Download ARVO 2015 Annual Meeting Abstracts 113 New techniques and

ARVO 2015 Annual Meeting Abstracts
113 New techniques and approaches
Sunday, May 03, 2015 8:30 AM–10:15 AM
Exhibit Hall Poster Session
Program #/Board # Range: 364–404/D0100–D0140
Organizing Section: Retina
Contributing Section(s): Anatomy/Pathology, Biochemistry/
Molecular Biology, Genetics, Immunology/Microbiology,
Multidisciplinary Ophthalmic Imaging, Physiology/Pharmacology
Program Number: 364 Poster Board Number: D0100
Presentation Time: 8:30 AM–10:15 AM
DaVinci Robot Assisted Sub-Retinal Injection
Martin Schardt, Jeffrey M. Naids, Matthew R. Knouse, William J.
Foster, Kumar Nadhan. Temple University, Philadelphia, PA.
Purpose: With the advent of gene therapy for hereditary retinal
diseases, and a growing interest in performing gene therapy on young
children, techniques to perform retinal injections that avoid creating a
hole in the retina are likely to be increasingly preferred. But transchoroidal, sub-retinal injections, which avoid making a retinal hole,
also require extreme precision positioning the needle. Therefore,
we have attempted to reproduce this procedure using the DaVinci
robot. Its motion scaling ability affords a 10:1 de-amplification of the
surgeon’s range of movement, which reduces complications related to
human tremor and yields increased precision.
Methods: A Da Vinci Si(R) Surgical Robot, was utilized to perform
trans-scleral sub-retinal injections in a porcine model. A Thornton
fixation ring was utilized to stabilize the globe, the retina was directly
visualized using only the Da Vinci camera, and ICG dye was injected
in a sub-retinal fashion and directly visualized with the ICG filters
on the Da Vinci system. The injection was made using a repurposed
Butterfly needle, which was grasped and maneuvered by the large
ridged forceps tool of the Da Vinci system. The injection itself
was made by hand using a 10cc syringe attached to the tube of the
Butterfly needle.
Results: Trans-scleral, sub-retinal injection with the Da Vinci robot is
possible, with appropriate modification of the instrumentation. There
is a significant learning curve to be able to successfully perform such
injections.
Conclusions: Our results suggest that robotically assisted, transchoroidal delivery of viral vectors for gene therapy may be effective
in reducing the incidence of retinal detachment and other adverse
outcomes. Stabilization of the eye is critical to successful sub-retinal
injection. Larger (thicker than 25 gauge) needles are better able to
penetrate the porcine sclera.
Commercial Relationships: Martin Schardt, None; Jeffrey M.
Naids, None; Matthew R. Knouse, None; William J. Foster, None;
Kumar Nadhan, None
Program Number: 365 Poster Board Number: D0101
Presentation Time: 8:30 AM–10:15 AM
Correlating phenotype with protein secretion profile based on
genotype in X-linked Retinoschisis
Rajiv Raman, Jayamuruga Pandian, Srividya Neriyanuri, Sudha D.
Sankara Nethralaya, Chennai, India.
Purpose: To correlate the protein secretion profile based on
genotypic characterization with phenotypic characteristics in
X-linked Retinoschisis in Indian population.
Methods: Phenotypic characterization was done in 21 patients with
X-linked retinoschisis. It included age of onset, visual acuities,
refraction, fundus findings, optical coherence tomography (OCT),
electroretinogram (ERG). All underwent screening for RS1 mutation.
Secretion profile of the novel mutant proteins were analysed by
transfecting the mutant constricts into COS7 cell line and studying
their expression pattern by western blot. Secretion prolife of known
mutants were taken from the literature. Data from both the eyes was
used for analysis. Statistical analysis was performed using SPSS 17
software. A p value of 0.05 was set as statistical significance.
Results: We identified 7 new mutations and 8 reported mutations.
85% (18 patients) had mutations lying in the discoidin domain of
the RS1 protein, 15% (3 patients) had mutations lying on the leader
sequence domain. There was a statistically significant association of
the location of the schisis (foveal/ foveal and peripheral) and ERG
features (b/a ratio and latency of the b wave) with protein secretion
pattern (p<0.05).
Conclusions: Protein secretion within and outside the cell may give
rise to the difference’s in the severity of the disease. This genotypephenotype correlations can help unraveling the disease mechanism.
Commercial Relationships: Rajiv Raman, None; Jayamuruga
Pandian, None; Srividya Neriyanuri, None; Sudha D, None
Program Number: 366 Poster Board Number: D0102
Presentation Time: 8:30 AM–10:15 AM
Development of an intraocular B-scan optical coherence
tomography (OCT)-guided micro-needle
Karen M. Joos, Jin H. Shen. Vanderbilt Eye Institute, Vanderbilt
University, Nashville, TN.
Purpose: Real-time intraoperative B-scan optical coherence
tomography (OCT) observation of intraocular surgery may enhance
precise surgical techniques such as gene therapy delivery or subretinal surgery. A micro-needle was combined with a forwardimaging 25-gauge OCT probe to perform real-time imaging of the
needle as it touches a retinal surface, perforates through the retina,
and injects sub-retinal fluid.
Methods: The forward-imaging OCT probe has a disposable
25-gauge tip. A 36-gauge needle was welded to the probe tip with its
end extending 3.5 mm with a smooth curve to permit imaging of the
needle tip. Silicone tubing with a saline syringe was attached to the
external proximal needle tubing. An electromagnetic controller was
embedded within the handpiece to drive the 125 mm single-mode
fiber optic actuator within the 25-gauge probe tip. A sealed 0.35 mm
diameter GRIN lens (Go!Foton, Somerset, NJ) within the probe tip
protected the fiber scanner and focused the scanning beam 3 to 4
mm distant. A VHR spectral-domain optical coherence tomography
(SDOCT) system (870 nm, Bioptigen, Inc. Durham, NC) produced
the B-scan images with the fiber optic oscillations matched to the
engine’s scanning rate. Real-time imaging trials of the needle tip as
it touched the porcine ex vivo retinal surface, perforated the retina,
and injected saline under the neurosensory retina to form a retinal
detachment were performed.
Results: A 36-gauge needle combined with the 25-gauge tip of
the forward-imaging OCT probe formed an instrument capable
of passing through a 23-gauge vitrectomy port. This probe has
an internal scanning system so it can be held steady to produce a
two-dimensional B-scan image. Unprocessed real-time OCT video
showed the needle tip touching the surface of an ex vivo porcine
retina without the aid of a surgical microscope. Reproducible guided
perforation through the retina to the subretinal space was observed.
Formation of a localized neurosensory retinal detachment coincided
with initiation of the saline injection.
Conclusions: Real-time intraocular B-scan OCT visualization and
feedback of surgical maneuvers can provide additional infromation
about the targeted retinal layers during surgical intervention. An
intraocular 25-gauge B-scan forward-imaging probe coupled with
a 36-gauge needle developed to pass through 23-gauge ports was
capable of real-time guidance of sub-retinal saline injection.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Commercial Relationships: Karen M. Joos, No. 8,655,431/
Vanderbilt University (P); Jin H. Shen, No. 8,655,431/ Vanderbilt
University (P)
Support: This study was supported by: NIH 1R21EY019752,
Joseph Ellis Family Research Fund, William Black Research
Fund, NIH P30EY008126 to Vanderbilt Vision Research Center,
and an Unrestricted Departmental Grant from Research to Prevent
Blindness, Inc., N.Y. to the Vanderbilt Eye Institute.
Program Number: 367 Poster Board Number: D0103
Presentation Time: 8:30 AM–10:15 AM
Metabolomics analysis of human vitreous in the disease of
Rhegmatogenous retinal detachment associated with choroidal
detachment
Zhifeng Wu. ophthalmology, Wuxi No.2 Hospital Afliated to Nanjing
Medical University, Wuxi, China.
Purpose: Our aim was to identify potential biomarkers and metabolic
pathways that may be used to diagnosis and treat rhegmatogenous
retinal detachment associated with choroidal detachment (RRDCD)
and to elucidate the etiology of RRDCD.
Methods: We used liquid chromatography-quadrupole-time of flight/
mass spectrometer (LC-Q-TOF/MS) to obtain the metabolome of
vitreous from patients with rhegmatogenous retinal detachment
(RRD) and RRDCD. The metaboloms from 29 vitreous samples (14
from RRD patients and 15 from RRDCD patients) were analyzed
by principal component analysis (PCA) and partial least squares
discriminant analysis (PLS-DA). A one-way analysis of variance
with a Bonferroni correction was used to test significance. Biofluid
Metabolites Database and Human Metabolome Database were used
to identify ions.
Results: Metabolites were used to completely discriminate between
RRD and RRDCD. PLS-DA identified 265 (VIP1) ions whose levels
were significantly different in vitreous from patients with RRD
and RRDCD. Among the 265 ions, 24 of them (23 observed in the
positive mode and 1 observed in the negative mode) were identified
by searching MS and MS/MS fragments in the Biofluid Metabolites
Database and Human Metabolome Database. The metabolites were
associated with pathways related to proliferation, inflammatory
reactions and hemodynamic changes.
Conclusions: Metabolites were found that discriminated between
RRDCD and RRD. Theses metabolites are likely to be involved in the
pathology of the diseases and may potentially be used to diagnosis
and treat RRDCD and RRD.
Commercial Relationships: Zhifeng Wu, None
Support: YGZZ1106
Clinical Trial: 14005142
Program Number: 368 Poster Board Number: D0104
Presentation Time: 8:30 AM–10:15 AM
Metabolomic analysis of urine in patients with age related
macular degeneration
Sreekumari Pushpoth1, 2, Martin Fitzpatrick2, James S. Talks3, Stephen
Young2, Yit C. Yang4, Graham R. Wallace2. 1Ophthalmology, The
James Cook University Hospital, Billingham, United Kingdom;
2
University of Birmingham, Birmingham, United Kingdom; 3Royal
Victoria Infirmary, Newcastle upon Tyne, United Kingdom; 4The
Wolverhampton Hospitals NHS Trust, Wolverhampton, United
Kingdom.
Purpose: Age related macular degeneration is a complex disease
where multiple factors show associations but do not explain the
full nature of the disease. We hypothesized that a systems approach
based on metabolomic analysis would be able to segregate diseases
types and provide insights into the pathology. Metabolomics assesses
the broad range of low molecular weight metabolites in biofluids
and, since these are influenced by important AMD-related factors
including diet, age and smoking, this approach may provide a useful
novel window into the AMD-disease process.
Methods: Serum and urine samples were collected from 104
patients with dry and wet macular degeneration. Serum samples
were centrifuged to remove cells, and 0.5ml aliquots stored at -80
degree C. After thawing, serum was filtered through 3kD MW cutoff
filter to remove proteins. The filtrate was made with 10% in D2O,
100mM phosphate 0.5mM TMSP and pH 7.00.One-dimensional 1H
spectra were acquired using a standard spin-echo pulse sequence on
a Bruker DRX 600MHz NMR spectrometer equipped with a 1.7mm
cryoprobe. 2D JRes spectra were also acquired to aid metabolite
identification. Spectra were be segmented into 0.005-ppm (2.5 Hz)
chemical shift ‘bins’ between 0.2 and 10.0 ppm, and the spectral area
within each bin integrated. Principal component analysis (PCA) and
partial least squares discriminant analysis (PLS-DA) of the processed
data was conducted using PLS Toolbox (Eigenvector Research)
within MATLAB.
Results: The serum samples from dry AMD patients cluster together
reasonably well based on their metabolomics profile. With regards to
serum samples from patients with wet AMD the clustering is far more
complex. Samples from patients with dry AMD show an increase
in arginine, and decreased glucose, lactate, glutamine and reduced
glutathione. Urine sample cluster also showed similar interestig
pattern
Conclusions: Metabolomic analysis showed clear separation between
body fluid samples from patients with wet and dry AMD. Several
samples from patients with wet AMD cluster with samples from dry
AMD strongly indicates that common pathways are involved in both
types of disease and that dry AMD can develop into the wet form.
Commercial Relationships: Sreekumari Pushpoth, None; Martin
Fitzpatrick, None; James S. Talks, Novartis, Bayer (S); Stephen
Young, None; Yit C. Yang, Novartis, Bayer (S); Graham R.
Wallace, None
Program Number: 369 Poster Board Number: D0105
Presentation Time: 8:30 AM–10:15 AM
Study of funduscopic and tomographic findings in the retina of
patients with Alzheimer disease
Mireia Garriga Beguiristain1, Miguel A. Zapata1, Laura N.
Distefano1, Jose Garcia-Arumi1, 2. 1Ophtalmology, Vall d’Hebron
Hospital, Barcelona, Spain; 2Institut de Microcirurgia Ocular,
Barcelona, Spain.
Purpose: To derminate if mental state of patients with initial
Alzheimer disease correlates with the subretinal depts (drusen-like),
even in peripheral and central retina.
Methods: Transversal, descriptive and unicentric study. 60 patients,
clinical diagnosticated of mild to moderate Alzheimer disease with
DSM-IV criteria, where assessed with MMSE, scale of Blessed,
MEC and DAD test. The ophthalmologic study was performed with
wide-camp Optos for the rethinography and autofluorescence and
OCT cirrus image.
Results: Peripheral drusen where detected in 45- 50% of the
right and left eyes respectively, and totally drusen in 63-60%. The
thickness average of CNFL was of 84.5 mm, and in GCL was of 74%.
The choroid thickness average detected was of 226 mm.
This results were studied jointly with de mental tests, finding a
significant statistical positive correlation between the results of the
tests and the quantification of peripheral drusen (i.e. more peripheral
drusen where detected in patients with worst punctuation in the test).
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Conclusions: The presence of peripheral subretinal drusen could be
associated to a worsen mental state in patients with mild to moderat
Alzheimer disease.
Commercial Relationships: Mireia Garriga Beguiristain, None;
Miguel A. Zapata, None; Laura N. Distefano, None; Jose GarciaArumi, None
Program Number: 370 Poster Board Number: D0106
Presentation Time: 8:30 AM–10:15 AM
Methods development for testing aqueous formulations of Met12,
a novel inhibitor of photoreceptor cell death, in vitro and in vivo
Lee Kiang1, Cagri Besirli1, Rae-Sung Chang2, Anna Schwendeman2,
David N. Zacks1. 1Ophthalmology, Kellogg Eye Ctr, Univ of
Michigan, Ann Arbor, MI; 2College of Pharmacy, University of
Michigan, Ann Arbor, MI.
Purpose: A significant consequence of retina-RPE separation is death
of photoreceptor (PR) cells. We previously reported that Met12, a
small molecule antagonistic of the Fas receptor, inhibits Fas-mediated
PR death in a rodent retinal detachment (RD) model. Those previous
studies all had Met12 dissolved in the organic solvent dimethyl
sulfoxide (DMSO). The purpose of this study was to develop an
assay to screen new aqueous solvents for Met12 that would measure
caspase inhibition in vitro and increase PR survival in vivo, with
the eventual goal to develop this drug for therapeutic application in
humans.
Methods: For this study, Met12 was dissolved in either DMSO
or citrate buffer at concentrations ranging between 0.07uM –
175uM. Cultured 661w cells incubated with Fas-ligand and various
concentrations of Met12-citrate buffer. Caspase 8 activity was
measured using Caspase-Glo 8 Assay (Promega). For in vivo testing,
RDs were created in Brown Norway rats as previously described,
followed by intravitreal administration of Met12, dissolved in either
DMSO or citrate buffer, in a 5 mL volume. Eyes were harvested at 24
hours for caspase 8 activity assays, 3 days for TUNEL staining or at 2
months for PR cell counts.
Results: Doses of 0.5, 5 and 50 mg of Met12-DMSO, administered
intravitreally, were equally effective at inhibiting caspase 8
activation, in vitro, with 5 mg being the lowest dose that provided
maximal suppression of photoreceptor TUNEL staining and
preservation of outer nuclear layer cell counts in vivo.
In Met12-citrate assays, 100uM Met12 yielded maximum inhibition
of Caspase 8 activity following Fas activation in vitro, with an IC50
of approximately 20uM. In vivo, Met12-citrate inhibited PR apoptosis
as measured by TUNEL staining, with a less potent effect than
Met12-DMSO.
Conclusions: Met12-DMSO inhibits PR apoptosis in a dosedependent fashion. Compared to Met12-DMSO, Met12-citrate was
a less potent inhibitor of caspase activity in vitro, and this translated
to a less effective inhibition of PR apoptosis in vivo. As such, we
successfully demonstrate the ability of our in vitro assay to predict
in vivo efficacy and the usefulness of our in vitro assay to screen
aqueous formulations of Met12.
Commercial Relationships: Lee Kiang, ONL Therapeutics (P);
Cagri Besirli, ONL Therapeutics (P); Rae-Sung Chang, ONL
Therapeutics (P); Anna Schwendeman, ONL Therapeutics (I), ONL
Therapeutics (P); David N. Zacks, ONL Therapeutics (C), ONL
Therapeutics (I), ONL Therapeutics (P)
Program Number: 371 Poster Board Number: D0107
Presentation Time: 8:30 AM–10:15 AM
Scleral Wound Healing and Crosslink Technique. Influence on
Sclerotomy Cicatrization on the Rabbit Eyes. Evaluation with
Histological and Morphological Analysis in the Scleral Tissue. A
Pilot Study.
Eduardo Damasceno1, Nadyr Damasceno1, Bryan Hudson Hossy2,
Ana Carolina Rio2, Nadia C. Miguel2. 1Ophthalmology, Universidade
Federal Fluminense, Rio de Janeiro, Brazil; 2Histology and
Morphology, Universidade Federal do Rio de Janeiro, Rio de Janeiro,
Brazil.
Purpose: Wound healing evaluation in the postoperative of
sclerotomy. It was performed after the crosslink technique on the
rabbit eyes. The hypothesis: Crosslink technique could stabilize the
sclerotomy incision more and improve the sclerotomy cicatrization.
<!—EndFragment—>
Methods: Study Design: Experimental. Therapy procedure chosen
for the Right eye (sclerotomy plus crosslink technique) and control
procedure chosen for the left eye (sclerotomy only). Animals Treated:
Three male New Zealand rabbits. The Crosslink technique: UVA
Emission of 370 nm with a surface irradiance of 3 mW/cm2 and
application of Riboflavin drops for 30 minutes. This procedure was
performed on the right eye at 2 mm from the limbus. Sclerotomy
procedure with 23 gauge incision was performed at 2mm from
the limbus in both right (on the treated area) and left eye. Thirty
days after surgery, an evaluation with slit lamp biomicroscopy and
morphological analysis were performed and compared, the right
sclerotomy cicatrization to the left sclerotomy.
The morphological Analysis: evaluation of sclerotomy wound healing
(closed or opened).
The histological analysis with hematoxylin-eosin (HE) and
Picrosirius-Red staining (PR). Final evaluation analyzed collagen
fibers density on the sclerotomy wound incision. Image pro plus
software was used.
Results: The Morphological Analysis: Right eye – All samples
presented sclerotomy closure. In two cases the incision edges
revealed almost no evidence. Left eye – All samples presented the
sclerotomy closure and the incision edges were evident .
The Histological Analysis: Right and Left eye - HE staining - in
the incision of all samples there were no evidence of immflamatory
cells. PR staining revealed no evidence of collagen fibers type III in
the wound healing process of sclerotomy in both eyes (figure 1 a,b).
Image pro plus revealed a higher collagen fiber density in the Right
eye .
Conclusions: The scleral wound healing happened in a tissue with
poor vascular support and with low incidence of new collagen fibers
in the scaring process thirty days after surgery. So, the crosslink
technique seems to stabilize the sclerotomy incision and help the
scaring process. It could be a good factor to improve sclerotomy
cicatrization.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Figure 1a. Right Eye
Figure 1b - Left Eye
Commercial Relationships: Eduardo Damasceno, None; Nadyr
Damasceno, None; Bryan Hudson Hossy, None; Ana Carolina
Rio, None; Nadia C. Miguel, None
Program Number: 372 Poster Board Number: D0108
Presentation Time: 8:30 AM–10:15 AM
Simultaneous sustained release of VEGF and bFGF from a
non-biodegradable implant in the supra-choroidal space induces
experimental choroidal neovascularization in both New Zealand
white and Dutch belt rabbits
Corinne G. Wong1, Lucy H. Young2, Kathy Osann3, Ricardo A.
Carvalho4, Timothy You5, TC Bruice6. 1SCLERA LLC, Carlsbad,
CA; 2Harvard, Cambridge, MA; 3Medicine, University of California
Irvine, Irvine, CA; 4Ophthalmology, University of California, Irvine,
CA; 5Orange County Retina Group, Santa Ana, CA; 6UCLA, Los
Angeles, CA.
Purpose: Previous studies have shown that intravitreal placement
of a VEGF/bFGF slow-release implant produces florid robust retinal
neovascularization in the Dutch belt rabbit but not in the New
Zealand white rabbit. The purpose of this study is to define whether
a difference exists in the response between New Zealand white
rabbit and Dutch belt rabbit following suprachoroidal implantation
of a VEGF/bFGF slow-release pellet for induction of experimental
choroidal neovascularization.
Methods: Adult New Zealand white (NZW) rabbits (N = 12) and
pigmented Dutch belts (N = 6) were implanted transclerally with a
non-biodegradable polymeric implant containing both VEGF and
bFGF. Negative controls of NZW (N =6) and Dutch belt rabbits (N
= 6) were given blank implants. Experimental CNV was assessed
over a four-week time period. Eyes were examined by indirect
ophthalmoscopy after surgery at 24 hrs, 48 hrs, 4 days, 7 days, and
28 days. Both color fundus and fluorescein angiography (FA) images
were captured by a TOPCON 50EX digital IMAGE net retinal
camera system at 1, 2, 3, and 4 weeks. CNV was graded between 0 to
3. P values were calculated by using either the Kruskal-Wallis nonparametric test or the Fisher’s exact test at the 4 time points.
Results: In all eyes implanted with VEGF/bFGF pellets (N =
18), experimental CNV was observed within 1 week after pellet
implantation and continued through weeks 2, 3, and 4. The
progression of experimental CNV was similar in both NZW and
Dutch belt rabbits. P values were calculated by using either the
Kruskal-Wallis non-parametric test or the Fisher’s exact test at the 4
time points.
Conclusions: These results demonstrate that experimental CNV is
induced rapidly in both types of animals by simultaneous sustained
release of VEGF and bFGF in a polymeric implant within the
supra-choroidal space. Pigmentation does not appear to change
experimental CNV in the rabbit. Progression of experimental rabbit
CNV provides a novel pre-clinical tool for rational quantitative
structure-activity relationship (QSAR) in retinal drug development,
sustained-release drug formulations, and drug delivery device
systems in amelioration of exudative AMD.
Commercial Relationships: Corinne G. Wong, None; Lucy H.
Young, None; Kathy Osann, None; Ricardo A. Carvalho, None;
Timothy You, None; TC Bruice, None
Program Number: 373 Poster Board Number: D0109
Presentation Time: 8:30 AM–10:15 AM
Ophthalmic Plasma Probe - A New Instrument for Delivery of
Non-thermal Plasma to the Ocular Surface and Sub-surface
David S. Pao1, Gregory Fridman2, Kristina Pao1. 1Wills Eye Hospital,
Levittown, PA; 2Drexel University, Philadelphia, PA.
Purpose: To utilize non-thermal plasma to the ocular surface for
prophylaxis against infection at a safe energy range for normal ocular
tissue safety. To explore the energy range for treatment of surface and
sub-surface pathological tissue with minimal damage to surrounding
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
tissue. To accomplish these goals, a new probe was designed and this
instrument is submitted for presentation.
Methods: Plasma produces reactive oxygen species (ROS),
predominately nitrogen and oxygen species in air. These have
antibacterial properties and may penetrate tissue surface with tissue
damage when energy is above a certain threshold.
To test the antibacterial property, live New Zealand white rabbit
corneas were innoculated with Staphylococcus aureus. Half were
treated with plasma and all cornea swab specimens were plated on
blood agar.
To test tissue damage, live rabbit and porcine eyes were treated with
plasma energy beyond the range of antibacterial effects to produce
visible damage and gradually the energy was reduced. Histological
sections for light microscopy were obtained to view any damage.
Results: Comparing bacterial colony counts of blood agar plates,
there was marked reduction from innoculated specimen swabs of
plasma treated corneas.
Histological examination of eye tissue showed no light microscopy
damage at energies used as bactericidal. Increasing above this energy
produced clinical and histological damage.
From these trials it was established that the probe will be the
dielectric barrier discharge (DBD) design type.
Conclusions: Non-thermal plasma has been shown to achieve blood
coagulation, wound sterilization, and treatment of some diseases
both in vitro and animal models. Plasma medicine is a new field
with many applications in ophthalmology due to its unique specialty.
Ophthalmology can be a prime contributor.
Antibacterial surface treatment may consist of a plasma probe over
the cataract wound and/or intravitreal injection site for surface wound
sterilization, reducing the incidence of endophthalmitis.
Surface and intraocular tumors may be susceptible to plasma
treatment, as the tissues penetrated are relatively thin.
This new probe will provide the instrument to conduct these intial
studies, particularly in microscopic surgery. The potential for “plasma
ophthalmology” is vast and it has just begun.
Commercial Relationships: David S. Pao, None; Gregory
Fridman, None; Kristina Pao, None
Support: Wills Eye Innovation Research Grant
Program Number: 374 Poster Board Number: D0110
Presentation Time: 8:30 AM–10:15 AM
In vitro and in vivo evaluation of an anti-VEGF controlled release
drug delivery system
Christian R. Osswald1, Micah J. Guthrie1, William F. Mieler2,
Jennifer J. Kang Mieler1. 1Biomedical Engineering, Illinois Institute
of Technology, Chicago, IL; 2Ophthalmology and Visual Sciences,
University of Illinois at Chicago, Chicago, IL.
Purpose: Current therapies for posterior segment diseases often
require monthly bolus injections of anti-vascular endothelial growth
factors (anti-VEGFs). The purpose of this study was to validate a
drug delivery system (DDS) capable of bioactive anti-VEGF release
for over 6 months.
Methods: The DDS consists of poly(lactic-co-glycolic acid)
microspheres suspended in a thermoresponsive poly(Nisopropylacrylamide)-based hydrogel. Microspheres were loaded
with ranibizumab or aflibercept. The MTS assay was used to
determine bioactivity of release samples on human umbilical vascular
endothelial cells (HUVECs) under VEGF-induced proliferation. A
rat model of choroidal neovascularization (CNV) was used whereby
CNV was induced in male Long-Evans rats using an Ar-green laser.
At 1, 2 and 4wk post-CNV induction/treatment, lesion areas were
measured using a multi-Otsu threshold technique. Dark-adapted
electroretinogram (ERG) intensity series were elicited with full-field
Ganzfeld stimulation (max flash intensity: 307 scotopic cd×s×m-2;
duration: 2ms). Rats treated with our DDS were compared to bolus
anti-VEGF injection counterparts, non-treated and negative control
(DDS without anti-VEGF) rats.
Results: Our DDS is capable of releasing ranibizumab and
aflibercept for ~200 days with an initial burst (first 24hr) of
21.0±2.0% and 20.1±0.8%, respectively, followed by controlled
release of 0.2μg/day and 0.07μg/day, respectively. No toxicity was
seen in HUVECs at any time for any treatment group; negative
control showed no inhibitory effect on HUVEC proliferation. Both
anti-VEGFs remained bioactive throughout release. In vivo results
confirmed our DDS is non-toxic with no significant changes in
ERG maximal response (Rmax) or half-saturation (σ) compared
to control measurements (Amax: -472±20μV; Bmax: 770±42μV; σa:
2.8±0.7cd×s×m-2; σb: 0.003±0.0005cd×s×m-2). Negative control
lesion areas (0.05±0.001mm2) were not significantly different
than non-treated at any time point (0.05±0.002mm2). At 4wk, rats
treated with our DDS had CNV lesion areas that were 37% and 32%
smaller than their bolus-injection counterparts for ranibizumab and
aflibercept, respectively (p<0.05).
Conclusions: The results indicate our DDS can deliver bioactive
anti-VEGFs with significant impacts in both in vitro and in vivo
models. Our DDS may provide a significant advantage over current
bolus injection therapies in the treatment of posterior segment
diseases.
Commercial Relationships: Christian R. Osswald, None;
Micah J. Guthrie, None; William F. Mieler, Genentech, Inc.
(C), ThromboGenics, Inc. (C); Jennifer J. Kang Mieler, US
20140065226 A1 (P)
Program Number: 375 Poster Board Number: D0111
Presentation Time: 8:30 AM–10:15 AM
New Clinical Biomarkers of Inflammatory Response after
Intravitreal Steroids in Center Involving Diabetic Macular
Edema
Stela Vujosevic1, Silvia Bini1, Marianna Berton1, Giulia Midena2,
Ferdinando Martini1, Annarita Daniele1, Porzia Pucci1, Edoardo
Midena1, 3. 1Ophthalmology, University of Padova, Padova, Italy;
2
Universita’ Campus Biomedico, Roma, Italy; 3Fondazione G.B.
Bietti, IRCCS, Roma, Italy.
Purpose: To asses, non invasively, early morphologic modifications,
as signs of (anti)inflammatory response, in the retina and choroid,
after intravitreal steroid treatment in centre-involving diabetic
macular edema (DME).
Methods: Retrospective analysis of images of 20 eyes (20 patients)
who underwent intravitreal dexamethasone implant. All patients
had good quality fundus color photo, spectral-domain (SD)-OCT
and fundus autofluorescence (FAF) before and at 2 months after
treatment. The following parameters were evaluated on SD-OCT:
number of hyper-reflective spots (HRS) in the area between 500μm
and 1500μm nasally and temporally to the fovea, on a linear 180°
scan, in three specific layers: from inner limiting membrane-(ILM)
to inner plexiform layer-(IPL), from inner nuclear layer-(INL) to
outer plexiform layer-(OPL) and in the outer nuclear layer-(ONL);
inner and outer retinal and choroidal thickness (RT, CT) in 5 specific
sites (fovea, 500μm and 1500μm both nasally and temporally to
the fovea); FAF images were evaluated for patterns of normal and
increased FAF in the fovea and for the extent of increased FAF. All
measurements were performed by 2 masked graders, independently.
Signed Rank test was used for statistical analysis.
Results: At baseline HRS were mainly located in the INL-OPL and
ILM-IPL. After treatment: there was a significant decrease in HRS in
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
the INL-OPL(-3.8+2.5, p=0.007) and ONL, (-1.8+2.3, p=0.04); mean
RT decreased in the fovea (612.9μm+157.3 vs 337.9μm +118.5,
p=0.003), and at almost all measurement sites, (p<0.03, et least for
all). CT increased in the fovea (201.4μm+51.3 vs 229.8μm+39.6,
p=0.03), temporally at 1500μm, (160.9 μm+43.3 vs 211.9 μm+46.1,
p=0.01) and nasally at 500μm, (170.8μm+50.4 vs 220.0μm+24.5,
p=0.01); FAF changed from increased to normal in 12 eyes (60%),
and decreased in extent in all 20 eyes (100%).
Conclusions: HRS and increased foveal FAF have been recently
proposed as signs of microglial cells activation in DME. Activated
microglia is considered responsible of retinal inflammation in
diabetics. SD-OCT and FAF show, non invasively, the decrease in
inflammatory signs and safety (increase in CT) after intravitreal
dexamethasone implant treatment. These parameters could become
new clinical biomarkers of inflammation in DME.
Commercial Relationships: Stela Vujosevic, None; Silvia Bini,
None; Marianna Berton, None; Giulia Midena, None; Ferdinando
Martini, None; Annarita Daniele, None; Porzia Pucci, None;
Edoardo Midena, None
Program Number: 376 Poster Board Number: D0112
Presentation Time: 8:30 AM–10:15 AM
Decanted Triamcinolone for Macular Edema in NonVitrectomized and Vitrectomized Eyes
Frank Tsai, William R. Freeman. UCSD Department of
Ophthalmology, Shiley Eye Center, La Jolla, CA.
Purpose: Intravitreal triamcinolone acetonide (IVTA) doses varying
from 1-4mg have traditionally been used. Higher doses of IVTA
may provide better treatment response or longer duration of effect.
We report the largest retrospective case series evaluating the safety
and efficacy of high-dose decanted IVTA for macular edema in
vitrectomized and non-vitrectomized eyes.
Methods: A retrospective chart review of 78 injections in 40
consecutive eyes treated with intravitreal injection of decanted 20mg
triamcinolone acetonide injected for macular edema was performed.
The non-vitrectomized group comprised of 15 eyes, while 25
eyes had prior vitrectomy. Change in best-corrected visual acuity
(BCVA), intraocular pressure, central macular thickness (CMT),
and complications were reviewed. Macular edema was associated
with diabetes, vein occlusion, uveitis, or post-operative surgery. All
patients had at least 3 months of follow-up with a mean of 9 months.
Reinjection of IVTA was performed on a PRN basis. Injection cost
was compared to dexamethasone intravitreal implant. Data analyses
was performed using JMP version 5 (SAS,Cary,NC).
Results: Improvement in BCVA was found in 12 of 15 eyes(80%)
at 2 months, 8 of 14 eyes(57%) at 6 months, and 8/12 eyes(67%)
at 12 months for non-vitrectomized eyes, compared with 13 of
25 eyes(52%), 14 of 25 eyes(56%), and 9 of 17 eyes(53%) of
vitrectomized eyes, respectively. The mean change in BCVA letter
score from baseline at 1, 2, and 3 months was 7.5, 9.8, and 5.86 in
the non-vitrectomized eyes, and 1.3, 6.8, and 0.9 in the vitrectomized
eyes, respectively. CMT improved in both groups and was
statistically significant at all time points. 40% of eyes developed IOP
>21mm of Hg. The rate of cataract requiring surgery was 50% after
1 year. Retinal detachment and glaucoma surgery occurred in 1 eye
each.
Conclusions: Decanted IVTA is safe and effective for treating
macular edema due to various etiologies. Although it appears
more effective in non-vitrectomized patients, decanted IVTA
provides statistically significant visual gains peaking at 2 months
after injection, but up to 12 months in some patients after a single
injection. The dose of IVTA can be reliably titrated by the decantation
method. The duration of decanted IVTA appears similar to Ozurdex,
which peaks at 2 months and generally lasts 3-6 months. The
acquisition cost of Kenalog was $75 per dose, compared to $1295 per
dose of Ozurdex.
Commercial Relationships: Frank Tsai, None; William R.
Freeman, None
Support: Support in part by NEI vision core grant P30EY022589
and RPB (Research to Prevent Blindness) Inc.
Program Number: 377 Poster Board Number: D0113
Presentation Time: 8:30 AM–10:15 AM
The increased stability of FpFs compared to monoclonal
antibodies
Hanieh Khalili1, 2, Steve Brocchini1, 2, Peng T. Khaw2, Ashkan Khalili2,
Garima Sharma2, 1. 1Pharmaceutics, UCL School of Pharmacy,
London, United Kingdom; 2NIHR Biomedical Research Centre
for Ophthalmology, Moorfields Eye Hospital & UCL Institute of
Ophthalmology, London, United Kingdom.
Purpose: Fab-PEG-Fab (FpF) molecules are new IgG mimetics
that are designed to be more stable than therapeutic IgG monoclonal
antibodies (MAbs). The accessible disulfides in MAbs are prone to
scrambling and the hinge region is susceptible to cleavage. The FpFs
have been designed to alleviate these intrinsic limitations of MAbs.
The aim of this study was to evaluate FpF stability to storage and to
freeze-drying.
Methods: Storage stability studies were conducted in glass vials at
37C with bevacizumab used in its pharmaceutical formulation and
the corresponding bevacizumab derived FpF in PBS (pH 7.4) at
double the bevacizumab concentration. For pre-formulation freezedrying studies, bevacizumab was eluded over a PD-10 column to
remove excipients and both the de-formulated bevacizumab and FpF
were subjected to freeze-drying. Primary drying was performed at
-20C for 12 hours at 100 μBar, followed by secondary drying at 20C
for 2 hours. Biacore binding analyses were performed using a VEGF
functionalised CM3 chip (92 RU).
Results: The FpF did not aggregate or display any light/heavy
chain dissociation after 30 days storage at 37C as liquid (0.250 mg/
mL[SB1] ). Chain dissociation and aggregation were observed for
the pharmaceutical formulation of bevacizumab (0.125 mg/mL) after
7 days (37C). Both the FpF and deformulated bevacizumab were
subjected to freeze-drying. SDS-PAGE analysis indicated there was
no loss of FpF, or the occurrence of de-PEGylation or aggregation
for the FpF molecule. Binding of FpF to VEGF was maintained after
reconstitution. In contrast there appeared to be loss of bevacizumab
as observed by decreased band intensity on SDS-PAGE. Particulates
were observed when bevacizumab was reconstituted.
Conclusions: The FpF is a MAb mimetic that appears to be less
prone to aggregation and precipitation than a MAb. The increased
stability of FpFs indicates they have the potential to be fabricated into
longer acting dosage forms than may be possible with MAbs.
Commercial Relationships: Hanieh Khalili, None; Steve
Brocchini, None; Peng T. Khaw, None; Ashkan Khalili, None;
Garima Sharma, None
Program Number: 378 Poster Board Number: D0114
Presentation Time: 8:30 AM–10:15 AM
Evaluation of a retinal detachment model for testing hydrophilic
vitreous substitute
Nele Schneider1, Kai Januschowski1, Jose Hurst1, Lisa Pohl1,
Maximilian Schultheiss1, Christine Hohenadl2, Charlotte Reither2,
Sven Schnichels1, Martin S. Spitzer1. 1Centre for Ophthalmology,
University Eye Hospital, Schleichstr. 12/1, D-72076 Tübingen,
Germany, Stuttgart, Germany; 2Croma Pharma, Leobendorf, Austria.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Purpose: Previously we showed that hydrophilic gels based on
UV-crosslinked hyaluronate are well tolerated in rabbit eyes and
thereby the lens usually remains clear. Consequently we created a
rabbit model of retinal detachment to evaluate the efficacy of the gel,
enabling us to evaluate this tamponade against silicone oil.
Methods: At first, we performed preoperative investigations on 2
groups consisting of 8 young rabbits (2 months old) using ERG,
OCT and examining both eyes with the slit lamp and measuring the
intraocular pressure. Thereafter, pars plana vitrectomy was performed
in the right eye. Retinal detachment of one quadrant was induced by
creating a small retinotomy near the vascular arcade and subretinal
injection of balanced salt solution (BSS). Retinal hemorrhage was
induced by a circumscribed vascular breach in the periphery of
the vascular arcade. The retina was reattached by increasing the
infusion pressure and the blood was washed out until the bleeding
stopped. The retinal tear was treated by endolaser photocoagulation.
At the end of the procedure the eye was either filled with 5000 cs
silicone oil (after fluid air exchange) or the respective hydrogel was
injected into the BSS filled eye until egress of the hydrogel of the
opposite sclerotomy. After surgery, the right eye was reexamined
at regular intervals and compared to the unoperated left eye
by slit lamp examination, funduscopy and intraocular pressure
measurements. The first OCT and ERG was made after one month.
Afterwards both eyes were removed and processed for histology and
immunohistochemistry with GFAP and Brn3a antibodies.
Results: Seven of eight rabbits that received silicon oil, developed a
partial or even total retinal detachment with pronounced proliferative
vitreoretinopathy within the first two weeks after surgery. The ERG
was completely extinguished in six rabbits, greatly reduced in one,
and stayed unchanged in only one rabbit. In contrast, in the hydrogel
group the retina stayed attached in 50% of the cases.
Conclusions: With the high rate of retinal detachments and
proliferative retinopathy this model provides a good prerequisite
for testing the efficacy of novel hydrogels as vitreous substitutes.
Preliminary tests in this model point towards superior efficacy of
cross-linked hyaluronate compared to silicone oil.
Commercial Relationships: Nele Schneider, None; Kai
Januschowski, Croma Pharma, Leobendorf, Austria (F); Jose Hurst,
None; Lisa Pohl, None; Maximilian Schultheiss, None; Christine
Hohenadl, Croma Pharma, Leobendorf, Austria (E); Charlotte
Reither, Croma Pharma, Leobendorf, Austria (E); Sven Schnichels,
Croma Pharma, Leobendorf, Austria (F), Croma Pharma, Leobendorf,
Austria (F), Croma Pharma, Leobendorf, Austria (F); Martin S.
Spitzer, Croma Pharma, Leobendorf, Austria (F)
Program Number: 379 Poster Board Number: D0115
Presentation Time: 8:30 AM–10:15 AM
Stainless steel micro needle for retinal vein cannulation.
Koen Willekens1, Andy Gijbels2, emmanuel Vander Poorten2,
Dominiek Reynaerts2, Peter Stalmans1. 1Ophthalmology, UZ Leuven,
Leuven, Belgium; 2Production Engineering, Machine Design and
Automation, KULeuven, Leuven, Belgium.
Purpose: The current management of retinal vein occlusion
consists mainly of treating its complications instead of the cause.
Although successful blood clot removal via retinal vein cannulation
using glass micropipettes has been reported, such glass design is
fragile and implicates significant restrictions in shape. Therefore, a
dedicated stainless steel micro needle was developed for retinal vein
cannulation.
Methods: A stainless steel 45° angled micro needle with an outer
diameter of 80 mm (approximately 44 Gauge) and a lumen of 35mm
was developed. Using perfused chorio-allantoic membrane vessels
of fertilized chicken eggs (aged 11 days) this needle was evaluated
for puncture and infusion capability proven by visual confirmation of
blood washout and bleeding after needle retraction.
Results: Out of 25 freehanded cannulation attempts, 20 punctures
were successfully executed by a junior vitreoretinal surgeon with
visual confirmation of blood washout and bleeding after needle
retraction. Vessel diameter ranged from 100mm to 250mm. There
were no double punctures noted although in one vessel a slit-like
injury to its wall was observed. Failure of puncture was related to
difficult freehanded alignment and rolling of the vessels. All vessels
were cannulated with the same needle that showed no signs of
wearing. Perfusion pressures below 30 psi rendered stable infusion
flow throughout the experiments.
Conclusions: Micro vessel puncture and infusion of liquid is possible
with this custom-built stainless steel 44 Gauge needle.
CAM vessel cannulation
1) TOP: vessel puncture
2) MID: blood washout
3) BOTTOM: Bleeding after needle retraction
Commercial Relationships: Koen Willekens, None; Andy Gijbels,
KULeuven R&D (P); emmanuel Vander Poorten, KULeuven R&D
(P); Dominiek Reynaerts, KULeuven R&D (P); Peter Stalmans,
None
Program Number: 380 Poster Board Number: D0116
Presentation Time: 8:30 AM–10:15 AM
Local Anesthesia with Blunt Subtenon Cannula vs. Sharp
Retrobulbar Needle for Vitreoretinal Surgery: A Retrospective,
Comparative Study
David Reichstein1, Clinton Warren2, Dennis P. Han2, William
Wirostko2. 1Tennessee Retina, Nashville, TN; 2Medical College of
Wisconsin, Milwaukee, WI.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Purpose: Local anesthesia using sharp retrobulbar needle can be
associated with complications. We compare safety and efficacy of
blunt subtenon cannula for administering local anesthesia before
vitreoretinal surgery to anesthetic delivered by a sharp retrobulbar
needle.
Methods: This was a retrospective, comparative study of all patients
undergoing local anesthesia before vitreoretinal surgery at the
Medical College of Wisconsin between August 2009 and November
2013. Local anesthesia administered either via blunt subtenon
cannula or sharp retrobulbar needle prior to vitreoretinal surgery. The
incidence of local and systemic anesthesia-related complications as
well as the incidence of inadequate local surgical anesthesia requiring
conversion to general anesthesia was recorded.
Results: Blunt subtenon cannula was used in 940 cases while sharp
retrobulbar needle was used in 771 cases for local anesthesia. Factors
associated with use of sharp retrobulbar needle over subtenon
cannula were presence of prior scleral buckle and inclusion of scleral
buckle placement in the procedure. Of 940 surgeries performed with
subtenon’s cannula, only 2 cases were not completed, due to ether
suprachoroidal hemorrhage (1 case) or sleep-apnea-related hypoxia
(1 case). Of 771 surgeries performed with sharp retrobulbar needle,
only 1 case was not completed, due to suprachoroidal hemorrhage (1
case). Intraoperative conversion to general anesthesia was required
after subtenon cannula in 9 patients and after retrobulbar needle
in 12 patients. No case of globe perforation, severe retrobulbar
hemorrhage, or severe conjunctival chemosis was observed in either
group.
Conclusions: Blunt subtenon cannula appears as effective and
safe as sharp retrobulbar needle for administering local anesthesia
prior to vitreoretinal surgery. Vitreoretinal surgeons may wish to
consider using a blunt subtenon cannula for local anesthesia during
vitreoretinal procedures.
Photograph of blunt subtenon cannula (Beaver Visitec, Waltham,
Massachusetts). The delivery of subtenon anesthesia is demonstrated.
Commercial Relationships: David Reichstein, None; Clinton
Warren, None; Dennis P. Han, None; William Wirostko, None
Support: Unrestricted grant from Research to Prevent Blindness
Program Number: 381 Poster Board Number: D0117
Presentation Time: 8:30 AM–10:15 AM
Extraocular non-invasive transscleral LED-Endoilluminator for
Eye Speculum Integration
Frank H. Koch1, Philipp Simon Koelbl3, Christoph Lindner3,
Maximillian Bader3, Svenja Deuchler1, Pankaj Singh1, Adonis Chedid
De Robaulx1, Christian Lingenfelder3, Martin Hessling2. 1Retina and
Vitreous Unit, University Eye Clinic Frankfurt / Main, Frankfurt
am Main, Germany; 2Department of Mechatronics and Medical
Engineering,, University of Applied Sciences Ulm, Ulm, Germany;
3
alamedics, Dornstadt, Germany.
Purpose: To introduce an extraocular non-invasive transcleral
eye speculum with integrated light-emitting diode (LED) Endoilluminator which increases the efficacy of performing pars
plana vitrectomy (PPV) procedures.
Methods: A white LED is incorporated as a light source. The
speculum which has the integrated LED applies appropriate pressure
to the sclera. A prototype has been developed incorporating the
relevant international standards for endoillumination. Tests were
performed with enucleated porcine eyes.
Results: This new instrument efficiently illuminates the vitreous
cavity in the porcine eyes. We could demonstrate that all parameters
are well within their limits required by the international standards for
endoillumination of the eye. Considering a distance of 18 mm (light
delivered through pars plana) and 1 mm (light delivered through
sclera from behind the retina) irradiance values of 6,22E-05 and
1,28E-03 (W/cm2) allow exposure time of 44 hrs respectively 2 hrs
with no phototoxicity risk from the short wavelength light.
The thermal relevant irradiance EVIS-R of 11,7mW/cm2 is significantly
lower than the clinically relevant threshold value of 0,7 W/cm2. The
measured maximum temperature of 27.8° C is far below the threshold
of between 47° C and 57° C which is considered to be relevant for
potential thermal damage.
Conclusions: Conventional chandelier endoilluminators for PPV
consist of a light-fibre which is introduced into the eye through an
incision. A novel extraocular LED-Endoilluminator was developed.
This illuminator is attached to a speculum and does not require
connection to a light source or a light fibre. The transmitted and
diffused light results in an incision free and complete illumination
of the vitreous cavity. Compared to standard illumination systems,
the possible exposure time to light of all wavelengths becomes
significantly longer. This is also true in cases when the illumination
is not applied at the PP but more posterior through the retina. Neither
the amount of ultraviolet radiation nor thermal effects are clinically
relevant.
Peyman, G. A. Improved vitrectomy illumination system. AJO 81:
99 –100 (1976). Koch F, et al. Multiport illumination system (MIS)
for panoramic bi-manual vitreous surgery. Graefe’s Arch 229:425429 (1991). Bashkatov, A et al. Optical properties of human sclera
in spectral range 370–2500 nm. Optics and Spectroscopy 109, 2,
197–204 (2010).
Commercial Relationships: Frank H. Koch, None; Philipp Simon
Koelbl, None; Christoph Lindner, alamedics (E); Maximillian
Bader, None; Svenja Deuchler, None; Pankaj Singh, None; Adonis
Chedid De Robaulx, None; Christian Lingenfelder, alamedics (P);
Martin Hessling, university ulm (P)
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Program Number: 382 Poster Board Number: D0118
Presentation Time: 8:30 AM–10:15 AM
Effect of symptom duration and extent of retinal detachment on
intravitreal cytokines and chemokines
Andreas Pollreisz1, Katharina Eibenberger1, Stefan Sacu1, Danijel
Kivaranovic2, Michael Georgopoulos1, Ursula Schmidt-Erfurth1.
1
Ophthalmology, Medical University Vienna, Vienna, Austria;
2
Section for Medical Statistics, Medical University Vienna, Vienna,
Austria.
Purpose: The aim of the study was to evaluate the effect of
duration and extent of retinal detachment (RD) on the expression of
intravitreal cytokines and chemokines.
Methods: 60 patients with rhegmatogenous RD and 20 age
and gender-matched control patients with idiopathic epiretinal
membranes were included in the study. Vitreous samples were taken
undiluted at the beginning of primary vitrectomy and immediately
stored at -80 °C. The following clinical parameters were assessed
from the RD patients prior to surgery: symptom duration (days);
number of quadrants detached; fovea-off – fovea-on (assessed by
optical coherence tomography); RD height and refractive power.
Concentrations of 40 different cytokines and chemokines were
measured in the vitreous of RD eyes by Luminex bead assay,
compared to control patients and correlated to clinical parameters in
patients with RD.
Results: 10 cytokines and chemokines were significantly upregulated in the vitreous of RD eyes compared to the control group
including tissue inhibitor of metalloproteinases (TIMP) 1 and 2,
macrophage inflammatory protein 1 alpha, monocyte chemoattractant
protein-1, interleukin 8 and 6, interferon gamma-induced protein-10
(IP-10), brain-derived neurotrophic factor, transforming growth factor
beta and platelet-derived growth factor AB/BB. Levels of TIMP-1,
a modulator of extracellular matrix (ECM), significantly decreased
with longer symptom duration of RD suggesting reduced protection
against ECM remodeling (p=0.0013). The concentrations of proinflammatory IL-8 and myeloperoxidase (MPO) were significantly
higher when 2 or more quadrants were involved in eyes with RD
(p=0.0004; p=0.0032). In eyes with detached fovea levels of the proinflammatory IP-10 were significantly elevated compared to fovea-on
eyes (p=0.0416;). Height of RD or refractive power did not cause any
significant changes of protein concentrations.
Conclusions: Our study results depict the alteration of intravitreal
cytokines and chemokines in eyes with RD and demonstrate that
duration and extent of RD influence their intraocular expression.
Hence, these data add fundamental understanding to reported
observations of poorer final visual acuity in eyes with longer and
more severe forms (fovea-off) of RD and may help to develop
potential therapeutics administered before or during retinal repair
surgery to facilitate the restoration of maximum visual acuity.
Commercial Relationships: Andreas Pollreisz, None; Katharina
Eibenberger, None; Stefan Sacu, None; Danijel Kivaranovic,
None; Michael Georgopoulos, None; Ursula Schmidt-Erfurth,
None
Program Number: 383 Poster Board Number: D0119
Presentation Time: 8:30 AM–10:15 AM
Change of Cytokines in Recurred Patients After Injection of
Intravitreal Ranibizumab for Branch Retinal Vein Occlusion
With Macular Edema
Hidetaka Noma1, Kanako Yasuda1, Hayate Nakagawa1, Ryosuke
Motohashi1, Osamu Kotake1, Hiroshi Goto2, Masahiko Shimura1.
1
Department of Ophthalmology, Tokyo Medical University Hachioji
Medical Center,, Tokyo, Japan; 2Department of Ophthalmology,
Tokyo Medical University, Tokyo, Japan.
Purpose: To investigate the relations among visual acuity, central
macular thickness (CMT), recurrent times, and changes of growth
factors, vascular endothelial growth factor (VEGF) receptor, and
inflammatory factors, including VEGF in recurred patients after
intravitreal ranibizumab (IVR) for branch retinal vein occlusion
(BRVO) with macular edema.
Methods: Thirty-one eyes with BRVO were performed IVR. After
initial IVR, needed injection (PRN) of IVR was done for 6months.
At the time of IVR, a mean volume of 0.1 mL of aqueous humor
was collected by anterior chamber limbal paracentesis. All patients
signed an informed consent form. Then the aqueous humor levels of
vascular endothelial growth factor (VEGF), soluble VEGF receptor
(sVEGFR)-1, sVEGFR-2, soluble intercellular adhesion molecule
(sICAM)-1, monocyte chemotactic protein (MCP)-1, plateletderived growth factor (PDGF)-AA, interleukin (IL)-6, and IL-8 were
measured by the suspension array method (xMAP; Luminex Corp.
Austin, TX). Macular edema was examined by optical coherence
tomography and its severity was determined from the CMT.
Results: An average of 1.7 times of additional IVR was required
from initial IVR for six months (0 times, 4 patients; 1 times; 7
patients; 2 times, 14 patients; 3 times, 6 patients). There were no
significant relations between improvement of visual acuity and
CMT or recurred times. There were significant correlations between
baseline levels of sVEGFR-1 and PDGF-AA or recurred times
(P=0.047, P=0.023, respectively). The aqueous humor levels of
sVEGFR-1 and PDGF-AA after IVR significantly decreased in
the first recurred time and second recurred time than baseline, but
increased in the third recurred time than second recurred times. On
the other hand, the aqueous humor levels of VEGF, PlGF, IL-6, and
IL-8 after IVR significantly decreased in the first recurred time, the
second recurred time, and the third recurred time than baseline (all
P<0.05).
Conclusions: These findings suggest that sVEGFR-1 and PDGF-AA
may contribute to the pathogenesis of BRVO in the patients who
repeat a recurrence after IVR.
Commercial Relationships: Hidetaka Noma, None; Kanako
Yasuda, None; Hayate Nakagawa, None; Ryosuke Motohashi,
None; Osamu Kotake, None; Hiroshi Goto, None (F); Masahiko
Shimura, None
Support: Grant(Novartis Pharma)
Clinical Trial: NCT02169648
Program Number: 384 Poster Board Number: D0120
Presentation Time: 8:30 AM–10:15 AM
Optical Characterization of Vitreous with Multi-Wavelength
Photon Correlation Spectroscopy (MWPCS)
Ashwin Sampathkumar1, Matin Khoshnevis2, 3, Jeffrey A. Ketterling1,
Alfredo A. Sadun3, 4, J Sebag2, 3. 1Biomedical Engineering, Riverside
Research, New York City, NY; 2VMR Institute for Vitreous Macula
Retina, Huntington Beach, CA; 3Doheny Eye Institute, Los Angeles,
CA; 4Ophthalmology, UCLA, Los Angeles, CA.
Purpose: Age-related vitreous degeneration with posterior vitreous
detachment and myopic vitreopathy are often accompanied by
symptomatic floaters due to altered vitreous morphology. While
floaters reduce contrast sensitivity and disturb vision, asteroid
hyalosis (AH) is often asymptomatic, even when advanced. It is
hypothesized that floaters are bothersome because of irregular surface
morphology, while asteroid bodies are not disturbing because of
smooth surfaces. This study developed instrumentation and analytics
to measure light scattering to test this hypothesis.
Methods: Vitreous was modeled using solutions of type II collagen
(Coll; .1%, .05%, .03%, .01%, .005%, .003%, .001%, and hyaluronan
(HA; .25, .15, .075, .025, .015, .0075, and .0025μg/μl). The two
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
were combined at concentrations of .01% Coll & .025μg/μl HA.
AH was modeled with solutions of 75mm and 150mm microspheres,
both individually and combined at concentrations ranging from .01%
to 2.5%. The on- (α) and off-axis (β) scattering coefficients of all
solutions were measured by MWPCS using broadband light that
was collimated to the samples in cuvettes in an integrating sphere.
The forward on-axis and lateral off-axis scattered light was collected
using two spectrometers. Peaks in the off-axis spectra indicate surface
irregularity of the structures in solution.
Results: On-axis scattering of Coll, HA, Coll+HA and microsphere
solutions declined with increasing concentration, up to a threshold
beyond which multiple scattering produced pronounced nonlinear trends (Fig 1). Off-axis measurements showed distinctive
scattering peaks in the spectra, indicative of surface irregularities in
vitreous model solutions, whereas measurements in the AH model
(microsphere) solutions showed less scattering and smoother trends.
Conclusions: This MWPCS system detects variable light scattering
as a function of concentration. Further, light scattering in model
vitreous solutions was consistent with the hypothesis that the surface
morphology of vitreous structures is irregular, influencing light
scattering in a manner that would interfere with vision. Microspheres,
however, had less pronounced effects owing to their smooth surface
morphology and spherical shape. These results suggest utility for this
approach in evaluating light scattering by various vitreous disorders.
Fig 1: On-axis scattering spectra of collagen solutions
Commercial Relationships: Ashwin Sampathkumar, None; Matin
Khoshnevis, None; Jeffrey A. Ketterling, None; Alfredo A. Sadun,
None; J Sebag, None
Program Number: 385 Poster Board Number: D0121
Presentation Time: 8:30 AM–10:15 AM
NEW PROTOTYPE OF ULTRASOUND HARMONICS
VITRECTOR (UHV)
FLUIDICS ANALYSIS: FIRST REPORT
Paulo E. Stanga1, 2, Salvador Pastor1, 2, Isaac Zambrano3, Paul
Carlin4. 1Manchester Royal Eye Hospital and Manchester
Academic Health Science Centre and Centre for Ophthalmology
and Vision Research, Institute of Human Development, University
of Manchester, Manchester, United Kingdom; 2Manchester Vision
Regeneration (MVR) Lab at NIHR/Wellcome Trust Manchester CRF,
Manchester, United Kingdom; 3Eye Bank, Manchester Royal Eye
Hospital, Manchester, United Kingdom; 4Operating Theatre Services,
Manchester Royal Eye Hospital, Manchester, United Kingdom.
Purpose: The liquefaction and excision of the vitreous body using
low power ultrasound harmonics (UH) is a promising new alternative
to pneumatic guillotine vitrectomy systems. The purpose of this
study is to evaluate the effect of gauge size, percentage of ultrasound
power (US) and aspiration settings on flow rate performance using
a prototype ultrasonic harmonics vitrector (UHV). We compare
our results using this new technology with those of a currently
commercially available pneumatic guillotine vitrector.
Methods: Performance of the UHV (Bausch + Lomb, St. Louis,
MO, USA) and currently commercially available pneumatic
guillotine vitrector using 23G and 25G gauge needles (Stellaris
PC® Vitrectomy system, Bausch + Lomb, St. Louis, MO, USA).
Pre-determined aspiration levels (50, 100, 200, 300, 400, 500 and
600 mmHg), cut rates for guillotine vitrector (0, 500, 1000, 1500,
2000, 3000, 4000 and 5000 cuts per minute (CPM)) and percentages
of US power for the 23G and 25G UHV needles (0, 10, 20, 30, 40,
50 %) were used in Balanced Salt Solution (BSS®) and porcine
vitreous. Porcine eyes were obtained within 12-24h of slaughter and
the anterior segment removed at the pars plana to allow for opensky vitrectomy surgery of undiluted vitreous. The vials with BSS®
and the sectioned eyes were weighed on a high-precision balance
(high-speed (2 samples/s) weight scale precise to 0.0001g). Two
independent observers performed six measurements of the mass
of BSS® and the vitreous: before (n=3) and after (n=3) vitrectomy
surgery. Results were converted to volume removed as a function
of time-flow rate (ml/min), using the arithmetic mean of the weight
calculations.
Results: There was no vitreous flow at zero cut rates or US% (off)
for both UH and pneumatic vitrectors. Needle gauge of UHV did not
affect flow rate when used with either BSS® or vitreous. However,
gauge size strongly affected flow rate when using the guillotine
vitrector (p<0.01), both in BSS® and vitreous. Wall thickness of
UHV needles did not affect flow rates neither in BSS® nor vitreous.
Conclusions: Flow rate is not affected by gauge of UHV needle for
similar ports and vacuums and this could allow for the use of smaller
gauge and port size, as well as lower infusion pressures than with a
guillotine vitrector. These new findings demonstrate a promising new
alternative to the currently commercially available technology for
vitrectomy systems.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
over time. Certain patterns of vascular anatomy seem to increase
vulnerability to capillary closure and development of macular edema.
There has been inadequate time to allow comparison of the model to
the actual clinically observed patterns of macular capillary closure
seen in individual patients. This would be the test of whether this
modelling approach can provide an important aspect of individualized
medicine in diabetic retinopathy.
Conclusions: The Compucell3D modelling of the development of
diabetic maculopathy yielded results highly comparable to those
seen in patients but further verification of clinical utility will require
longitudinal studies.
Commercial Relationships: Thomas Gast, None; Fu Xiao, None;
John Scott Gens, None; Lucie Sawides, None; Yuen Ping Toco
Chui, None; James Glazier, None; Stephen A. Burns, None
Support: Indiana University Office Vice-President of Research,
EY04395, P30-EY019008
Commercial Relationships: Paulo E. Stanga, Bauch & Lomb (C),
Bauch & Lomb (F), Bauch & Lomb (R); Salvador Pastor, Bauch &
Lomb (F); Isaac Zambrano, None; Paul Carlin, None
Program Number: 386 Poster Board Number: D0122
Presentation Time: 8:30 AM–10:15 AM
A Compucell3D Model of Diabetic Maculopathy Applicable to
Individual Patients
Thomas Gast1, Fu Xiao2, John Scott Gens2, Lucie Sawides1, Yuen Ping
Toco Chui1, James Glazier2, Stephen A. Burns1. 1Optometry, Indiana
University, Bloomington, IN; 2Physics, Biocomplexity, Indiana
University, Bloomington, IN.
Purpose: Replication of diabetic maculopathy based on a model
initiated with normal macular capillaries from adaptive optics
scanning laser ophthalmoscopy imaging. Using the model with actual
capillary anatomy, to test if relatively simple physiologically based
assumptions can predict the pathology seen in diabetic patients.
Giving a deeper understanding of the actual pathophysiology of
ischemic and edematous diabetic maculopathy. To apply the model
to vascular networks determined in individual patients allowing
prediction of natural history and response to therapeusis with antiVEGF agents.
Methods: With a multiscale Compucell3D model using input
vascular structure of multiple subjects’ macular capillaries from
AOSLO, models were made which included a)flow and pressure
calculations for each point on the vascular map from the arterioles,
through the capillaries, to the venules, b)an underlying cellular
field with a basal VEGF secretion which increases with hypoxia
and declines with cellular VEGF uptake, c)oxygen distributions
determined by diffusion from the vessels and consumption by the
cellular field,d)a probabilitic model of capillary closure based on
VEGF levels modelling ‘stickiness’ of leukocytes and resultant
retinal capillary closure,e)recurrent iterations of the resultant oxygen
distribution and consequent VEGF distribution subsequent to each
capillary closure.
Results: The application of the Compucell3D model to the vascular
anatomy of a number of subjects yielded results highly similar to
those seen in actual patients. A pattern is seen in which the loss
of a given capillary increases the susceptibility of surrounding
capillaries to closure. It does this through the intermediary of local
VEGF concentrations. Areas of ischemia and edema increase in size
Program Number: 387 Poster Board Number: D0123
Presentation Time: 8:30 AM–10:15 AM
The Novel Use of Fluorescein Angiography to Evaluate Retinal
Hemorrhages in Abusive Head Trauma
Audrey C. Ko, Daniel Choi, Nathan W. Blessing, Sander R. Dubovy,
Audina M. Berrocal. Ophthalmology, University of Miami/Bascom
Palmer Eye Institute, Miami, FL.
Purpose: The evaluation of retinal hemorrhages in the setting of
suspected abusive head trauma (AHT) has important social and
legal implications. However, many of these patients have a delayed
presentation after trauma, resulting in partially or fully resolved
retinal hemorrhages at the time of initial evaluation. This severely
hampers the evaluation and attribution of the retinal hemorrhages
to AHT. We report the novel use of fluorescein angiography (FA) to
evaluate retinal pathology secondary to AHT.
Methods: Part A: Retrospective case series of initial and follow-up
FA findings in three cases of suspected and proven AHT. Part B: IRB
approved prospective study of initial and follow-up FA findings in
cases of suspected and proven AHT.
Results: Part A: Case 1. A 5 month old female born at full term
presented with facial bruising, left subdural hematoma, seizure, and
retinal hemorrhages secondary to proven abuse. Four days later,
she underwent a bedside FA showing peripheral nonperfusion (Fig
1A,B). Repeat FA 4 months later showed persistence of findings.
Case 2. An 8 month old male born at 38 weeks gestation presented
with focal seizure and right subdural hemorrhage. Ophthalmic exam
showed the presence of retinal hemorrhages. Three months after
presentation, the patient underwent an exam under anesthesia with
FA that did not show areas of peripheral retinal ischemia (Fig 2C,D).
Due to this finding, it was eventually discovered that the patient had
von Willebrand Disease and the etiology of the exam findings was
more likely to be secondary to minor accidental head trauma in the
setting of a bleeding disorder. Case 3. A 6 week old male born at
full term presented with bilateral subdural hematomas, subarachnoid
hemorrhage, ischemic infarct, new onset seizures, and retinal
hemorrhage. Two days later, he underwent a bedside FA showing
peripheral nonperfusion (Fig 3E,F). Repeat FA 3 months later showed
persistence of findings. Part B: data collection ongoing and expected
to be completed in Spring 2015.
Conclusions: Although retinal hemorrhages are found in the majority
of children with AHT, these findings are not specific for abuse and
can represent mimickers of AHT. Fluorescein angiography is useful
for the evaluation of acute and remote retinal trauma, even after
resolution of retinal hemorrhages. The finding of bilateral peripheral
nonperfusion and remodeling in the setting of abuse may suggest
AHT.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Results: After a mean follow-up of 6.09 ± 0.71 months, mean bestcorrected visual acuity was significantly increased (p: 0.003) from
20/100 to 20/50, mean central retinal thickness decreased from 391 ±
54.5 mc to 335 ± 49.32 mc (p: 0.000), and mean intraocular pressure
changed (p: 0.000) from 12.5 ± 1.06 to 16.7 ± 3.01 In no case
postoperative complication was observed.
Conclusions: Injection of dexamethasone intravitreal implant
resulted effective in the treatment of refractory macular edema
secondary to combined cataract extraction and vitrectomy for
macular epiretinal membrane removal.
Commercial Relationships: Audrey C. Ko, None; Daniel Choi,
None; Nathan W. Blessing, None; Sander R. Dubovy, None;
Audina M. Berrocal, None
Program Number: 388 Poster Board Number: D0124
Presentation Time: 8:30 AM–10:15 AM
Intravitreal dexamethasone implant for macular edema
secondary to vitrectomy for epiretinal membrane
Carlos A. Abdala, Melvin Cabreja, Maria A. Izquierdo. Retina
& Vitreous, Unidad Laser Clinica Oftalmologica, Barranquilla,
Colombia.
Purpose: To report the efficacy of 0.7 mg dexamethasone intravitreal
implant in vitrectomized eyes with refractory macular edema
secondary to combined vitrectomy with epiretinal membrane removal
and phacoemulsification with IOL implantation.
Methods: We report a cases series of 12 eyes with refractory
macular edema secondary to combined 23-gauge vitrectomy for
epiretinal membrane removal with internal limiting membrane
peeling without dying and cataract extraction with IOL implantation.
These patients were previously treated for 3 months with NSAIDs,
topical corticosteroid and periocular corticosteroid injections and
the injection of the 0.7 mg dexamethasone implant was performed
if macular edema persisted despite topical treatment. Best-corrected
visual acuity, central retinal thickness measured by CIRRUS spectral
domain optical coherence tomography, fundus examination and
intraocular pressure were evaluated at baseline, 1 month, 2, 3 and 6
months. Cases of ERM secondary to other pathologies (retinal and
choroid inflammation, vascular occlusion, ocular trauma, diabetes
mellitus, retinal detachment, severe myopia, uveitis,) and age macular
degeneration were excluded from the study.
Commercial Relationships: Carlos A. Abdala, None; Melvin
Cabreja, None; Maria A. Izquierdo, None
Program Number: 389 Poster Board Number: D0125
Presentation Time: 8:30 AM–10:15 AM
Retinal imaging biomarkers for early diagnosis of Alzheimer’s
disease
Heather E. Whitson1, Sina Farsiu2, Sandra Stinnett2, Sung Lee2, Leon
Kwark2, Guy Potter3, James Burke4, Scott W. Cousins2, Eleonora
Lad2. 1Medicine & Ophthalmology, Duke University, Durham, NC;
2
Ophthalmology, Duke University, Durham, NC; 3Psychiatry and
Behavioral Sciences, Duke University, Durham, NC; 4Neurology,
Duke University, Durham, NC.
Purpose: This project is based on the concept that retinas of people
with Alzheimer’s Disease (AD) may experience neuroinflammation
similar to the brain. Inflammatory injury may cause atrophy of the
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
ganglion cell layer (GCL) in the retina and its axonal projections
which may manifest during the prodromal stage of AD. In this
interim analysis of an ongoing case-control study, our objective is
to determine whether GCL or nerve fiber layer (NFL) thickness may
serve as non-invasive, inexpensive biomarkers to help diagnose AD.
A second objective is to create retinal imaging processing software
useful for the development of novel retinal biomarkers of AD.
Methods: NCT01937221 is enrolling 18 patients with mild cognitive
impairment (MCI)/prodromal AD, 18 patients with mild-to-moderate
AD and 18 cognitively normal, age-matched adults. All behavioral,
imaging, and laboratory data are reviewed by a neurologist and
neuropsychologist for consensus diagnosis regarding assignment
to cognitive group. We carefully exclude any eyes with major
eye diseases and diagnoses that may cause GCL or NFL thinning
(e.g. normal tension glaucoma). Study participants undergo a full
ophthalmic examination, ultra-high-resolution spectral domain
optical coherence tomography (SD-OCT), wide-field fundus color
and autofluorescence photography and stereo disc photography.
Location-specific NFL and GCL thicknesses are measured using the
Duke Optical Coherence Tomography Retinal Analysis Program
(DOCTRAP) software, which has been validated in numerous largescale clinical trials. The extent of peripheral drusen and amyloid
plaques are graded by blinded investigators and quantified using the
DOCTRAP software.
Results: Preliminary analysis showed a statistically significant
difference between neurocognitive status of 14 control subjects and
10 mild-moderate AD subjects but did not reveal a reduction in NFL
or GCL in AD patients. Analysis of all data collected from control,
MCI and AD subjects will be presented.
Conclusions: Careful exclusion of normal tension glaucoma may
account for our preliminary, negative findings, which controvert
a previous group’s finding of thinner NFL among persons with
moderate-to-severe AD. Comparison of retinal images between
normal subjects and subjects with different stages of cognitive
impairment, prodromal and mild-moderate AD, will allow evaluation
of the most promising retinal-based imaging biomarkers for
diagnosing early AD.
Commercial Relationships: Heather E. Whitson, None; Sina
Farsiu, None; Sandra Stinnett, None; Sung Lee, None; Leon
Kwark, None; Guy Potter, None; James Burke, None; Scott W.
Cousins, None; Eleonora Lad, None
Support: Alzheimer’s Association NIRG-13-282202; Duke Institute
for Brain Sciences; NIH P30 EY-005722
Clinical Trial: NCT01937221
Program Number: 390 Poster Board Number: D0126
Presentation Time: 8:30 AM–10:15 AM
Influence of the blood glucose level on FLIO measurements at the
human retina in healthy volunteers
Matthias Klemm1, Dietrich Schweitzer2. 1Biomedical Engineering
& Informatics, Technische Universität Ilmenau, Ilmenau, Germany;
2
Experimental Ophthalmology, University of Jena, Jena, Germany.
Purpose: The detection of metabolic changes in the retina is the
goal of fluorescence lifetime ophthalmoscopy (FLIO). Changes in
the blood sugar level (BSL) may cause short-term alterations in
the cellular metabolism and thus affect FLIO measurements. We
performed an observational clinical study in young healthy volunteers
to learn about the changes in the fluorescence lifetimes during an oral
glucose tolerance test.
Methods: In 10 healthy volunteers (28.9±3.9 years) timeresolved retina auto-fluorescence was measured (scanning laser
ophthalmoscope: 30° of fundus, 34mm resolution; excitation: diode
laser with pico-second pulses, 473nm, 80MHz repetition rate;
detection: spectral channels 500-560nm (ch1) and 560-720nm (ch2),
time-correlated single photon counting method). All subjects had a
crystalline lens. The pupil was not dilated. The BSL was measured
based on blood samples taken from the finger using an Accu-Chek®
Aviva self-monitoring device. Volunteers were instructed to fast for
10–16 hours prior to the study. A baseline measurement of BSL and
FLIO was taken before the volunteers were drinking 300ml solution,
which contained 75g of glucose (Accu-Chek® Dextrose O.G.T.).
Every 15 minutes BSL and FLIO were measured. A modified
3-exponential approach was applied to determine the fluorescence
lifetimes. The ETDRS grid was applied to the FLIO measurements,
its inner ring was used to compare the fluorescence lifetime before
and after glucose intake.
Results: Results are mean ± standard deviation over all volunteers in
ch1. The baseline BSL was 5.3±0.4 mmol/l (SI unit), τ1: 57±5ps, τ2:
307±49ps, τ3: 1086±120ps and the τmean: 148±13ps. The highest
BSL of 8.4±1.1 mmol/l was observed 30min after glucose intake.
The corresponding fluorescence parameters are: τ1: 51±5ps, τ2:
282±41ps, τ3: 1043±103ps and τmean: 140±13ps. The measurements
after 90 minutes resulted in: BSL: 6.3±1.4 mmol/l and τ1: 48±5ps,
τ2: 278±40ps, τ3: 1104±106ps and the τmean: 139±13ps. Results of
ch2 are show even smaller deviations in the fluorescence lifetimes.
Conclusions: The fluorescence lifetime changes during the oral
glucose tolerance test are within the standard deviation. Thus, FLIO
measurement in young healthy volunteers are mostly independent of
short-term changes in the blood sugar level. Volunteers don’t have to
fast to obtain reliable fluorescence lifetimes from the retina.
Commercial Relationships: Matthias Klemm, None; Dietrich
Schweitzer, None
Support: German Federal Ministry of Education and Research
(Grant No. 03IPT605A; http://www.unternehmen-region.de); German
Research Council (DFG HA 2899/19-1; www.dfg.de)
Program Number: 391 Poster Board Number: D0127
Presentation Time: 8:30 AM–10:15 AM
En face imaging of retinal pathology using multicolor confocal
scanning laser ophthalmoscopy
Henry Feng1, Sumit Sharma2, Sanjay G. Asrani2, Sandra Stinnett2,
Prithvi Mruthyunjaya2. 1Rutgers Robert Wood Johnson Medical
School, Piscataway, NJ; 2Duke Eye Center, Durham, NC.
Purpose: Multicolor confocal scanning laser ophthalmoscopy
(cSLO) is a novel imaging modality that utilizes blue, green, and
infrared (IR) lasers to generate a composite multicolor image. Our
study correlates multispectral laser reflectance patterns with findings
on SD-OCT in order to evaluate the ability of multicolor cSLO to
detect sub-, intra-, and epi-retinal pathology.
Methods: Retrospective review of 159 eyes that underwent posterior
pole imaging using multicolor cSLO and SD-OCT on the Heidelberg
Spectralis (Heidelberg Retina Angiograph-Optical Coherence
Tomography, Heidelberg Engineering, Heidelberg, Germany) in a
single session. Multicolor cSLO was acquired using 820nm, 518nm,
and 488nm lasers with a scan angle of 30°, laser power of 25%,
and automatic real time image averaging of 25. Positive percent
agreement (PPA) and negative percent agreement (NPA) were
calculated for each finding on each en face cSLO image using SDOCT as the reference standard.
Results: Multicolor reflectance demonstrated equal or greater PPA
compared to IR, green, and blue reflectance images in the detection
of choroidal lesions (1.00), chorioretinal scars (0.90), peripapillary
atrophy (1.00), drusen (0.77), and pigment epithelial detachment
(0.77). RPE atrophy was most effectively detected on IR reflectance
with a PPA of 0.92. NPA was 0.98 or greater for these subretinal
and choroidal findings on all reflectance images. For intraretinal and
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
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ARVO 2015 Annual Meeting Abstracts
subretinal fluid, multicolor reflectance demonstrated the highest PPA
at 0.49 and 0.54, respectively, while IR reflectance demonstrated the
lowest PPA at 0.08 and 0.25, respectively. Intraretinal and subretinal
hemorrhage was detected most often on multicolor cSLO with NPA
at 0.87 and 0.97, respectively, although PPA could not be calculated
because zero cases were detected on SD-OCT (Figure 1). Epiretinal
membrane was equally detected on green, blue, and multicolor
reflectance with a PPA of 0.94, and least detected on IR reflectance
with a PPA of 0.40 (Figure 2).
Conclusions: Multicolor cSLO, compared to SD-OCT, effectively
visualizes sub- and epi-retinal pathology, choroidal lesions, and
hemorrhages at various fundal depths. This modality may be
clinically useful for detecting en face aberrations in retinal contour
and may be combined with SD-OCT to improve diagnostic accuracy.
Subretinal hemorrhage on SD-OCT and multicolor cSLO
Epiretinal membrane on SD-OCT and multicolor cSLO
Commercial Relationships: Henry Feng, None; Sumit Sharma,
None; Sanjay G. Asrani, Heidelberg Engineering (R); Sandra
Stinnett, None; Prithvi Mruthyunjaya, Allergan, Inc. (C)
Program Number: 392 Poster Board Number: D0128
Presentation Time: 8:30 AM–10:15 AM
An improved method of centrifuge concentration of
triamcinolone acetonide (Triesence) for intravitreal injection
Christy M. Cunningham1, Karl N. Becker2, Jamie Keen3, Neerav
Lamba1, Norbert M. Becker1, 3. 1Ophthalmology, Cook County
Hospital, Chicago, IL; 2University of Illinois College of Medicine,
Chicago, IL; 3Chicago Medical School, RFUMS, North Chicago, IL.
Purpose: A typical intravitreal injection dose of triamcinolone
acetonide injectable suspension (Triesence, Alcon, Ft. Worth, Texas),
an FDA approved preservative-free suspension, is 4.0 mg based on a
0.1 mL injection from a concentration of 40 mg/mL. A prior study has
shown an increased duration of effect with increased concentration
of injectant, and there is a theoretical decrease in complications
with decreased volume of injectant. We describe an experimental
drug preparation method for improved centrifuge concentration of
triamcinolone.
Methods: Four separate trials were conducted, in which 1.0 mL of
triamcinolone acetonide injectable suspension (Triesence) was drawn
from well-agitated vials into 1.0 mL syringes. The plunger was cut at
the point where it protrudes from the syringe at the flanges, the needle
was replaced with a sterile cap, and the syringes were placed into
a balanced Quest Diagnostics Mini E 642E centrifuge with flanges
down for either one or two minutes at 3380 RPM. This centrifuge
allows for complete horizontal positioning of the tubes during
centrifugation. After removal of syringes, separation of supernatant
from medication pellet was noted and volume of each sample was
measured. Using a 30-gauge needle, the supernatant was then ejected,
followed by the concentrated drug pellet.
Results: After two minutes of centrifugation, a constant volume
of approximately 0.12 mL of medication pellet was achieved with
1.0 mL of triamcinolone, which equated to 40 mg of triamcinolone.
This method of centrifugation resulted in a horizontal precipitate/
supernatant interface. This allowed for easy ejection of supernatant
liquid while minimizing disruption of the drug pellet, which could
then be separately ejected from the syringe.
Conclusions: For physicians trying to concentrate triamcinolone
from a 40 mg/mL preparation, centrifugation of triamcinolone using
our methods for intravitreal use may be an effective technique for
administering the drug in higher concentrations. Our method resulted
in an improved technique from a prior study, in that the precipitate/
supernatant interface was horizontal, which allows for estimation of
injected drug dose and easier removal of the supernatant liquid.
Commercial Relationships: Christy M. Cunningham, None;
Karl N. Becker, None; Jamie Keen, None; Neerav Lamba, None;
Norbert M. Becker, None
Program Number: 393 Poster Board Number: D0129
Presentation Time: 8:30 AM–10:15 AM
In Vivo and In vitro Force Testing of a New Guarded Injection
Device
Alexander M. Eaton1, Gabriel M. Gordon1, Dave Booth3, Hussein
Wafapoor1, Robert L. Avery2, Dyson Hickingbotham1. 1Retina Health
Center, Fort Myers, FL; 2California Retina Consultants, Santa
Barbara, CA; 3Invengen, West Milford, NJ.
Purpose: A novel guarded injection device (GID) designed to reduce
the incidence of contamination during an intravitreal injection (IVI)
was recently shown to reduce the time needed for an IVI with a trend
towards making injections more comfortable. This GID has been
redesigned by a major needle manufacturer for more efficient mass
production. We hypothesize that insertion of the 30G GID will not
require more work than insertion of a standard 30G needle.
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Methods: The total work in Newtons (N) for ten 30G GID needles
was compared to that of ten standard 30G needles in freshly
explanted porcine eyes as well as nine of each in synthetic test media
similar to the human sclera. Cross head speeds for the test set were
kept constant at 100 microns per second to provide good needle tip
and cutting edge force resolution and a force profile with 1000 data
points was generated for analysis. Work was compared after insertion
to 25 and 50% of the total needle length.
Results: There was no statistically significance difference in the total
work after insertion of 25% of the total GID needle length compared
to 25% of a standard 30G needle in both the synthetic test media
(137.48 N vs 149.93 N, p=0.74) and the porcine eyes (26.50 N vs
35.15 N, p=0.10). There was a statistically significance difference
in the total work after insertion of 50% of the GID needle length
compared to 50% of a standard 30G needle in both the synthetic test
media (273.69 N vs 196.08 N, p<0.001) and the porcine eyes (216.63
N vs 176.93 N, p=0.04).
Conclusions: Insertion of the needle of the new 30G GID does
not require significant additional work than a standard 30G needle
during insertion of 25% of its length and minimal additional work at
insertions of 50% of its length. Understanding the mechanical forces
surrounding the use of the GID should help physicians optimize use
of the device.
Commercial Relationships: Alexander M. Eaton, I-TECH JV
DEVELOPMENT COMPANY, LLC (I); Gabriel M. Gordon, None;
Dave Booth, I-TECH JV DEVELOPMENT COMPANY, LLC (I);
Hussein Wafapoor, I-Tech JV Development Company, LLC (I);
Robert L. Avery, I-TECH JV DEVELOPMENT COMPANY, LLC
(I); Dyson Hickingbotham, I-TECH JV DEVELOPMENT, LLC (I)
Program Number: 394 Poster Board Number: D0130
Presentation Time: 8:30 AM–10:15 AM
NEW PROTOTYPE OF ULTRASOUND HARMONICS
VITRECTOR (UHV)
HISTOPHATOLOGICAL FINDINGS: First Report
Salvador Pastor1, 2, Richard Bonshek3, Irion Luciane3, Isaac
Zambrano4, Paul Carlin5, Paulo E. Stanga1, 2. 1Manchester Royal
Eye Hospital and Manchester Academic Center for Ophthalmology
and Vision Research, Institute of Human Development, University
of Manchester, Manchester, United Kingdom; 2Manchester Vision
Regeneration (MVR) Lab at NIHR/Wellcome Trust Manchester CRF,
Manchester, United Kingdom; 3Ocular Histopathology, Manchester
Royal Eye Hospital, Manchester, United Kingdom; 4Eye Bank,
Manchester Royal Eye Hospital, Manchester, United Kingdom;
5
Operating Theatre Services, Manchester Royal Eye Hospital,
Manchester, United Kingdom.
Purpose: The purpose of this study was to evaluate the effect of
gauge size, percentage of ultrasound power (US) and aspiration
settings on the structural components of the different retinal layers
and the crystalline lens using a new prototype of ultrasonic harmonics
vitrector (UHV). We compared our results with those obtained with a
currently commercially available guillotine vitrector.
Methods: Vitrectomy surgery (PPV) was performed using the UHV
(Bausch + Lomb, St. Louis, MO, USA) and a currently commercially
available pneumatic guillotine vitrector and 23G needles with both
technologies (Stellaris PC® Vitrectomy system, Bausch + Lomb,
St. Louis, MO, USA). Pre-determined aspiration levels (50, 100,
200, 300, 400, 500 and 600 mmHg) for both systems, cut rates
for guillotine cutter (3000 and 5000 cuts per minute (CPM)) and
percentages of US power for the UHV needles (10, 20, 30, 40, 50 %)
were used in 14 consecutive cadaveric porcine eyes obtained within
12-24hs of slaughter. Six (6) closed PPV and eight (8) open-sky
PPV (after removal of the anterior segment at the pars plana) were
performed. The UHV and guillotine vitrector were held at 3 to 5 mm
in front of the macula. The effects of US and guillotine technology
were also tested in 4 cadaveric porcine crystalline lenses submerged
in Balanced Salt Solution (BSS®) and by touching the posterior
capsule with both the UH and the guillotine vitrector. The retinal
specimens and the crystalline lens were evaluated using microscopy
and histochemical methods.
Results: There were no macroscopic retinal defects associated with
the use of neither the UH nor the guillotine vitrectors at 10, 20, 30,
40, 50% US and 3000 and 5000 cut rates. Neither needle gauge nor
wall thickness of the UHV affected the molecular structure of the
retina layers. Disruption of the posterior capsule and structural lens
defects were found after two min contact with guillotine vitrectors.
However, small disruptions of the posterior capsule or structural lens
defects were found after two min contact with the UHV.
Conclusions: The retinal structure does not seem to be affected by
gauge of the UHV needle for similar ports and vacuums. The use of
an UHV in the middle of the vitreous cavity and at 3 to 5 mm from
the retina and at the described US % of power may be safe. UHV is
a promising new alternative to the currently commercially available
guillotine-based technology for vitrectomy systems.
Commercial Relationships: Salvador Pastor, Bausch & Lomb (F);
Richard Bonshek, None; Irion Luciane, None; Isaac Zambrano,
None; Paul Carlin, None; Paulo E. Stanga, Bausch & Lomb (C),
Bausch & Lomb (F), Bausch & Lomb (P), Bausch & Lomb (R)
Program Number: 395 Poster Board Number: D0131
Presentation Time: 8:30 AM–10:15 AM
Metabolomics in AMD – identifying metabolic pathways and
biomarkers for improved prediction and prevention
Eiko de Jong1, Yara Lechanteur1, Nicolas Schauer2, Tina Schick3,
Sascha Fauser3, Carel C. Hoyng1, Anneke I. Den Hollander1.
1
Ophthalmology, Radboudumc, Nijmegen, Netherlands;
2
Metabolomic Discoveries, Berlin, Germany; 3University Hospital of
Cologne, Cologne, Germany.
Purpose: A metabolomics study was performed on serum samples to
discover novel biomarkers for (dry) AMD and to uncover clinically
relevant metabolic pathways.
Methods: 132 subjects were categorized into four groups based on
their genetic risk profile (CFH and ARMS2 alleles) and their disease
status: 1) healthy controls with low genetic risk, 2) AMD patients
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
with low genetic risk, 3) AMD patients with high genetic risk, 4)
healthy controls with high genetic risk. Groups were matched for
age, sex, smoking, BMI and levels of activated complement. For all
subjects, serum was collected and analyzed using a metabolomics
approach, employing the parallel application of gas chromatographymass spectrometry and liquid chromatography- mass spectrometry.
Results: On average, approximately 1200 metabolites per subject
were detected. In each of the groups, on average 5 unique metabolites
were detected that were not present in any other group. By comparing
groups 1 and 2, we identified 17 metabolites significantly associated
with AMD, independent of CFH/ARMS2 genotypes. By comparing
high risk groups 3 and 4, we identified 32 differentially expressed
metabolites. Within the groups, several associated metabolites
showed a high degree of correlation with each other. Random forest
analysis could predict the groups based on their composition with 5879% accuracy. Not all metabolites could be annotated directly.
Conclusions: By comparing groups 1 and 2, potentially novel
metabolic networks underlying AMD have been identified. By
comparing groups 3 and 4, metabolites have been identified that
potentially could protect against AMD in the context of high
genetic risk. Because there is a high degree of correlation between
metabolites in each group, these results are likely to represent
meaningful metabolic networks rather than false positives. Further
characterization of metabolites that are not yet annotated is required
to determine the exact nature of these networks.
Commercial Relationships: Eiko de Jong, Bayer (F); Yara
Lechanteur, Bayer (F); Nicolas Schauer, Metabolomic Discoveries
(E); Tina Schick, None; Sascha Fauser, None; Carel C. Hoyng,
Bayer (F); Anneke I. Den Hollander, Bayer (F)
Support: Global Ophthalmology Awards Program, Bayer
Program Number: 396 Poster Board Number: D0132
Presentation Time: 8:30 AM–10:15 AM
Reduced Vitreous Traction Associated with 23-Gauge Dual
Pneumatic High Speed Cutters
Dina Joy K. Abulon, Tingting Wang. Medical Affairs, Alcon Labs,
Lake Forest, CA.
Purpose: To quantify and compare traction forces between 2500
cuts per minute (cpm), 5000cpm, and 7500cpm with dual pneumatic
vitrectomy probes in an ex- situ porcine eye model.
Methods: A mechanical force measurement system was developed to
quantify traction forces applied to the vitreous by a vitrectomy cutter.
Traction measurements were performed in an ex-situ test set up using
vitreous harvested from fresh porcine eyes. To minimize variability
of vitreous tissue, traction was measured with the same vitreous
mass by alternating between three cut rates (2500cpm, 5000cpm, and
7500cpm) and maintaining the same aspiration level (300mmHg),
under 50/50 duty cycle. A total of 6 UltraVit® 23-Gauge (Ga)
vitrectomy cutters and 30 porcine eyes (N=30) were tested. Peak
traction forces and mean traction forces for each cut rate were
averaged from each test. Significant differences in traction forces
between cut rates were determined using a paired t-test and analysis
of variance with p<0.05 as the statistical significance level.
Results: The 2500cpm cut rates created the highest vitreous traction
peak force of 1.17±0.36mN, and a mean force of 0.47±0.11mN.
5000cpm created less traction with a peak force of 0.86±0.27mN, and
a mean force of 0.46±0.10mN. 7500cpm generated the least traction
among the three with a peak force of 0.66±0.21mN, and a mean force
of 0.39±0.09mN. Peak and mean traction forces of 7500cpm were
significantly smaller than 5000cpm (-0.20mN, p<0.0001; -0.06mN,
p<0.001), and 2500cpm (-0.51mN, p<0.0001; -0.08mN, p<0.0001).
When comparing the relative reduction in force, the peak force of
7500cpm was 74% and 54% of 5000cpm and 2500cpm respectively
(p<0.001); the mean force of 7500cpm was 85% and 80% of
5000cpm and 2500cpm(p<0.001) respectively. Pure vitreous flow
tests revealed increased vitreous flow at 7500cpm as compared to
5000cpm and 2500cpm.
Conclusions: Compared to lower cut rates, the high speed dual
pneumatic probes operating at 7500cpm may improve vitrectomy
surgery by generating less vitreous traction and more efficient
vitreous removal.
Commercial Relationships: Dina Joy K. Abulon, Alcon Research,
Ltd. (E); Tingting Wang, Alcon Research, Ltd. (E)
Support: n/a
Program Number: 397 Poster Board Number: D0133
Presentation Time: 8:30 AM–10:15 AM
Sealing retinal breaks with FocalSeal® in experimental
rhegmatogenous retinal detachment in rabbit eyes
Sujin Hoshi1, Fumiki Okamoto1, Yoshimi Sugiura1, Genichiro
Kishino1, Tomoya Murakami1, Mikki Arai2, 4, Tatsuo Hirose3, 4, Tetsuro
Oshika1. 1Department of Ophthalmology, Faculty of Medicine,
University of Tsukuba, Tsukuba, Japan; 2Arai Eye Clinic, Fukuoka,
Japan; 3The Schepens Eye Research Institute, Harvard Medical
School, Boston, MA; 4Boston Eye Group, Brookline, MA.
Purpose: FocalSeal® is an absorbable polyethylene glycol-based
synthetic hydrogel sealant. This liquid is polymerized under
visible xenon illumination, and forms clear, flexible, and firmly
adherent hydrogel. This study was conducted to examine the
effect of FocalSeal® for sealing retinal breaks in experimental
rhegmatogenous retinal detachment in rabbit eyes.
Methods: Experimental retinal detachment with a break was made
during a 25-gauge vitrectomy in six rabbit eyes. After performing
fluid-air exchange, FocalSeal® was applied to entirely cover
iatrogenic retinal breaks, and was polymerized with a 60-second
application of xenon light in three rabbit eyes (FS group). Air-fluid
exchange was then performed, and operations were finished without
intraocular tamponade. In other three eyes, the same procedures
were performed without FocalSeal® application (control group).
Funduscopic examination and optical coherence tomography (OCT)
was carried out in both groups at 1 and 7 days, and 1 and 3 months
postoperatively.
Results: The funduscopic examination showed that the retina was
reattached in all eyes of the FS group. Meanwhile all three eyes of the
control group resulted in proliferative vitreoretinopathy. FocalSeal®
was observed on the retinal breaks at 1 and 7 days, but was not
observed after 1 month postoperatively.
Conclusions: In our study, FocalSeal® successfully sealed
retinal breaks, and repaired experimental rhegmatogenous retinal
detachment without intraocular tamponade. FocalSeal® was found
to be beneficial to seal retinal breaks during and after vitrectomy for
rhegmatogenous retinal detachment.
Commercial Relationships: Sujin Hoshi, None; Fumiki Okamoto,
None; Yoshimi Sugiura, None; Genichiro Kishino, None; Tomoya
Murakami, None; Mikki Arai, None; Tatsuo Hirose, None;
Tetsuro Oshika, None
Support: KAKENHI 26462631
Program Number: 398 Poster Board Number: D0134
Presentation Time: 8:30 AM–10:15 AM
A Role of Phosphatidylinoitol 5-Phosphate 4-Kinases in
Proliferative Vitreoretinopathy
Hetian Lei, Andrius Kazlauskas. Schepens Eye Research Institute/
MEEI/Harvard Medical School, Boston, MA.
Purpose: Proliferative vitreoretinopathy (PVR) is the major cause
of failure in retinal detachment (RD) surgery. The p53 codon
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
polymorphism (rs1042522) is associated with PVR, and the T309G
in the MDM2 gene, a key negative regulator of p53, is also associated
with a higher risk of developing PVR in patients undergoing a
RD surgery. In addition, we recently found that vitreous suppress
expression of p53 by engaging platelet-derived growth receptor
(PDGFR)a/PI3K/Akt pathway. Intriguingly, reducing the level
of p53 is required for experimental PVR. However, molecularly
suppressing p53 does not fully phenocopy stimulation with vitreous.
Thus it appears that vitreous does more than suppress p53 to promote
cellular responses that are associated with PVR. The viability of
tumor cells in which p53 is suppressed depends on elevation of
phosphatidylinositol 5-phosphate 4-kinases (PI5P4Ks). Thus we
hypothesized that vitreous-mediated elevation of PI5P4K(s) was an
essential event in PVR pathogenesis.
Methods: Vitreous-induced expression of PI5P4Ks in ARPE19,
ARPE19 with PDGFRa overexpression (ARPE19a), and ARPE19
with PDGFRa silence (ARPE19KD) was examined using Western
blot. Knockdown of p53 or PDGFRa in ARPE19 cells was achieved
by shRNA. Overexpression of PI5P4Ks or PDGFRa in ARPE19
cells was accomplished by using a retrovirus-based approach. The
resulting cells were subjected to proliferation, survival and collagen
gel contraction assays.
Results: Normal rabbit vitreous, which contains a variety of growth
factors but not PDGFs, induced PI5P4Ks expression in a PDGFRadependent manner. In addition, we found that knockdown of p53
caused collagen gel contraction, but the extent of contraction was less
than what occurred in response to vitreous. The extent of contraction
in p53 knockdown cells increased to the level attained in response
to vitreous by overexpressing PI5P4Ks. Finally, overexpression of
PI5P4Ks in the p53 knockdown cells promoted additional PVRassociated cellular response including proliferation and survival.
These data indicate that experimental PVR may require not only a
knockdown in p53, but also an increase in PI5P4Ks.
Conclusions: These findings suggest that inhibition of PI5P4Ks and
activation of p53 are novel approaches to prevent PVR.
Commercial Relationships: Hetian Lei, None; Andrius
Kazlauskas, None
Program Number: 399 Poster Board Number: D0135
Presentation Time: 8:30 AM–10:15 AM
Phenotypic variability and utility of red-free and wide-field
autofluorescence imaging in incontinentia pigmenti patients
Katherine E. Talcott, Shizuo Mukai. Ophthalmology, Massachusetts
Eye and Ear Infirmary, Boston, MA.
Purpose: Incontinentia pigmenti (IP) is a rare syndrome with ocular
manifestations including peripheral retinal telangiectasias, avascular
zones, and neovascularization (NV). Fluorescein angiography (FA) is
the current gold standard in diagnosing and guiding treatment. This
study assesses retinal phenotypic variability in IP and evaluates the
utility of red-free and wide-field autofluorescence (AF) imaging as an
adjuvant to FA.
Methods: Consecutive, retrospective case series of IP patients with
retinal involvement seen between 2007 and 2014.
Results: Seven IP patients (median age 5 months at initial exam)
were followed longitudinally. All patients had skin changes while
neurologic manifestations (29%), family history (29%), and known
NEMO mutations (14%) were less common. 13 eyes (one eye had
total retinal detachment) underwent 30 imaging sessions, including
28 FA. All eyes had peripheral avascular retinal zones and 12 eyes
had vessel tortuosity and retinal telangiectasias. 6 eyes had active
NV on FA that was treated with laser (100%) and intravitreal
bevacizumab (17%). 2 eyes with NV also had development
abnormalities—one had residual anterior tunica vasculosa lentis,
optic atrophy, and hypoplastic fovea with extensive retinal
avascularity and posterior NV stabilized with serial bevacizumab
injections and laser while another had optic nerve misdevelopment
with extensive NV treated with laser. 17 FA were accompanied by
RetCam red-free imaging and 4 by Optos AF imaging. Peripheral
avascular zones, vessel tortuosity, and telangiectasias were detectable
on all images. However, vitreous hemorrhage or NV was only seen in
a minority (11% of red-free images, 50% of AF images).
Conclusions: Among IP patients with known retinal involvement,
there is a high incidence of NV that can be managed with serial
FAs, laser and intravitreal bevacizumab. While FA is superior for
surveillance of active NV, obtaining red-free or AF images from
retinal periphery may identify avascular zones, vessel tortuosity and
telangiectasias without the need for fluorescein administration in
young patients.
Commercial Relationships: Katherine E. Talcott, None; Shizuo
Mukai, None
Program Number: 400 Poster Board Number: D0136
Presentation Time: 8:30 AM–10:15 AM
Robot-assisted ab-externo drug delivery
Thijs H. Meenink2, 1, Maarten Beelen2, Gerrit Naus2, Sicco H.
Popma3, Maarten Steinbuch1, Marc D. de Smet4, 2. 1Mechanical
Engineering, Technische Universiteit Eindhoven, Eindhoven,
Netherlands; 2PRECEYES Medical Robotics, Eindhoven,
Netherlands; 3Janssen R&D, Padnor, PA; 4Retina and Inflammation,
MIOS, Lausanne, Switzerland.
Purpose: Drugs can be delivered to the sub-macular region via abexterno delivery using a microcatheter [de Smet 2012]. In a previous
study, it was demonstrated that robot assistance can reduce the risk
of retinal penetrations during the required choroidotomy phase [de
Smet 2014]. The purpose of this study is to demonstrate that robot
assistance for the catheter insertion phase enables aiming of the
catheter for targeted delivery and a high level of control in pausing
and resuming procedural steps, which is difficult to achieve manually.
Methods: Equatorial sclerotomies down to the choroidal vessels
are performed in in-vivo porcine eyes. The choroidotomy is made,
while injecting a visco-elastic fluid to create the sub-retinal bleb.
Finally the catheter is inserted and fed to the submacular region.
The choroidotomy is assisted by the PRECEYES Surgical System
[Meenink 2012]. For catheter insertion, a novel approach using a
guiding instrument is devised, allowing for a controlled, automated
insertion. Using robot assistance, this instrument is inserted into
the choroidotomy, and aimed towards the macula. The instrument
is oriented such that its tip is near tangential to the choroid. The
catheter is introduced through this guiding instrument, and fed to the
submacular region, under visual inspection via an endoscope.
Results: In the in-vivo animal tests, a total of 22 catheter insertions
were performed. The incidence of retinal perforations during catheter
insertion converged to 18%, which is comparable to manual surgery
[de Smet 2012]. The study shows that robot assistance provides a
high level of control during the procedure. This gives the possibility
to anticipate complications when seen with the endoscope and
corrective actions can be taken. This allows for example to detect
and to resolve high subretinal bleb tension prior to continuing the
catheter insertion, which is very difficult manually. Furthermore,
robot assistance allows re-aiming of the catheter for targeted
delivery, which cannot be safely performed manually. Automation of
procedural steps reduced the average catheter insertion time (100s),
compared to manual surgery.
Conclusions: Using robot assistance, a high level of control
in pausing and resuming steps in the procedure is achieved. In
combination with a guiding instrument, robot assistance also allows
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
for targeted delivery of the drug by re-aiming of the catheter. This is
especially useful if injection into a critical direction is required.
Commercial Relationships: Thijs H. Meenink, PRECEYES (E);
Maarten Beelen, PRECEYES (E); Gerrit Naus, PRECEYES (E);
Sicco H. Popma, None; Maarten Steinbuch, None; Marc D. de
Smet, PRECEYES (C)
Program Number: 401 Poster Board Number: D0137
Presentation Time: 8:30 AM–10:15 AM
Ultra-wide field fundus fluorescein angiography in
rhegmatogenous retinal detachment
Bhushan R. Wadekar, Rohan Chawla, Koushik Tripathy, Raj Pal, Yog
Raj Sharma, Harsh Inder Singh, Babulal Kumawat, Ravi Bypareddy,
Pradeep Venkatesh. OPHTHALMOLOGY, ALL INDIA INSTITUTE
OF MEDICAL SCIENCES, Aurangabad, India.
Purpose: Ultra-wide field fundus (UWF) imaging allows assessment
of 2000 fundus thereby facilitating analysis of peripheral retina.
We sought to assess UWF fluorescein angiographic (UWFA)
characteristics of rhegmatogenous retinal detachments (RRD), a
predominantly peripheral retinal pathology.
Methods: We enrolled 30 consecutive eyes clinically diagnosed with
RRD and studied their angiographic features using UWFA (200Tx™,
Optos PLC, Dunfermline Scotland, United Kingdom).
Results: The age range of patients was 8 to 64 years with 17
(56.67%) having RRD duration of less than 8 days and 13 (43.33%)
with history of more than 8 days. 14 (46.67 %) patients had previous
history cataract surgery, 6 (20%) were myopes, 2 (6.67%) had giant
retinal tears, 3 (10%) were post-traumatic, 2 (6.67%) had iridofundal
coloboma and remaining 3 (10%) had miscellaneous associations.
Salient angiographic findings included paravascular changes such
as venous dilatation (100%), paravascular leak (100%), peripheral
capillary non-perfusion (CNP) (100%), vascular tortuosity (73%),
minimal disc leak (46.67%) and neovascularisation elsewhere
(26.7%). The edge of break could be identified by multiple small
hyperfluorescent knob-like points along it (“street-lamps along a
city-road appearance”). Additionally sub-retinal proliferative vitreoretinopathy (PVR) changes (23.33%) were noted as blocked patterns
of hypo-fluorescence.
Conclusions: UWF technology efficiently images peripheral retina.
Angiographic features such as venous dilatation, paravascular leak,
peripheral CNP are seen in almost all RRD cases. Vascular tortuosity,
neovascularisation, minimal disc leak, patterned break and subretinal PVR, though relatively inconsistent are other associated
important features of RRD. These features help in understanding
the pathophysiology of detached retina and would provide valuable
inputs while analyzing and managing exudative retinal detachments.
Ultra-wide field colour fundus photograph patient no 14. (age 14
years, duration 16 days) showing total rhegmatogenous retinal
detachment with a large temporal break.
Ultra-wide field fundus fluorescein angiographic image at 3 minutes
in patient no.14 demonstrating vascular changes (venous dilatation,
vascular tortuosity, paravascular leak, peripheral capillary nonperfusion areas). The retinal break is highlighted by knob-like
vascular hyperfluorescent ends: “street-lamps along a city-road
appearance”.
Commercial Relationships: Bhushan R. Wadekar, None; Rohan
Chawla, None; Koushik Tripathy, None; Raj Pal, None; Yog Raj
Sharma, None; Harsh Inder Singh, None; Babulal Kumawat,
None; Ravi Bypareddy, None; Pradeep Venkatesh, None
Program Number: 402 Poster Board Number: D0138
Presentation Time: 8:30 AM–10:15 AM
Safe intravitreal injection in eyes with retinoblastomadevelopment of a model
Gopal Lingam1, Valencia H. Foo2. 1Vitreoretinal, National University
Hospital, Singapore, Singapore; 2National University of Singapore,
Singapore, Singapore.
Purpose: To develop a model of safe intravitreal injection that can be
adopted for eyes with retinoblastoma
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
Methods: This experiment was conducted on pig eyes obtained from
slaughter house. The eye was stabilised on a holder and the screw
tightened till the intra ocular pressure was in the range of 12-15mm/
hg as measured by tonopen. A 30 gauge needle entry track was made
1mm internal to the limbus till the anterior chamber was entered.
The needle was withdrawn. A 36 gauge needle atttached to a syringe
with fluorescein stained saline is threaded through this track and
then antero posteriorly through the iris root and zonules into the
vitreous cavity. 0.1 ml of the dye stained saline was injected and
the needle was withdrawn. Flurotron (ocumetrics, Mountain view,
CA) was used to measure fluorescence in the anterior chamber. As a
reference fluorescein was applied on corneal surface after epithelial
debridement. The cornea and the lens were excised to visibly confirm
the presence of significant fluorescein in the vitreous gel.
Results: No fluorescence was observed in the anterior chamber on
measurement with Fluorotron. The vitreous was found to be intensely
stained with fluorescein on direct examination.
Conclusions: 36 gauge needle entry through the iris root and zonules
does not permit intra vitreous contents to migrate into the anterior
chamber. A 2 step approach of entering anterior chamber through
the relatively tough cornea with 30 gauge needle and the subsequent
entry through relatively soft iris and zonules with a finer 36 gauge
needle, can potentially prevent any exit of tumor cells extra ocularly
in eyes with retinoblastoma.
Fluorotron measurement of fluorescence reveals high spike from
cornea (reference) and absence of fluorescence from anterior chamber
Commercial Relationships: Gopal Lingam, None; Valencia H.
Foo, None
Program Number: 403 Poster Board Number: D0139
Presentation Time: 8:30 AM–10:15 AM
Design of an intuitive motion controller for robot-assisted
vitreoretinal surgery
Marc D. de Smet1, 2, Gerrit Naus2, Thijs H. Meenink2, Maarten
Beelen2, Nicky de Jonge2, Ron Hendrix3, Maarten Steinbuch3.
1
Ophthalmology, MIOS, Lausanne, Switzerland; 2PRECEYES,
Eindhoven, Netherlands; 3Mechanical Engineering, Eindhoven
University of Technology, Eindhoven, Netherlands.
Purpose: In vitreoretinal surgery, limited manual precision curtails
development of new, precision-enabled surgeries. Robot assistance
can support surgeons in performing existing vitreoretinal procedures
with higher reproducibility and enable new procedures.
Intuitive control of any robotic system is an important prerequisite.
An intuitive motion controller allowing the appropriate transfer of
information and feedback between a surgeon and a robotic assistant
are assessed for vitreoretinal surgical applications.
Methods: The motion controller mimics the instrument by: (1)
having a pen like grip held at the tip as if manipulating the tip of an
intraocular instrument;
(2) having a remote center of motion comparable to the instrument
usage, resulting in control in ‘eye coordinates’ as opposed to standard
control in cartesian coordinates; (3)being able to automatically mirror
motion to the same degree as instruments entering through the pars
plana.
Position encoders measure the motion input, from which control
algorithms determine the motion output to the instrument
manipulator, These algorithms further, enable tremor filtering, motion
scaling and advanced support in e.g. semi automated motion patterns
such as piercing motions.
The motion controller is positioned within close reach of the
surgical area, enabling hybrid control, i.e., using one hand to hold
a manual instrument and the other to control the robot assisted
instrument, while using the conventional surgical setting including
the microscope. Coupling of manipulator and motion controller is
insured by pressing a button on the side of the controller.
Results: 13 surgeons have evaluated the motion controller. The
kinematic design and “eye coordinates” were seen as intuitive.
Hybrid surgery, with one hand using the motion controller and the
other a light pipe was experienced as inherently feasible. Being able
to couple or uncouple the motion controller from the manipulator,
or the ability to pause a task by releasing the actuating button was
considered most valuable.
Conclusions: Mimicking the instrument motion by using ‘eye
coordinates’ was seen as intuitive and desireable. Use in a mixed
setting (robotic assistance) was easily integrated in existing
procedures and with little interference during the non robotic portion
of the surgery. Access to the surgical field was not hindered. Further
development will focus on optimizing the gripper ergonomy.
Commercial Relationships: Marc D. de Smet, PRECEYES (C);
Gerrit Naus, Preceyes (E); Thijs H. Meenink, Preceyes (E);
Maarten Beelen, Preceyes (E); Nicky de Jonge, Preceyes (E); Ron
Hendrix, None; Maarten Steinbuch, Preceyes (C), Preceyes (P)
Program Number: 404 Poster Board Number: D0140
Presentation Time: 8:30 AM–10:15 AM
Genetic association of polymorphisms in the p53 and LTA genes
with proliferative vitreoretinopathy after retinal detachment
surgery in Mexican population
Natalia P. Quiroz Casian1, José L. Rodríguez3, Salvador LopezRubio3, Antonio Miranda Duarte4, Juan C. Zenteno1, 2. 1Department
of Genetics-Research Unit, Institute of Ophthalmology “Conde de
Valenciana” Mexico City, Mexico City, Mexico; 2Biochemistry,
Faculty of Medicine UNAM, Mexico City, Mexico; 3Retinal
Department, Institute of Ophthalmology “Conde de Valenciana”
Mexico City, Mexico City, Mexico; 4Department of Genetics,
Instituto Nacional de Rehabilitación (INR), Mexico City, Mexico.
Purpose: Proliferative vitreoretinopathy (PVR) is a leading blinding,
multifactorial disease, which is often associated with ocular trauma,
rhegmatogenous retinal detachment and diabetic retinopathy and
remains as the major cause of failure after retinal detachment (RD)
surgery. Recent studies have suggested that polymorphisms in genes
related to inflammatory pathways increase the risk of developing
PVR. The aim of this investigation was to perform a case-control
study to determine the possible association between PVR and
polymorphisms in the p53 and lymphotoxin-alpha (LTA) genes in
Mexican patients.
Methods: One hundred and eighty two unrelated Mexican subjects
were enrolled in the study, including 82 patients with PVR after
RD, and 100 ethnically matched patients with a history of RD
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].
ARVO 2015 Annual Meeting Abstracts
without PVR. Genomic DNA was isolated from blood leukocytes in
each subject and genotyping of the non-synonymous coding SNPs
rs1042522 (p53 gene, codon 72) and rs2229094 (LTA gene, codon
13) was performed using polymerase chain reaction amplification
and direct nucleotide sequencing. Allele frequencies, genotype
frequencies, and Hardy-Weinberg equilibrium were assessed with the
HaploView software.
Results: The C allele of rs1042522 in the p53 gene was found to be
more frequent in PVR patients compared to non-PVR controls (odds
ratio [95% confidence intervals] = 1.3 [0.8-2.0]; p = 0.3). Specifically,
the homozygous CC genotype of this SNP was found to be strongly
associated with the development of PVR (odds ratio [95% confidence
intervals] = 2.2 [0.7-7.6.5]; p = 0.1). In addition, in patients with
PVR, the homozygous CC genotype of LTA rs2229094 was found in
higher frequency compared to non-PVR subjects (odds ratio [95%
confidence intervals] = 1.2 [0-96.9], p= 0.9).
Conclusions: This is the first study performed in Latin American
populations associating specific gene polymorphism and PVR after
RD surgery. Our results replicate those observed in Caucasian ethnic
groups and confirm that the C allele of p53 SNP rs1042522 is a
significant genetic risk factor for PVR after RD surgery. Subjects
carrying the homozygous genotype CC have a 2.2 fold risk to
develop this complication. Further studies are needed to evaluate the
role of this polymorphism in the development of PVR in additional
populations.
Commercial Relationships: Natalia P. Quiroz Casian, None;
José L. Rodríguez, None; Salvador Lopez-Rubio, None; Antonio
Miranda Duarte, None; Juan C. Zenteno, None
©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission
to reproduce any abstract, contact the ARVO Office at [email protected].