(Investigative Ophthalmology and Visual Science. 2000;41:1507-1512.)
© 2000
by The Association for Research in Vision and Ophthalmology, Inc.
Squalamine Improves Retinal Neovascularization
Rosemary D. Higgins1,
Raymond J. Sanders2,
Yun Yan1,
Michael Zasloff3 and
Jon I. Williams3
1 From the Department of Pediatrics, Division of Neonatology, Georgetown University Medical Center, Washington, DC; the
2 Department of Pediatrics, Neonatal Program, New York University Medical Center, New York; and
3 Magainin Pharmaceuticals, Inc., Plymouth Meeting, Pennsylvania.
 |
Abstract
|
|---|
PURPOSE. Modalities for inhibiting neovascularization may be one avenue to the
development of effective therapies for retinopathy. The effect of
squalamine, an antiangiogenic amino sterol, on oxygen-induced
retinopathy (OIR) was assessed in a mouse model.
METHODS. OIR was induced in C57BL6 mice by a 5-day exposure to 75% oxygen from
postnatal day (P)7 through P12. Squalamine (25 mg/kg,
subcutaneous)-treated animals received either daily doses for five days
from P12 to P16 or one dose just after removal from oxygen on P12. Each
set of animals was killed at P17 to P21. Retinopathy was assessed with
a retinopathy scoring system evaluation of retinal wholemounts and by
quantification of neovascular nuclei on retinal sections.
RESULTS. Animals receiving 5 days of squalamine after a 5-day exposure to oxygen
had total retinopathy scores (expressed as median score with 25th and
75th quartiles in parentheses) of 4(3, 5) versus oxygen-only–reared
animals with scores of 8(7, 9; P < 0.001). Animals
reared in room air and animals exposed to squalamine only had similar
retinopathy scores: 1(1, 2) and 1(0, 2). Oxygen-reared animals
receiving single-dose squalamine also showed improvement, with a median
retinopathy score of 4(4, 6.75) versus oxygen-only–reared animals with
median retinopathy score of 9(7, 10; P < 0.001).
There was a decreased number of neovascular nuclei extending beyond the
inner limiting membrane on retinal sections in animals treated with 5
days (P < 0.01) and 1 day (P < 0.001) of squalamine.
CONCLUSIONS. Squalamine significantly improved retinopathy and may be a novel agent
for effective treatment of ocular
neovascularization.
|
Introduction
|
|---|
Squalamine is a broad-spectrum aminosterol antibiotic originally
isolated from the dogfish shark Squalus
acanthias.1
It has recently been reported to inhibit
tumor-induced angiogenesis and tumor growth.2
3
Squalamine
also improved tissue oxygenation in rats bearing the 13762 mammary
carcinoma.3
On a cellular level, squalamine inhibits
growth factor–mediated endothelial cell proliferation and migration,
including that stimulated by vascular endothelial cell growth factor
(VEGF) at concentrations that are not toxic to endothelial cells in
vitro.2
Vasoproliferative retinopathy is a leading cause of blindness and
occurs primarily because of overexpression of VEGF.4
5
Steroids have been previously described to inhibit retinal
neovascularization in the mouse,6
inhibit preretinal
neovascularization in a pig model,7
prevent
hyperoxia-induced neovascularization in the rabbit,8
and
inhibit subretinal neovascularization in a primate model.9
The goal of this study was to determine whether squalamine, a newly
described antiangiogenic steroid with no mineralocorticoid or
glucocorticoid function, could inhibit retinal neovascularization in a
mouse model of oxygen-induced retinopathy (OIR).
|
Methods
|
|---|
Mouse Model
C57BL6 mice were obtained from Taconic Laboratories (Germantown,
NY). Infant mice were placed in an infant incubator (Ohmeda, Columbia,
MD) with 75% oxygen at P7 through P12 with their nursing mothers as
previously described.6
10
11
12
The oxygen was delivered at
75% ± 2%, measured with an oxygen analyzer (Hudson Ventronics,
Temecula, CA), and checked at least twice daily during the oxygen
exposure. Individual litters were either room air or oxygen reared.
Within most litters, animals were divided into no treatment, treatment
with squalamine, and/or vehicle treatment. On P12, the animals were
returned to room air to induce relative retinal hypoxia. Animals were
treated with vehicle or squalamine injections that were initially given
for 5 days from P12 to P16. Subsequent experiments were performed using
a single dose of squalamine on P12. Vehicle-treated animals were
administered diluent (sterile water) for either 5 days or 1 day in
parallel to the squalamine-treated animals. Animals were killed by
lethal pentobarbital injection from P17 to P21, because retinal
neovascularization is at its maximum and is consistent at that
time.10
The protocol was reviewed and approved by the New
York University Medical Center Institutional Animal Care and Use
Committee for the Division of Laboratory Animal Resources and the
Georgetown University Animal Care and Use Committee and adhered to the
ARVO Statement for the Use of Animals in Ophthalmic and Vision
Research.
Squalamine Administration
Squalamine was administered subcutaneously in the nape of the neck
at 25 mg/kg · d in a single daily injection. This dose was selected
because prior effective doses of 20 to 40 mg/kg · d of squalamine
were used in oncologic studies in rats.2
3
Initial pilot
experiments were performed on several litters, by using a 5-day regimen
of squalamine after oxygen exposure from P12 through P16. Five-day
squalamine experiments were then performed using room air– or
oxygen-reared animals with intralitter assignments, as described. All
animals were used in the data analysis. Subsequent experiments were
performed using a single dose of squalamine at P12.
Retinal Perfusion
Fluorescein-conjugated dextran perfusion of the retinal vessels
was performed as previously described13
using
high-molecular-weight fluorescein conjugated dextran in 4%
paraformaldehyde in phosphate-buffered saline (PBS). Briefly, animals
were given a lethal dose of pentobarbital sodium (120 mg/kg) and when
deep anesthesia was obtained, a median sternotomy was performed. The
left ventricle was identified, and 1 ml of a 50-mg/ml solution of
fluorescein-conjugated dextran was injected. Eyes were enucleated and
placed in 4% paraformaldehyde for 3 to 24 hours. The retinas were
dissected using light microscopy and flat mounted. In the 5-day
squalamine treatment experiments the following numbers of animals were
used: the room air (control) group, n = 19 from four
litters (16 killed at P17 and 3 killed at P19); room air + squalamine
group, n = 17 from 4 litters (9 killed at P17 and 8
killed at P19); oxygen group, n = 20 from 4 litters (17
killed at P17, 2 at P18, and 2 at P19); and oxygen + squalamine group,
n = 25 from 8 litters (25 killed at P17). For the
single-dose squalamine experiments the numbers of animals were as
follows: the room air (control) group, n = 13 from 5
litters (1 killed at P17, 2 at P18, 4 at P19, and 6 at P20); room air +
squalamine group, n = 19 from 6 litters (3 killed at
P17, 2 at P18, 8 at P19, 2 at P20, and 4 at P21); oxygen group,
n = 21 from 6 litters ((13 killed at P18, 6 at P19, and
2 at P21); and oxygen + squalamine group, n = 22 from 4
litters (2 killed at P18, 17 at P19, and 3 at P21).
Retinae were scored in a masked fashion using a previously validated
retinopathy scoring system11
as shown in Table 1
. The minimum score according to this method is 0, and the
maximum score is 17. Maximal vasoproliferation in this mouse model has
previously been reported to occur from P17 to P21.10
In
addition, our laboratory has previously reported concordance of scores
in mice killed from P17 to P20.12
Quantification of Extraretinal Neovascularization
After perfusion of animals with 4% paraformaldehyde, eyes were
removed, placed in optimal cutting temperature embedding compound, and
frozen at -70°C. Serial sections (7–10 µm) were cut through the
cornea parallel to the optic disc using a cryostat. Tissue sections
were stained with periodic acid–Schiff stain and hematoxylin. For the
5-day treatment with squalamine, there were the following numbers of
animals: room air (control) group, n = 7 from 5 litters
(6 killed at P17 and 1 at P18); room air + squalamine group,
n = 6 from 2 litters (all 6 killed at P17); oxygen
group, n = 8 from 4 litters (4 killed at P17, 2 at P18,
and 2 at P19); and oxygen + squalamine, n = 5 from 2
litters (all five killed at P17). In the single-dose squalamine
experiments, the animal numbers were as follows: the room air (control)
group, n = 10 from 6 litters (3 killed at P17, 1 at
P19, and 6 at P20); room air + squalamine group, n = 8
from 4 litters (4 killed at P17, 2 at P19, and 2 at P20); oxygen group,
n = 15 from 11 litters (1 killed at P17, 8 at P18, 5 at
P19, and 1 at P20); and oxygen + squalamine group, n =
9 from 6 litters (1 killed at P18, 7 at P19, and 1 at P21). Multiple
sections were scored in a masked fashion by counting the number of
nuclei extending beyond the inner limiting membrane into the vitreous,
as previously described10
and used in our
laboratory.6
11
12
A minimum of eight sections at least 50
µm apart over a minimum distance of 450 µm were counted and
averaged for each eye. The average number for each eye was pooled and
averaged across replicates for each treatment condition, and these
averages were used in the statistical data analysis.
Animal and Organ Weights
Animals and individual organs were weighed on a standard
laboratory balance. In addition, a log of animal death was kept
throughout the course of the experiments.
Statistical Analyses
Analysis of variance using the Kruskal–Wallis test was performed
to test for differences between the treatment groups. The Mann–Whitney
test was used to compare the total retinopathy scores and retinopathy
subscores between individual groups. Student’s t-tests
assuming unequal variance were performed to compare the number of
nuclei in the retinal sections, weights, and organ-to-body weight
ratios. Significance was defined as P < 0.05.
|
Results
|
|---|
Five-Day Regimen of Squalamine
Retinopathy scores are expressed as median with 25th and 75th
quartiles in parentheses. Animals that received 5 days of squalamine
from P12 through P16 after exposure to a hyperoxic environment had a
lower retinopathy score than their counterparts that were exposed to
oxygen only: 4(3, 5) and 8(7, 9), respectively (Fig. 1A ). Squalamine-only–treated animals reared in room air had a
retinopathy score similar to that of the untreated control group: 1(0,
1) and 1(1, 2), respectively. Vehicle-treated animals (sterile water
diluent) had retinopathy scores of 1(0, 1) in the room air–reared
group (n = 6 from three litters) and 8(7, 9) in the
oxygen-reared group (n = 2 from two litters), which
were similar to the scores of untreated animals in both groups. Within
each of the groups, day of sacrifice did not affect the scores. Retinal
neovascularization was notably improved after the squalamine treatment
in the specific categories of blood vessel tuft formation, extraretinal
neovascularization (ERNV), and blood vessel tortuosity (Table 2)
. Squalamine-treated eyes had less clock hours or circumferential
disease with regard to all parameters that were scored. Control animals
had slightly positive retinopathy scores, because mice are born with an
immature retinal vasculature that is sometimes not fully mature by P17
to P21. Vessel remodeling occurs into adulthood14
and may
also help explain the small positive values noted in the retinopathy
scores for room-air–reared mice.
View larger version (17K):
[in this window]
[in a new window]
|
Figure 1. (A) Median total retinopathy scores (error
bar denotes 75th quartile) in 5-day treatment with squalamine.
Vehicle-treated animals (data not shown) had median (25th, 75th
quartile) retinopathy scores of 1(0, 1) in the room air group and 8(7,
9) in the oxygen-treated group that were similar to those of untreated
animals in both groups. By Kruskal–Wallis test, P < 0.001 for the entire group; by Mann–Whitney test,
P < 0.001 for oxygen versus control, oxygen versus
control + squalamine, and oxygen versus oxygen + squalamine groups.
(B) Neovascular nuclei per retinal section in animals
treated with a 5-day regimen of squalamine. For oxygen versus oxygen +
squalamine, P < 0.01 by Student’s
t-test.
|
|
In confirmation of the retinopathy score observations,
squalamine-treated, oxygen-reared animals had less ERNV as measured by
neovascular nuclei (12.3 ± 4.7) counts on retinal sections
compared with oxygen-reared animals (56.0 ± 24.4) as shown in
Figure 1B . Control animals showed few neovascular nuclei, as has
been previously described in this model.6
10
11
12
This
observation in room-air–reared animals may again be explained as
vessel remodeling into adulthood.14
There were no observed
histologic differences (other than ERNV) between oxygen-exposed and
squalamine-treated retinal sections when compared with control
sections.
Single Dose of Squalamine
Subsequent experiments were performed to test the hypothesis that
a single dose of squalamine on P12, the day the animals are removed
from a hyperoxic environment, would be sufficient to alter the
development of retinal neovascularization. Results again showed an
improvement in retinopathy as measured by total retinopathy scores
(Fig. 2A
) and subscores in categories of blood vessel tuft formation, ERNV, and
blood vessel tortuosity (Table 3)
. Vehicle-treated animals had median retinopathy scores of 0(0, 1) in
the room air group (n = 9 from 4 litters) and 10(9, 10)
in the oxygen-reared group (n = 8 from 2 litters),
which were similar to the untreated animals in both groups. Further, a
decrease in the number of neovascular nuclei was found in oxygen +
single-dose squalamine–treated animals (16.3 ± 6.8) compared
with oxygen-only–reared animals (46.2 ± 24.1) as shown in Figure 2B
.
View larger version (17K):
[in this window]
[in a new window]
|
Figure 2. (A) Median total retinopathy scores (error
bar denotes 75th quartile) for single-dose treatment of
squalamine. Vehicle-treated animals (data not shown) had median
retinopathy scores of 0(0, 1) in the room air group and 10(9, 10) in
the oxygen-treated group that were similar to those of untreated
animals in both groups. By Kruskal–Wallis test, P < 0.001 for entire group. By Mann–Whitney test P < 0.001 for oxygen versus room air (control), oxygen versus room air +
squalamine, and oxygen versus oxygen + squalamine groups.
(B) Neovascular nuclei per retinal section in animals
treated with a 1-day regimen of squalamine. For oxygen versus oxygen +
squalamine, P < 0.001 by Student’s
t-test.
|
|
The degree of suppression of neovascular nuclei in
hyperoxia-exposed mice after treatment with a single dose of squalamine
(64%) was comparable with that seen after 5 days of squalamine
treatment (78%), suggesting that a long-lived biologic response to
squalamine occurs. Representative retinal wholemounts are shown in
Figure 3 from both sets of experiments. Although there is some variability
within the room-air–reared and oxygen-reared groups (see error bars
for quartiles in Figs. 1A
and 2A
and error bars for SDs in Figs. 1B
and 2B
), retinopathy developed in all animals exposed to 75% oxygen, and
squalamine consistently improved the retinopathy. Squalamine did not
affect normal vascularization or development of the retina as measured
by the retinopathy scoring system or by histologic observation of
retinal sections.
View larger version (129K):
[in this window]
[in a new window]
|
Figure 3. Representative retinal wholemounts showing control (A),
oxygen-treated (B), 5-day squalamine + oxygen–treated
(C), and single-dose squalamine + oxygen–treated
(D) animals. Note the smooth vascular pattern in the control
group (A) compared with the loss of central vasculature and
presence of multiple blood vessel tufts at the junction of the loss of
central vasculature and the remainder of the retinal blood vessels in
the oxygen-treated retina (B). Both squalamine-treated
retinae (C, D) had some loss of central
vasculature as did the retina in (B), but few blood vessel
tufts, ERNV, and less tortuous vessels than the oxygen-only–reared
retina. Magnification, x4.5.
|
|
Effect of Squalamine on Growth
Systemic administration of squalamine to the mice did not have any
gross adverse developmental effects when assessed by animal weight gain
and individual organ weight-to-body weight ratios (Table 4)
. Normal growth of animals and individual organs suggests that
squalamine does not alter normal vessel growth while inhibiting
pathologic neovascularization during the period from P12 to P21. There
were no deaths in the room-air–reared group with 5 days of squalamine
treatment, the oxygen-reared group with 5 days of squalamine treatment,
or the room-air–reared vehicle group. There was one death each in the
room-air–reared group, the room air + single-dose squalamine group,
and the oxygen-reared vehicle group. There were two deaths in the
oxygen-reared group and oxygen-reared + single dose squalamine group.
|
Discussion
|
|---|
This is the first study that has examined inhibition of
angiogenesis by squalamine in a noncancerous model—i.e., a mouse model
of oxygen-induced retinal neovascularization. The measurements of
retinal neovascularization collected in this study show that squalamine
significantly reduces retinal neovascularization in a mouse model of
oxygen-induced proliferative retinopathy, as measured by a previously
validated retinopathy scoring system and by quantitation of
extraretinal nuclei on retinal sections. Systemic squalamine given
either during the 5 days from P12 through P16 or as a single dose on
P12 significantly decreased retinopathy. There was less
neovascularization when measured by blood vessel tuft formation, ERNV,
and blood vessel tortuosity in the squalamine + oxygen–treated retinas
when compared with the oxygen-only–treated retinas. Thus, squalamine
is able to inhibit or decrease the neovascular response seen after
exposure of the neonatal mice to 75% oxygen. In addition no
deleterious systemic side effects of squalamine such as weight loss or
impaired organ growth were found in this study. Squalamine did not
affect normal vascular development as assessed by the retinopathy score
or retinal histology. There may be a more favorable therapeutic index
for squalamine in treating retinopathy than for an antiangiogenic agent
that suppresses all vessel growth. Further experiments are required to
define the basis for squalamine’s selectivity with respect to
inhibition of abnormal blood vessel growth.
Both single and 5-day regimens of squalamine had similar effects. This
may be explained by postulating a long half-life for squalamine—that
is, the single dose lasts long enough to inhibit neovascularization in
this model. The presence or persistence of squalamine in the neonatal
eye was not measured in these studies. There also may be a critical
time (i.e., day 12 and shortly thereafter) when retinal hypoxia
triggers a maximal response and VEGF-stimulated proliferation is at its
peak in the mouse model.
We speculate that squalamine broadly inhibits growth factor–stimulated
endothelial cell growth, leading to inhibited neovascularization in the
mouse retina. A previous report provided evidence that squalamine
inhibited in vivo angiogenesis in a tumor model.2
In
addition, squalamine has been reported to inhibit mitogen (including
VEGF)-stimulated proliferation of endothelial cells.2
Although it was beyond the scope of this study to assess squalamine
concentrations in the retina, squalamine may also improve retinal
oxygenation as it did in a tumor model3
and thereby may
suppress hypoxia-mediated signaling for VEGF or other growth factor
production at a critical time in the development of retinopathy.
Alternatively, if squalamine decreases VEGF production or downregulates
VEGF receptor expression, then the metabolic demands of the tissue may
be lower, and tissue oxygenation may be improved by a lower metabolic
rate in the retinas of animals treated with squalamine. Specific target
cells for squalamine in this model of OIR are not known.
Dexamethasone has been reported to inhibit retinal neovascularization
in a primate model,9
mouse,5
and
rabbit.8
Triamcinolone acetate has been shown to inhibit
preretinal and optic nerve head neovascularization in the
pig.7
The exact mechanism of corticosteroid-induced
inhibition of retinopathy is unclear but may be related to several
factors including decreasing inflammation and inhibiting angiogenic
growth factors.5
6
7
8
9
Concerns regarding the long-term effects of angiogenesis inhibitors are
valid, particularly in the context of patients with multiple medical
problems. For instance, inhibiting retinal neovascularization in
retinopathy of prematurity or diabetic retinopathy may be important but
may have effects on other systemic diseases (i.e., hernia surgery,
bronchopulmonary dysplasia, decubitus ulcer). In this regard, it is
meaningful that short-term treatment (i.e., a single dose) with
squalamine in this animal model of oxygen-induced retinopathy led to a
marked improvement of retinal neovascularization. Intermittent
treatment of vasoproliferative retinopathy with squalamine may
therefore be a preferred way to reduce possible side effects.
A short-term therapy that may prevent blindness could prove clinically
sightsaving in ocular diseases such as diabetic retinopathy and
retinopathy of prematurity. Squalamine is currently in phase II trials
for patients with late-stage non–small-cell lung cancer. If squalamine
is shown to be efficacious in preventing or inhibiting tumor growth or
metastasis in human cancers, it may also be useful as a potential
treatment to prevent human retinal neovascularization. Squalamine’s
ability to block angiogenesis in a noncancerous model such as this
mouse oxygen-induced retinopathy model is a significant finding, and it
may ultimately become a new agent for use in the treatment of ocular
neovascularization.
|
Footnotes
|
|---|
Supported in part by Magainin Pharmaceuticals, Inc., and National Eye Institute Grant EY-00330 (RDH).
Submitted for publication March 26, 1999; revised September 2 and November 11, 1999; accepted December 30, 1999.
Commercial relationships policy: C5(RDH, RJS, YY); E(MZ, JIW).
Corresponding author: Rosemary D. Higgins, Department of Pediatrics, Division of Neonatology, Georgetown University Medical Center, 3800 Reservoir Road, NW M3400, Washington, DC 20007. higginsr1{at}gunet.georgetown.edu
|
References
|
|---|
-
Moore, KS, Wehrli, S, Roder, H, et al (1993) Squalamine: an amino sterol antibiotic from the shark Proc Natl Acad Sci USA 90,1354-1358[Abstract/Free Full Text]
-
Sills, AK, Williams, JI, Tyler, BM, et al (1998) Squalamine inhibits angiogenesis and solid tumor growth in vivo and perturbs embryonic vasculature Cancer Res 58,2784-2792[Abstract/Free Full Text]
-
Teicher, BA, Williams, JI, Takeuchi, H, Ara, G, Herbst, RS, Buxton, D. (1998) Potential of the amino-sterol squalamine in combination therapy in the rat 13762 mammary carcinoma and the murine Lewis Lung carcinoma Anticancer Res 18,2567-2574[Medline][Order article via Infotrieve]
-
Pierce, EA, Avery, RL, Foley, ED, Aiello, LP, Smith, LEH (1995) Vascular endothelial growth factor/vascular permeability factor expression in a mouse model of retinal neovascularization Proc Natl Acad Sci USA 92,905-909[Abstract/Free Full Text]
-
Young, TL, Anthony, DC, Pierce, E, Foley, E, Smith, LEH (1997) Histopathology and vascular endothelial growth factor in untreated and diode laser-treated retinopathy of prematurity J Am Assoc Pediatr Ophthalmol Strabismus 1,105-110
-
Rotschild, T, Nandgaonkar, BN, Yu, K, Higgins, RD (1999) Dexamethasone reduces oxygen induced retinopathy in a mouse model Pediatr Res 46,94-100[Medline][Order article via Infotrieve]
-
Danis, RP, Bingaman, DP, Yang, Y, Ladd, B. (1996) Inhibition of preretinal and optic nerve head neovascularization in pigs by intravitreal triamcinolone acetate Ophthalmology 103,2099-2104[Medline][Order article via Infotrieve]
-
Lawas–Alejo, PA, Slivka, S, Hernandez, H, Bry, K, Hallman, M. (1999) Hyperoxia and glucocorticoid modify retinal vessel growth and interleukin-1 receptor antagonist in newborn rabbits Pediatr Res 45,313-317[Medline][Order article via Infotrieve]
-
Ishibashi, T, Miki, K, Sorgente, N, Patterson, R, Ryan, SJ (1985) Effects of intravitreal administration of steroids on experimental subretinal neovascularization in the subhuman primate Arch Ophthalmol 103,708-711[Abstract/Free Full Text]
-
Smith, LEH, Wesolowski, E, McLellan, A, et al (1994) Oxygen-induced retinopathy in the mouse Invest Ophthalmol Vis Sci 35,101-111[Abstract/Free Full Text]
-
Higgins, RD, Yu, K, Sanders, RJ, Nandgaonkar, B, Rotschild, T, Rifkin, DB (1999) Diltiazem reduces retinal neovascularization in a mouse model of oxygen induced retinopathy Curr Eye Res 18,20-27[Medline][Order article via Infotrieve]
-
Nandgaonkar, BN, Rotschild, T, Yu, K, Higgins, RD (1999) Indomethacin improves oxygen induced retinopathy in the mouse Pediatr Res 46,184-188[Medline][Order article via Infotrieve]
-
D’Amato, R, Wesolowski, E, Smith, LE (1993) Microscopic visualization of the retina by angiography with high-molecular-weight fluorescein-labeled dextrans in the mouse Microvasc Res 46,135-142[Medline][Order article via Infotrieve]
-
Connolly, SE, Hores, TA, Smith, LEH, D’Amore, PA (1988) Characterization of vascular development in the mouse retina Microvasc Res 36,275-290[Medline][Order article via Infotrieve]
This article has been cited by other articles:
|
|
|
|
 
M. N. Chernova, D. H. Vandorpe, J. S. Clark, J. I. Williams, M. A. Zasloff, L. Jiang, and S. L. Alper
Apparent receptor-mediated activation of Ca2+-dependent conductive Cl- transport by shark-derived polyaminosterols
Am J Physiol Regulatory Integrative Comp Physiol,
December 1, 2005;
289(6):
R1644 - R1658.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. J. Franklin, T. L. Jetton, C. L. Kuchemann, S. R. Russell, and E. C. Kohn
CAI Is a Potent Inhibitor of Neovascularization and Imparts Neuroprotection in a Mouse Model of Ischemic Retinopathy
Invest. Ophthalmol. Vis. Sci.,
October 1, 2004;
45(10):
3756 - 3766.
[Abstract]
[Full Text]
[PDF]
|
|
|
Top
Abstract
Introduction
