ARVO 2012 Annual Meeting Abstracts
229 Corneal Biomechanics I
Monday, May 7, 2012, 8:30 AM - 10:15 AM
Hall B/C Poster Session
Program #/Board # Range: 1510-1532/A466-A488
Organizing Section: Cornea
Program Number: 1510 Poster Board Number: A466
Presentation Time: 8:30 AM - 10:15 AM
Effect of Intraocular Pressure on Corneal Elasticity Measurement using
Acoustic Radiation Force
Raksha Urs1, Ronald H. Silverman1,2. 1Ophthalmology, Columbia University
Medical Center, New York, NY; 2Frederic L. Lizzi Center for Biomedical
Engineering, Riverside Research Institute, New York, NY.
Purpose: Measurement of intraocular pressure (IOP) by applanation techniques is
known to be affected by corneal thickness and elasticity. The purpose of this study
is to determine the effect of IOP on corneal elasticity values determined from
stress/strain relationships resulting from application of acoustic radiation force
(ARF) to the cornea.
Methods: A cannula was inserted into the optic nerve of intact porcine eye globes
(n=5) immersed in 0.9% saline. The cannula was attached to a saline bag, whose
height was varied, to vary the IOP. A pressure sensor with digital output (Omega
PX154; Omega Engineering, Inc., Stamford, CT) was used to monitor the IOP.
Pressure was increased from 5 to 40 mmHg and then decreased to 5 mmHg in steps
of 5 mmHg. At each pressure increment or decrement, an ARF scan of the cornea
was recorded with a 25 MHz ultrasound transducer (Olympus SN 781943). The
ARF scan consisted of 20 pushing pulses applied at 25% duty cycle, at a 1 kHz
pulse repetition rate, with imaging impulses interleaved between push pulses, to
allow radiofrequency (RF) data acquisition during the push mode. After the push
sequence, the cornea was imaged for another 400 ms. RF data were sampled at 400
MHz (12 bits/sample). Displacement of the front and back surfaces of the cornea,
during and after the push, was computed with a spline-based algorithm (Viola et al,
2005), and change in corneal thickness and strain were calculated. Strain during
ARF application was fit to the Voigt model to determine the elastic modulus at
each IOP level.
Results: The average elastic modulus ranged from 20 to 40 kPa. Elastic modulus
increased with IOP (p<0.0001), at a rate of 0.15 kPa/mmHg. The slope was higher
during the increasing-IOP cycle (0.21 kPa/mmHg; p<0.0001) compared to the
decreasing-IOP cycle (0.11 kPa/mmHg; p=0.02).
Conclusions: Elastic modulus of the cornea, determined by ARF, is influenced by
IOP, with corneas measuring progressively stiffer with increasing IOP. Stress-strain
curves are thus non-linear over the range of IOP values. For ex-vivo porcine eyes,
over the range of 5-40 mmHg, the variation is 13-26% of the average elasticity
measured over the full range of IOP values. An IOP-based correction is required
when corneal elasticity is measured in vivo, with ARF, in potential applications
such as detection of keratoconus, and evaluation of corneal cross-linking therapy.
Commercial Relationships: Raksha Urs, None; Ronald H. Silverman, None
Support: NIH Grant R01EY010955; Research to Prevent Blindness;
Program Number: 1511 Poster Board Number: A467
Presentation Time: 8:30 AM - 10:15 AM
Changes in Corneal Biomechanics after Penetrating Keratoplasty in
Keratoconus
Brian E. Goldhagen1, Richard Y. Hwang2, Anthony N. Kuo1, Natalie A. Afshari1.
1
Ophthalmology, Duke University Eye Center, Durham, NC; 2Ophthalmology,
Vanderbilt University, Nashville, TN.
Purpose: To investigate corneal biomechanical changes in keratoconus after
penetrating keratoplasty.
Methods: The study was conducted at Duke University as a retrospective chart
review. Twenty one keratoconus eyes were analyzed. Control eyes did not undergo
surgical interventions. Cases were keratoconus eyes that underwent penetrating
keratoplasty. Eyes with conditions that might affect corneal biomechanical
measurements, such as a history of corneal infection or trauma, were excluded.
Main outcome measures were taken with the Ocular Response Analyzer (ORA;
Reichert; Depew, NY): corneal hysteresis (CH), corneal resistance factor (CRF),
Goldman correlated intraocular pressure (IOPg) and compensated intraocular
pressure (IOPcc).
Results: Control keratoconus eyes had average CRF, CH, IOPg and IOPcc values
of 6.02+/-0.71, 7.05+/-0.61, 10.35+/-1.17, and 15.18+/-1.01 respectively.
Keratoconus eyes with penetrating keratoplasty had statistically increased values of
CRF, CH and IOPg: 10.33+/-1.28 (p=0.0003), 10.20+/-0.80 (p<0.0001), and
15.8+/-3.30 (p=0.02) respectively. IOPcc was not statistically affected with
penetrating keratoplasty: 16.49+/-2.85 (p=0.43).
Conclusions: Eyes with keratoconus that underwent penetrating keratoplasty had
an increase in corneal hysteresis (71%) and corneal resistance factor (45%),
approaching normal corneal values.
Commercial Relationships: Brian E. Goldhagen, None; Richard Y. Hwang,
None; Anthony N. Kuo, None; Natalie A. Afshari, None
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.
Support: Research to Prevent Blindness
Program Number: 1512 Poster Board Number: A468
Presentation Time: 8:30 AM - 10:15 AM
Influence of Internal Pressure and Distance from Air Nozzle on Maximum
Deformation Depth under an Air Puff of Pig Corneas
Sue Shiao1A, Ashraf M. Mahmoud1B,1C, Jun Liu1C,1B, David Lee1C, Kimberly
Metzler1C, Chris Minning2, Cynthia J. Roberts1B,1C. ACollege of Medicine,
B
Ophthalmology, CBiomedical Engineering, 1Ohio State University, Columbus,
OH; 2Internal Medicine, Riverside Methodist Hospital, Columbus, OH.
Purpose: To investigate the effect of both internal pressure and distance from the
air nozzle on maximum corneal deformation induced by an air puff in pig corneas
with an artificial anterior chamber mount.
Methods: The CorVis ST (Oculus, Wetzlar, Germany) is a new device that
captures approximately 140 Scheimpflug images in a single horizontal meridian
during a 30ms air puff. Deformation data for 8 pig corneas mounted in a Barron
disposable artificial anterior chamber were acquired and analyzed. Each cornea was
tested at pressures of 10, 20, 30, and 40 mmHg, and each pressure setting was
tested with distances of 0.5, 1.0, and 1.5 mm from the cornea to the air nozzle on
the Corvis head. A 0mm distance was set as the distance at which the anterior
cornea surface was captured just within the top border of the image window on the
Corvis. One scan at each setting was acquired. A repeated measures Multivariate
Analysis of Variance (MANOVA) on pressure and distance was performed to
evaluate the dependent variable of maximum corneal deformation with a p value of
0.05 as the significance threshold.
Results: At distances 0.5, 1.0, and 1.5 mm, the means of corneal deformation were
0.672 0.198, 0.761 0.267 , 0.660 0.273 (at pressure 10 mmHg and), 0.791
0.228, 0.676 0.212, 0.660 0.156 (at pressure 20 mmHg), 0.708 0.232, 0.665
0.169, 0.662 0.186 (at pressure 30 mmHg), 0.634 0.134, 0.624 0.159, and
0.763 0.244 mm (at pressure 40 mmHg), respectively. The MANOVA showed no
pressure or distance effect as single variables, but a significant interaction effect of
pressure and distance on maximum corneal deformation (p = 0.0179).
Conclusions: The internal pressure and distance of the cornea from the air nozzle
affects the maximum corneal deformation in pig corneas mounted on a rigid
artificial anterior chamber. Future studies will investigate the influence pressure
and distance will have on maximum deformation in a whole globe mount.
Commercial Relationships: Sue Shiao, None; Ashraf M. Mahmoud, None; Jun
Liu, None; David Lee, None; Kimberly Metzler, None; Chris Minning, None;
Cynthia J. Roberts, Oculus (C, R)
Support: Ohio Lions Eye Research Foundation
Program Number: 1513 Poster Board Number: A469
Presentation Time: 8:30 AM - 10:15 AM
A Mechanical Modelling Approach To Porcine Eye Cornea
Julien Bullet1,2, Morgane Bauer2, Vincent Borderie1, Stphane Roux2, Laurent
Laroche1. 1CHNO des Quinze-Vingts, Paris, France; 2Mechanics Laboratory, Ecole
Normale Suprieure de Cachan, Cachan, France.
Purpose: Overall corneal biomechanical parameters commonly used today have
limited reproducibility and clinical reliability. The present work aims at proposing
a different methodology of measurement for corneal mechanical properties based
on a precise finite element model. It is limited to porcine enucleated eyes
Methods: A freshly enucleated pig eye is mounted on a support with anterior
chamber perfusion, in order to control intraocular pressure. To check endothelial
viability, central corneal thickness is measured and remains stable during
experiment. 3D geometrical characterization of cornea is achieved at rest and under
different load conditions (IOP variation) through a rotating slit illumination
combined with a Scheimpflug camera system (Oculus Pentacam).
From detailed anterior and posterior surface maps and central corneal thickness at
rest reference state, a mechanical model of the cornea using the finite element
method (Abacus code) is built. Regular elastic mechanical laws are used to deduce
final displacement field with increasing IOP load. Comparing this calculated
displacement with measured corneal 3D map, boundary conditions as well a
biomechanical corneal characteristics such as Poisson coefficient and Young
modulus are estimated (least square optimization with Matlab software). To
minimize the number of unknowns to be identified, only few Fourier series
decomposition terms of boundary conditions are computed.
Results: Figure 1 shows the deformed geometry of a cornea after a slight increase
in intra-ocular pressure, and Figure 2 is a comparison between predicted and
measured model after optimization of boundary conditions.
Conclusions: Results presented in Figure 2 show that the difference between
measured and calculated geometry is in the tens of micrometer range, or about 2%
of the dynamics. However, cornea is here considered as purely elastic with simple
mechanical laws. Viscosity can be excluded from model as only dynamically
balanced states are simulated.
This first simple mechanical model of cornea demonstrates that the chaining of
experiment, data acquisition, and modeling works. Other more sophisticated
models including other load conditions, viscosity or other imaging techniques are
ARVO 2012 Annual Meeting Abstracts
to be developed.
Commercial Relationships: Julien Bullet, None; Morgane Bauer,
None; Vincent Borderie, None; Stphane Roux, None; Laurent Laroche, None
Support: None
Program Number: 1514 Poster Board Number: A470
Presentation Time: 8:30 AM - 10:15 AM
Aliphatic -nitroalcohols For Therapeutic Corneoscleral Cross-linking:
Chemical Stability Studies Using 1H-NMR
David C. Paik1A, Xia Li1B, Yongjun Li1B, Stephen L. Trokel1A, Nicholas J. Turro1B.
A Spectroscopic Measurements on Cadaveric Cornea During the Cross-Linking
Ophthalmology, BChemistry, 1Columbia University, New York, NY.
Purpose: The ability to selectively enhance the biomechanical properties of a given
tissue in vivo could have widespread clinical utility. Our recent studies suggest that
aliphatic -nitro alcohols (BNAs) may represent a useful class of compounds for
use as in vivo therapeutic corneoscleral cross-linking agents. In addition, higher
order nitroalcohols (HONAs), which include a nitrodiol and nitrotriol, exhibit much
greater tissue cross-linking efficacy than their nitromonol counterparts. Thus, the
current study was undertaken in order to determine the chemical stability of these
compounds under long-term storage conditions.
Methods: Two mono-nitroalcohols (2-nitroethanol=2ne and 2-nitro-1-
propanol=2nprop) and two HONAs, a nitrodiol (2-methyl-2-nitro-1,3-
propanediol=MNPD), and a nitrotriol (2-hydroxymethyl-2-nitro-1,3-
propanediol=HNPD) were monitored for chemical stability by 1H-NMR for up to 7
months. Each compound was studied at two concentrations (1% and 10%) either in
unbuffered H2O or 0.2 M NaH2PO4/Na2HPO4 (pH=5), for a total of 4 conditions for
each compound. The 1H-NMR spectra for the starting material was compared to
subsequent spectra. The formation of new peaks over time indicated decomposition
of the starting material and hence chemical instability.
Results: Typical 1H-NMR spectra were identified for each of the compounds tested
and were in agreement with existing literature. Unbuffered solutions maintained a
pH in the range of 7.0- 7.14. Under all 4 of the conditions studied, both the
nitrodiol (MNPD) and nitrotriol (HNPD) were stable for the duration of 7 months
showing no change in the 1H-NMR spectrum. 2nprop was stable under all
conditions at 10 days but became unstable under all conditions at 3 months. 2ne
was the most unstable of all the compounds tested, becoming unstable in
unbuffered solution at 10 days. No effort was made to identify decomposition
products as this was not the intent of the study.
Conclusions: HONAs exhibit excellent chemical stability under long-term storage
conditions. In contrast, the nitromonols tested are significantly less stable. Because
compound stability is important from a pharmaceutical standpoint, these findings
are relevant to translation of this technology into clinical use.
Commercial Relationships: David C. Paik, Patent pending (P); Xia Li,
None; Yongjun Li, None; Stephen L. Trokel, patent pending (P); Nicholas J.
Turro, None
Support: NIH Grant EY020495 and Research to Prevent Blindness
Program Number: 1515 Poster Board Number: A471
Presentation Time: 8:30 AM - 10:15 AM
Biomechanical Properties Of The Cornea In An Adult Population: The
Alienor Study
cedric schweitzer1, lemy agbessi1, melanie le goff2, florence malet1, jean-francois
korobelnik1, marie B. rougier1, marie-noelle delyfer1, jean-Francois dartigues2A,
Cecile DelCourt3, joseph colin1. 1ophthalmology, university hospital pellegrin,
bordeaux, France; Aophthalmology, 2Univ. Bordeaux, ISPED, Centre INSERM
U897-Epidemiologie-Biostatistique, bordeaux, France; 3Universite Bordeaux
Segalen, INSERM, U897, Bordeaux, France.
Purpose: To analyze biomechanical properties of the cornea in elderly patients.
Methods: The ALIENOR (Antioxydants, Lipides Essentiels, Nutrition and
maladies OculaiRes) Study is a population-based epidemiological study on age-
related eye diseases. In 2009-2010, 625 subjects, aged 75 years or more, had an eye
examination, including intraocular pressure and central corneal thickness (CCT)
measurements, and an evaluation of the biomechanical properties of the cornea
using the ocular response analyser (ORA). Patients were classified in 3 groups
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.
of age (Group1: 75-79 years old (yo), Group2: 80-84 yo, Group3: > 84yo). Corneal
hysteresis (CH), corneal resistance factor (CRF), corneal compensated intraocular
pressure (IOPcc), Goldmann correlated intraocular pressure (IOPg), air pressure
curve and infrared curve parameters were analyzed and compared between groups.
Results: 436 patients had complete data. Mean age was 82.2+/-4.2 years, and 165
patients were included in the group 1 (37.8%), 164 in the group 2 (37.6%), 107 in
the group 3 (24.5%). The mean CCT was 549.7+/-36.4m and was not statistically
different between groups (p=0.76). The mean CH was 9.9+/-1.8 mmHg and was
significantly different between the 3 groups (Group 1: 10.1+/-1.7mmHg, Group 2:
9.9+/-1.9mmHg, Group 3: 9.4+/-1.7mmHg,p=0.008). The mean CRF was 10.2+/-
1.8 mmHg and was significantly different between the 3 groups (group1:10.4+/-
1.8mmHg, Group 2:10.4+/-1.8mmHg, Group 3:9.8+/-1.8mmHg,p=0.02). CH and
CRF were not associated with gender.
Conclusions: CH and CRF mean values significantly decrease with age in elderly
patients. Biomechanical properties of the cornea change with advancing age and
may influence intraocular pressure measurements.
Commercial Relationships: cedric schweitzer, None; lemy agbessi,
None; melanie le goff, None; florence malet, None; jean-francois korobelnik,
None; marie B. rougier, None; marie-noelle delyfer, None; jean-Francois
dartigues, None; Cecile DelCourt, None; joseph colin, None
Support: Laboratoires Tha, Fondation Voir et Entendre
Program Number: 1516 Poster Board Number: A472
Presentation Time: 8:30 AM - 10:15 AM
Procedure
Liliane Ventura1, Victor A. Lincoln1, Marcio M. Mello1, Daniela Sampaio2, Sidney
J. Faria e Sousa2. 1Electrical Engineering SEL/EESC/USP, Universidade de Sao
Paulo EESC, S Carlos - SP, Brazil; 2Ophthalmology, University of Sao Paulo -
FMRP, Ribeirao Preto - SP, Brazil.
Purpose: Study of the UVA transmission through the eye during the collagen cross
linking procedure (CCP) and corneal dehydration influence on the method.
Methods: The 31 human corneas preserved in Optisol-GS (average of 5 days post-
mortem and 571m thickness) were washed with saline, deepithelized and cross-
linked with the currently used protocol. Riboflavin solution (0.1% and 400 mOsm)
was applied prior to and during exposure. The UV beam - 365nm at 3mW/cm2 -
was focused directly onto the corneal stroma. Two groups were tested: G1 - 10
corneas - the optical fiber (probe from the USB2000 Ocean Optics
Spectrophotometer) was placed immediately after the endothelium; G2 - 21 corneas
- probe was placed 10mm away from the endothelium (crystalline lens position)
and the transmission of the UVA light was measured every 4 minutes after each
instillation.
Results: The measured average transmittance of the cornea without Riboflavin was
64.1% 5.1% for G1; and 63.2% 7.5% for G2. Preceding the irradiation but after
6 applications of Riboflavin at 5min intervals (total of 30min) transmittance
decreased to 21.1% 3.1% for G1; and 33.0% 8.4% for G2. The 30min of
irradiation were then accompanied by an additional 6 applications of Riboflavin at
5min intervals, resulting in a further decrease in transmittance to 12.2% 5.1% for
G1; and 21.5% 8.2% for G2. Three additional corneas were evaluated with the
probe 1.5mm away from the endothelium, but without going under the CCP or
Riboflavin instillation. Measurements of the UVA transmission were collected for
60min, every 1min, in order to study the influence of the dehydration of the cornea.
An average of 3% of the transmitted UVA light was detected.
Conclusions: The average energy reaching the endothelium during irradiation
fluctuated from 0.59 to 0.38mW/cm2, which is beyond the limits established by
literature. The dehydration of the cornea that occurs during our performed
measurements has no influence on the measurements. The results suggest that the
endothelium safe limit for humans might be higher than established on literature,
since no endothelial damage has been observed on the patients submitted to CCP.
Also, concentration of UVA radiation reaching the crystalline lenses should be
considered, since the cornea focus the beam on the lens, due to its high refractive
power and may cause early cataract.
Commercial Relationships: Liliane Ventura, None; Victor A. Lincoln,
None; Marcio M. Mello, None; Daniela Sampaio, None; Sidney J. Faria e
Sousa, None
Support: FAPESP and CNPq
Program Number: 1517 Poster Board Number: A473
Presentation Time: 8:30 AM - 10:15 AM
Outcomes of Epithelium-On Crosslinking in Older Adults
William Trattler1, Roy Rubinfeld2, Gabriela Perez3, John Talamo4, Kathryn M.
Hatch4, Jodi Luchs5, Gaston Lacayo1, Gregg Berdy6. 1Cornea, Center For
Excellence in Eye Care, Miami, FL; 2Washington Eye Physicians and Surgeons,
Chevy Chase, MD; 3Ctr for Excellence in Eye Care, Miami, FL; 4Talamo Laser
Consultants, Waltham, MA; 5Southshore Eye Care, Wantagh, NY; 6The Cornea
and Laser Vision Institute, St. Louis, MO.
ARVO 2012 Annual Meeting Abstracts
Purpose: To evaluate patient outcomes following Epithelium-On CXL in patients
over the age of 35.
Methods: This prospective, non-randomized trial evaluated patients who
underwent epithelial-ON CXL. A subgroup of patients over the age of 35 was
evaluated for this analysis. Post-Op measures included corneal hysteresis (CH) and
corneal resistance factor (CRF), Pentacam data, and uncorrected and best-spectacle
corrected visual acuities. Eyes evaluated had a minimum follow-up visit of 3
months and a maximum follow up visit of 12.8 months.
Results: 90 eyes met the entry criteria (65 male, 22 female), with a follow up range
of 3 months minimum to 12.8 months maximum. The mean follow up was 7.9
months. 67% of these eyes had a pre-op diagnosis of keratoconus, 20% had post-
LASIK Ectasia, and 3% had pellucid marginal degeneration. Average CH and CRF
values increased post-Operatively from 7.7 to 8.0 and 6.5 to 8.0, respectively, at the
latest follow up visit. Average refractive cylinder decreased from 4.59 D to 4.15D.
On Pentacam the average 2mm K Astigmatism decreased from 5.94 to 4.71 and
Holladay Astigmatism decreased from 3.31 to 2.26. 34.8% of eyes experienced no
change in the number of lines of BSCVA and 46% of eyes gained 1 or more lines
of BSCVA. Average BSCVA pre-Op was 20/163 and post-Op was 20/109.
Conclusions: Epithelial-on cross-linking appears to be safe and effective for
patients of age 35 and over with ectatic corneal disease.
Commercial Relationships: William Trattler, CXLUSA (C); Roy Rubinfeld,
CXLUSA (E); Gabriela Perez, None; John Talamo, None; Kathryn M. Hatch,
None; Jodi Luchs, None; Gaston Lacayo, None; Gregg Berdy, None
Support: None
Clinical Trial: http://www.clinicaltrials.gov, NCT01024322
Program Number: 1518 Poster Board Number: A474
Presentation Time: 8:30 AM - 10:15 AM
Comparative Stress Strain Measurements Of Human Corneas After
Transepithelial UV-A Induced Cross-linking: Impregnation With
Iontophoresis, Different Riboflavin Solutions And Irradiance Power
Riccardo Vinciguerra1, Eberhard Spoerl2, Mario R. Romano1, Pietro Rosetta1,
Paolo Vinciguerra1. 1Opthalmology, Istituto Clinico Humanitas, Milan, Italy;
2
Ophthalmology, University of Dresden, Dresden, Germany.
Purpose: To compare the change in biomechanical properties of human cadaver
corneas after standard transepithelial cross-linking (CXL-TE) versus CXL-TE
using iontophoresis, different solutions and UV-A power.
Methods: Twelve human cadaver corneas were divided in 4 different groups
according to methods of impregnations and UV-A power used: Group A (three
corneas, treated with CXL-TE using an irradiance power of 3 mW/cm2 for 30
minutes and riboflavin solution with 15 % dextrane and Tromethamine); Group B
(three corneas treated with CXL-TE using an irradiance power of 3 mW/cm2 for 30
minutes and riboflavin solution with Tromethamine); Group C (three corneas
treated with CXL-TE using an irradiance power of 10 mW/cm2 for 10 minutes and
riboflavin solution with Tromethamine); Group D (three corneas treated with an
irradiance power of 10 mW/cm2 for 10 minutes, the impregnation was obtained
with the aid of iontophoresis and a riboflavin solution with Tromethamine). After
cross- linking, static stress-strain measurements of the corneas were performed
using a microcomputer-controlled biomaterial tester with a pre-stress of 5x103 Pa.
Stress stain curves were fitted with an exponential function and the Young's
modulus was calculated. Thickness of the cornea was measured with an ultrasound
pachymeter.
Results: Stress strain measurement showed an increase in corneal rigidity after
cross-linking compared to standard CXL-TE, indicated by a rise in strain and in
Youngs modulus calculated at 10% strain. Considering group A as standard of
comparison, group B showed an increase by a factor of 1.45, group C by a factor of
1.26, group D by a factor of 1.81. Mean corneal thickness was: 627 m for group
A, 628 m for group B, 527 m for group C, 665 m for group D.
Conclusions: CXL-TE is able to increase mechanical rigidity in human corneas in
selected groups. Stress strain results showed a maximal effect in the iontophoresis
group, probably due to the increased riboflavin concentration in the stroma. Stress
strain measurement in the other groups showed a better results using riboflavin
solution without dextrane and 3 mW/cm2 of irradiance power.
Commercial Relationships: Riccardo Vinciguerra, None; Eberhard Spoerl,
None; Mario R. Romano, None; Pietro Rosetta, None; Paolo Vinciguerra,
SOOFT (C)
Support: None
Program Number: 1519 Poster Board Number: A475
Presentation Time: 8:30 AM - 10:15 AM
Changes In Corneal Air Puff Deformation Parameters After Cross-Linking
From Dynamic OCT
Carlos Dorronsoro, Daniel Pascual, Pablo Prez-Merino, Sabine Kling, Susana
Marcos. Instituto de Optica, CSIC, Madrid, Spain.
Purpose: To retrieve quantitative corneal biomechanical properties in-vivo and
non-invasively, from high-speed OCT images of the dynamic response of the
cornea to an air puff inducing a deformation.
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.
Methods: A spectral OCT instrument combined with an air tonometer in a non-
collinear configuration was used to image the corneal deformation over full corneal
cross-sections, as well as to obtain high speed measurements of the temporal
evolution of the corneal apex. With this new technique the entire deformation
process can be dynamically visualized. A quantitative analysis allows direct
extraction of several deformation parameters. The potential of the technique is
demonstrated in vitro on porcine corneas under controlled IOP for different
conditions (untreated, de-epithelialized, after Riboflavin instillation, and after UV-
cross-linking, CXL), as well as in vivo in humans.
Results: Corneal deformation dynamics varied significantly across species and
treatments. Despite its lower thickness, the living human cornea showed lower
deformation amplitude (0.56 mm) than porcine untreated corneas (0.85 mm),
shorter deformation durations (14.3 vs 17.3 ms) and reduced recovery speeds (0.11
vs 0.21 mm/ms). CXL (and, to a minor extent, Riboflavin) decreased corneal
thickness by 50% and reduced several deformation parameters in comparison to
those of virgin corneas: deformation amplitude from 0.85 to 0.61 mm, deformation
diameters from 5.7 to 4.4 mm, displaced volumes from 10.1 to 4.0 mm3, and
recovery speed from 0.21 to 0.12 mm/ms. CXL also produced changes in the
temporal symmetry of the deformation, advancing the peak deformation time.
These differences are consistent with increased corneal stiffness following CXL.
Conclusions: High-speed OCT in combination with air-puff allowed one- and two-
dimensional tracking and quantification of corneal deformation events in vivo and
in vitro. The differences in the corneal response across treatments (i.e. Riboflavin
vs CXL) to the same deformation stimulus are likely the result of changes in the
microscopic structure of the cornea, and not a consequence of changes in corneal
thickness, geometry or IOP across conditions. Combinations of these measurements
with finite element modelling will allow quantification of the biomechanical
properties of corneal tissue, at an individual level and in vivo, to improve diagnosis
and prognosis of diseases and treatments.
Commercial Relationships: Carlos Dorronsoro, None; Daniel Pascual,
None; Pablo Prez-Merino, None; Sabine Kling, None; Susana Marcos, None
Support: MICINN FIS2008-02065 and FIS2011-25637; EURHORCs-ESF
EURYI-05-102-ES; ERC-2011-AdG-294099 to SM.
Program Number: 1520 Poster Board Number: A476
Presentation Time: 8:30 AM - 10:15 AM
Effect Of Corneal Collagen Crosslinking On Intraocular Pressure
Measurement In Patients With Keratoconus: Analysis By Goldmann And
Pascal Tonometers
Eduardo H. Araujo, Hellmann D. Cavalcanti, Ronald F. Cavalcanti, Diego N.
Gadelha, Bernardo M. Cavalcanti, Ana Ceclia S. Escario. Retina, Fundacao
Altino Ventura, Recife, Brazil.
Purpose: To evaluate, using Goldmann and Pascal tonometers, the effect of
collagen corneal crosslinking on intraocular pressure (IOP) measurement in
patients with keratoconus.
Methods: A prospective interventional study was done. It was evaluated 12
patients (17 eyes) with keratoconus, of whom eight (66.7%) were females and four
(33.3%) males. The mean age was 24.49.1 year-old. It was analyzed the
intraocular pressure by Goldmann and Pascal tonometers, and central corneal
pachymetry before and one month after the corneal crosslinking (CXL) with
riboflavin 0.1% and ultraviolet A for 30 minutes.
Results: There was statistically significant increase in the mean IOP one month
after CXL, with both tonometers. The increase in the means were 2.40.4 mmHg
and 2.90.7 mmHg for Goldmann (P= 0.0053) and Pascal (P=0.0001), respectively.
There was no statistically significance on pachymetry in both moments analyzed,
with the means of 481,142,8 m on the first moment, and 481,637,4 m one
month after the corneal crosslinking (P=0,8852).
Conclusions: The increased IOP could be caused by an increase in corneal rigidity.
However, one cannot exclude the possibility that the true IOP increased after
CXL. This possibility could be verified through comparison between these
tonometers and an invasive method of IOP measurement in eyes with keratoconus
after corneal crosslinking. Although the crosslinking be considered a safe method,
one can not exclude the possibility of a trabecular meshwork collagen crosslinking
ARVO 2012 Annual Meeting Abstracts
as a possible etiology.
Commercial Relationships: Eduardo H. Araujo, None; Hellmann D.
Cavalcanti, None; Ronald F. Cavalcanti, None; Diego N. Gadelha,
None; Bernardo M. Cavalcanti, None; Ana Ceclia S. Escario, None
Support: None
Program Number: 1521 Poster Board Number: A477
Presentation Time: 8:30 AM - 10:15 AM
Ora Waveform Analysis Of Biomechanical Properties Of The Cornea In
Normal Tension Glaucoma
Alice Grise-Dulac, Jr., Alain Saad, Damien Gatinel. Rothschild Fundation, Paris,
France.
Purpose: To analyze the Ocular Response Analyzer (ORA) waveform of
biomechanical properties of the corneas in normal tension glaucoma (NTG)
patients and compare them with those of patients with primary open angle
glaucoma (POAG), isolated ocular hypertension (OHT) and normal subjects (N).
Methods: 37 parameters of ORA waveform, caracterizing peak 1 and 2, were
measured, among wich : area (p1 and p2area, p1and p2 area1), aspect ratio
(aspect1, 2, 11 and 21), downslope (dslope1, 2, 11 and 21), width (w1 and w2),
height (h1 and 2), dive (dive1 and 2) High frequency noise in region between
peaks (aplhf) was also quantified. All parameters were provided by ORA device for
58 eyes of 29 patients with NTG, 85 eyes of 43 patients with chronic POAG, 57
eyes of 29 eyes with OHT and 44 eyes of 22 N subjects. Ultrasonic central corneal
thickness (CCT) was also measured for each eye. ANOVA test was used for
statistical analysis. A p value lower than 0.05 was considered statistically
significant.
Results: CCT was not significantly different between the 4 groups. NTG subjects
had significant lower aspect1, aspect11, dive1, dslope1, dslope11, h1 and h11 but
higher aplhf than normal subjects. POAG subjects had significant higher aplhf,
aspect11, w1 and lower path1 than normal subjects. NTG subjects had significant
lower dive1, h1, h11, p1area, p1area1, p2area and p2area1 than POAG patients.
OHT subjects had similar results to normal subjects.
Conclusions: NTG and POAG subjects were associated with significant
differences on peak 1 and higher frequency noise (aplhf) compared to normal
subjects. ORA waveform analysis could be useful in early diagnosis of NTG and
POAG.
Commercial Relationships: Alice Grise-Dulac, Jr., None; Alain Saad,
None; Damien Gatinel, None
Support: None
Program Number: 1522 Poster Board Number: A478
Presentation Time: 8:30 AM - 10:15 AM
Corneal Thickness Changes During Corneal Collagen Cross-linking With
Ultraviolet-a Irradiation And Hypo-osmolar Riboflavin Solution In Thin
Corneas
Belquiz A. Nassaralla1, Joao J. Nassaralla, Jr.2. 1Cataract Cornea & Refractive
Surgery, Instituto de Olhos de Goiania, Goiania, Brazil; 2Retina and Vitreous,
Instituto de Olhos de Goiania and UnB, Goiania, Brazil.
Purpose: To evaluate the corneal pachymetric variations during and after corneal
collagen cross-linking (CXL) treatment with ultraviolet-A irradiation (UVA) and
hypo-osmolar riboflavin solution in thin corneas.
Methods: Eighteen eyes of 18 patients, 11 men and 7 women, with progressive
keratoconus and thinnest corneal thickness (TCT) less than 400m. After the
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.
epithelium removal, iso-osmolar riboflavin was applied to the cornea every 3
minutes (30min). Hipo-osmolar riboflavin was then applied every 20 seconds until
the TCTreached 400m. UVA irradiation was performed for 30 min. Pachymetry
was recorded at the thinnest point of the cornea preoperatively, after epithelial
removal, after iso-osmolar riboflavin, after hypo-osmolar riboflavin, after UVA
irradiation, and at 1, 6 and 12 months.
Results: Mean patient age was 24 years. Mean preoperative TCT was 380m.
After epithelial removal, mean TCT was 341m. After iso-osmolar riboflavin
instillation it was 330m. After hypo-osmolar riboflavin it was 418 m. After UVA
irradiation it was 384m. At 1, 6 and 12 months it was 372m, 381m, and
379m, respectively. After 12 months, all corneas remained clear.
Conclusions: Hypo-osmolar 0.1% riboflavin solution seems to be safe and
effective for swelling thin corneas and allowing a CXL for the treatment of
progressive keratoconus.
Commercial Relationships: Belquiz A. Nassaralla, None; Joao J. Nassaralla,
Jr., None
Support: None
Clinical Trial: http://www.clinicaltrials.gov, NCT01485211
Program Number: 1523 Poster Board Number: A479
Presentation Time: 8:30 AM - 10:15 AM
Effect of Corneal Crosslinking on Anterior and Posterior Stromal Elasticity
Assessed Ex Vivo by Atomic Force Microscopy
Janice Dias, Noel M. Ziebarth. Dept. of Biomedical Engineering, University of
Miami, Coral Gables, FL.
Purpose: To quantify the anterior and posterior stromal elasticity after crosslinking
with ultraviolet light and riboflavin.
Methods: Four pairs of porcine eyes (< 2 days postmortem) were used for this
study. The eyes were retrieved from an abattoir, placed in a bag filled with saline,
and shipped to the laboratory overnight. Upon arrival, the epithelium was removed
using a cotton swap, and the cornea was excised with a generous scleral rim and
placed in 20% Dextran overnight to restore the cornea to physiological levels (550-
800m). One eye of each pair underwent crosslinking treatment and the
contralateral eye served as the control. For the treated eye, one drop of 0.1%
riboflavin solution (10mg riboflavin-5-phosphate in 10mL Dextran 20% solution)
was placed on the cornea every 5 minutes for 30 minutes. The cornea was then
irradiated for 30 minutes using UVA light (=378nm) with an irradiance of
3mW/cm2. During irradiation, one drop of the riboflavin solution was placed on
the cornea every 5 minutes. Manual delamination of Bowmans membrane was
performed to expose the anterior stroma. Elasticity was then measured with a
custom-built Atomic Force Microscope designed for mechanical testing of
ophthalmic tissues. Afterwards, delamination was continued to access the posterior
stromal region and elasticity measurements were repeated. Youngs modulus was
calculated using the Hertz model for a spherical indenter.
Results: Youngs modulus of the untreated anterior and posterior stroma ranged
from 16kPa to 393kPa and 8.4kPa to 57.1kPa, respectively. Youngs modulus of
the treated anterior and posterior stroma ranged from 20.2kPa to 333kPa and
13.4kPa to 85.7kPa, respectively. The anterior stroma is consistently stiffer than the
posterior stroma for both untreated and treated eyes. Corneal crosslinking resulted
in a much greater increase in posterior stromal stiffness than anterior.
Conclusions: The effect of corneal crosslinking on stromal elasticity appears to be
depth dependent.
Commercial Relationships: Janice Dias, None; Noel M. Ziebarth, None
Support: NIH Ruth L. Kirschstein National Research Service Awards for
Individual Predoctoral Fellowships to Promote Diversity in Health-Related
Research (1F31EY021714-01)
Program Number: 1524 Poster Board Number: A480
Presentation Time: 8:30 AM - 10:15 AM
Measurement of Varying Elasticity Between Central and Peripheral Human
Cornea With Acoustic Radiation Force Elasticity Microscope
Eric R. Mikula1A, James Jester1B, Tibor Juhasz1B. ABiomedical Engineering, BGavin
Herbert Eye Institute, 1University of California, Irvine, Irvine, CA.
Purpose: The non-homogeneous organization of collagen across the cornea
suggests a unique distribution of elastic moduli. Specifically, a difference in
elasticity is expected between the central and peripheral cornea due to underlying
structural differences. Using acoustic radiation force elastic microscopy (ARFEM),
we measure the distribution of elastic properties across the cornea, both radially
and axially, with the goal of constructing a cross-sectional map of elasticity.
Methods: Corneas from human cadaver eyes (San Diego Eye Bank, San Diego
California) were excised from the globe leaving a 2 mm scleral rim intact. The
corneal samples were suspended in collagen gelatin (10% w/w) within a water tank
filled with deionized, degassed water. The water tank was attached to a 3-D
mechanical stage allowing for precise control of cavitation bubble placement within
the cornea. Femtosecond laser pulses induced optical breakdown and produced
cavitation at points across the cornea. A confocal ultrasonic transducer applied a
2ms train of 200s acoustic radiation force-chirp bursts to the bubble at 1.5 MHz
ARVO 2012 Annual Meeting Abstracts
while monitoring bubble position using pulse-echoes at 20 MHz. A cross-
correlation method was used to calculate bubble displacements. Maximum bubble
displacements are inversely proportional to the Youngs modulus.
Results: Initial results indicate that the central cornea in the human eye is stiffer
than the peripheral cornea in the direction orthogonal to the corneal surface.
Conclusions: Our non-invasive ARFEM results show the central cornea has a
higher elastic modulus than the peripheral cornea and that elasticity varies
throughout the cornea.
Commercial Relationships: Eric R. Mikula, None; James Jester, None; Tibor
Juhasz, None
Support: NIH Grant EY018665
Program Number: 1525 Poster Board Number: A481
Presentation Time: 8:30 AM - 10:15 AM
The Cell Fade Of Cultured Corneal Endothelial Cells On The Genipin-
crosslinking Descemet Membrane
Lung-Kun Yeh1,2, Shih-Chun Huang3, Hsin-Ta Wang3. 1Ophthalmology, Chang
Gung Mem Hosp, linko, Taipei, Taiwan; 2Chang-Gung University College of
Medicine, Taiwan, Linko, Taiwan; 3Institute of Organic and Polymeric Materials,
National Taipei University of Technology, Taiwan, Institute of Organic and
Polymeric Materials, National Taipei University of Technology, Taiwan, Taipei,
Taiwan.
Purpose: Genipin, a nature crosslinking material extracted from the fruits of
Gardenia jasminoides, has been widely used in many biological applications. The
Descement membrane is easy rolled and easy teard after peeling from corneal
tissue. In this study, we try to study the phenotypes of cultured endothelial cells on
the genipin-crosslinker Descement membrane.
Methods: The crosslinking process between Descement membrane.and genipin
was performed before cell plating. The crosslinking characteristics of the genipin-
fixed Descemet membrane was investigated. Furthermore, the mechanical strength
and resistance were compared between the genipin-fixed Descemet membrane and
non-genipin-fixed Descemet membrane. The proliferation rate of cultured bovine
corneal endothelial cells were study by MTT assay. Furthermore, the
immunohistochemistry study and Scanning EM were also investigated.
Results: The mechanical strength and resistance were increased in the group of the
genipin-fixed Descemet membrane as compared with the group of non-genipin-
fixed Descemet membrane. The genipin-fixed Descemet membrane was easy
handled, more smooth surface, and not easy teared. The SEM analysis showed the
normal endothelial cells phenotype between the two groups. The MTT assay
showed the proliferation rate was increased in the group of genipin-fixed Descemet
membrane. The immunohistochemistry staining showed the ZO-1 positive and Na-
K ATPase positive cultured cells on the genipin-fixed Descemet membrane.
Conclusions: The results demonstrated that genipin can form stable crosslinked
products and is an effective crosslinking agent for Descemet membrane fixation.
Therefore, the non-toxic nature crosslinker genipin may be useful for the
application on the reconstruction of the ocular surface.
Commercial Relationships: Lung-Kun Yeh, None; Shih-Chun Huang,
None; Hsin-Ta Wang, None
Support: None
Program Number: 1526 Poster Board Number: A482
Presentation Time: 8:30 AM - 10:15 AM
Keratometric effect of Genipin, a non UV crosslinking system: In vivo Efficacy
Studies
Mauricio E. Narvez, Sr., Marcel Y. Avila, Sr.. Ophthalmology, Universidad
Nacional de Colombia, Bogota, Colombia.
Purpose: To evaluate the effects of genipin crosslinking in corneal curvature in
rabbit eyes.
Methods: Genipin crosslinking was induced in 12 rabbit eyes (Adult New Zealand
albino rabbits, 3 to 3.5 kg). Baseline keratometry was performed in both eyes using
an automatic keratometer. One cornea was crosslinked with genipin 0.25% for 20
minutes , the other eye was used as a control sample. Keratometric measurements
were done before the crosslinking and 30 and 60 days after the treatment. All
procedures were done according to the ARVO statement for use of animals in
research.
Results: Average baseline keratometric values were 49.70.6 diopters (control) vs.
49.20.8 (genipin), at day 30 values were 47.90.3 (control) vs. 47.10.8(genipin)
and at day 60 values were 46.280.5 (control) vs. 44.80.4 (genipin) (p 0.001) .
Reduction of the steepest values were calculated and correlated between both eyes
(control vs. treated). Reduction of 2.52 (control) vs. 5.1 0.5 (genipin) diopters
were found with a p =0.005. None of the rabbits showed any complication.
Conclusions: Genipin crosslinking induces a significant corneal flattening of 2
diopters in average and about 2.5 diopters in the steepest meridian at 60 days post-
treatment, in vivo, suggesting that this exogenous non UV crosslinking could be
useful in the treatment of corneal ectasia.
Commercial Relationships: Mauricio E. Narvez, Sr., None; Marcel Y.
Avila, Sr., None
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.
Support: Universidad Nacional /Facultad de medicina Direccion de investigacion /
Grant #203010016838
Program Number: 1527 Poster Board Number: A483
Presentation Time: 8:30 AM - 10:15 AM
Biomechanical Study Of Corneal Cross-linking With Riboflavin
William A. Eddington, Marc D. Friedman, Evan A. Sherr, David Muller. Avedro,
Waltham, MA.
Purpose: To determine the effects of varying riboflavin concentration, presoak
time, UVA irradiance, and duration of UVA exposure on corneal cross-linking with
riboflavin measured by total corneal stiffness change using strip extensiometry.
Methods: Fresh whole rabbit eyes were obtained <24 hours postmortem in saline
on ice from Pel-Freez biologicals (Arkansas). Eyes were brought to room
temperature, epithelium was removed with a dull blade, intraocular pressure of
15mmHg was applied using a water column, and corneal thickness readings were
taken through pachymetry. The eyes were subjected to varying riboflavin
concentration, presoak time, UVA irradiance, and duration of UVA exposure. The
eyes were left to rest for at least one hour before being excised leaving ~2mm
scleral boundary. A 4mm strip was cut using parallel cutting blades from each
cornea paying careful attention to orientation. Strips were placed in an
extensiometer using rake like attachments with 5 tines (length 3mm) which were
secured to the sclera on each side of the cornea. Strips were pulled at a constant rate
of 10m/min from a position where zero load was applied until either sample
failure or the force limit of the load cell was reached.
Results: The increase in corneal stiffness for varying test conditions was consistent
with expected predictions. Corneal stiffness increased proportionally with
increasing riboflavin concentration, increasing soak time and increasing total
energy dose.
Conclusions: The effects of varying riboflavin concentration, presoak time, UVA
irradiance, and duration of UVA exposure on corneal cross-linking with riboflavin
measured by total corneal stiffness change using strip extensiometry are consistent
with expected predictions from our internal 2-D finite element analysis and
photochemical kinetics models.
Commercial Relationships: William A. Eddington, Avedro Inc. (E); Marc D.
Friedman, Avedro Inc. (E); Evan A. Sherr, Avedro Inc. (E); David Muller,
Avedro Inc. (E)
Support: None
Program Number: 1528 Poster Board Number: A484
Presentation Time: 8:30 AM - 10:15 AM
Continuous Measurement Of Physical Forces During Human Primary
Corneal Fibroblast Migration
Dhananjay Tambe1, Ramin Zareian2, Dimitrios Karamichos3, James D. Zieske3,
Jeffrey W. Ruberti2. 1Department of Environmental Health, Harvard school of
public health, Boston, MA; 2Mechanical Engineering, Northeastern University,
Boston, MA; 3Department of Ophthalmology, Schepens Eye Research Institute,
Harvard medical school, Boston, MA.
Purpose: : Little is known about the mechanobiology of matrix production by
corneal fibroblasts which have been shown to self organize and produce
extracellular matrix in vitro. During coordinated migration, epithelial and
endothelial cells apply forces to the substrate as well as to their neighbors [Trepat
et al 2009 Tambe et al 2011]. If such forces are generated by fibroblasts during
matrix production, they may influence the deposition and retention of collagenous
matrix [Zareian et al 2010]. However it is very difficult to follow fibroblast
behavior while measuring forces on time scales long enough to permit matrix
production. Here we present an in vitro mechanobiological assay capable of
directly observing the behavior of PHCF over relatively long time scales while
simultaneously measuring intrinsic (cell-cell and cell-substrate) physical forces.
Methods: We combine the mechanobioreactor described in [Paten et al 2011] with
a 100um thick, collagen coated, calibrated polyacrylamide gel [E=1250 Pa Yeung
et al 2005] to observe the mechanobiology of PHCF culture. Forces applied to the
substrate are interrogated continuously via embedded 0.5um fluorescent beads.
Using Fourier transform traction microscopy the gel deformation is transformed
into a continuous traction vector field applied by the cells to the substrate. Using
monolayer stress microscopy the traction field is transformed into stresses applied
by the cells to their neighbors.
Results: Over the time course of this experiment, the corneal fibroblasts actively
migrate over the gel surface and begin to form a confluent monolayer. We observe
that cells gradually build up tractions. When they self organize, the traction at a
point has contributions from two sources, contraction generated by cells attached at
that location, and contractions generated by cells far from that location but
transmitted through cell-cell contacts. Magnitude of this traction is on the order of
150Pa.
Conclusions: The combination of the mechanobioreactor with the traction force
microscopy method permits direct measurement PHCF physical forces during
migration and growth to confluence. The system will ultimately enable dynamic,
continuous and quantitative observation of the mechanobiology of matrix
ARVO 2012 Annual Meeting Abstracts
production.
Commercial Relationships: Dhananjay Tambe, None; Ramin Zareian,
None; Dimitrios Karamichos, None; James D. Zieske, None; Jeffrey W.
Ruberti, None
Support: NIH NIAMS R21 AR053551-01
Program Number: 1529 Poster Board Number: A485
Presentation Time: 8:30 AM - 10:15 AM
Live, Long-Term Observation of the Mechanobiology of Confluent Primary
Human Corneal Fibroblast Cultures Subjected to Applied Force
Ramin Zareian1, Dimitrios Karamichos2, Jeffrey A. Paten1, James D. Zieske2,
Jeffrey W. Ruberti1. 1Mechanical & Industrial Engineering, Northeastern
University, Boston, MA; 2Department of Ophthalmology, Schepens Eye Research
Institute, Harvard medical school, Boston, MA.
Purpose: : It has now been clearly established that mechanics plays a critical role
in the initial templating, subsequent growth and maintenance of mesenchymal
tissue. Mechanics strongly influences both the differentiation state and general
behavior of mesenchymal cells (stem and fibroblasts). In addition, the interaction
systems of associated molecules have shown exquisite mechanosensitvity. We have
employed a custom mechanobioreactor to observe changes in the dynamic
organizational behavior of Primary Human Corneal Fibroblasts (PHCFs) exposed
to uniaxial load.
Methods: Our device was sterilized under aseptic conditions and loaded with a
dense, disorganized collagen scaffold (DDCS) comprising type I bovine collagen.
PHCFs from human donors were extracted, cultured, and seeded into the device
where they grew both on the DDCS and on the coverslip. The culture medium was
continuously fed to the cells at a fixed flow rate (8ul/min). After reaching
confluence the DDCS culture was exposed to a mechanical stretch of 6% (N=4). In
one of the four experiments PHCFs were exposed to the live nuclear stain to permit
cell tracking. Fluorescent and DIC images were taken every hour for two weeks.
PIV (Particle Imaging Velocimetry) was used to track cell while FFT (Fast Fourier
Transform) methods were used directly on the DIC images to extract local and
global orientation of cells.
Results: All experiments exhibited similar organizational behavior indicating that
the live stain did not alter the culture significantly. Continuous movies reveal a
highly dynamic culture system with rapidly moving PHCFs which initially display
random orientation. Confluence led to local organization both on the DDCS and on
the glass coverslip. However, loading the DDCS led to consistent global alignment
of the PHCF at a relatively fixed angle to the applied force.
Conclusions: Mechanical load has a profound effect on the migration and
organizational behavior of PHCFs in culture and could likely be used to influence
the organization of synthesized matrix.
A) Live nuclear stain of PHCFs, green and white arrows show the direction of
motion and load respectively, B) Median velocity angle of cells on substrate
(green) and coverslip (red) C) FFT analysis of the cells.
Commercial Relationships: Ramin Zareian, None; Dimitrios Karamichos,
None; Jeffrey A. Paten, None; James D. Zieske, None; Jeffrey W. Ruberti,
None
Support: NIH NIAMS R21 AR053551-01
Program Number: 1530 Poster Board Number: A486
Presentation Time: 8:30 AM - 10:15 AM
Corneal Biomechanical Properties And Intraocular Pressure In High
Anisometropia
Nour Maya N. Haddad, Alain Saad, Hana Landoulsi, Florence Cabot, Damien
Gatinel. Cataract and Refractive Surgery, Fndtn Ophthal Adolphe de Rothschild,
Paris, France.
Purpose: To compare corneal biomechanical properties and intraocular pressure in
myopic anisometropia
Methods: In order to understand changes of the anterior segment in high myopia,
we compared corneal biomechanical properties, IOP and CCT between the less and
more myopic eye of 25 anisometropic patients. Subjects (spherical equivalent
between +3.25D and -21.75 D) were enrolled. Corneal biomechanical properties
and intraocular pressures provided by the Ocular Response Analyser (ORA) were
Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.
compared between the more (MM) and less myopic eyes (LM) and correlated to
central corneal thickness (CCT) and axial length (AL).
Results: In both groups: corneal hysteresis (CH) was highly correlated to CCT and
AL as was AL to Goldmann-correlated IOP (IOPg) and corneal corrected IOP
(IOPcc) (p<0.001).No significant differences of CH, CRF,IOPg and IOPcc were
found between MM and LM.
Conclusions: Higher AL was correlated with a lower CH which indicates that
some aspects of corneal biomechanical properties may be altered in high myopic
eyes. A higher AL was highly correlated to IOPg. IOP may be implicated in the
pathogenesis of myopic eyes which showed altered biomechanical properties
Commercial Relationships: Nour Maya N. Haddad, None; Alain Saad,
None; Hana Landoulsi, None; Florence Cabot, None; Damien Gatinel, None
Support: None
Program Number: 1531 Poster Board Number: A487
Presentation Time: 8:30 AM - 10:15 AM
Amylase Reduces The Biomechanical Stiffness Of The Cornea
Eberhard Spoerl, Naim Terai, Frederik Raiskup, Lutz E. Pillunat. Ophthalmology,
University of Dresden, Dresden, Germany.
Purpose: As the concentration of amylase is enhanced in keratoconus patients, the
aim of the present study was to investigate the effect of amylase on the
biomechanical stiffness of the cornea in vitro.
Methods: Corneal strips were prepared from 20 fresh enucleated porcine eyes. Ten
strips were treated with amylase solution for 48 hours, 10 other strips served as
controls. Stress-strain measurements using a microcomputer-controlled biomaterial
tester (MiniMat; Rheonetric Scientific GmbH, Benshaim, Germany) were
performed to determine the stiffness of the corneal strips.
Results: In contrast to the control corneas, the amylase treated corneas showed a
reduced corneal thickness (CCT = 632.159.6 m, P=0.0001). At 10 % strain the
stress in the control group was 213.4 53.1 kPa and in the amylase group 96.527.0
kPa which was statistically significantly different between both groups (P=0.0001).
The Youngs modulus in the control group was 3.8 MPa and in the amylase group
2.2 MPa.
Conclusions: The present study showed a reduction of the corneal stiffness by the
effect of amylase leading to a dissolving of the ground substance from the stroma.
It is the first study showing the influence of the ground substance on the
biomechanical properties of the cornea. As the amount of amylase in the lacrimal
gland is enhanced in keratoconus, the reduced corneal stiffness might contribute to
the pathogenesis of keratoconus.
Commercial Relationships: Eberhard Spoerl, None; Naim Terai,
None; Frederik Raiskup, None; Lutz E. Pillunat, None
Support: None
Program Number: 1532 Poster Board Number: A488
Presentation Time: 8:30 AM - 10:15 AM
Precision of ORA (Ocular Response Analyzer)
Susanne Goebels1A, Berthold Seitz1A, Achim Langenbucher1B. ADepartment of
Ophthalmology, BDepartment of Experimental Ophthalmology, 1University of
Saarland, Homburg/Saar, Germany.
Purpose: To evaluate the repeatability of corneal hysteresis and corneal resistance
measurements as well as the consistency of the 4 shots within each measurement
using the ORA (Ocular Response Analyzer, Reichert Ophthalmic Instruments,
USA) and to generate a pool of data of a normal population.
Methods: A total of 45 eyes from 45 healthy volunteers without ocular pathologies
and normal visual acuity were enrolled in this study. A sequence of five
consecutive measurements was performed with each patient with the ORA. The
biomechanical properties of the cornea in terms of corneal hysteresis (CH) and
corneal resistance factor (CRF) were recorded, as well as the Goldmann-correlated
IOP (IOPg) and cornea correlated IOP (IOPcc). The trend in each measurement
sequence was analysed and Cronbachs alpha was derived for the repeatability. The
average of 4 shots within each measurement was compared with the best signal
value (BSV) provided by the internal data processing of the ORA.
Results: Mean value for CH was 11.58, BSV was 11.55. For CRF mean value was
11.21 and BSV was 11.28. No significant difference was found between average
value and BSV of one measurement (p>0.05). No significant difference was found
between average value and BSV of the five consecutive measurements, only IOPg
showed a significant difference (p=0,017), average value of IOPg is higher than
BSV. Within a sequence of 5 measurements for each individual, the average values
of the four shots per measurement for IOPg and IOPcc are decreasing by 1.19
mmHg and 1.05 mmHg on average. The repeatability test revealed good results for
CH, CRF (all are higher than 0.9).
Conclusions: The ORA provides beside intraocular pressure additional information
about the biomechanical properties of the cornea such as hysteresis and resistance.
It proofs to yield good repeatability for corneal hysteresis and resistance in normal
subjects.
Commercial Relationships: Susanne Goebels, None; Berthold Seitz,
None; Achim Langenbucher, None
ARVO 2012 Annual Meeting Abstracts

Support: None

Copyright 2012 by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. For permission to reproduce any abstract, contact the ARVO Office at pubs@arvo.org.