Download ARVO 2016 Annual Meeting Abstracts 374 Ocular Surface Health

ARVO 2016 Annual Meeting Abstracts
374 Ocular Surface Health and Disease
Tuesday, May 03, 2016 3:45 PM–5:30 PM
Exhibit/Poster Hall Poster Session
Program #/Board # Range: 3853–3896/A0001–A0044
Organizing Section: Cornea
Program Number: 3853 Poster Board Number: A0001
Presentation Time: 3:45 PM–5:30 PM
APR-246/PRIMA-1Met inhibits and reverses squamous metaplasia
in human conjunctival epithelium
Cheng Li, Wei Li, ZUGUO LIU, JIng Li. Ophthalmology and Visual
Sciences, Eye Inst & Affiliated Xiamen Eye Ctr, Xiamen, China.
Purpose: Squamous metaplasia is a common pathological condition
in ocular surface diseasesfor which there is no therapeutic medication
in clinic. In this study, we investigated the effect of a small
molecule APR-246/PRIMA-1Met on squamous metaplasia in human
conjunctival epithelium.
Methods: Human conjunctival explants were cultured for up to
12 days under airlift conditions. Epithelial cell differentiation
and proliferation were assessed based on K10, K14, K19, Pax6,
MUC5AC, and p63 immunostaining patterns. β-catenin and TCF-4
immunofluorescent staining and real-time PCR characterized Wnt
signaling pathway involvement. Pterygium clinical samples were
cultured under airlift conditions with or without APR-246 for 4 days.
p63, K10, β-catenin and TCF-4 expression in pterygial epithelium
was determined by immunofluorescent staining and real-time PCR.
Results: Airlifting conjunctival explants resulted in increased
stratification and intrastromal epithelial invagination. Such pathology
was accompanied by increases in K10, K14 and p63 expression
whereas K19 and Pax6 levels declined compared to those in freshly
isolated tissue. On the other hand, APR-246 reversed all of these
declines in K10, K14, and p63 expression. Furthermore, K19 and
Pax6 increased along with rises in goblet cell density. These effects
of APR-246 were accompanied by near restoration of normal
conjunctival epithelial histology. APR-246 also reversed squamous
metaplasia in pterygial epithelium that had developed after 4 days ex
vivo culture.
Conclusions: Reductions in squamous metaplasia induced by APR246 suggest it may provide a novel therapeutic approach in different
squamous metaplasia associated ocular surface diseases.
Commercial Relationships: Cheng Li; Wei Li, None;
ZUGUO LIU, None; JIng Li, None
Program Number: 3854 Poster Board Number: A0002
Presentation Time: 3:45 PM–5:30 PM
The Effect of Betadine on Vision
Caren Oquindo1, William H. Ridder1, Kavita Dhamdhere2,
James A. Burke2. 1Southern California College of Optometry,
Marshall B. Ketchum University, Fullerton, CA; 2Allergan, Inc.,
Irvine, CA.
Purpose: Betadine is instilled topically on the eye prior to ocular
surgical procedures to decrease the risk of endophthalmitis. Its effects
on functional vision and the cornea have not been determined. The
purpose of this study was to determine the effects of 5% betadine on
vision function, corneal integrity and subjective complaints.
Methods: Twenty subjects were chosen to participate in this study
(Ten young: 25.8 +/- 2.94; and ten old: 58.2 +/- 5.59). LogMAR
acuity, contrast sensitivity, corneal fluorescein staining, and the
Schein dry eye questionnaire were measured before and after 60 µl
of 5% betadine was applied to one eye, randomly chosen, (baseline,
5, 30, and 60 minutes and 4 and 24 hours post-application). Contrast
sensitivity at 14 cpd was determined with a spatial two-alternative,
forced choice procedure (BeethovenTM software). The NEI grid was
used to grade corneal staining. Subjective complaints were monitored
using the Schein dry eye questionnaire.
Results: The data were analyzed with an ANOVA (linear mixedeffects model). The logMAR acuity was significantly reduced
from baseline at the 30 and 60 minute visits (all p values < 0.05)
and contrast sensitivity was reduced from baseline at 5, 30, and 60
minutes after betadine application (all p values < 0.0001). Total
corneal staining, maximum NEI sector staining, and the Schein dry
eye questionnaire were significantly different from baseline at every
visit (all p values < 0.05).
Conclusions: There was an increase in corneal staining and
subjective complaints that lasted 24 hours and a decrease in
functional vision that lasted 1 hour after 5% betadine instillation.
The central corneal staining cleared more rapidly than the peripheral
cornea; therefore, visual acuity recovered more quickly than overall
corneal staining and subjective complaints. This may have resulted
from pooling of Betadine at the limbus and lid margins. 5% Betadine
application significantly affects corneal integrity which decreases
functional vision and increases subjective complaints.
Commercial Relationships: Caren Oquindo, Allergan, Inc.
(F); William H. Ridder, Allergan, Inc. (F); Kavita Dhamdhere,
Allergan, Inc.; James A. Burke, Allergan, Inc.
Support: Allergan Inc
Program Number: 3855 Poster Board Number: A0003
Presentation Time: 3:45 PM–5:30 PM
The Effect of Betadine on Vision and on Cornea in Rabbits
Kavita Dhamdhere, Alexandra S. Almazan, Michael Engles,
James A. Burke. Allergan Inc, Irvine, CA.
Purpose: Betadine is instilled topically on the eye prior to most
ocular surgical procedures to decrease the risk of endophthalmitis.
Its effects on vision and the cornea have not been determined. The
purpose of this study is to evaluate the effects of Betadine® 5%
sterile ophthalmic prep solution (Alcon, Fort Worth, TX) on vision
(function) and corneal integrity (structure) in rabbits.
Methods: Twenty-four female Dutch-Belted rabbits with normal
vision were included in this study (Mean age: 2.5 ± 0.94 yrs), divided
in two groups: vision group (n=18) and structure group (n=6). The
study eye in each group was randomized and was treated with a drop
of tetracaine hydrochloride ophthalmic solution USP 0.5% (Bausch
and Lomb, Tampa, FL) for topical anesthesia, then with 50 μl of
Betadine. Betadine was washed out with sterile saline after 2 mins
of exposure. Monocular visual acuities (VA) and corneal fluorescein
staining (CFS) were measured at baseline and at 1, 4 and 24 hours
post treatment in conscious rabbits. OptoMotor™ system (Prusky
et al, 2004; Douglas et al, 2005) was used to collect VA (spatial
thresholds) and the total number of punctate stains were used to grade
CFS. The effect of Betadine on vision and cornea was evaluated as
a function of change from the baseline. The data is presented in the
form of mean ± SD.
Results: The data were analyzed with an ANOVA for multiple
comparisons. Mean baseline VA and CFS was respectively 0.789 ±
0.017 cpd and 3 ± 0.8 total punctate stains in the study eye. The VA
was significantly reduced post betadine dosing at each follow up time
point (0.470 ± 0.17, 0.511 ± 0.17 and 0.609 ± 0.19 respectively at 1,
4 and 24 hrs; p < 0.0001). Total CFS was significantly increased from
baseline at each follow up time point (80 ± 4.4, 72 ± 10 and 43 ± 5.1
respectively at 1, 4 and 24 hrs; p < 0.0001). Two thirds of the animals
had conjunctival hyperemia and/or blepharospasm up to 24 hrs post
Betadine application.
Conclusions: There was a decrease in VA and an increase in corneal
staining that lasted for 24 hours after 5% betadine instillation. The
central corneal staining cleared more rapidly than the peripheral
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
cornea. This may have resulted from pooling of Betadine at the
limbus and lid margins.
Commercial Relationships: Kavita Dhamdhere;
Alexandra S. Almazan, None; Michael Engles, None;
James A. Burke, None
Program Number: 3856 Poster Board Number: A0004
Presentation Time: 3:45 PM–5:30 PM
Hoxc8 misexpression prevents embryonic eyelid fusion and
transforms cornea and conjunctiva into keratinized skin
Lara S. Carroll. Moran Eye Center, University of Utah, Salt Lake
City, UT.
Purpose: Hox gene expression is excluded from the developing
eye, although several members of this gene family are expressed in
fetal and adult epidermis. We found that misexpression of Hoxc8
throughout mouse embryonic ectoderm results in pups born with
their eyes open (eyes open at birth-EOB), and the cornea and
conjunctiva transformed into a skin-like, keratinized epidermis. We
examined expression of essential genes for eyelid fusion and corneal
and conjunctival development to determine how Hoxc8 expression
hijacks these specific epithelial programs to initiate epidermal
development.
Methods: Mice carrying a conditional CAGGSHoxc8iresEGFP
cassette were bred to Wnt6cre mice to create double heterozygous
offspring that misexpress Hoxc8 throughout the Wnt6 domain,
which is expressed broadly across the embryonic ectoderm. Pregnant
dams were sacrificed to collect offspring between E11.5 and birth
for phenotypic characterization. Tissue was collected for RNA and
immunohistochemical analysis of genes involved in eyelid fusion
pathways and corneal differentiation, including: Wnt, Bmp, Shh,
Fgf10, ERK, and retinoic acid signaling, as well as Pax6 and various
cytokeratin markers.
Results: Newborn H&E staining showed sloughing squamous
epithelium covering the cornea and conjunctiva of conditional
mutants, with aberrant hair follicles present in the conjunctiva.
Examination of earlier timepoints showed that mutants fail to develop
an eyelid fusion front with associated migrating periderm and aligned
F-actin fibers. Mutant eyelid tips lacked localized expression of
p-cJun, p-ERK, Shh, Bmp4, and failed to downregulate Wnt signaling
in contrast to age-matched control embryos. Conversely, mutant
corneal epithelium failed to upregulate Wnt signaling, as evidenced
by the lack of Lef1. Pax6, which is essential for maintaining corneal
epithelium, was absent in mutant corneas, while skin specific
keratins, K1 and K10 were upregulated throughout the corneal and
conjunctival epithelium.
Conclusions: Emerging evidence suggests that Hox genes play
a key role in regionalization of the skin and epidermal organs.
Aberrant expression of Hoxc8 in the developing eye field ectoderm
aborts Pax6 expression in corneal epithelium, leading to corneal
and conjunctival keratinization, and disrupts most known signaling
pathways for eyelid fusion, the last major morphogenetic event to
occur in mammalian eye development.
Commercial Relationships: Lara S. Carroll
Support: 2RO1EY017182
Program Number: 3857 Poster Board Number: A0005
Presentation Time: 3:45 PM–5:30 PM
Identification and Characterization of Long Noncoding RNA
Contributions to Mouse Corneal Development
Weiwei Chen1, 2, Shuai Yang1, 2, Zhonglou Zhou1, 2, Xiaoting Zhao1, 2,
Dongsheng Yan1, 2. 1State Key Laboratory of Ophthalmology,
Wenzhou Medical University, Wenzhou, China; 2School of
Ophthalmology and Optometry, Wenzhou Medical University,
Wenzhou, China.
Purpose: Long noncoding RNAs (lncRNAs) are important regulators
of cellular functions. However an extensive in-depth analysis of their
expression profile and function in mouse corneal development is still
lacking. Here we investigated lncRNAs profiles of the developing
mouse cornea by microarray and bioinformatics.
Methods: Affymetrix mouse transcriptome 1.0 assay identified
corneal lncRNA profile changes occurring between birth and 8 weeks
in mice. Real-time RT-PCR analysis validated array findings. Gene
ontology (GO) and KEGG pathway mapping of protein-coding genes
adjacent to signature lncRNAs clarified potential lncRNA roles in
regulating corneal development.
Results: In newborn and 8-week-old mice, 41,987 protein-coding
and noncoding gene transcripts were identified. In these two subsets,
19, 623 of this ensemble are lncRNAs annotated in public data banks.
During development, 1,272 lncRNAs underwent ≧ two-fold changes
in expression levels. qPCR analysis confirmed gene microarray
analysis results since there was 90% agreement between the two
procedures in identifying lncRNAs contributing to this process. GO
analysis of protein-coding genes proximal to lncRNA signatures
resolved numerous neighboring protein coding genes regulating cell
division and adhesion as well as collagen and cytokine production,
suggesting a role for signature lincRNAs in controlling corneal
development.
Conclusions: Time dependent changes in lncRNA expression
patterns occurring during mouse corneal development suggest that
they play an important role in regulating this process.
Commercial Relationships: Weiwei Chen, None; Shuai Yang,
None; Zhonglou Zhou, None; Xiaoting Zhao, None;
Dongsheng Yan, None
Support: 973 Project (2012CB722303) from the Ministry of Science
and Technology of China, and Science Foundation of Wenzhou
Medical University QTJ11020
Program Number: 3858 Poster Board Number: A0006
Presentation Time: 3:45 PM–5:30 PM
IKZF1 Expression and Upregulation by polyI:C in Human
Ocular Surface Epithelial Cells
Mayumi Ueta1, Hiromi Nishigaki1, Katsuhiko Shinomiya2,
Norihiko Yokoi2, Chie Sotozono2, Shigeru Kinoshita1. 1Department of
Frontier Medical Science and Technology for Ophthalmology, Kyoto
Prefectural University of Medicine, Kyoto, Japan; 2Department of
Opthalmology, Kyoto Prefectural University of Medicine, Kyoto,
Japan.
Purpose: Stevens-Johnson syndrome (SJS) / toxic epidermal
necrolysis (TEN) are acute inflammatory vesiculobullous reactions
of the skin and mucosa, such as the ocular surface and oral cavity. In
genome wide association study (GWAS), we found the association
between IKAROS Family Zinc Finger 1 (IKZF1) and Cold medicine
related SJS/TEN with severe ocular complication (CM-SJS/TEN
with SOC). Ikaros is a transcription factor which regulates numerous
biological events, but the function of it was reported only in the
immune cells such as lymphocytes. On the other hands, we have
suggested that epithelium might be contribute to the pathobiology
of CM-SJS/TEN with SOC, because EP3, the protein of PTGER3,
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
which SNPs significantly associated with CM-SJS/TEN with SOC,
was dominantly expressed on the ocular surface epithelium and EP3
on the epithelium negative regulated ocular surface inflammation.
Furthermore, we also reported that TLR3, which SNPs significantly
associated with CM-SJS/TEN with SOC in Japanese, also strongly
expressed on the ocular surface epithelium and TLR3 positively
regulated ocular surface inflammation.
In this study, we examined the expression of IKZF1 in the human
ocular surface (conjunctival and corneal) epithelial cells, and
investigated whether it could be upregulated by TLR3 ligand.
Methods: For quantitative RT-PCR, primary human conjunctival
epithelial cells were harvested from conjunctival tissue obtained
at conjunctivochalasis surgery and cultured using the previously
described method. Primary human corneal cells were harvested
from the rest of the limbus of import donor cornea tissue after using
for corneal transplantation. For RT-PCR and immunohistology,
conjunctival tissue obtained at conjunctivochalasis surgery was used.
Results: We found that human conjunctival epithelium expressed
IKZF1 mRNA by RT-PCR and using quantitative RT-PCR, we found
that it could be upregulated by TLR3 ligand, polyI:C. We also found
that IKZF1 mRNA expression in corneal epithelial cells could be
upregulated by polyI:C. Immunohistology showed that IKZF1 protein
also expressed in conjunctival epithelium as same as CD3 positive T
cells.
Conclusions: Our results might suggest that IKZF1 in the ocular
surface could contribute to the ocular surface inflammation in the
SJS/TEN.
Commercial Relationships: Mayumi Ueta, None;
Hiromi Nishigaki, None; Katsuhiko Shinomiya, None;
Norihiko Yokoi, None; Chie Sotozono, None; Shigeru Kinoshita,
None
Support: This work was supported by grants-in-aid from the
Ministry of Education, Culture, Sports, Science and Technology of
the Japanese government (BioBank Japan Project), and by the JSPS
Core-to-Core Program, A. Advanced Research Networks and also
partly supported by grants-in-aids for scientific research from the
Japanese Ministry of Health, Labor and Welfare, and a research grant
from the Kyoto Foundation for the Promotion of Medical Science
and the Intramural Research Fund of Kyoto Prefectural University of
Medicine.
Program Number: 3859 Poster Board Number: A0007
Presentation Time: 3:45 PM–5:30 PM
Ocular Surface Adverse Events of Systemic Inhibitors of the
Epidermal Growth Factor Receptor (EGFRi): a Prospective Trial
Alejandro Saint-Jean1, 2, Ana Eixarch2, Noemi Reguart4,
Nuria Pardo4, Cristina Castella2, 3, Maite Sainz De La Maza2,
Alfredo Adan Civera2, Bernardo Sanchez Dalmau2, Marta Aldea5,
Ruben Torres2. 1Ophthalmology, CHU Nantes, Nantes, France;
2
Ophthalmology, Hospital Clinic Barcelona, Barcelona, Spain;
3
Clinique Sourdille, Nantes, France; 4Oncology Department, Hospital
Clinic, Barcelona, Spain; 5Unit of Infections and Cancer (UNIC),
Cancer Epidemiology Research Programme, Institute of Oncology,
IDIBELL, Barcelona, Spain.
Purpose: Controversy exists regarding the safety of agents that
systemically inhibits epidermal growth factor receptor (EGFRi) in
the ocular surface for oncologic patients. We performed a prospective
observational clinical study to compare the ocular surface toxicity of
systemic EGFRi between a case group and a control group without
EGFRi.
Methods: Patients with lung or colon cancer (34 lung and 18
colorectal adenocarcinoma) were divided in two groups: 26 patients
treated with systemic EGFRi wether erlotinib for lung cancer or
panitumumab for colon cancer (18 lung and 8 colon cancer, mean
age 73.4+/-7.3 years old) and 26 patients in a control group without
EGFRi treatment matched by the stage of the disease (16 lung and
10 colon cancer, mean age 63.5+/-8.6 years old). Patients in both
groups patients were chemotherapy naive (de novo treatment). Four
visits were scheduled in a one year period comparing signs and
symptoms in terms of symptom questionnaires (SIDEQ, OSDI and
AVS), corneal fluorescein staining (Oxford test), tear production
(Shirmer’s test), and a quantitative evaluation of conjunctival
chemosis and hyperemia. Basal epithelial cell density (BECD) and
corneal subepithelial nerve fiber density (SNFD) were measured
and compared using confocal microscopy (Heidelberg Ingeneering,
Germany). The differences in each variable between the 2 groups
were compared with the analysis of variance (ANOVA) (quantitative
variables). P value <0.05 was considered significant for all
comparisons.
Results: Statistically significant differences were found between
patients under EGFRi and the age-matched controls in terms of
conjunctival chemosis and Oxford test in all visits (P< 0.05). When
cases and controls were evaluated separatedly, the case group showed
a significant worse evolution in symptoms (SIDEQ and OSDI),
Oxford test and Shirmer’s test (all P <0.05). However the control
group did not show significant differences among visits. We did not
find significant differences in terms of ECD and SNFD between cases
and age/type of cancer matched controls.
Conclusions: Systemic EGFRi may increase conjunctival chemosis
and corneal fluorescein staining. Oncologic patients treated
with EGFR should be monitored closely with ophthalmologic
examinations to detect dry eye
Commercial Relationships: Alejandro Saint-Jean, None;
Ana Eixarch; Noemi Reguart, None; Nuria Pardo, None;
Cristina Castella, None; Maite Sainz De La Maza, None;
Alfredo Adan Civera, None; Bernardo Sanchez Dalmau, None;
Marta Aldea, None; Ruben Torres, None
Clinical Trial: ASJ-PAN-2012-01
Program Number: 3860 Poster Board Number: A0008
Presentation Time: 3:45 PM–5:30 PM
A role of primary cilia in Anterior Segment Dysgenesis disorders
Carlo Iomini, Grisanti Laura, Ekaterina Revenkova. Ophthalmology,
Icahn School of Medicine at Mount Sinai, New York, NY.
Purpose: The development of the anterior segment (AS) of the
mammalian eye involves tightly coordinated cellular interactions
between tissues of different embryonic origins. Defects in
this process lead to severe ocular disorders including aniridia,
microcornea and primary congenital glaucoma of unclear
pathophysiology. Interestingly, dysgenesis of the AS was reported in
patients affected by ciliopathies. Here we investigate the role of cilia
during development of the AS in mice.
Methods: Phenotypic analysis of the AS was performed using light
and transmission electron microscopy. Expression of ciliary-mediated
pathways target genes or transcription factors implicated in AS
morphogenesis were assessed using qRT-PCR and immunostaining
approaches. Cell proliferation of periocular mesenchyme was
assessed by BrdU incorporation and immunostaining.
Results: Null mutations that prevent cilia assembly such as Ift88-/are lethal at E9.5-10.5. In Ift88-/- embryos the optic vesicle, although
defective, is present. In contrast, at E10.5 the optic cup do not form
as observed in wild-type embryos. To overcome mid-gestation
lethality generated a conditional KO mouse Wnt1-Cre;IFT88flox/(Ift88 cKO) in which the Ift88 gene is excised in all migrating
mesenchymal cells of neural crest origin. cKO mice display strong
craniofacial defects and die at postnatal day P0. Early eye patterning
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
appears normal. However, later in development cKO mice display
significant reduction of the anterior chamber with occasional fusion
of the endothelium to the lens epithelium, a thinner stromal ECM and
corneal endothelial malformations. Interestingly, although migration
of neural crest-derived cells appears normal, we detected reduction of
mesenchymal cells at the angle between the stroma and the optic cup
at E17.5. Gene expression analysis revealed reduction of Hedgehog
pathway target genes and Ptx2 but not Foxc1, both encoding for
transcription factors linked to the Axenfeld-Rieger Syndrome.
Immunofluorescence analysis confirmed reduction of PTX2
expression at the iridocorneal angle and detected mislocalization of
cells expressing FOXC1.
Conclusions: We have uncovered a pivotal role of primary cilia in
the development of the AS and highlighted their involvement in the
etiology of ocular disorders derived from AS dysgenesis. We have
also provided an accessible paradigm to address the role of primary
cilia in complex tissue morphogenesis.
Commercial Relationships: Carlo Iomini, None; Grisanti Laura,
None; Ekaterina Revenkova, None
Support: NIH Grant EY022639 and RPB Dolly and Unrestricted
Awards
Program Number: 3861 Poster Board Number: A0009
Presentation Time: 3:45 PM–5:30 PM
PROSE lens use for exposure keratopathy in trigeminal and
facial nerve palsies
Elizabeth Marlow, Jessica Ciralsky, Michelle Lee, Gary Lelli.
Ophthalmology, Weill Cornell Medical College, New York, NY.
Purpose: For patients with severe ocular surface disease, prosthetic
replacement of the ocular surface ecosystem (PROSE) devices have
shown efficacy in improving visual acuity and the ocular surface.
Individuals with facial and trigeminal nerve palsies are at particularly
high risk for exposure keratopathy, making PROSE a potentially
beneficial tool in the management of these cases. This retrospective,
observational clinical study reviewed the effects of initiating PROSE
wear on nine eyes affected by facial (VII), trigeminal (V1), or both
(VII and V1) cranial nerve palsies.
Methods: Best corrected visual acuity (BCVA) at the initial PROSE
fitting, three months after initiating PROSE wear, and BCVA
achieved at any time after starting PROSE wear were compared. In
addition, the quality of the ocular surface before starting PROSE and
at the time of longest PROSE wear was compared.
Results: After three months of PROSE wear, patients gained an
average of 0.9 ± 1.4 lines on the Snellen eye chart compared to
initial BCVA. The average maximal gain in lines on the Snellen eye
chart achieved at any time after starting PROSE was 1.9 ± 1.3 lines,
which occurred an average of 5.3 ± 5.4 months after starting PROSE
wear. The ocular surface quality improved in 8 out of 9 eyes with
the resolution of punctate epithelial erosions (n = 7), reduced corneal
haze (n = 2), and reduced mucus strands (n = 2) being the most
commonly sited changes.
Conclusions: These findings show that the PROSE device can be an
effective tool in the management of exposure keratopathy secondary
to trigeminal (V1) and facial nerve palsies.
Commercial Relationships: Elizabeth Marlow, None;
Jessica Ciralsky, None; Michelle Lee, None; Gary Lelli, None
Support: Unrestricted support from Research to Prevent Blindness
Program Number: 3862 Poster Board Number: A0010
Presentation Time: 3:45 PM–5:30 PM
Impact of conjunctival autograft on pterygium treatment:
evaluation of related symptoms and patients satisfaction after
surgery
Bruna Duarte Moron de Andrade, Giovana Capecci Siqueira,
Paulo Dechichi Neto, Augusto Terra Baccega, Marina Gonçalves
Monteiro Viturino, Ana Luiza Mylla Boso, Daniella de Paiva
Almeida, Monica Alves. Department of Ophthalmology, UNICAMP,
Campinas, Brazil.
Purpose: Pterygium is a fibrovascular condition of the ocular
surface that can cause a broad range of irritative and visual
symptoms. Controversy exists regarding the pterygium mechanisms,
management, surgical techniques, adjuvant approaches and its impact
on patients’ quality of life. We performed a retrospective survey to
better understand the impact of pterygium related symptoms before
surgery and patients satisfaction after excision surgery followed by
conjunctival autograft transplantation with fibrin glue.
Methods: All patients underwent surgery that consisted of an
extensive removal of the pterygium fibrovascular tissue, followed
by an autologous conjunctival graft fixed with fibrin glue to cover
the bare scleral area, performed by the same surgeon. A total of 215
patients were contacted by phone call and asked to answer a simple
2 questions survey. First, a graduation of the symptoms related
to pterygium before surgical intervention, such as pain, irritation,
tearing, red eye, photophobia, burning, body sensation, scaled from
0-10 (0 asymptomatic and 10 very important symptoms). We then
classified as mild (0-3), moderate (4-7) and severe (8-10). Then, the
patients were asked about their satisfaction on surgery results, scaled
from 0-10 (ranging from dissatisfied to fully satisfied).
Results: Patients mean age was 41.1 (min-19/max-82) years old and
the mean of days after surgery was 967.2 days (min-347/max-2155).
Symptoms were referred as severe (71.8%), moderate (23.9%) and
mild (4.3%). After surgery most patients were fully satisfied and the
mean grade was 9.4; 0.9% (0-3), 1.9% (4-7) and 97.1% (8-10).
Conclusions: The present study shows that pterygium excision
surgery using conjunctival autograft transplantation and fibrin glue
improved quality of life related to pterygium symptoms on patients
affected by the condition with high rates of satisfaction. However,
there is still lack of understanding on pterygium mechanisms, no
consensus on best surgical technique and the recurrence rates remain
a challenge, justifying the need of further research on this area.
Commercial Relationships: Bruna Duarte Moron de Andrade,
None; Giovana Capecci Siqueira, None; Paulo Dechichi Neto,
None; Augusto Terra Baccega, None; Marina Gonçalves Monteiro
Viturino; Ana Luiza Mylla Boso, None; Daniella de Paiva
Almeida, None; Monica Alves, None
Support: Fapesp grant #2014/19138-5
Program Number: 3863 Poster Board Number: A0011
Presentation Time: 3:45 PM–5:30 PM
Correlation between Meibomian Gland Dysfunction and
Keratitis in Patients with Demodex Brevis Infestation
Lingyi Liang1, Hongmin Ke1, Scheffer C. Tseng2. 1Cornea, Zhongshan
Ophthalmic Center, Guangzhou, China; 2Ocular Surface Research
and Education Center, Miami, FL.
Purpose: To investigate whether there is any correlation between
meibomian gland dysfunction (MGD) and keratitis in patients with
ocular demodicosis.
Methods: Prospective, cross sectional, comparative and controlled
case series. Sixty study patients with ocular demodicosis and 45
gender- and age- matched dry eye patients as the control without
demodicosis. All patients were younger than 35 years-old. Diagnosis
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
of demodicosis was based on microscopic counting of D. folliculorum
and D. brevis of epilated lashes. Severity of keratitis and MGD was
graded by photography and meibography, respectively, in a masked
fashion. Statistical correlation between Demodex infestation, MGD
and keratitis were analyzed.
Results: MGD defined by meibography and meiboscore was
significantly more prevalent and more severe in the study group than
the control group (both P<0.05). Meibomian gland loss was worse
in the upper lid in study patients but similar in upper and lower lids
in control patients. Superficial punctate keratitis was found in both
groups, while limbitis, stromal infiltration, vascularization, scarring,
and perforation were only noted in study patients. Both univariate and
multivariate analyses showed that the meiboscore was significantly
correlated with higher D. brevis count and more severe keratitis,
but not with age, gender, history course, and D. folliculorum count.
Bayesian network models tested by R package bnlearn suggested
that D. brevis counts contributed to both keratitis and MGD. Rapid
resolution of keratitis and blepharoconjunctivitis was accompanied
by significant reduction of the demodex count in 48 patients receiving
lid scrub using tea tree oil, which had been reported to kill Demodex
mites.
Conclusions: Ocular demodicosis, especially by D. brevis, may play
a causative role in MGD and keratitis in young patients. Its potential
pathogenic role in MGD in the older population cannot be ignored.
Commercial Relationships: Lingyi Liang, None; Hongmin Ke,
None; Scheffer C. Tseng, Biotissue (P)
Support: National Eye Institute, the National Institutes of
Health (R44 EY019586); National Natural Science Foundation
of China(81300739); Ministry of Education of China
(20110171120104); Technological Project Foundation of Guangdong
Province(2011B031800274, 2012B031800456)
Program Number: 3864 Poster Board Number: A0012
Presentation Time: 3:45 PM–5:30 PM
The Evaluation of Worldwide Distribution of Adenoviral
Genotypes in Acute/Epidemic Keratoconjunctivitis and
Adenoviral-negative Keratoconjunctivitis with Next Generation
Sequencing
Cecilia S. Lee1, Aaron Y. Lee1, Lakshmi Akileswaran1,
David W. Stroman2, Kathryn Najafi-Tagol2, Steve Kleiboeker4,
Anna Wald3, Russell N. Van Gelder1. 1Ophthalmology, University
of Washington, Seattle, WA; 2NovaBay Pharmaceuticals, Inc.,
Emeryville, CA; 3Infectious Diseases, University of Washington,
Seattle, WA; 4ViraCor-IBT Laboratories, Inc, Lee's Summit, MO.
Purpose: Conjunctivitis is among the most common outpatient
conditions and causes significant ocular morbidity worldwide. The
most severe and common form of viral conjunctivitis is epidemic
keratoconjunctivitis (EKC), caused by certain strains of adenovirus
(Ad). However, the geographic distribution of other Ad genotypes
and the role of the ocular surface microbiome (OSM) in EKC have
not been established. We investigated the worldwide distribution
of Ad genotypes and the OSM associated with keratoconjunctivitis
(KC).
Methods: A variety of analyses were conducted on samples obtained
in a large international clinical trial (NV-422 Phase IIB, NovaBay
Pharmaceuticals, Inc). The inclusion criteria were clinical signs
and symptoms consistent with KC and evidence of Ad using the
rapid screening test (Rapid Pathogen Screening Inc., Saratosa, FL).
Bilateral conjunctival swabs were obtained on day 1, 3, 6, 11, and
18. Ad genotype was determined by sequencing of Ad hexon gene
(ViraCor-IBT Laboratories, Inc, Lee’s Summit, MO) and the OSM
was assessed using next generation sequencing (NGS) on selected
samples. The difference in the distribution of Ad genotypes was
assessed with Fisher exact.
Results: Overall, 5000 samples were collected from 500 patients.
111, 200, 84, and 103 patients were recruited from US, India, Sri
Lanka, and Brazil, respectively. Mean age was 35 and 58% were
men. The most common Ad subgenera were subgroup D (316, 63%),
followed by E, B, then C. A significant portion of patients (110,
22%) lacked PCR evidence of Ad infection. There was a significant
geographic bias for Ad subgenera and serotype distribution (Fisher
exact, p-value <10-6). The odds ratio (OR) of having serotype 4 vs.
others in the US was 7.87 times higher than in non-US sites (95%CI
3.55, 17.35), while the OR of having serotype 8 vs. others in the
US was significantly lower (0.01, 95%CI 0.003, 0.043). Among the
samples that underwent NGS, Ad was the dominant virus in 10 of 16
Ad-positive samples, while there was no consistent, dominant virus
in 16 Ad-negative samples. Two Ad-negative samples contained
significant amounts of torque teno virus.
Conclusions: This study reveals unexpected geographic diversity in
Ad genotypes associated with KC, and suggests that nearly one fourth
of apparently viral KC may be due to agents other than adenovirus.
.
Commercial Relationships: Cecilia S. Lee, None; Aaron Y. Lee,
None; Lakshmi Akileswaran, None; David W. Stroman,
NovaBay Pharmaceuticals, Inc; Kathryn Najafi-Tagol, NovaBay
Pharmaceuticals, Inc; Steve Kleiboeker; Anna Wald, None;
Russell N. Van Gelder, None
Support: NH Grant K23EY024921, R01EY022038, K24 AI071113,
P30EY001730, Richard and Maude Charitable Foundation, Ferry
Research Grant, Latham Vision Science Research Grant
Clinical Trial: NCT01532336
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Program Number: 3865 Poster Board Number: A0013
Presentation Time: 3:45 PM–5:30 PM
Safety and efficacy study to evaluate wiping the lid margins with
tap water alone or in combination with lid hygiene shampoo in
subjects with normal meibomian glands
Hirotaka Tanabe, Minako Kaido, Motoko Kawashima, Reiko Ishida,
Kazuo Tsubota. Ophthalmology, Keio Univ School of Medicine,
Tokyo, Japan.
Purpose: We performed a prospective clinical study to evaluate the
safety and efficacy of using tap water alone or in combination with
lid hygiene shampoo to wipe lid margins with normal meibomian
glands.
Methods: Fourteen eyes of 7 subjects (6 males and 1 female aged
28 to 56 years old [40.8 ± 11.4 years old]) with normal meibomian
glands were evaluated in the Ophthalmology Department of Keio
University Hospital before and after wiping the lid margins using
tap water alone at first and then tap water in combination with
lid hygiene shampoo (Eye Shampoo [MediProduct, Japan]) one
week later. The ethics committee of the Keio University School of
Medicine approved the protocol. We used t-tests to analyze the tear
break-up time (TBUT) (grades 0 to 10) and Wilcoxon signed-rank
tests to analyze corneal and conjunctival fluorescein/lissamine green/
rose bengal staining scores (grades 0 to 9), lid margin lissamine
green staining scores (grades 0 to 3), subjective symptoms (grades
0 to 100) (as assessed via the visual analog scale [VAS]), and tear
lipid layer interference (grades 1 to 5) (as assessed using a DR-1 tear
interference camera [Kowa Co., Nagoya, Japan]).
Results: A significant exacerbating change (P<0.05) was not detected
after either method was applied in terms of the VAS score, the TBUT,
corneal and conjunctival staining, lid margin lissamine green staining,
and tear lipid layer interference. For the VAS score, foreign body
sensation and eye discharge recognition were significantly improved
(P<0.05) only in the group using lid hygiene shampoo (9.71±14.0 to
3.92±9.02, P=0.031; 4.96±5.52 to 1.14±2.28, P=0.022, respectively),
and eyestrain was significantly improved only in the group using
tap water alone (17.0±15.3 to 1.57±4.01, P=0.008). Although there
were no significant differences in the other VAS parameters, certain
changes indicating improvements (P<0.1) in the parameters related
to stimulation, i.e., pain and smarting pain, were noted only in the
group using lid hygiene shampoo (3.64±6.18 to 0.64±1.64, P=0.098;
3.28±5.42 to 0.64±1.73, P=0.098, respectively).
Conclusions: The use of tap water alone or in combination with
lid hygiene shampoo to wipe lid margins appears to be safe for
maintaining lid hygiene in normal subjects. The efficacy of lid
hygiene shampoo seems greater than tap water alone based on the
VAS parameters.
Commercial Relationships: Hirotaka Tanabe, None;
Minako Kaido, None; Motoko Kawashima; Reiko Ishida, None;
Kazuo Tsubota, MediProduct (F)
Support: MediProduct
Clinical Trial: http://www.umin.ac.jp/ctr/index.htm,
UMIN000016906
Program Number: 3866 Poster Board Number: A0014
Presentation Time: 3:45 PM–5:30 PM
Histatin-1 as a peptide based therapy for human corneal
epithelial wound healing
Dhara Shah, Zeeshan Pasha, Marwan Ali, Vinay K. Aakalu.
Department of Ophthalmology, University of Illinois at Chicago,
Chicago, IL.
Purpose: Purpose: Histatins are histidine-rich peptides that are
present in human saliva and in human lacrimal epithelium. Previous
experiments have shown that Histatin-1 can promote epithelialization
in multiple tissue types. We investigated the effects of applying
histatin-1 to a human corneal epithelial wound model in vitro.
Methods: Methods: Human corneal epithelial (HCE) cells were
utilized to perform a standard scratch based wound healing assay.
A confluent monolayer of HCE cells were wounded manually by
scraping with a 200ul pipette tip. Cells were then exposed to different
concentrations of Histatin-1 in a medium essential media containing
2% serum. Cell migration was followed for 24 hours after scraping
and imaged using time-lapse microscopy. Images were analyzed
using Neuro Image-J. Effects of Histatin-1 on cell proliferation
and cell death were assessed using colorimetric assays-lactate
dehydrogenase (LDH) assay and an MTT assay.
Results: Results: Our findings demonstrate that cell migration rates
were increased with increasing Histatin-1 concentration. We also
noted no significant change in LDH levels or MTT levels at tested
levels, suggesting a non-proliferative, non-toxic mechanism for
wound healing enhancement.
Conclusions: Conclusion: Histatin-1 peptides promote human
corneal epithelial migration and wound healing in vitro. Future
studies to investigate these findings in vivo will be necessary to
develop therapeutic applications for Histatin peptides.
Human Corneal Epithelial cells 4 hour post scratch (Control)
Human Corneal Epithelial cells 4 hour post scratch with Histatin-1
(10uM/ml)
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Commercial Relationships: Dhara Shah, None; Zeeshan Pasha,
None; Marwan Ali, None; Vinay K. Aakalu, None
Support: NIH—NEI Grant (K08EY024339), a seed grant from
Illinois Society to Prevent Blindness, a Research Grant from Midwest
Eye Banks, a Grant-in-Aid from Fight for Sight and Departmental
Support through an NIH-NEI Core grant (2P30EY001792-36A1) and
an Unrestricted Grant from Research to Prevent Blindness (NY,NY).
Program Number: 3867 Poster Board Number: A0015
Presentation Time: 3:45 PM–5:30 PM
Non-invasive technique for dynamic measurement of tear film
surface quality based on interferometry
Dorota H. Szczesna-Iskander, Slawomir Drobczynski. Dept. of Optics
and Photonics, Wroclaw University of Technology, Wroclaw, Poland.
Purpose: To demonstrate the development of a new generation
Lateral Shearing Interferometer (LSI) for non-invasive assessment of
tear film surface quality (TFSQ) dynamics.
Methods: The LSI technique allows for objective in vivo evaluation
of the tear film (TF) surface in the central area of a cornea or a
contact lens by analyzing the shape of interference pattern. High
sensitivity of this technique for subtle changes in TF dynamics has
been demonstrated with a prototype (Szczesna et al. OVS 2009)
whose several limitations hindered its potential to achieve full
clinical utility. Recently, a new compact LSI was built. A HeNe
laser (633nm) light source illuminates an area of 6 mm in diameter.
The design of the interferometer meets the safety standards (ANSI
Z136.1-2000). The LSI is fixed to a slit lamp base, the fixation target
for the measured eye has been incorporated into the system, and a
special head rest has been designed to minimize the head movements.
Temporal dynamics of TF is recorded by a CCD camera with
frequency of 29 frames per second.
Results: Measurements on artificial eyes of radius from 7.8 to 8.5
mm (with and without astigmatism) as well as on over 50 subjects
have been performed with the new LSI system as well as the
previously used prototype. The slit-lamp based operation of the new
LSI is more user friendly comparing to its predecessor. The new
fixation target aligned with the measured eye rather than the fellow
eye substantially improved eye’s stability during the measurement.
The larger coverage area (6 mm vs 4 mm) and higher frequency of
fringes determined by the optical wedge allow for more detailed
analysis of the TFSQ and accurate assessment of TF post-blink
stabilization phase and evolution of the TF break-up. The upward
movement of the post-blink deterioration features is clearly observed
(see figure).
Conclusions: The new improved generation of LSI enhances its
clinical utility for non-invasive assessment of TFSQ and its dynamic
properties on cornea and contact lenses. The new LSI provides basis
for studying relationships between TF post-blink stabilization phase
and TF related eye diseases.
Figure. Examples of interference patterns recorded with the new
generation LSI: 0.10s (A); 0.34s (B); 0.68s (C); 1.00s (D) post-blink.
The frames show the upward movement of the tear film features
within the first second after the blink.
Commercial Relationships: Dorota H. Szczesna-Iskander, None;
Slawomir Drobczynski
Support: National Science Center of Poland, Program SONATA,
grand no: 2011/03/D/ST7/02512
Program Number: 3868 Poster Board Number: A0016
Presentation Time: 3:45 PM–5:30 PM
Stimulation of Corneal Nociceptors Results in Pro-inflammatory
Molecular and Cellular Responses
Yashar Seyed-Razavi1, 2, Pedram Hamrah3, 4. 1Ophthalmology, Tufts
Medical Center, Boston, MA; 2Ophthalmology, Schepens Eye
Research Institute/Massachusetts Eye and Ear, Harvard Medical
School, Boston, MA; 3Ophthalmology, Schepens Eye Research
Institute/Massachusetts Eye and Ear, Cornea & Refractive Surgery
Service, Harvard Medical School, Boston, MA; 4Ophthalmology,
Cornea Service, New England Eye Center, Tufts Medical Center,
Boston, MA.
Purpose: We have recently shown immediate changes in
neuropeptides levels following CO2 stimulation of corneal
nociceptors. The peripheral nervous system, in part through
neuropeptides, is able to alter the balance towards a pro- or antiinflammatory environment by influencing the activity of resident
antigen presenting cells (APCs) such as dendritic cells (DCs),
modulating their chemotaxis, maturation and T-cell stimulatory
capacity. We therefore hypothesized that CO2-induced nociceptor
stimulation will result in alterations in downstream inflammatory
cytokines and adhesion molecules, consistent with, and resulting in
pain-induced neurogenic inflammation.
Methods: CO2 gas was applied to the central cornea of C57BL/6
and BALB/c mice in a series of 3 pulses every hour over 4 hours,
and collected 1 hour following the final burst. Corneas were then
excised, and gene and protein analysis for pro-inflammatory
cytokines and adhesion markers, as well as whole-mount staining for
CD11b (leukocyte marker), CD11c (DCs) and MHC class II (antigen
presenting cells), was performed. Naïve un-stimulated and airstimulated corneas served as controls.
Results: Long-term stimulation of corneal nociceptors resulted
in increased expression of the pro-inflammatory neuropeptides
substance P (7-fold, p<0.001) and CGRP (3-fold, p<0.05), as well as
the anti-inflammatory neuropeptide urocortin (70 fold, p<0.001) in
C57BL/6 mice. IL-1β was up-regulated (3-fold, p<0.05), and whilst
IL-6 expression increased, it was not significant (p=0.12) compared
to controls. Increased expression of adhesion molecules P- and
E-Selectin (p=0.059 and p<0.05, respectively) as well as elevated
ICAM-1 expression (p<0.05) was found. Analysis of BALB/c mice
revealed a similar increase in inflammatory cytokines (p=0.06 and
p<0.01 for IL-1β and IL-6, respectively) and adhesion markers
(p=0.053 and p<0.05 for E-Selectin and ICAM, respectively).
Fractalkine (CX3CL1) mRNA levels increased (p<0.05) following
CO2 stimulation. Finally, nociceptor stimulation resulted in alterations
in resident APC density and maturation levels.
Conclusions: Our study demonstrates downstream changes in
inflammatory cytokines, chemokines and adhesion markers,
ultimately leading to alterations in resident APC populations
following nociceptor activation. To our knowledge, this is the first
report to reveal direct neurogenic effect of pain-induced corneal
nociceptor activation.
Commercial Relationships: Yashar Seyed-Razavi;
Pedram Hamrah, None
Support: NIH R01-EY022695
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Program Number: 3869 Poster Board Number: A0017
Presentation Time: 3:45 PM–5:30 PM
Crosstalk Between Selective Autophagy and Nrf2-ARE Pathway
Through p62/SQSTM1 Confers Limbal Stem Cell Resistance to
Ultraviolet-A Irradiation
Ying-Ting Chen1, Maria Laggner1, Florian Gruber2,
Erwin Tschachler2, Ursula Schmidt-Erfurth1, Andreas Pollreisz1.
1
Department of Ophthalmology, Medical University of Vienna,
Vienna, Austria; 2Department of Dermatology, Medical University of
Vienna, Vienna, Austria.
Purpose: Recently we reported a cytoprotective role for autophagy
and Nrf2-ARE pathway in physiological UVA-stressed limbal stem
cells (LSC). The current study sought to identify the mechanistic
underpinnings linking these two pathways.
Methods: Krt14-Cre:Atg7f/f transgenic mice and global Nrf2-KO
mice on C57BL/6 background were used to generate Atg7-/- and Nrf2/LSCs, respectively. 3-MA was used to inhibit autophagy in cultured
Nrf2-/- LSCs for creating autophagy-deficient Nrf2-KO LSCs. UVA
irradiation at a low dose (10 J/cm2) was applied to the cultured LSCs,
pre-treated with either ROS-scanvenger NAC or drug vehicle. CMH2DCFDA live staining and Caspase3/7 CytoEvent were used to
detect intracellular ROS levels and cellular apoptosis, respectively.
Autophagic activity was measured by CytoID live staining. Dual
immunofluorescence was used to study the cellular kinetics of Nrf2
and p62/SQSTM1. A nuclear localization of Nrf2 was used as a
surrogate readout for Nrf2-ARE activity.
Results: Confocal microscopy using CM-H2DCFDA live staining
did not detect changes of ROS level in 10 J/Cm2-irradiated Atg7f/f
LSCs. In contrast, a 2-3 fold elevation in ROS level was observed in
Atg7-/-, Nrf2-/- and autophagy-deficient Nrf2-/- LSCs (all p<.05).
Compared to the perspective non-irradiated baseline, Caspase3(+)
apoptotic cells was increased by 3.5% (p>.05), 21.7% (p<.05), 29.3%
(p<.05), and 18.4% (p<.05) in Atg7f/f, Atg7-/-, Nrf2-/- and autophagydeficient Nrf2-/-LSC at 24-hr post-irradiation. The radiation-induced
apoptosis was successfully rescued by NAC pre-treatment. Dual
immunofluorescence revealed a diffuse cytosolic distribution for Nrf2
and p62 in unstressed LSCs, with a low-degree co-localization. After
irradiation, Nrf2 nuclear translocation was observed exclusively in
Atg7f/f cells with distinct p62 perinuclear speckle formation. Such a
Nrf2/p62 redistribution was not seen in any other irradiated groups.
Conclusions: Our data collectively suggest that autophagy activation
in LSCs under physiological UVA stress functions as a non-canonical
antioxidant defense mechanism. The p62/SQSTM1-mediated
enhancement of Nrf2-ARE response enables LSC to have a tighter
control in cellular redox. New therapeutics targeting p62/SQSTM1
might thus supplement the current antioxidant-based treatment for
corneal degenerative diseases associated chronic oxidative stress.
Commercial Relationships: Ying-Ting Chen, None;
Maria Laggner; Florian Gruber, None; Erwin Tschachler, None;
Ursula Schmidt-Erfurth, None; Andreas Pollreisz, None
Program Number: 3870 Poster Board Number: A0018
Presentation Time: 3:45 PM–5:30 PM
Ocular surface stories after cataract surgery
Rodrigo M. Torres1, 2, Pablo G. Lódolo1, 2. 1Ophthalmology, Centro
de Ojos Dr Lodolo, Colonia Avellaneda, Argentina; 2Asociación
Entrerriana de Oftalmología, Paraná, Argentina.
Purpose: “I can see but I’m felling bad”. When cataract surgeons
heard something like that and they couldn’t find real problems in
those eyes, usually the ocular surface (OS) service take place. This
work review and analyze this topic.
Methods: A retrospective clinical study was performed, evaluating
the clinical charts from patients referred to the OS service with
problems after cataract surgeries, during the present year (2015).
The follow data was analyzed: age, sex, time after surgery, visual
acuity, the kind of intraocular lens (IOL), post-surgical refraction,
the patient complaints/symptoms, their work or occupation, previous
topical treatments (dry eye/glaucoma), the discovered signs and their
therapeutic approach.
Results: A total of 15 patients operated (both eyes) by 5 different
surgeons were evaluated. The mean age was 67 years all, with 9
women and 6 men. All of the cases were referred between 2 to 3
months after cataract surgery (sutureless clear corneal incision of
2.0 mm to 2.5 mm, topical anesthesia, without intraocular surgical
or post-surgical complications). Binocular visual acuity without
correction in all of the patients were between 20/25 to 20/20, but 12
of them with 1 eye for near sight (spherical refraction: -1.0 to -1.5).
3 patients with far sight/both eyes. Symptoms: pain (14), foreign
body sensation (15), burning (12), itching (4); fear about blindness
(15). Work/occupation: retired (10) professional work (5: architect/
physician/agronomic engineering/politic/bank employee). Non of the
patients have history about glaucoma topical treatment but 3 have
history about OS problems (dry eye:2; Stevens Johnson: 1). Signs:
blepharitis (8), corneal edema around incision (4), epithelial keratitis
(4), entropion (3). symblepharon (3). 5 patients from all (33.3%, 3
women, 2 men) don’t have signs about OS problems. Artificial tears,
topical loteprednol and lid hygiene, were the therapeutic approach
for most of them but five with oral minocycline 50mg and topical
cyclosporin for three (two were pseudo-pemphigoid and one was the
Stevens Johnson).
Conclusions: Most women than men were referred to the OS service
because complaints after cataract surgery. OS problems were detected
in 66.6%, but fear about blindness was present in all the cases.
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Nonspecific pain was another common symptom. It is time to include
a psychologist to evaluate pre-op patients and manage them at postop time, as part of the ophthalmology team?
Commercial Relationships: Rodrigo M. Torres, None;
Pablo G. Lódolo
Program Number: 3871 Poster Board Number: A0019
Presentation Time: 3:45 PM–5:30 PM
A Structural Equation Model (SEM) Relating Eye Pain (EP) and
Ocular Surface Measurements (OSMs)
Richard A. Bilonick1, 2, Anat Galor3. 1UPMC Eye Center, Eye and
Ear Institute, Ophthalmology and Visual Science Research Center,
Department of Ophthalmology, University of Pittsburgh School of
Medicine, Pittsburgh, PA; 2Biostatistics, University of Pittsburgh
Graduate School of Public Health, Pittsburgh, PA; 3Bascom Palmer
Eye Institute, University of Miami, Miami, FL.
Purpose: Goals were to 1) parsimoniously describe the relationship
between subjective EP (measured by questionnaire with 7 ordinal
responses) and combination of objective OSMs and demographic
data, and 2) use data from both eyes and describe the ordinal EP
responses using a small number of latent factors.
Methods: 190 patients underwent ocular surface assessment,
including tear osmolarity (OSM), tear evaporation measured via tear
breakup time (TBUT), corneal epithelial cell disruption measured via
corneal staining (STN), and tear production measured via Schirmer’s
strips with anesthesia (SCH). Subjects rated the intensity of their EP
for right now (RN), worst 1 wk (W1W), average 1 wk (A1W), worst
3 months (W3M), average 3 months (A3M), worst 1 yr (W1Y), and
average 1 yr (A1Y) using an ordinal rating scale anchored at 0 for no
pain and 10 for most intense pain imaginable. 4 common factor (CF)
pure measrement models (1-factor, 2-non-overlapping factors [2NF],
2 overlapping factors, and a 3-factor model) for EP responses were
compared and the best measurement model for EP was chosen using
root mean square error of approximation (RMSEA). OSM, TBUT,
STN, SCH were measured in both eyes and were each represented
by a latent factor with the left and right measurements as indicators.
This approach accounted for random measurement error that would
otherwise bias effect estimates. These latent factors along with age,
sex and race were included as exogenous variables that could affect
EP. R packages lavaan and OpenMx were used to estimate path
coefficient and other SEM parameters.
Results: 2NF CF model with latent factors STP (short term pain)
and LTP (long term pain) was best according to RMSEA. The figure
shows the path diagram for the resulting SEM. Correlations between
each EP question response and corresponding STP and LTP all
exceeded 0.9. RMSEA for the SEM was 0.04, 75% lower than for the
2NF CF pain model. A 100 mOsm/L increase in OSM resulted in a
1.5 standard deviation (SD) drop in LTP (p<0.001) and 0.5 SD drop
in STP (p=0.016). While not SS, a 10 mm increase in SCH resulted in
a 0.2 SD drop in LTP (p=0.144) and a 0.1 SD drop in STP (p=0.599).
Conclusions: OSM was shown to affect both STP and LTP but in a
differential manner. No evidence was found for effects due to TBUT,
STN, age, sex or race. Possibly SCH had an effect which was also
differential.
Sem path diagram relating EP, OSMs, and demographics.
Commercial Relationships: Richard A. Bilonick; Anat Galor,
None
Support: P30-EY008098; Eye and Ear Foundation (Pittsburgh,
PA); Research to Prevent Blindness (New York, NY); Department of
Veterans Affairs, Veterans Health Administration, Office of Research
and Development; Clinical Sciences Research and Development’s
Career Development Award CDA-2-024-10S (Dr. Galor); NIH Center
Core Grant P30EY014801.
Program Number: 3872 Poster Board Number: A0020
Presentation Time: 3:45 PM–5:30 PM
Human blinking ‘eye-on-a-chip’
Jeongyun Seo1, Woo Y. Byun1, Andrea Frank1,
Mina Massaro -Giordano2, Vivian Lee2, Vatinee Y. Bunya2,
Dongeun Huh1. 1Bioengineering, University of Pennsylvania,
Phildelphia, PA; 2Scheie Eye Institute, Department of
Ophthalmology, Perelman School of Medicine at the University of
Pennsylvania, Philadelphia, PA.
Purpose: The structural and environmental complexity of the
ocular surface (OS) poses critical technical challenges to in vitro
investigation of its physiology and pathology using traditional cell
culture models. As a result, research in this area has relied heavily
on expensive and time-consuming in vivo animal studies. To realize
more physiological in vitro models, we have developed an ‘eye-ona-chip’ microdevice that recapitulates the three-dimensional (3D)
tissue structure, dynamic mechanical environment, and physiological
functionality of the human OS.
Methods: Our microdevice (Fig. A) contains a 3D dome-shaped
porous cell culture scaffold with a similar curvature to that of the
human cornea (Fig. B). The porous scaffold was impregnated with
human keratocytes (HKs) and collagen gel to mimic the stroma.
We also developed a novel microengineering technique to precisely
deposit human corneal epithelial cells (HCECs) and conjunctival
epithelial cells (HCjEs) on the 3D scaffold surface to replicate the
concentric epithelial patterns in vivo. These microengineered ocular
tissues were integrated with a biomimetic hydrogel eyelid, and the
eyelid was actuated to slide along the curved scaffold surface to
replicate eye blinking.
Results: The 3D tissue structures of the OS were reconstituted in
our microdevice. The HKs were cultured within the porous scaffold
to replicate the stroma (Fig. C). Our cell patterning technique
allowed us to generate a circular corneal epithelium surrounded
by a conjunctival epithelium on the curved scaffold surface (Fig.
D). These ocular epithelia exhibited differentiated phenotypes as
evidenced by stratification, tight junction formation, and increased
cytokeratin/mucin protein expressions. We also demonstrated tear
fluid dynamics in our microdevice by showing spreading of artificial
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
tear fluid and resultant wetting of the scaffold surface due to eyelid
actuation (Fig. E). Finally, we explored the possibility of leveraging
this microdevice to simulate pathological features of dry eye disease
and to examine key biological processes mediating the development
and progression of the disease.
Conclusions: Our ‘eye-on-a-chip’ holds great potential to serve as an
innovative modeling platform that represents a predictive and lowcost alternative to conventional cell culture and animal models.
Commercial Relationships: Jeongyun Seo, None; Woo Y. Byun,
None; Andrea Frank, None; Mina Massaro-Giordano, None;
Vivian Lee, None; Vatinee Y. Bunya, None; Dongeun Huh
Program Number: 3873 Poster Board Number: A0021
Presentation Time: 3:45 PM–5:30 PM
A disease model of aqueous deficient dry eye in a microengineered
ocular surface tear film platform
Nicole Qiaozhi Lu, Michael P. Grant, Jennifer Elisseeff. Wilmer Eye
Institute, Johns Hopkins University, Baltimroe, MD.
Purpose: Dry eye is an ocular surface disease affecting a large
population worldwide, and an in vitro model, recapitulating the
function of ocular surface, is in need for drug development studies.
Previously we have demonstrated the efficacy of cocultured lacrimal
gland (LG) and conjunctival epithelial cells (CECs) in mimicking the
tear secretory function of healthy ocular surface. Here we explored
the possibility of utilizing this microengineered platform as a disease
model of aqueous deficient dry eye.
Methods: Rabbit LGs were digested, and spheroids were formed
in a shaking flask. CECs were isolated by dispase II digestion.
CECs were seeded on top of the Transwell insert and allowed to
stratify under airlifting condition. LG spheroids were encapsulated
in Matrigel and added to the periphery of the membrane (Fig.
1). Dry eye was introduced by the addition of either IL-1β or
hyperosmolarity. Cyclosporin A (CsA) and dexamethasone (DEX)
were used as treatments. The systems were maintained for 10 days
in vitro. Samples were harvested and processed for the analyses of
gene expression by qPCR, mucin secretion of CECs by texas-red
conjugated dextran staining, LG secretion by β-hexosaminidase
assay, and immunohistochemistry.
Results: Exposure to IL-1β and hyperosmolarity increased the
mRNA level of proinflammatory cytokines and chemokines (TNFα,
IL-6 and MMP9), while treatments with CsA and DEX were able to
reduce it. The model also reflected a change in epithelial morphology
as shown in the H&E staining (Fig. 2a). The induced inflammation
in the model significantly decreased epithelial stratification and led
to more keratinization, and DEX was more effective in reversing this
effect. Immunohistochemistry with mucin 5AC supported that mucin
expression was also affected, and both CsA and DEX could restore
the mucin level comparable to control (Fig. 2b). Similar trends were
observed in LG and mucin secretions.
Conclusions: This study has demonstrated that dry-eye like
symptoms were observed in the in vitro model after the induction
of inflammation, and administering of treatments were proven to
be effective as well. We mimicked the physiological conditions of
aqueous deficient dry eye in the cocultured system with CECs and
LGs. Potentially this microengineered ocular surface platform could
be further investigated for the screening of new dry eye therapeutics.
Fig1
Fig2
Commercial Relationships: Nicole Qiaozhi Lu; Michael P. Grant,
None; Jennifer Elisseeff, None
Support: KKESH-Wilmer Research grant; Research to Prevent
Blindness
Program Number: 3874 Poster Board Number: A0022
Presentation Time: 3:45 PM–5:30 PM
Contralateral investigation of postoperative refractive surgery
inflammation: Small-incision Lenticule Extraction vs LASIK
Marianthi Stergiou1, A. J. Kanellopoulos1, 2, George Asimellis1, 3.
1
LaserVision.gr Eye Institute, Athens, Greece; 2Ophthalmology, NYU
Medical School, NY, NY; 3Kentucky College of Optometry, Pikeville,
KY.
Purpose: The purpose of this study was to comparative investigate
potential impact of inflammation following myopic correction. The
established current gold standard of femtosecond-assisted LASIK
was compared to an all femto-second laser Small Incision refractive
Lenticule Extraction (SMILE). No study has investigated the
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
potential differences between SMILE and LASIK from the standpoint of postoperative inflammation.
Methods: This is a contralateral, perspective stud employing 10
consecutive myopic patients, in which one eye (group-A) was treated
with the SMILE, while the other eye (group-B) with LASIK. The
LASIK procedure employed the Alcon Refractive surgery platform
(Alcon Surgical, Ft. Worth, TX) comprised of the FS200 femtosecond
and the EX500 excimer laser. The SMILE procedure employed the
500 kHz VisuMax® femtosecond laser (Carl Zeiss Meditec AG, Jena,
Germany).
InflammaDry (Rapid Pathogen Screening, Inc., Sarasota, FL) is
an in-office test that detects matrix metalloproteinase MMP-9, an
inflammatory marker that is consistently elevated in the tears of
patients with dry eye disease. Using direct sampling microfiltration
technology, InflammaDry identifies elevated levels of MMP-9
protein in tear fluid samples taken from the palpebral conjunctiva.
Levels of MMP-9 were tested 1-month, 3-months and 6-months
postoperatively, facilitated with the use of InflammaDry solution.
Inclusion criteria: 18 years of age or older. Exclusion Criteria:
Allergy to cornstarch or Darcon, Allergy to topical anesthetic or
fluorescein dye.
Results: MMP-9 levels in group-A (LASIK) were 65±15 ng/ml,
55±12 ng/ml, 45±9 ng/ml at 1-, 3-, and 6- months, respectively.
MMP-9 levels in group-B (SMILE) were 45±13 ng/ml, 42±12 ng/
ml, 37±8 ng/ml at 1-, 3-, and 6- months, respectively. All groups
displayed statistically significant differences at the perspective
follow-up periods examined.
Conclusions: Potential differences between LASIK and SMILE
include increased inflammation levels postoperatively up to 6 months.
Commercial Relationships: Marianthi Stergiou, None;
A. J. Kanellopoulos, i-Optics (C), ISP Surgical (C), Keramed (C),
Avedro (C), Allergan (C); George Asimellis, None
Program Number: 3875 Poster Board Number: A0023
Presentation Time: 3:45 PM–5:30 PM
First Report of Simultaneous Double-Headed Pterygiectomy with
Conjunctival Autograft, Amniotic Membrane and Fibrin Glue
Jenny Ha1, Jesus A. Martinez1, 2, Yasaman Hosseini1,
Michael Korchak2, Jayson Koppinger3, 2, Sandra L. Cremers1.
1
Visionary Ophthalmology, Rockville, MD; 2Ophthalmology, Medstar
Georgetown University Hospital / Washington Hospital Center,
Washington, DC; 3Georgetown University School of Medicine,
Washington, MD.
Purpose: There is a paucity of data regarding the safety of
simultaneous double-headed pterygiectomy (SDP). We present a
new suture and cautery free technique that uses both conjunctival
autograft (CAG) and amniotic membrane graft (AM) over
conjunctival defects. We evaluated the outcomes of these SDPs.
Methods: This retrospective study examined the outcomes of 45
consecutive eyes of 43 patients who received SDP by one surgeon
(JAM). Patients received 5 mg diazepam and intraoperative
local anesthesia, in an office-based operating suite without an
anesthesiologist. Pterygia bases were excised, extensive tenonectomy
performed, and medial and lateral recti cleaned. The cornea was
polished with a diamond burr and Mitomycin C 0.025% was applied
for 2 min under the conjunctival edges using 6-8 soaked corneal
shields. Over the temporal defect, a CAG measuring 1-2mm greater
than the width (W) and anteroposterior (AP) of the defect was placed.
A cryopreserved AM (AmnioGraft, Bio-Tissue) was used over the
nasal defect. Both grafts were secured with fibrin glue (TISSEEL,
Baxter Inc). Cauterization and sutures were avoided. Postoperative
complications were recorded.
Results: Of the 45 eyes, 2 were recurrent double headed pterygia
at the time of surgery, previously performed by a different surgeon,
and 1 eye was a recurrent nasal and primary temporal pterygia. The
average nasal and temporal conjunctival defects were 7.4mm (limbal
defect, L), 10.8mm (AP) and 11.2mm (W) and 7.3mm (L), 12.6mm
(AP), and 12.0mm (W), respectively. One patient’s CAG was
supplemented with AM over the remaining temporal defect due to
insufficient superior bulbar conjunctiva; this patient did not develop
recurrence. The mean follow up time was 39.6 weeks (range 4.6142.9w). There was 1 nasal recurrence (1.1%) from a primary case at
57w requiring a second CAG. One dellen (1.1%) and 4 granulomas
(4.5%) were resolved with topical steroid drops and artificial tears.
Conclusions: To our knowledge, this the largest study of
simultaneous double-headed pterygiectomy. The use of conjunctival
autograft over the temporal defect, cryopreserved amniotic membrane
over the nasal defect, and fibrin glue in SDP eliminates trauma from
sequential surgery. This procedure is safe and results in the one of the
lowest recurrence rates demonstrated in the literature.
a) Preoperative double headed pterygium. b) Postoperative outcome
at 3 months.
Commercial Relationships: Jenny Ha, None; Jesus A. Martinez,
None; Yasaman Hosseini, None; Michael Korchak, None;
Jayson Koppinger, None; Sandra L. Cremers, None
Program Number: 3876 Poster Board Number: A0024
Presentation Time: 3:45 PM–5:30 PM
The impact of meibomian gland dysfunction on the ocular
surface parameters of glaucoma patients on long-term topical
hypotensive medications
Mehmet C. Mocan, Enes Uzunosmanoglu, Sibel Kocabeyoglu,
Murat T. Irkec. Department of Ophthalmology, Hacettepe Univ
School of Medicine, Ankara, Turkey.
Purpose: Meibomian gland dysfunction (MGD) is a prevalent
inflammatory eyelid condition that adversely affects tear stability and
may potentially exacerbate ocular surface inflammation in patients on
chronic topical glaucoma medications. The purpose of this crosssectional, observational study was to evaluate the impact of MGD on
the ocular surface parameters of medically treated glaucoma patients.
Methods: Seventy eyes of 70 glaucoma subjects with a mean age
of 65.0±9.9 years (range=46-86 years) who were on long-term
(>12 months) topical hypotensive medication were prospectively
recruited. Patients with a history of ocular or periocular surgery,
cicatrizing conjunctivitis and contact lens use were excluded. MGD
was defined as the presence of signs consistent with meibomian gland
terminal duct obstruction with or without accompanying resistance to
meibomium expressibility. MGD was categorized between grades 1-5
according to clinical severity. Ocular surface disease index (OSDI)
questionnaire was completed at the time of enrollment. Ocular
surface tests consisting of tear break-up time (BUT), ocular surface
staining with lissamine green (LG) and Schirmer test with anesthesia
were employed. Student’s t test, chi-square test and Mann-Whitney
U test were used in statistical comparisons. Forty-five healthy control
subjects with no ocular disease were also included.
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Results: The mean duration of glaucoma was 12.0±9.0 years
(range=1-35 years). Patients were on average 1.7±0.8 classes of
glaucoma medication. MGD was detected in 56 (80.0%) glaucoma
subjects. Of these 47 (83.9%) had signs consistent with mild to
moderate MGD. The ocular surface test results of both glaucoma
patients with MGD and those without MGD fared significantly
worse (p<0.001) for all investigated parameters compared to those
of healthy controls. However there were no significant differences
between OSDI scores (p=0.912), BUT (p=0.635), LG staining
scores (p=0.248), Schirmer test values (p=0.991), between glaucoma
patients MGD versus those without MGD.
Conclusions: Mild to moderate MGD is frequently encountered in
medically treated glaucoma patients. However the presence of MGD
does not appear to have an additional detrimental effect on the ocular
surface to that already induced by chronic topical medication use.
Commercial Relationships: Mehmet C. Mocan, None;
Enes Uzunosmanoglu, None; Sibel Kocabeyoglu, None;
Murat T. Irkec, None
Program Number: 3877 Poster Board Number: A0025
Presentation Time: 3:45 PM–5:30 PM
Microfluidic system based high throughput toxicity and efficacy
screening system for eyedrops in ocular surface experiments
Kyong Jin Cho2, Jongil Ju3, Rina Lee1, Dae Yu Kim5, 4,
Jeongyun Kim1. 1Biomedical Science, Dankook University Graduate
School, Cheonan, Korea (the Republic of); 2Ophthalmology, College
of Medicine, Dankook University, Cheonan, Korea (the Republic
of); 3Biomedical Engineering, College of Health Science, Korea
University, Seoul, Korea (the Republic of); 4Biomedical Engineering,
College of Medicine, Dankook University, Cheonan, Korea (the
Republic of); 5Beckman Laser Institute Korea, Cheonan, Korea (the
Republic of).
Purpose: Toxicity and efficacy tests of eye drops is complex because
the administration times and wash out duration of eyedrop should
be considered. In addition, using a pipet tip on the corneal wound
healing experiments, the size of the wound is not uniform and a
physical injury is caused. We have developed a fully automated high
throughput screening system based on the microfluidic cell culture
array to evaluate toxicity and efficacy of eyedrops.
Methods: The present micro fluidic chip used in the experiment is
high throughput drug screening system and we can observe 8 step’
concentrations at the same time. These process are operated by fully
automatic pneumatic controller that is engaged by the programmed
LabVIEW software. We evaluated the toxic effect of benzalkonium
chloride (BAK) and wound healing effect of ursolic acid on
immortalized human corneal epithelial cell line (HCE-Ts) in this
system.
Results: 1. Micro fluidic high throughput screening (μHTDS)
system for cytotoxicity of preservatives in eye drop : Cytotoxicity
of BAK on HCE-Ts cells was increased depend on treatment time
with comparing same dose conditions. Live cells with 10 minutes
treatment are quickly decrease from 0.002% comparing to 5 minutes
treatment results. The result in this study was similarity between
plate experiment of conventional method and μHTDS system. 2.
Micro fluidic high throughput screening system for wound healing
(proliferation and invasion) model : 64 cell chambers which are
arranged 8x8 forms, it means that each 8 concentrations can be
observed on 8 chambers at same concentration. 7uM was most
effective concentration of ursolic acid on migraion and invasion of
HCE-Ts cells.
Conclusions: We have developed a micro fluidic high throughput
drug screening (μHTDS) system for cytotoxicity and efficacy
screening of eyedrops with 8 different drug concentrations. Our
μHTDS system can obtain more progressed cytotoxicity and efficacy
results with minimizing the consumption of reagent, time, and labor.
This system can be useful for cytotoxicity and efficacy screening the
new drug against diverse cell type instead of human eye.
Commercial Relationships: Kyong Jin Cho; Jongil Ju, None;
Rina Lee, None; Dae Yu Kim, None; Jeongyun Kim, None
Support: This work was supported by grants (NRF2013R1A1A1010734) from Ministry of Education.
Program Number: 3878 Poster Board Number: A0026
Presentation Time: 3:45 PM–5:30 PM
Signal detection theory examination of ocular surface sensory
processing of corneal pneumatic stimuli
Varadharajan Jayakumar, Trefford L. Simpson. University of
Waterloo, Waterloo, ON, Canada.
Purpose: To evaluate the feasibility of using signal detection
theory (SDT) to examine the ocular surface sensory processing of
corneal pneumatic mechanical stimuli using the Waterloo Belmonte
aesthesiometer.
Methods: Ten asymptomatic participants were recruited in this
study. All participants were naïve to the SDT part of the experiment,
but 6 participants were familiar with the other experiments using
esthesiometer. The pneumatic stimulus was at eye temperature
(approx. 33°C) and to change strength, flow was systematically
varied. The stimulus intensities for the SDT experiment were
obtained using the ascending method of limits, in which three
ascending runs were averaged to estimate the threshold. Then, to
keep the signal constant for each participant, flow was set at 1.5X
threshold. 100 trials (demarcated using auditory cues) were presented
with a signal probability of 0.4 (i.e., 60 catch trials with stimulus
intensity of zero) and with signals and catch trials in random order.
Responses (“yes” (there was a signal) or “no” (there was no signal))
were recorded using a button box with auditory feedback for each
response. Breaks occurred after approx. 50 trials. Participants were
instructed to blink normally between stimuli, and training was
provided before the data were used. Detectability (d’), criterion (c)
and likelihood ratio (β) were calculated using false alarms and hit
rates.
Results: The average (± SE) d’ was 0.52 (± 0.12), criterion was 0.22
(± 0.10) and β was 1.11 (± 0.06). The area under the curve obtained
was 0.64 (±0.03). In addition, non-parametric SDT metrics were
calculated. The average A’ was 0.67 (±0.03) and the non-parametric
response bias β” was 0.07 (±0.03).
Conclusions: The experiment shows the feasibility of using a SDT
approach to examine ocular surface sensory processing with less
concern about the potentially important impact of participant criterion
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
changing threshold and masking meaningful effects. The data suggest
that participants in this experiment chose a relatively conservative
strategy (reporting ‘no’ to trials more commonly) but this might be
anticipated considering we designed the experiment with a relatively
large proportion of catch trials. The data also suggested that some
of the assumptions about classical SDT (underlying homoscedastic
Gaussian ‘signal + noise’ and ‘noise’ distributions) need to be
examined.
Commercial Relationships: Varadharajan Jayakumar, None;
Trefford L. Simpson, None
Support: NSERC Canada, CFI
Program Number: 3879 Poster Board Number: A0027
Presentation Time: 3:45 PM–5:30 PM
Ocular Involvement In Sjogren’s syndrome – The Singapore
Sjogren’s Syndrome Study
Rachel Lim, Dinesh Gunasekeran, Jeggrey Kam, Bernard Thong,
Rupesh V. Agrawal. Tan Tock Seng Hospital, Singapore, Singapore.
Purpose: Sjogren’s syndrome may present with both ocular and
extraocular manifestations. This study aims to compare and contrast
ocular and extraocular manifestations in consecutive patients
with Sjogren’s Syndrome (SS) recruited from rheumatology and
ophthalmology outpatient clinics in Singapore.
Methods: Computerized Physician Order Entry records of patients
with physician-diagnosed SS between 1993 and 2013 were
retrospectively analyzed. Patients were grouped into different
categories from 1 to 10 based on the number of organ-systems
involved including ocular involvement.
Results: A total of 355 patients were included, with a majority being
females(94.6%); and Chinese (84.5%). Mean duration of disease
was 8±4 years. While 135 (38.0%) fulfilled American European
Consensus Group (AECG) criteria for diagnosis, only 24 (6.8%)
fulfilled Sjögren’s International Collaborative Clinical Alliance
(SICCA) criteria for SS. Primary-SS comprised 76.9%, secondarySS 23.1%; with the most prevalent associations of rheumatoid
arthritis (11.8%) and systemic lupus erythematosus (11.3%). The
most common ocular manifestations were keratoconjunctivitis
sicca (KCS) (91.0%), anterior uveitis (2.54%), episcleritis (2.54%),
lacrimal gland enlargement (1.69%) and retinal vasculitis (0.28%).
The most common extraocular manifestations were musculoskeletal
(75.2%), lymphoreticular (47.6%) and constitutional (41.7%). All
patients had at least 1 other extraocular organ-system involved,
with median number of systems affected being 4 in 26% patients,
and maximum of 10 organ-systems affected in 1 (0.28%) patient.
Ocular-musculoskeletal involvement was the most common
combination (244 patients, 68.73%). Anti nuclear antibody (ANA)
was tested positive in 298/343 (86.9%) with titres of ≥ 1:320 in
193/298 (64.8%), rheumatoid factor (RF) was positive in 191/280
(68.2%), anti-Ro in 241/350 (68.9%), and anti-La in 90/314 (41%).
Hydroxychloroquine (83.4%), methotrexate (14.4%) and azathioprine
(12.4%) were the most commonly used therapies.
Conclusions: Keratoconjunctivitis sicca is the commonest ocular
manifestation of Sjogren’s syndrome. 90% of the patients had two
or more organ-systems involved, with musculoskeletal involvement
being the most common.
Commercial Relationships: Rachel Lim, None;
Dinesh Gunasekeran, None; Jeggrey Kam, None; Bernard Thong,
None; Rupesh V. Agrawal, None
Program Number: 3880 Poster Board Number: A0028
Presentation Time: 3:45 PM–5:30 PM
HLA class I genes associated with Cold Medicine related StevensJohnson Syndrome with Severe Ocular Complications in the
Brazilian population
Tais H. Wakamatsu1, Mayumi Ueta2, Katsushi Tokunaga3,
Yukinori Okada4, Renata R. Loureiro1, Karita A. Costa1,
Juliana M. Sallum1, José Milhomens1, Chie Sotozono5, Jose
Alvaro P. Gomes1, Shigeru Kinoshita5. 1Federal University of Sao
Paulo, Sao Paulo, Brazil; 2Department of Frontier Medical Science
and Technology for Ophthalmology, Kyoto Prefectural University of
Medicine, Kyoto, Japan; 3Department of Human Genetics, University
of Tokyo, Tokyo, Japan; 4Human Genetics and Disease Diversity,
Tokyo Medical and Dental University, Tokyo, Japan; 5Department of
Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto,
Japan.
Purpose: Stevens-Johnson syndrome (SJS) and toxic epidermal
necrolysis (TEN) are immune-complex mediated hypersensitivity
diseases that typically involve the skin and mucous membranes. Here
we investigated etiologic factors such as causative drugs and the
association between human leukocyte antigen (HLA) class I genes
and cold medicine-related SJS/TEN (CM-SJS/TEN) with severe
ocular complications (SOC) in Brazilian patients.
Methods: We enrolled 74 Brazilian patients with SJS/TEN with
SOC and 135 healthy Brazilian volunteers. The patients’ ethnicity
was Pardo (51%), European ancestry (40%) and others. All were
interviewed with respect to possible etiologic factors.
HLA class I (HLA-A, -B, -C) were examined to determine whether
there was a genetic predisposition for CM-SJS/TEN with SOC.
Results: We identified cold medicine as the main causative
drug (39 patients - 53%). HLA-A*66:01, HLA-B*44:03 and
HLA-C*12:03 were associated, and HLA-A*11:01, HLA-B*08:01,
and HLA-B*51:01 were inversely associated with Brazilian CMSJS/TEN with SOC. After dividing in Pardo and European ancestry,
HLA-A*66:01 was associated with among individuals both Pardo
and European ancestry; HLA-B*44:03 and HLA-C*12:03 among
individuals with only European ancestry.
Conclusions: In conclusion, our findings suggested that
HLA-A*66:01 might be a marker in Pardo and European ancestry
and HLA-B*44:03 and HLA-C*12:03 might be markers in only
European ancestry. Moreover, HLA-A*11:01 might be a marker of
resistance to CM-SJS/TEN with SOC.
Commercial Relationships: Tais H. Wakamatsu, None;
Mayumi Ueta, None; Katsushi Tokunaga, None; Yukinori Okada,
None; Renata R. Loureiro, None; Karita A. Costa, None;
Juliana M. Sallum, None; José Milhomens, None; Chie Sotozono,
None; Jose Alvaro P. Gomes, None; Shigeru Kinoshita, None
Program Number: 3881 Poster Board Number: A0029
Presentation Time: 3:45 PM–5:30 PM
EVALUATION OF CHANGES IN OCULAR SURFACE AFTER
VITRECTOMY SURGERY
Chiara Del Noce, Francesca Bruzzone, Silvio Lai,
Maurizio . ROLANDO, Carlo E. Traverso. DINOGMI, University of
Genoa Eye Clinic, Genoa, Italy.
Purpose: To evaluate the relationship between vitrectomy surgery
and the alteration of ocular surface homeostasis.
Methods: A total of 25 phakic patients with vitreo-retinal interface
pathologies and without ocular surface diseases were enrolled. All
underwent the conventional 25/23 gauge pars plana vitrectomy
surgery. Evaluation were made using a VAS symptoms questionnaire
(subjective parameters scored from 0 to 10) and a slit lamp (blink
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
rate, palpebral and bulbar conjunctival irritation, BUT, meniscus,
fluorescine and lissamine green staining).
Results: One year after surgery, according to the VAS classification
the ranking resulted: desire to keep the eyes closed, foreign body
sensation, burning, pain, roughness and heat. Blink was incomplete
and more frequent in 32 % of patients. Approximately 98% of
patients revealed both palpebral and conjunctival irritation. BUT
was reduced in 80% of patients. The meniscus was irregular and
thinned in 88% of patients. According to the Oxford scale, fluorescein
staining was positive in 60% of patients: 73% Grade I and 27%
Grade II while lissamine green staining was positive in 84% of
patients: 52% of grade I, 28% grade II and 20% of grade III.
Conclusions: Vitrectomy surgery 23/25 gauge with transconjunctival
approach might give ocular surface alterations and ocular discomfort.
Commercial Relationships: Chiara Del Noce,
None; Francesca Bruzzone, None; Silvio Lai, None;
Maurizio ROLANDO, None; Carlo E. Traverso, None
Program Number: 3882 Poster Board Number: A0030
Presentation Time: 3:45 PM–5:30 PM
Deletion of the Vitamin D Receptor Affects Meibomian Gland of
Mice
Kai Jin1, Motoko Kawashima1, Masataka Ito2, Kokoro Sano1,
Kazuo Tsubota1. 1Ophthalmology, Keio University, school of
medicine, Tokyo, Japan; 2National Defense Medical College,
Saitama, Japan.
Purpose: The vitamin D receptor (VDR) and vitamin D metabolic
associated enzymes have been determined in various kinds of cells
in both mouse eyes and human eyes. In this study, we examined the
effects of VDR knockout on the pathological changes of meibomian
gland (MG), eyelid tissue, and the secretion of tear fluid.
Methods: Eyelids of VDR knockout mice aged 3 months (n=10) and
7 months (n=10) were investigated and compared with 20 normal
eyelids from age-matched wild type C57/B6J mice. Microscopy
examination and fluorescein staining were performed to check the
ocular surface. Cotton thread test, MILLIPLEX® MAP and qRTPCR were used to analyze the tear fluid and eyelid tissue. We also
examined the haemotoxylin and eosin staining of the eyelid and
lacrimal gland (LG), and observed the immunofluorescent staining
for cytokeratins CK1, CK5, CK6.
Results: Keratinous cysts of MG, partial dropout of MG, and
thinning and whitening of the cilia were shown in VDR knockout
mice aged 3 months or 7 months. With aging, MG cysts progressively
grew while the obstruction, lump or nodule of MG on the
conjunctival surface can be observed. The secretion of tear fluid from
VDR knockout mice significantly increased (P<0.05) and mRNA
expression of mucin and some inflammatory markers in eyelid tissue
were also upregulated significantly (P<0.05). The lid margin of VDR
knockout mice showed more expression of CK1. In addition, there
were no significant differences of corneal fluorescein staining or
haemotoxylin and eosin staining of the lacrimal gland (LG).
Conclusions: Vitamin D receptor knockout influences the
pathogenesis of MG, hyper-keratinization of the eyelid, and secretion
of tear fluid.
Commercial Relationships: Kai Jin; Motoko Kawashima, None;
Masataka Ito, None; Kokoro Sano, None; Kazuo Tsubota, None
Program Number: 3883 Poster Board Number: A0031
Presentation Time: 3:45 PM–5:30 PM
Calculating the health economic burden of microbial keratitis
(MK) admission in a tertiary referral centre in the UK
Jasvir Virdee1, George Moussa1, Nicholas Gooch2, Jesse Kigozi2,
Cristina Penaloza2, Saaeha Rauz1. 1Academic Unit of
Ophthalmology, The University of Birmingham, Birmingham, United
Kingdom; 2Health Economics Unit, The University of Birmingham,
Birmingham, United Kingdom.
Purpose:
The UK health care system (National Health Service, NHS) provides
medical care that is available to all and free for all. MK is the
commonest ophthalmic emergency admission in the developed world,
with large cost burden to the NHS and few health economic studies.
This study was designed to provide a quantification of direct costs of
inpatient care for MK versus income generated though coding in a
supra-regional tertiary centre in the UK.
Methods:
Extensive clinical, demographic and economic data were collected
retrospectively for a period of twelve months (Jan-Dec 2013) for
101 consecutive patients admitted with MK on a validated electronic
proforma. Direct cost of admission (COA) was calculated using
national reference costs for individual patients for various parameters
including length of stay in days (LOS), topical medication, cost of
microbiological services and cost of an ophthalmic hospital bed,
together with health economic analytical assumptions to generate
profit/deficit profiles based upon actual income and estimated
expenditure A one-way ANOVA analysis was performed to compare
groups.
Results:
The total income generated through discharge coding for all patients
was £267,028, whilst calculated cost of admission was £382,473,
giving an overall deficit of £115,445 per annum. The median
individual deficit was £779 (interquartlie range £1880). The most
critical factor driving the cost deficit was length of stay with median
cost neutrality achieved between days 5 and 6 (table 1). Severity of
microbial keratitis (graded according to Keay et al) was not found to
be a significant factor in driving costs (Table 2). Surgical intervention
(corneal gluing, evisceration) drove up costs of care.
Conclusions:
Health economic analysis shows that length of stay is the key driver
for direct costs of care for patients admitted with microbial keratitis
with the pivotal LOS of 4 days. The microbial keratitis treatment
pathway should encourage discharge of patients who are able to selfadminister treatment after the sterilisation phase to enable financial
parity. Prospective data collection is required to refine direct cost
analyses and to evaluate the clinical and financial burden (indirect
costs) of the impact of visual morbidity on quality of life.
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Commercial Relationships: Jasvir Virdee, None; George Moussa,
None; Nicholas Gooch, None; Jesse Kigozi; Cristina Penaloza,
None; Saaeha Rauz, None
Program Number: 3884 Poster Board Number: A0032
Presentation Time: 3:45 PM–5:30 PM
Using corneal chemical detection thresholds to select subjects for
ocular surface discrimination sensory panels
Trefford L. Simpson4, 1, Nancy J. Keir2, William Ngo3, 1,
Yunwei Feng4, 1. 1University of Waterloo, Waterloo, ON, Canada;
2
Cooper Vision, Pleasanton, CA; 3School of Optometry & Vision
Science, Centre for Contact Lens Research, Waterloo, ON, Canada;
4
School of Optometry & Vision Science, Waterloo, ON, Canada.
Purpose: To determine if we can we select sensitive (‘good’) and less
sensitive (‘bad’) participants in a sensory panel examining interocular
judgments of contact lens comfort, using Belmonte esthesiometry.
Methods: 49 subjects (adapted contact lens wearers) had chemical
detection thresholds (% added CO2) measured with the Waterloo
Belmonte Esthesiometer, on 3 occasions (at least a day apart),
using the ascending method of limits. From the averages of these
3 measurements, 25 subjects were grouped as more sensitive
(thresholds at or below median) or less sensitive (thresholds above
median). Subjects also interocularly matched the discomfort of
3 optimally fit contact lenses using the mechanical or chemical
stimuli of the esthesiometer and scaled discomfort using magnitude
estimation. We tested a simple hypothesis: Subjects in the sensitive
group would be better at interocular matching - regression pneumatic
stimulus match to rated discomfort would be linear with a positive
slope, but the control group would not perform well, with random
slopes (positive, 0 and negative). Bayesian mixed modeling (with
vague priors) estimated the group slopes (among others) and the
distributions of these estimates were compared in the sensitive and
control groups defined by chemical detection thresholds. R and rjags
were used for the data analysis.
Results: For the ‘bad’ group the regression slopes mode was 0.05
and the 95% HDI was -0.526-0.731. The figure (of the posterior
distribution of the slope estimates) shows that for the ‘good’ group
the mode was shifted and the 95% HDI did NOT include zero.
Conclusions: Chemical thresholds can be used to separate sensitive
and insensitive subjects who can match contact lens discomfort in a
‘well behaved’ way and therefore pneumatic esthesiometry may be
used to select sensory panels.
Commercial Relationships: Trefford L. Simpson, Alcon (F);
Nancy J. Keir, None; William Ngo, None; Yunwei Feng
Support: NSERC Canada, CFI, Alcon IIS.
Program Number: 3885 Poster Board Number: A0033
Presentation Time: 3:45 PM–5:30 PM
Ocular Surface Homeostatic Surveillance by Th17 Cells in the
Closed Eye
Cameron K. Postnikoff, Kelly K. Nichols. Vision Sciences, University
of Alabama at Birmingham, Birmingham, AL.
Purpose: T cell infiltration of the ocular surface is commonly
only associated with inflammation and diseases such as dry eye.
Neutrophils are well known to populate the closed eye during eyelid
closure, and have been shown to signal the adaptive immune response
in other organ systems. The purpose of this investigation was to
catalogue and phenotype various leukocytes of the closed eye.
Methods: Eleven healthy participants were recruited and were
trained to wash the ocular surface with phosphate buffered saline for
at-home self-collection of tear film leukocytes following a full night
of sleep. Cells were isolated and counted and were incubated with
fluorescently-labeled antibodies against CD45, CD14, CD15, CD16,
CD11b to identify neutrophils and monocytes. Antibodies against
CD45, CD3, CD4, CD8, CD196, and CD161 were used to identify
T cells. A Becton Dickinson (BD; San Jose, CA) proprietary fixable
viability stain was used to exclude dead cells from analysis. A BD
LSR II flow cytometer was used for all analysis.
Results: Neutrophils were the most dominant of all cell populations,
representing roughly 58% of all leukocytes. Monocytes were
not readily observed, representing only a small portion of cells
(<2%). Both CD4+ and CD8+ T cells were found in the ocular
surface eyewash after a full night of sleep. CD4+ were more
abundant, representing approximately 7% of the total leukocyte
population, while CD8+ cells were roughly 1.5% of the total
leukocyte population. Finally, Th17 cells, as identified as being
CD196+CD161+ represented, on average, 20% of the total CD4+ T
cells.
Conclusions: This investigation demonstrated T cell surveillance
of the ocular surface, during sleep. Combined neutrophil and T
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
cell infiltration suggests that closed eye homeostasis is complex,
and future studies seek to identify the mechanisms of leukocyte
interaction and activation, and ultimately determine how
dysregulation may contribute to disease.
Commercial Relationships: Cameron K. Postnikoff, None;
Kelly K. Nichols, None
Support: Thank you to Dr. Karen Ersland for her assistance
with flow cytometry. The authors also acknowledge the UAB
Comprehensive Flow Cytometry Core, which is supported by NIH
P30 AR048311 and NIH P30 AI27667.
Program Number: 3886 Poster Board Number: A0034
Presentation Time: 3:45 PM–5:30 PM
GRADING BULBAR REDNESS USING THE KERATOGRAPH
5M. CORRELATIONS WITH EFRON AND MCMONNIES
SCALES
Néstor Ventura Abreu1, Francisco Pérez Bartolomé1, Claudia Sanz
Pozo1, Jose María Martínez de la Casa1, Javier Moreno-Montanes2.
1
Ophthalmology, Hospital Clínico San Carlos, Madrid, Spain;
2
Ophthalmology, Clínica Universitaria de Navarra, Pamplona, Spain.
Purpose: To examine interobserver reproducibility of the Efron and
McMonnies bulbar redness (BR) scales and establish correlations
between these scales and the new Keratograph 5M.
Methods: This was an observational cross- sectional study.
203 eyes of 203 subjects (50 controls, 153 under treatment with
topical hypotensive drugs for glaucoma) with varying degrees of
conjunctival hyperaemia were scored automatically for BR using
the Keratograph 5M. These scores were then correlated with the
gradings provided by two image-based subjective scales: Efron and
McMonnies. The interobserver reproducibility of both scales was also
evaluated.
Results: Excellent reproducibility was observed for both the Efron
(Weighted K= 0.897, 95% CI 0.823 – 0.904) and McMonnies
(Weighted K= 0.783, 95% CI 0.752 – 0.795) scales. Keratograph
BR scores (overall redness) and the scores obtained with both Efron
(Spearman’s Rho= 0.43, P<0.001) and McMonnies (Spearman’s
Rho= 0.48, P< 0.001) were significantly correlated. Redness scores
provided for the bulbar and limbar nasal and temporal quadrants also
correlated well with the two subjective scales.
Conclusions: The Keratograph 5M method of assessing BR seems to
be a good alternative to the subjective interpretation of image-based
BR scales.
Commercial Relationships: Néstor Ventura Abreu, None;
Francisco Pérez Bartolomé, None; Claudia Sanz Pozo, None; Jose
María Martínez de la Casa, None; Javier Moreno-Montanes,
None
Program Number: 3887 Poster Board Number: A0035
Presentation Time: 3:45 PM–5:30 PM
Novel pterygium animal model using White New Zealand rabbit
and murine fibroblasts (NIH 3T3 cell line)
Julio C. Hernandez2, Jorge E. Valdez2, Judith Zavala2,
Jorge Valenzuela1. 1Tecnologico de Monterrey, School of Medicine,
Monterrey, Mexico; 2Ophthalmology and Visual Sciences
Research Chair, Tecnologico de Monterrey School of Medicine,
MONTERREY, Mexico.
Purpose: To develop an animal model for pterygium using
subconjunctival injection of murine fibroblast cells (NIH 3T3 cell
line) in white New Zealand rabbits
Methods: Under general and topical anesthesia with tetracaine
hydrochloride, six 3 month-old white New Zealand rabbits received
subconjuntival injection of 20,000 murine fibroblast cells (NIH
3T3 cell line) and 5 µL of Matrigel(Corning Inc, NY, US) on the
perilimbal temporal bulbar conjunctiva of the right eye using a 1ml
30G needle. Left eyes were injected with 5 µL of Matrigel under
the perilimbal temporal bulbar conjunctiva (control). Eyes were
photographed under a magnification of 16x using a 12 megapixel
digital camera attached to the microscope (WPI, Inc. FL, US) on day
1, 3 and 7. Conjunctival vascularization was measured on day 1,3
and 7 by analyzing images using Adobe PhotoshopCS5 (Adobe Inc,
San Jose, CA) color histograms to measure red pixel saturation on a
6x6 mm area on the site of injection. Area of corneal and conjunctival
fibrovascular tissue formation was assessed by analyzing the images
on day 3 and 7 using area measurement software (Adobe Photoshop
CS5). Statistical analysis was performed using t-test for mean
comparison, statistical significance was considered with a p value
<.05
Results: Red pixel saturation for right and left eyes was
102.53±11.07pxs and 104.84±8.34pxs (p=.609), 176.15±6.35px and
168.29±18.89pxs (p=.325), 221.58±33.85pxs and 168.70±17.75pxs
(p=.006) on day 1, 3 and 7 respectively. Area of fibrovascular tissue
on the site of injection in the right and left eyes was 14.32±3.47mm2
and 8.41±2.08mm2 (p=.002), 41.18±4.40 mm2 and 14.69±2.81 mm2
(p<.001) on day 3 and 7 respectively. Significant difference was
observed between day 3 and 7 on right eyes (p<.001) and left eyes
(p=.002) when comparing the area of fibrovascular growth. Red
pixel saturation was higher for both eyes on day 7 when compared to
day 3, but was only the different was only significant for right eyes
(p=.017)
Conclusions: Subconjunctival injection of murine fibroblast cells
and Matrigel in white New Zealand rabbits was more effective for
inducing conjunctival vascularization and fibrovascular tissue growth
on the site of injection and surrounding cornea than subconjunctival
Matrigel alone. A novel animal model for pterygium is presented with
potential use for research on molecular mechanisms of pathogenesis
and new treatment strategies.
Commercial Relationships: Julio C. Hernandez, None;
Jorge E. Valdez, None; Judith Zavala, None; Jorge Valenzuela,
None
Program Number: 3888 Poster Board Number: A0036
Presentation Time: 3:45 PM–5:30 PM
Lax Eyelid Syndrome, Obstructive Sleep Apnea Syndrome
(OSAS), and Ocular Surface Inflammation
Mackenzie Becker1, Clayton Kirk1, Ramsudha Narala2,
Sunita Kumar1, William Adams1, Charles S. Bouchard1.
1
Ophthalmology, Loyola University Medical Center, Maywood, IL;
2
Kresge, Detroit, MI.
Purpose: Lax eyelid syndrome (LES) describes the association of
distensible “floppy” eyelids and chronic papillary conjunctivitis that
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
has a well-known association with OSAS. Eyelid histopathology
suggests that LES may result from overexpression of matrix
metalloproteinase 9 (MMP-9) related elastin degradation. However,
the relationship between the degree of eyelid laxity and OSAS and
elucidation of MMP-9 levels in LES subjects has not been clearly
demonstrated. The purpose of this study therefore was: 1) establish
the prevalence and degree of eyelid laxity in patients with newly
diagnosed OSA; 2) to evaluate the presence of MMP-9 in the tear
film of patients with LES; and 3) to compare the current LES grading
systems with a newly designed “laxometer.”
Methods: Following OSAS evaluation by polysomnography, subjects
were referred for eyelid laxity determination and an MMP tear
assay. Measurements of the degree of eyelid laxity were determined
for each eye using the following four methods: 1) degree of tarsal
conjunctiva exposure; 2) ease of upper eyelid eversion; 3) degree of
medial canthal tendon laxity; and 4) distensability of the laxometer.
The MMP-9 tear film assay was conducted using a commercially
available assay.
Results: There was a small but non-significant positive association
between laxometer measurement and duration of eyelid eversion (τ
= 0.16, p = .29). Conversely, there was a small but non-significant
negative association between laxometer measurement and duration of
eyelid eversion (τ = -0.19, p = .25). A nonparametric Kreskas-Wallis
test was used to assess whether eyelid elasticity varied as a function
of sleep apnea severity. Although the study was unpowered for
statistical significance, a positive trend was found between degree of
sleep apnea and eyelid laxity determined by laxometer measurements.
For the MMP-9 levels, 14 of 16 eyes (89%) with LES had a positive
MMP-9 assay (p< .001).
Conclusions: Although our current study sample is small, the
laxometer data suggests an association between the severity of
LES and the severity of OSAS that may have clinical relevance.
The findings of elevated MMP-9 supports a notion that MMP plays
a role in elastin degradation that may be associated with chronic
conjunctivitis and reactive ocular surface typically found in patients
with LES. We are currently recruiting additional patients to better
assess the relationship between lid laxity and OSAS.
Commercial Relationships: Mackenzie Becker, None;
Clayton Kirk, None; Ramsudha Narala, None; Sunita Kumar,
None; William Adams, None; Charles S. Bouchard, None
Support: Richard A. Perritt Charitable Foundation
Program Number: 3889 Poster Board Number: A0037
Presentation Time: 3:45 PM–5:30 PM
A novel murine behavioral model of ocular neuropathic pain
Romulo Albuquerque1, Jooyoung Cho1, Bradley Taylor2,
Jayakrishna Ambati1. 1Ophthalmology, University of Kentucky,
Lexington, KY; 2Physiology, University of Kentucky, Lexington, KY.
Purpose: Neuropathic pain is a complex chronic state arising from
injured and dysfunctional nerve tissue that is poorly responsive
to analgesic drugs. Neuropathic pain also affects the eye. Ocular
neuropathic pain, disguised as ocular surface disease, is often
unrecognized and undertreated by clinicians (Rosenthal and Borsook
2015). We tested the hypothesis that alkali burn injury to the ocular
surface leads to neuropathic changes in the ocular-trigeminal system
and can be used as a molecular and behavioral model to study ocular
neuropathic pain.
Methods: Wild-type C57Bl6 mice were used. The alkali burn injury
model was performed as previously shown (Anderson et al. 2014).
Satellite glia activation and neuronal sensitization were studied in
the trigeminal ganglia by immunohistochemistry using glial fibrillary
acidic protein (GFAP) and neuropeptide-Y (NP-Y). Ocular pain
behavior was assessed by eye wiping behavior after application of
hypertonic (5M) saline solution to the ocular surface. Ocular pain
behavior was tested before and after intra-peritoneal gabapentin
(100mg/kg) administration.
Results: Alkali injury to the ocular surface induced expression
of GFAP and NP-Y in trigeminal ganglion satellite glial cells
and neurons respectively. Alkali injury also increased eye wiping
behavior in the injured eye, but not in the contralateral eye up to one
week after injury. Increased eye wiping behavior was significantly
reduced by treatment with intra-peritoneal gabapentin.
Conclusions: Our model combines two previous ophthalmology
models: a surface alkali burn injury to model ocular inflammation
(Anderson, Zhou et al. 2014), together with an ocular pain testing
involving topical application of hypertonic saline to elicit an eyewiping response, a behavioral index of pain. Here we show that eyewiping behavior on the eye ipsilateral but not contralateral to alkali
burn was augmented, suggesting sensitization of ocular nociception.
These behavior changes were paralleled by increased levels of GFAP
and NP-Y in the trigeminal ganglia satellite glial cells and neurons.
Eye wiping behavior was abrogated by systemic administration of
gabapentin, an antiseizure drug used for the treatment of neuropathic
pain. The ability of an analgesic drug like gabapentin to decrease
augmented eye wiping suggests that this response is neuropathic
in nature and provides a novel model to study the molecular
mechanisms of ocular neuropathic pain.
Commercial Relationships: Romulo Albuquerque; Jooyoung Cho,
None; Bradley Taylor, None; Jayakrishna Ambati, iVeena
Pharmaceuticals (I), Inflammasome Therapeutics (I), iVeena
Holdings (I), iVeena Pharmaceuticals (S), Inflammasome
Therapeutics (S), University of Kentucky (P), iVeena Holdings (P),
iVeena Pharmaceuticals (P), iVeena Holdings (S), Allergan (R), Olix
Pharmaceuticals (F), iVeena Delivery Systems (I), iVeena Delivery
Systems (S), iVeena Delivery Systems (P)
Program Number: 3890 Poster Board Number: A0038
Presentation Time: 3:45 PM–5:30 PM
Prosthetic Replacement of the Ocular Surface Ecosystem
Therapy for Sjogren’s Syndrome Patients
Billy X. Pan, Gloria B. Chiu, Martin Heur. Ophthalmology, USC Eye
Institute, Los Angeles, CA.
Purpose: Sjogren’s syndrome is an autoimmune disease
that predominantly affects the salivary and lacrimal glands.
Immune-mediated destruction of the lacrimal gland produces
keratoconjunctivitis sicca that can be difficult to manage with
conventional dry eye therapies. Prosthetic Replacement of the Ocular
Surface Ecosystem (PROSE) therapy utilizes a custom-designed
scleral device that vaults the entire cornea and limbus, bathing the
ocular surface in preservative-free sterile saline. PROSE therapy
has been shown to be beneficial in numerous other ocular surface
diseases. The objective of this study was to evaluate the utility of
PROSE therapy for patients with Sjogren’s related dry eye disease
with outcomes based on visual acuity and function.
Methods: The University of Southern California Institutional Review
Board approved this study. A retrospective review from July 2009 to
October 2015 of patients referred to USC Eye Institute for PROSE
therapy consultation identified 37 patients with Sjogren’s syndrome
related dry eye disease. Eight patients were excluded because they
did not complete the fitting process for reasons such as being lost
to follow-up, inability to obtain insurance authorization, difficulty
with insertion and removal of the lens, or desire to continue with
topical therapy. One additional patient was excluded because he was
previously fit with PROSE at another institution. Of the 28 remaining
patients, visual acuity, before and after PROSE fitting, was assessed
using a Snellen chart under standardized conditions. Visual function
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
was assessed before and after PROSE fitting using the Ocular Surface
Disease Index (OSDI) questionnaire. Wilcoxon signed-rank test was
used to evaluate changes in visual acuity and OSDI scores.
Results: Fifty-six eyes from 27 female patients and 1 male patient
were included in this initial study. The average age was 52.3 years
with a range of 30 to 65 years. The average visual acuity improved
from 0.18 ± 0.20 logMAR (approximately 20/30) pre-PROSE to 0.08
± 0.15 logMAR (approximately 20/25) post-PROSE (Z = -4.42; p <
0.01). Nine patients completed both the pre- and post-PROSE OSDI
questionnaire. The average OSDI score improved from 66.9 ± 23.1
pre-PROSE to 31.4 ± 12.6 post-PROSE (Z = -3.24; p < 0.01).
Conclusions: The data suggest that PROSE therapy is an effective
treatment for patients suffering from Sjogren’s related dry eye disease
Commercial Relationships: Billy X. Pan, None; Gloria B. Chiu,
None; Martin Heur, None
Program Number: 3891 Poster Board Number: A0039
Presentation Time: 3:45 PM–5:30 PM
Prevalence and risk factors of superior limbal
keratoconjunctivitis in Graves’ disease
Anfal Almazrooei1, Omnia Hamam1, Laurence Dupasqiuer2,
Michel Berche1, Emmanual Heron2, Christophe Baudouin1,
Antoine Labbé1. 1Service d’Ophtalmologie, CHNO des QuinzeVingts, Paris, France; 2Department of Internal Medicinel, CHNO des
Quinze-Vingts, Paris, France.
Purpose:
To evaluate the prevalence and risk factors of superior limbal
keratoconjunctivits (SLK) in patients with Graves’ disease.
Methods:
Thirty-five patients diagnosed as having SLK from a cohort of
459 patients with Graves’ disease were retrospectively evaluated.
SLK was defined as a superior ocular surface fluorescein staining
associated with superior conjunctival laxity and inflammation of the
superior tarsal and bulbar conjunctiva. Demographic data and clinical
features were evaluated and all patients had a complete evaluation
of the ocular surface. Graves’ orbitopathy was diagnosed on the
basis of the criteria of the European Group On Graves’ orbitopathy
(EUGOGO) and the disease activity was quantified according to the
Clinical Activity Score (CAS) and Ocular Surface Disease Index
(OSDI) questionnaires.
Results:
Within the 35 patients with SLK, 75.8% were female. Most of the
patients were Caucasian (48.5%) followed by African (27,3%) and
Asian (18,2%). Mean age was 45.48 ± 13.2 years. Considering
smoking habit, 42.4% of patients never smoked while 39.4% were
current smokers. Diabetes was found in 3.2% of patients. Bilateral
SLK was observed in 54.8% of cases. Although there were good
correlations between CAS score and antibodies against TSH receptor
(TRAK) (p=0.03) and OSDI (p<0.05), there was no significant
correlation between CAS score and the presence of SLK.
Conclusions:
SLK is not unusual in patient with Graves’ disease however it is not
related to its severity. Careful ocular surface evaluation has to be
performed in Graves’ orbitopathy to diagnose SLK.
Commercial Relationships: Anfal Almazrooei, None;
Omnia Hamam; Laurence Dupasqiuer, None; Michel Berche,
None; Emmanual Heron, None; christophe baudouin, None;
Antoine Labbé, None
Program Number: 3892 Poster Board Number: A0040
Presentation Time: 3:45 PM–5:30 PM
Modulating the toxic effects of benzalkonium chloride on human
corneal epithelial cells in vitro
Elham Ghahari, Mohammadreza Ghahari, Sanaz Gidfar,
Behrad Y. Milani, Sara Sanjari, Medi Eslani, Thasarat S. Vajaranant,
Ahmad A. Aref, Ali R. Djalilian. ROI, University of Illinois at
Chicago, Chicago, IL.
Purpose: Many topical ophthalmic medications have documented
ocular surface toxicity attributed to preservatives, most commonly
benzalkonium chloride (BAK). The main purpose of this study is to
determine the critical cytotoxic BAK concentration for telomerizedimmortalized Human Corneal Epithelial Cells (HCECs) using a series
of concentrations and different exposure time, and find a way to
prevent or treat this damage.
Methods: HCECs was plated in 96-well plates and incubated for 1
day. Culture medium was then replaced by basal media with different
BAK concentrations (0.0005% to 0.005% in 0.0005 increment steps),
incubated for 5, 10, or 15 minutes. In another set of experiments,
the cells were incubated in a specific conditioned media 30 minutes
before BAK application. Cell proliferation/viability was assessed by
MTT assay.
Results: BAK treatment time of 10 minutes was selected. BAK toxic
concentration was 0.0015. Our results suggest that conditioned media
has some degrees of protective effects against BAK ocular toxicity.
HCECs pre-treated with the specific conditioned media demonstrated
71% cell viability in comparison to 56% for un-conditioned media.
Conclusions: These results indicate that BAK induced cell damage
might be reduced using a specified conditioned media. This may be
used clinically to decrease the adverse effect of topical eye drops.
Commercial Relationships: Elham Ghahari;
Mohammadreza Ghahari, None; Sanaz Gidfar, None;
Behrad Y. Milani, None; Sara Sanjari, None; Medi Eslani,
None; Thasarat S. Vajaranant, None; Ahmad A. Aref, None;
Ali R. Djalilian, None
Program Number: 3893 Poster Board Number: A0041
Presentation Time: 3:45 PM–5:30 PM
Corneal bioimpedance evaluation in an animal model of anterior
surface inflammation
Mario Crespo-Moral1, Alfredo Holgueras-López1, Anton Guimerà2, 3,
Rosa Villa2, 3, Miguel Maldonado1. 1IOBA - University of Valladolid,
Valladolid, Spain; 2Institut de Microelectrònica de Barcelona IMBCNM (CSIC), Barcelona, Spain; 3CIBER-BBN, Networking Center
on Bioengineering, Biomaterials and Nanomedicine, Zaragoza,
Spain.
Purpose: Changes in the corneal barrier function can be shown by
measuring the corneal bioimpedance. We evaluated if we were able
to charactericise the damage produced in an experimental model of
anterior surface inflammation (ASI) using a tetrapolar impedance
meter.
Methods: ASI was produced in 14 female New Zealand white rabbits
by two different methods. In the first one (n=4) 40ul of 3% croton oil
diluted in 2-ethoxyethanol was administered into the fornix of one of
the eyes from each animal while the vehicle was administered into
the other one. In the second one (n=10) 20ul of 3% croton oil diluted
in dimethyl sulfoxide (DMSO) were administered into the fornix of
one of the eyes from each animal while the vehicle was administered
into the other one. Bioimpedance measures, mucous secretion (MS),
chemosis (CH), neovascularization (NV) and corneal staining (CS)
were assessed at each time point evaluation (baseline, 6, 24, 48
and 148 hours after exposure). Corneal tissue was extracted at the
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
endpoint, evaluated by light microscopy, and inmunostained for ZO-1
and ZO-2.
Results: In the first group we show that MS, CH and CS were present
in the 6, 24 and 48 hours evaluations in 3/4 of the cases while only
MS and CS were present in the remaining case after 6 and 24 hours.
In the second group we show MS in every animal after 6 hours and
CH in 90% of the cases after 6 hours and 80% after 24 hours, CS
is not present at any time point of the evaluation. Bioimpedance
measures show low impedance values after 6 and 24 hours in the first
group while there were no differences with the basal values at other
times. In the second group higher impedance values were observed 6
and 24 hours after the administration in 70% of the cases while there
were no differences at anyother time point.
Conclusions: We had been able to reproduce a short time animal
model of ASI and measure its bioimpedance at different time points.
The combination of croton oil 3% in DMSO is the best way of the
two methods to study the effect of inflammation over the ocular
surface because it do not damage corneal epithelium integrity, even
though it produces shorter time effects. The bioimpedance measures
reflect damage in the corneal barrier function at the same times that
corneal staining is observed.
Commercial Relationships: Mario Crespo-Moral, None;
Alfredo Holgueras-López, None; Anton Guimerà, Laboratorios
SALVAT S.A. (P); Rosa Villa, Laboratorios SALVAT S.A. (P);
Miguel Maldonado, Laboratorios SALVAT S.A. (P)
Program Number: 3894 Poster Board Number: A0042
Presentation Time: 3:45 PM–5:30 PM
Reproducibility and Outcomes of Simple Limbal Epithelial
Transplantation (SLET) technique
Alexandra Abdala, Arturo J. Ramirez-Miranda, Alejandro Lichtinger,
Alejandro Navas, Enrique O. Graue-Hernandez. Instituto de
Oftalmologia Conde de Valenciana, Mexico City, Mexico.
Purpose: To report the results of simple limbal epithelial
transplantation (SLET) for limbal stem cell deficiency (LSCD) at an
ophthalmological institution in Mexico City.
Methods: Five patients (5 eyes) with LSCD due to chemical
injuries (3 eyes), thermal injury (1 eye) and failure of tectonic
sclerokeratoplasty after corneal melting of tectonic patch graft
secondary to rheumatoid arthritis perforation (1 eye). All patients had
unilateral involvement and received an autologous graft of limbal
stem cells from the contralateral healthy eye, technique described by
Sangwan. Preoperative best-corrected visual acuity, postoperative
best-corrected visual acuity, previous surgeries, side effects on the
donor eye, time from injury to surgery, recurrence and postoperative
complications were evaluated. Paired t test was used for statistical
analysis.
Results: Preoperative mean best-corrected visual acuity was
1.55LogMAR (1 to 2.3LogMAR). The mean follow-up was 10.2
months (6 to 14 months), with postoperative mean best-corrected
visual acuity of 0.33LogMAR (0.17 to 0.47LogMAR). The ocular
surface was improved in all patients, however focal recurrence with
superficial corneal vascularization was observed on the affected eye.
The donor eye did not have complications.
Conclusions: The result of our study suggests that SLET is a safe and
effective treatment option for the management of unilateral LSCD,
improving the ocular surface for future corneal procedures.
Commercial Relationships: Alexandra Abdala, None;
Arturo J. Ramirez-Miranda, Carl Zeiss Meditec (C);
Alejandro Lichtinger, None; Alejandro Navas, Alcon Laboratories
Inc (C); Enrique O. Graue-Hernandez, None
Program Number: 3895 Poster Board Number: A0043
Presentation Time: 3:45 PM–5:30 PM
Topical Interferon Alpha-2b In Giant Squamous Ocular Surface
Neoplasia
Erick Hernandez-Bogantes, Andrew Olivo-Payne, Alexandra Abdala,
Juan Carlos Serna-Ojeda, Enrique O. Graue-Hernandez,
Alejandro Navas, Arturo J. Ramirez-Miranda. Cornea and Refractive
Surgery, Insituto de Oftalmologia Fundacion Conde de Valenciana,
Mexico D.F, Mexico.
Purpose: To report a case series of 8 patients clinically
diagnosed with giant ocular squamous neoplasia (OSSN) who
underwent medical therapy with topical interferon alpha-2b for
immunoreduction and immutherapy purposes
Methods: Patients received monthly dosis of topical interferon
alpha-2b (1 million IU/mL) for a total of 3 months or with a
subconjunctival injection (3 million IU/mL) single dose per week
monthly until tumor resolution.
Results:
A total of eight eyes of 8 patients (mean age of 69 y/o) with a clinical
diagnosis of unilateral giant OSSN were managed with interferon
alpha-2b. Six patients received topical therapy (1 million IU/mL) 4
times per day for a minimum of 3 months for immunotherapeutic
purposes and 2 patients received subconjunctival therapy (3 million
IU/mL) single dose per week for 1 month for immunoreduction.
The median time to resolution was 5 months (range, 1.5-6 months)
and the median follow-up was 22 months (range, 6-28 months).
Despite the dramatic clinical response, all patients continued with
topical interferon alpha-2b at least 1 month beyond complete
clinical resolution. The two cases in which immunoreduction was
successfully achieved, were scheduled for a lesion excision with a
no-touch technique followed by cryotherapy and amniotic membrane
to cover the large excision area. There has been no recurrence during
follow-up.
Conclusions: In cases of giant OSSN interferon alpha-2b can
successfully reduce or achieve complete resolution, thus avoiding the
patient a more extensive surgical procedure with its known risks and
complications.
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.
ARVO 2016 Annual Meeting Abstracts
Results: MSCs were isolated and their specific markers, such as
CD29, CD44, CD105 and CD90, were expressed at high levels,
while CD31, CD34, CD45 and MHC-DR could not be detected.
Rat conjunctiva showed mild hyperemia at the injection site on the
second day and diminished on day 3. All the injected cells remained
in the engrafted regions and showed no migration. Majority of the
MSCs survived 3 days after implantation. However, the number of
cells gradually decreased thereafter, and completely disappeared
on day 9 after implantation. Using Ki67 as a marker, we could only
observe quite a number of positive staining cells surrounding the
cell mass. Immunostaining against a-SMA was negative from day 3
to day 9, indicating that the injected cells did not differentiate into
myofibroblasts. TUNEL positive cells were present in the injected
cell mass. The quantities of infiltrated CD11b+, CD45+ and PMN+
cells were increased in the injected eyes at day 3, and gradually
reduced to normal basal levels at day 9.
Conclusions: MSCs were gradually eliminated from the injection site
without cell migration, proliferation and differentiation. However,
MSCs activated proliferation of host cells and recruited immune cells
to the microenvironment.
Commercial Relationships: Juan Li, None; Chengyou Zuo, None;
Shangkun Ou, None; Sanming Li, None; Liying Zhang, None;
Changkai Jia, None; Zuguo Liu, None; Wei Li, None
Support: Chinese National Key Scientific Research Project
(2013CB967003)
Commercial Relationships: Erick Hernandez-Bogantes, None;
Andrew Olivo-Payne, None; Alexandra Abdala, None; Juan
Carlos Serna-Ojeda, None; Enrique O. Graue-Hernandez, None;
Alejandro Navas, None; Arturo J. Ramirez-Miranda, None
Program Number: 3896 Poster Board Number: A0044
Presentation Time: 3:45 PM–5:30 PM
The fate of mesenchymal stem cells after subconjunctival
implantation
Juan Li, Chengyou Zuo, Shangkun Ou, Sanming Li, Liying Zhang,
Changkai Jia, Zuguo Liu, Wei Li. Xiamen University Medical
College, Eye Institute of Xiamen University, Xiamen, China.
Purpose: Subconjunctival injection of mesenchymal stem cells
(MSCs) has been applied in the treatment of ocular surface chemical
burn, while the fate of MSCs in the local microenvironment after
injection remains unknown. In this study, we investigated the
survival, migration, proliferation, differentiation and paracrine
of human MSCs, as well as their impact on the ocular surface
microenvironment after subconjunctival implantation in rat.
Methods: MSCs were isolated from umbilical cord characterized
by flow cytometry on the basis of published criteria. Overall, 27 SD
rats were subjected to MSCs injection. For each rat, the left eye was
injected subconjunctivally with 2X105 MSCs suspended in 50ml
DMEM, and the right eye was served as a control. On the 3rd, 6th
and 9th day after injection, the rats were examined by slit-lamp to
evaluate conjunctival edema and hyperemia. Immunohistochemistry
was performed to investigate the expression patterns of Vimentin,
a-SMA, Ki67 and PMN. TUNEL assay was performed to detect cell
apoptosis. Furthermore, the proportion of CD11b+ and CD45+ cells
in conjunctiva were quantified by flow cytometry.
These abstracts are licensed under a Creative Commons Attribution-NonCommercial-No Derivatives 4.0 International License. Go to http://iovs.arvojournals.org/
to access the versions of record.