal wound healing. Exp
Eye Res 2004; 78: 579-558.
[115] Folkman, J., O'Reilly, M., Cao, Y., Sim, K.: US20090075369A1
(2009).
[116] Soff, G., Gately, S.T., Twardowski, P.: US20060099671A1 (2006).
[117] O'Reilly, M.S., Folkman, M. J., Cao, Y.: US20040023877A1
(2004).
[118] Folkman, M. J., O'Reilly, M. S., Cao, Y., Sim, K.L.:
US20030064926A1 (2003).
[119] O'Reilly, M.S., Folkman, M.J., Cao, Y.: US20010029246A1
(2001).
[120] O’Reilly MS, Boehm T, Shing Y, et al. Endostatin: An endogenous
inhibitor of angiogenesis and tumor growth. Cell 1997; 88: 277-
285.
[121] Nyberg P, Heikkila P, Sorsa T, et al. Endostatin inhibits human
tongue carcinoma cell invasion and intravasation and blocks the
activation of matrix metalloprotease-2, - 9, and -13. J Biol Chem
2003; 278(25): 22404-22411.
[122] Kim YM, Hwang S, Pyun BJ, et al. Endostatin blocks vascular
endothelial growth factor-mediated signaling via direct interaction
with KDR/Flk-1. J Biol Chem 2002; 277: 27872-27879.
[123] Hanai J, Dhanabal M, Karumanchi SA, et al. Endostatin causes G1
arrest of endothelial cells through inhibition of cyclin D1. J Biol
Chem 2002; 277: 16464-16469.
[124] Dhanabal M, Ramchandran R, Waterman MJ, et al. Endostatin
induces endothelial cell apoptosis. J Biol Chem 1999; 274: 11721-
11726.
[125] Urbich C, Reissner A, Chavakis E, et al. Dephosphorylation of
endothelial nitric oxide synthase contributes to the antiangiogenic
effects of endostatin. FASEB J 2002; 16: 706-708.
[126] Abdollahi A, Hahnfeldt P, Maercker C, et al. Endostatin’s
antiangiogenic signaling network. Mol Cell 2004; 13: 649-663.
[127] Ferreras M, Felbor U, Lenhard T, et al. Generation and degradation
of human endostatin proteins by various proteinases. FEBS Lett
2000; 486: 247-251.
[128] Maatta M, Heljasvaara R, Pihlajaniemi T, et al. Collagen
XVIII/endostatin shows a ubiquitous distribution in human ocular
tissues and endostatin-containing fragments accumulate in ocular
fluid samples. Graefes Arch Clin Exp Ophthalmol 2007; 245(1):
74-81.
[129] Lai LJ, Xiao X, Wu JH. Inhibition of corneal neovascularization
with endostatin delivered by adeno-associated viral (AAV) vector
in a mouse corneal injury model. J Biomed Sci 2007; 14(3): 313-
322.
[130] Niu XG, Wang W, Shi WY, et al. Inhibition of corneal
neovascularization by liposomes mediated plasmid encoding human
endostatin. Zhonghua Yan Ke Za Zhi 2005; 41(3): 260-264.
[131] Zhang P, Wu D, Ge J, et al. Experimental inhibition of corneal
neovascularization by endostatin gene transfection in vivo. Chin
Med J 2003; 116(12): 1869-1874.
[132] Folkman, J., Javaherian, K., Robert, T.T.: US20060251699A1
(2006).
[133] Folkman, M.J., O'Reilly, M.: US20050282253A1 (2005).
[134] O'Reilly, M., Folkman, M. J.: US20020123458A1 (2002).
[135] Wilson SE, He YG, Lloyd SA. EGF, EGF receptor, basic FGF,
TGF beta-1, and IL-1 alpha mRNA in human corneal epithelial
cells and stromal fibroblasts. Invest Ophthalmol Vis Sci 1992;
33(5): 1756-1765.
[136] Shams NB, Sigel MM, Davis RM. Interferon-gamma,
Staphylococcus aureus, and lipopolysaccharides/ silica enhance
interleukin-1 beta production by human corneal cells. Reg Immunol
1989; 2: 136-148.
[137] Sakamoto S, Inada K, Chiba K, et al. Production of IL-6 and IL-1a
by human corneal epithelial cells. Acta Soc Ophthalmol Jpn 1991;
95: 728-732.
[138] Sakamoto S, Inada K. Human corneal epithelial, stromal and
endothelial cells produce interleukin-6. Acta Soc Ophthalmol
Jpn1992; 96: 702-709.
[139] Cubitt CL, Lausch RN, Oakes JE. Differences in interleukin-6 gene
expression between cultured human corneal epithelial cells and
keratocytes. Invest Ophthalmol Vis Sci 1995; 36: 330-336.
[140] Cubitt CL, Tang Q, Monteiro CA, et al. IL-8 gene expression in
cultures of human corneal epithelial cells and keratocytes. Invest
Ophthalmol Vis Sci 1993; 34: 3199-3206.
[141] Dana R. Comparison of topical interleukin-1 vs tumor necrosis
factor-alpha blockade with corticosteroid therapy on murine corneal
inflammation, neovascularization, and transplant survival (an
American Ophthalmological Society thesis). Trans Am Ophthalmol
Soc 2007; 105: 330-343.
[142] Hong JW, Liu JJ, Lee JS, et al. Proinflammatory chemokine
induction in keratocytes and inflammatory cell infiltration into the
cornea. Invest Ophthalmol Vis Sci 2001; 42: 2795-2803.
[143] Saika S, Miyamoto T, Yamanaka O, et al. Therapeutic effect of
topical administration of SN50, an inhibitor of nuclear factor-B, in
treatment of corneal alkali burns in mice. Am J Pathol 2005; 166:
1393-1403.
[144] Nakao S, Hata Y, Miura M, et al. Dexamethasone inhibits
interleukin-1
-induced corneal neovascularization role of nuclear
factor-B-activated stromal cells in inflammatory angiogenesis. Am
J Pathol 2007; 171(3): 1058-1065.
[145] Eigler A, Sinha B, Hartmann G, et al. strategies to restrain this
proinflammatory cytokine. Immunol Today 1997; 18: 487-492.
[146] Henricson BE, Benjamin WR, Vogel SN. Differential cytokine
induction by doses of lipopolysaccharide and monophosphoryl lipid
A that result in equivalent early endotoxin tolerance. Infect Immun
1990; 58: 2429-2437.
230 Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3
[147] Yoshida S, Ono M, Shono T, et al. Involvement of interleukin-8,
vascular endothelial growth factor, and basic fibroblast growth
factor in tumor necrosis factor alpha-dependent angiogenesis. Mol
Cell Biol 1997; 17: 4015-4023.
[148] Niederkorn JY, Peeler JS, Mellon J. Phagocytosis of particulate
antigens by corneal epithelial cells stimulates interleukin-1
secretion and migration of Langerhans cells into the central cornea.
Reg Immunol 1989; 2: 83-90.
[149] Staats HF, Lausch RN. Cytokine expression in vivo during murine
herpetic stromal keratitis. Effect of protective antibody therapy. J
Immunol 1993; 151: 277-283.
[150] Elner VM, Strieter RM, Pavilack MA, et al. Human corneal
interleukin-8 IL-1 and TNF-induced gene expression and secretion.
Am J Pathol 1991; 139: 977-988.
[151] Dekaris I, Zhu SN, Dana MR. TNF-alpha regulates corneal
Langerhans cell migration. J Immunol1999; 162: 4235-4239.
[152] Dorrell M, Uusitalo-Jarvinen H, Aguilar E, et al. Ocular
neovascularization: Basic mechanisms and therapeutic advances.
Surv Ophthalmol 2007; 52: 3-19.
[153] Schreiber AB, Winkler ME, Derynck R. Transforming growth
factor-alpha: A more potent angiogenic mediator than epidermal
growth factor. Science 1986; 232(4755): 1250-1253.
[154] Cursiefen C, Rummelt C, Kuchle M. Immunohistochemical
localization of vascular endothelial growth factor, transforming
growth factor alpha, and transforming growth factor beta1 in human
corneas with neovascularization. Cornea 2000; 19(4): 526-533.
[155] Pepper MS. Transforming growth factor-beta: Vasculogenesis,
angiogenesis, and vessel wall integrity. Cytokine Growth Factor
Rev 1997; 8(1): 21-43.
[156] Sakamoto T, Ueno H, Sonoda K, et al. Blockade of TGF-beta by
in vivo gene transfer of a soluble TGF-beta type II receptor in the
muscle inhibits corneal opacification, edema and angiogenesis.
Gene Ther 2000; 7(22): 1915-1924.
[157] Pasquale LR, Dorman-Pease ME, Lutty GA, et al.
Immunolocalization of TGF-beta 1, TGF-beta 2, and TGF-beta 3 in
the anterior segment of the human eye. Invest Ophthalmol Vis Sci
1993; 34(1): 23-30.
[158] Nishida K, Kinoshita S, Yokoi N, et al. Immunohistochemical
localization of transforming growth factor-beta 1, -beta 2, and -beta
3 latency-associated peptide in human cornea. Invest Ophthalmol
Vis Sci 1994; 35(8): 3289-3294.
[159] Joyce NC, Zieske JD. Transforming growth factor-beta receptor
expression in human cornea. Invest Ophthalmol Vis Sci 1997;
38(10): 1922-1928.
[160] Hayashi K, Frangieh G, Wolf G, et al. Expression of transforming
growth factor-beta in wound healing of vitamin A-deficient rat
corneas. Invest Ophthalmol Vis Sci 1989; 30: 239-247.
[161] Pertovaara L, Kaipainen A, Mustonen T, et al. Vascular endothelial
growth factor is induced in response to transforming growth factor-
beta in fibroblastic and epithelial cells. J Biol Chem 1994; 269(9):
6271-6274.
[162] Baer JC, Foster CS. Corneal laser photocoagulation for treatment of
neovascularization. Efficacy of 577 nm yellow dye laser.
Ophthalmology 1992; 99(2): 173-179.
[163] Nirankari VS, Baer JC. Corneal argon laser photocoagulation for
neovascularization in penetrating keratoplasty. Ophthalmology
1986; 93: 1304-1309.
[164] Ahmad N, Mukhtar H. Mechanism of photodynamic
therapyinduced cell death. Methods Enzymol 2000; 319: 342-358.
[165] Sheppard Jr, Epstein RJ, Lattanzio Jr, et al. Argon laser
photodynamic therapy of human corneal neovascularization after
intravenous administration of dihematoporphyrin ether. Am J
Ophthalmol 2006; 141(3): 524-529.
[166] Yoon KC, You IC, Kang IS, et al. Photodynamic therapy with
verteporfin for corneal neovascularization. Am J Ophthalmol 2007;
144(3): 390-395.
[167] Clark, A.F.: US6297228B1 (1999).
[168] BenEzra D, Griffin BW, Maftzir G, et al. Topical formulations of
novel angiostatic steroids inhibit rabbit corneal neovascularization.
Invest Ophthalmol Vis Sci 1997; 38: 1954-1962.
[169] Phillips K, Arffa R, Cintron C, et al. Effects of prednisolone and
medroxyprogesterone on corneal wound healing, ulceration, and
neovascularization. Arch Ophthalmol 1983; 101: 640-643.
[170] Planck SR, Rich LF, Ansel JC, et al. Trauma and alkali burns
induce distinct patterns of cytokine gene expression in the rat
cornea. Ocul Immunol Inflamm 1997; 5(2): 95-100.
Shakiba et al.
[171] Haynes WL, Proia AD, Klintworth GK. Effect of inhibitors of
arachidonic acid metabolism on corneal neovascularization in the
rat. Invest Ophthalmol Vis Sci 1989; 30(7): 1588-1593.
[172] Pleyer U, Dannowski H, Volk HD, et al. Corneal allograft rejection:
Current understanding. I. Immunobiology and basic mechanisms.
Ophthalmologica 2001; 215(4): 254-262.
[173] Nauck M, Karakiulakis G, Perruchoud AP, et al. Corticosteroids
inhibit the expression of the vascular endothelial growth factor gene
in human vascular smooth muscle cells. Eur J Pharmacol 1998;
341(2-3): 309-315.
[174] Crum R, Szabo S, Folkman J. A new class of steroids inhibits
angiogenesis in the presence of heparin or a heparin fragment.
Science 1985; 230(4732): 1375-1378.
[175] Gillies MC, Kuzniarz M, Craig J, et al. Intravitreal triamcinolone-
induced elevated intraocular pressure is associated with the
development of posterior subcapsular cataract. Ophthalmology
2005; 112(1): 139-143.
[176] Jonas J, Heatley G, Spaide R, et al. Intravitreal triamcinolone
acetonide and secondary ocular hypertension. J Glaucoma 2005;
14(2): 168-171.
[177] Jonas JB, Kreissig I, Hugger P, et al. Intravitreal triamcinolone
acetonide for exudative age related macular degeneration. Br J
Ophthalmol 2003; 87(4): 462-468.
[178] Clark, A. F.: US5770592A (1998).
[179] Clark, A. F., Conrow R. E.: EP1236471A2 (2002).
[180] Clark AF. Mechanism of action of the angiostatic cortisene
anecortave acetate. Surv Ophthalmol 2007; 52: S26-34.
[181] Gately S. The contributions of cyclooxygenase-2 to tumor
angiogenesis. Cancer Metastasis Rev 2000; 19: 19 -27.
[182] Form DM, Auerbach R. PGE2 and angiogenesis. Proc Soc Exp Biol
Med 1983; 172: 214-218.
[183] Harada S, Nagy JA, Sullivan KA, et al. Induction of vascular
endothelial growth factor expression by prostaglandin E2 and E1 in
osteoblasts. J Clin Invest 1994; 93: 2490-2496.
[184] Pakneshan P, Birsner AE, Adini I, et al. Differential suppression of
vascular permeability and corneal angiogenesis by nonsteroidal
anti-inflammatory drugs. Invest Ophthalmol Vis Sci 2008; 49(9):
3909-3913.
[185] Jones MK, Wang H, Peskar BM, et al. Inhibition of angiogenesis
by nonsteroidal anti-inflammatory drugs: insight into mechanisms
and implications for cancer growth and ulcer healing. Nat Med
1999; 5: 1418-1423.
[186] Tsujii M, Kawano S, Tsuji S, et al. Cyclooxygenase regulates
angiogenesis induced by colon cancer cells. Cell 1998; 93: 705-716.
[187] Amico C, Yakimov M, Catania MV, et al. Differential expression
of cyclooxygenase-1 and cyclooxygenase-2 in the cornea during
wound healing. Tissue Cell 2004; 36(1): 1-12.
[188] Shibuya M. Structure and function of VEGF/VEGF-receptor system
involved in angiogenesis. Cell Struct Funct 2001; 26(1): 25-35.
[189] Hurwitz H, Fehrenbacher L, Novotny W, et al. Bevacizumab plus
irinotecan, fluorouracil, and leucovorin for metastatic colorectal
cancer. N Engl J Med 2004; 350: 2335-2342.
[190] Bock F, König Y, Kruse F, et al. Bevacizumab (Avastin) eye drops
inhibit corneal neovascularization. Graefes Arch Clin Exp
Ophthalmol 2008; 246(2): 281-284.
[191] Mackenzie SE, Tucker WR, Poole TR. Bevacizumab (avastin) for
corneal neovascularization--corneal light shield soaked application.
Cornea 2009; 28(2): 246-247.
[192] You IC, Kang IS, Lee SH, et al. Therapeutic effect of
subconjunctival injection of bevacizumab in the treatment of
corneal neovascularization. Acta Ophthalmol 2008; 87(6): 653-658.
[193] Han YS, Lee JE, Jung JW, et al. Inhibitory effects of bevacizumab
on angiogenesis and corneal neovascularization. Graefes Arch Clin
Exp Ophthalmol 2009; 247(4): 541-548.
[194] Hosseini H, Nejabat M, Mehryar M, et al. Bevacizumab inhibits
corneal neovascularization in an alkali burn induced model of
corneal angiogenesis. Clin Exp Ophthalmol 2007; 35(8): 745-748.
[195] Qian CX, Bahar I, Levinger E, et al. Combined use of superficial
keratectomy and subconjunctival bevacizumab injection for corneal
neovascularization. Cornea 2008; 27(9): 1090-1092.
[196] Carrasco MA. Subconjunctival bevacizumab for corneal neovas-
cularization in herpetic stromal keratitis. Cornea 2008; 27(6): 743-
745.
[197] D'amato, R.J., Green, S.J., Swartz Jr., G. M., Shah, J.H., Madsen,
J.: US20050004087A1 (2005).
[198] D'amato, R.: US6071948 (2000).
Corneal Neovascularization Recent Patents on Inflammation & Allergy Drug Discovery 2009, Vol. 3, No. 3 231
[199] Benelli U, Bocci G, Danesi R, et al. The heparan sulfate
suleparoide inhibits rat corneal angiogenesis and in vitro
neovascularization. Exp Eye Res 1998; 67: 133-142.
[200] Joussen AM, Germann T, Kirchhof B. Effect of thalidomide and
structurally related compounds on corneal angiogenesis is
comparable to their teratological potency. Graefes Arch Clin Exp
Ophthalmol 1999; 237(12): 952-961.
[201] Bocci G, Danesi R, Benelli U, et al. Inhibitory effect of suramin in
rat models of angiogenesis in vitro and in vivo. Cancer Chemother
Pharmacol 1999; 43: 205-212.
[202] Fotsis T, Pepper M, Adlercreutz H, et al. Genistein, a dietary-
derived inhibitor of in vitro angiogenesis. Proc Natl Acad Sci USA
1993; 90: 2690-2694.
[203] Wu PC, Liu CC, Chen C, et al. Inhibition of experimental
angiogenesis of cornea by somatostatin. Graefe’s Arch Clin Exp
Ophthalmol 2003; 241: 63-69.
[204] Demir T, Celiker UO, Kukner A, et al. Effect of Octreotide on
experimental corneal neovascularization. Acta Ophthalmol Scand
1999; 77: 386-390.
[205] Reis A, Reinhard T, Sundmacher R, Braunstein S, Godehardt E. A
comparative investigation of FK506 and cyclosporin A in murine
corneal transplantation. Graefes Arch Clin Exp Ophthalmol 1998;
236: 785-789.
[206] Joussen AM, Kruse FE: Topical application of methotrexate for
inhibition of corneal angiogenesis. Graefe’s Arch Clin Exp
Ophthalmol 1999; 237: 920-927.
[207] Kwon YS, Hong HS, Kim JC, Shin JS, Son Y. Inhibitory effect of
rapamycin on corneal neovascularization in vitro and in vivo. Invest
Ophthalmol Vis Sci 2005; 46: 454-460.
[208] Sloper CM, Powell RJ, Dua HS. Tacrolimus in the management of
high risk corneal and limbal grafts. Ophthalmology 2001; 108:
1838-1844.